PROBING THE EFFECT OF ACTIVE EDIBLE COATING ON SHELF STABILITY OF CHEESE AND BUTTER By Rizwan Arshad 2007-ag-1129 M.Sc. (Hons.) Food Technology A thesis submitted in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY IN FOOD TECHNOLOGY i
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PROBING THE EFFECT OF ACTIVE EDIBLE COATING ON SHELF STABILITY OF CHEESE AND
BUTTER
By
Rizwan Arshad
2007-ag-1129
M.Sc. (Hons.) Food Technology
A thesis submitted in partial fulfillment of the requirements for the degree of
DOCTOR OF PHILOSOPHY
IN
FOOD TECHNOLOGY
NATIONAL INSTITUTE OF FOOD SCIENCE & TECHNOLOGY
FACULTY OF FOOD, NUTRITION & HOME SCIENCESUNIVERSITY OF AGRICULTURE, FAISALABAD
i
PAKISTAN2019
ii
Dedication!
To my parents, who are brave enough to hope!
To myself as “I did it my way”.
Dedicated to everyone who wonders if I’m writing about them!!
I am.
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ACKNOWLEDGEMENTSWith the pen in hand, I proud to think that what words do justice to express my thanks to Almighty
Allah for His unlimited kindness. The Omnipotent, the Omniscient, the Merciful and Beneficent who
is the entire source of all knowledge and wisdom to mankind. He who created the universe and
bestowed the mankind with knowledge and wisdom to search for the secrets. I am greatly obligate to
Almighty Allah by Whom grace I have been able to complete this task successfully. All praises for
Holy Prophet Muhammad (PBUH), the city of knowledge, the illuminating torch and the rescuer of
humanity from going astray.
My heartfelt mention goes to Prof. Dr. Masood Sadiq Butt, Dean Faculty of Food, Nutrition
and Home Sciences University of Agriculture Faisalabad and Prof. Dr. Tahir Zahoor Director
General National Institute of Food Science and Technology, University of Agriculture, Faisalabad for
their skilful help and guidance during study course.
I am grateful to my supervisor Dr. Aysha Sameen, Assistant Professor, National Institute of
Food Science and Technology, University of Agriculture, Faisalabad for her supervision in planning,
execution and scholarly ideas that beautified the scientific nature of the research work performed. She
always directed to enlighten the ways of life as well.
I am thankful to the member of my supervisory committee, Prof. Dr. Nuzhat Huma,
National Institute of Food Science and Technology and Dr. Muhammad Anjum Zia, Associate
Professor, Department of Biochemistry, University of Agriculture Faisalabad for their kind help,
technical guidance, moral support and prayers to accomplish this study.
I grateful to my beloved Parents and My Brothers (Adeel Arshad, Imran Arshad and
Hamza Arshad) and Sister whom I am really indebted to, for their prayers and perpetual
encouragement. Before I finish, I would also like to thank my friends and lab fellows for their
consistent care and encouragement.
Rizwan Arshad
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TABLE OF CONTENTS
CHAPTER-1 INTRODUCTION 1
CHAPTER-2 REVIEW OF LITERATURE 5
2.1 Edible coating 5
2.2 Introduction to natural antimicrobials and antioxidants 6
2.3 Factors influencing the antimicrobial and antioxidant activity of natural products 8
2.4 Role of antimicrobial and antioxidant agents in edible coating 9
2.5 Historical use of essential oil 10
2.6 Essential oils 11
2.7 Current use of essential oil 15
2.8 Role of Edible coating in dairy products 17
2.9 Cytotoxicity and legal aspects for the use of Eos 19
CHAPTER-3 MATERIAL AND METHODS 22
3.1 Procurement of raw materials 22
3.2 Preparation of active edible coating for SC 23
3.2.1 Analysis of active edible coating 23
3.2.1.1 pH 23
3.2.1.2 Acidity 23
3.2.1.3 Viscosity 23
3.2.1.4 Water activity 24
3.3 Preparation of soft cheese 24
a. Milk fat standardization 24
b. Physicochemical analysis of milk 24
3.4 Quality assessment of soft cheese 25
3.4.1 Physico-chemical analysis of soft cheese 25
3.4.1.1 Moisture content 25
3.4.1.2 Fat 26
3.4.1.3 Protein 26
3.4.1.4 pH 27
3.4.1.5 Acidity 27
3.4.1.6 Ash 27
3.4.2 Color Analysis 27
3.4.3 Water Activity 27
3.4.4 Weight Loss 28
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3.4.5 Assessment of proteolysis by RP-HPLC 28
3.5 Butter 29
3.5.1 Physicochemical analysis of cream 29
3.5.1.1 Free Fatty Acids of cream 29
3.5.2 Preparation of active edible coating of butter 30
3.5.3 Manufacturing of butter 30
3.5.4 Quality assessment of butter 31
3.5.4.1 Physico-chemical analysis of butter 31
3.5.5 Free Fatty Acids of butter 31
3.6 Textural profile analysis of soft cheese and butter 31
3.7 Total Viable Count in soft cheese and butter sample 31
3.8 Antioxidant analysis of soft cheese and butter 32
3.8.1 2, 2-Diphenyl-1-picrylhdrazyl (DPPH) scavenging activity 32
3.8.2 Peroxide value 32
3.8.3 Measurement of Thiobarbituric acid reactive substance (TBARS) value 33
3.9 Sensory evaluation of soft cheese and butter 33
3.10 Statistical analysis 34
CHAPTER-4 RESULTS AND DISCUSSION 35
4.1 Physico-chemical analysis of milk 35
4.2 Analysis of edible coating 35
4.3 Physico-chemical analysis of soft cheese 37
4.3.1 Moisture 37
4.3.2 Fat 40
4.3.3 Protein 43
4.3.4 Ash 46
4.3.5 pH 46
4.3.6 Acidity 51
4.3.7 Water activity 54
4.3.8 Color Analysis 57
4.4 Texture analysis 59
4.4.1 Hardness 62
4.4.2 Cohesiveness 65
4.4.3 Springiness 65
4.4.4 Gumminess 70
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4.4.5 Chewiness 73
4.5 Weight Loss 76
4.6 Antioxidant tests 79
4.6.1 2, 2-Diphenyl-1-picrylhdrazyl (DPPH) scavenging activity 79
4.6.2 Peroxide value 79
4.6.3 Measurement of TBARs value 84
4.7 Proteolysis 87
4.8 Microbiological Analysis 91
4.9. Sensory Evaluation 94
4.9.1 Flavor 94
4.9.2 Appearance 97
4.9.3 Body/Texture 100
4.9.4 Overall Acceptability 103
Butter 106
4.10 Physico-chemical analysis of cream 106
4.11 Analysis of edible coating of butter 106
4.12 Physicochemical Analysis of butter 107
4.12.1 Moisture 107
4.12.2 pH 110
4.12.3 Acidity 110
4.12.4 Fat 115
4.12.5 Ash 118
4.12.6 Water activity 122
4.12.7 Color 125
4.13 Texture Analysis 129
4.13.1 Hardness 129
4.13.2 Cohesiveness 129
4.13.3 Springiness 134
4.13.4 Gumminess 137
4.13.5 Chewiness 137
4.14 Weight loss 142
4.15 Free fatty acid 145
4.16 Anti-oxidative analysis 148
4.16.1 2, 2-Diphenyl-1-picrylhdrazyl (DPPH) scavenging activity 148
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4.16.2 Peroxide value 151
4.16.3 Measurement of TBARs value 154
4.17 Microbiological Analysis 157
4.18 Sensory Evaluation 160
4.18.1 Flavor 160
4.18.2 Appearance 163
4.18.3 Body/Texture 163
4.18.4 Overall Acceptability 168
5. SUMMARY 173
LITERATURE CITED 177
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LIST OF TABLES
Table. No. Title Page No.
Table 3.1 Treatment plan for soft cheese 22Table 3.1 Treatment plan for butter 29Table 4.1 Compositional analysis of milk 36 Table 4.2 Analysis of edible coating of soft cheese 36
Table 4.3.1a Mean sum of squares for moisture of soft cheese 38
Table 4.3.1b Effect of treated SC on the % moisture during storage 39Table 4.3.2a Mean sum of squares for fat of soft cheese 41Table 4.3.2b Effect of treated SC on the % fat during storage 42
Table 4.3.3a Mean sum of squares for protein of soft cheese 44Table 4.3.3b Effect of treated SC on the % protein during storage 45Table 4.3.4a Mean sum of squares for ash of soft cheese 47Table 4.3.4b Effect of treated SC on the % ash during storage 48Table 4.3.5a Mean sum of squares for pH of soft cheese 49Table 4.3.5b Effect of treated SC on the pH during storage 50Table 4.3.6a Mean sum of squares for acidity of soft cheese 52Table 4.3.6b Effect of treated SC on the % acidity during storage 53Table 4.3.7a Mean sum of squares for water activity of soft cheese 55Table 4.3.7b Effect of treated SC on the water activity during storage 56Table 4.3.8a Mean sum of squares for color of soft cheese 58Table 4.3.8b Effect of treated SC on the L*, a* and b* value 61Table 4.4.1a Mean sum of squares for hardness of soft cheese 63Table 4.4.1b Effect of treated SC on the hardness (g) during storage 64Table 4.4.2a Mean sum of squares for cohesiveness of soft cheese 66Table 4.4.2b Effect of treated SC on the cohesiveness during storage 67Table 4.4.3a Mean sum of squares for springiness of soft cheese 68Table 4.4.3b Effect of treated SC on the springiness during storage 69Table 4.4.4a Mean sum of squares for gumminess of soft cheese 71Table 4.4.4b Effect of treated SC on the gumminess (g) during storage 72Table 4.4.5a Mean sum of squares for chewiness of soft cheese 74Table 4.4.5b Effect of treated SC on the chewiness (g) during storage 75Table 4.5a Mean sum of squares for weight loss of soft cheese 77Table 4.5b Effect of treated SC on the % weight loss during storage 78
Table 4.6.1a Mean sum of squares for DPPH activity of soft cheese 80Table 4.6.1b Effect of treated SC on the % DPPH during storage 81Table 4.6.2a Mean sum of squares for POV of soft cheese 82Table 4.6.2b Effect of treated SC on the POV(meq/kg) during storage 83Table 4.6.3a Mean sum of squares for TBARs value of soft cheese 85
Table 4.6.3b Effect of treated SC on the TBARs value (mg MDA/kg) during storage 86
Table 4.8.1a Mean sum of squares for TVC of soft cheese 92
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Table 4.8.1b Effect of treated SC on the TVC (x 104cfu/g) during storage 93
Table 4.9.1a Mean sum of squares for flavor of soft cheese 95 Table 4.9.1b Effect of treated SC on the flavor during storage 96
Table 4.9.2a Mean sum of squares for appearance of soft cheese 98Table 4.9.2b Effect of treated SC on the appearance during storage 99Table 4.9.3a Mean sum of squares for body/texture of soft cheese 101Table 4.9.3b Effect of treated SC on the body/texture during storage 102
Table 4.9.4a Mean sum of squares for overall acceptability of soft cheese 104
Table 4.9.4b Effect of treated SC on the overall acceptability during storage 104
Table 4.10 Physico-chemical analysis of cream 106Table 4.11 Physico-chemical analysis of edible coating of butter 106
Table 4.12.1a Mean sum of squares for moisture contents of butter 108Table 4.12.1b Effect of treated butter on the % moisture during storage 109Table 4.12.2a Mean sum of squares for pH of butter 111Table 4.12.2b Effect of treated butter on the pH during storage 112Table 4.12.3a Mean sum of squares for acidity of butter 113Table 4.12.3b Effect of treated butter on the % acidity during storage 114Table 4.12.4a Mean sum of squares for fat of butter 116Table 4.12.4b Effect of treated butter on the % fat during storage 117Table 4.12.5a Mean sum of squares for ash of butter 120Table 4.12.5b Effect of treated butter on the % ash during storage 121Table 4.12.6a Mean sum of squares for water activity of butter 123Table 4.12.6b Effect of treated butter on the water activity during storage 124Table 4.12.7a Mean sum of squares for color parameter of butter 126Table 4.12.7b Effect of treated butter on the L*, a* and b* during storage 128Table 4.13.1a Mean sum of squares for hardness of butter 130Table 4.13.1b Effect of treated butter on the hardness (g) during storage 131Table 4.13.2a Mean sum of squares for cohesiveness of butter 132Table 4.13.2b Effect of treated butter on the cohesiveness during storage 133Table 4.13.3a Mean sum of squares for springiness of butter 135Table 4.13.3b Effect of treated butter on the springiness during storage 136Table 4.13.4a Mean sum of squares for gumminess of butter 138
Table 4.13.4b Effect of treated butter on the gumminess (g) during storage 139
Table 4.13.5a Mean sum of squares for chewiness of butter 140
Table 4.13.5b Effect of treated butter on the chewiness (g) during storage 142
Table 4.14 a Mean sum of squares for weight loss of butter 143
Table 4.14 b Effect of treated butter on the % weight loss during storage 144
Table 4.15a Mean sum of squares for free fatty acid (FFA) of butter 142Table 4.15b Effect of treated butter on the % FFA during storage 147
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Table 4.16.1a Mean sum of squares for DPPH activity of butter 149Table 4.16.1b Effect of treated butter on the % DPPH during storage 150Table 4.16.2a Mean sum of squares for POV of butter 152
Table 4.16.2b Effect of treated butter on the POV(meq/kg) during storage 153
Table 4.16.3a Mean sum of squares for TBARs value of butter 155
Table 4.16.3b Effect of treated butter on the TBARs value (mg MDA/kg) during storage 156
Table 4.17.1a Mean sum of squares for TVC of butter 158
Table 4.17.2b Effect of treated butter on the TVC (x 104cfu/g) during storage 159
Table 4.18.1a Mean sum of squares for flavor of butter 161
Table 4.18.1b Effect of treated butter on the flavor during storage 162
Table 4.18.2a Mean sum of squares for appearance of butter 164Table 4.18.2b Effect of treated butter on the appearance during storage 165Table 4.18.3a Mean sum of squares for body/texture of butter 166Table 4.18.3b Effect of treated butter on the body/texture during storage 167Table 4.18.4a Mean sum of squares for overall acceptability of butter 169
Table 4.18.4b Effect of treated butter on the overall acceptability during storage 170
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LIST OF FIGURES
Fig. No. Title Page No.
Fig. 3.1 Flow line for manufacturing of soft cheese 24
Fig. 3.2 Flow line for manufacturing of butter 30
Fig 4.1 Effect of edible coating and essential oils on the moisture contents (%) of soft cheese 38
Fig. 4.2 Effect of edible coating and essential oils on the fat contents (%) of soft cheese 41
Fig. 4.3 Effect of edible coating and essential oils on the protein contents (%) of soft cheese 44
Fig. 4.4 Effect of edible coating and essential oils on the ash contents (%) of soft cheese 47
Fig. 4.5 Effect of edible coating and essential oils on the pH of soft cheese 49
Fig.4.6 Effect of edible coating and essential oils on the acidity (%) of soft cheese 52
Fig.4.7 Effect of edible coating and essential oils on the water activity (%) of soft cheese 55
Fig. 4.8 Effect of edible coating and essential oils on the L*value of soft cheese 58
Fig. 4.9 Effect of edible coating and essential oils on the a*value of soft cheese 60Fig. 4.10 Effect of edible coating and essential oils on the b*value of soft cheese 60
Fig.4.11 Effect of edible coating and essential oils on the hardness of soft cheese 63
Fig. 4.12 Effect of edible coating and essential oils on the cohesiveness of soft cheese 66
Fig. 4.13 Effect of edible coating and essential oils on the springiness of soft cheese 68
Fig. 4.14 Effect of edible coating and essential oils on the gumminess of soft cheese 71
Fig. 4.15 Effect of edible coating and essential oils on the chewiness of soft cheese 74
Fig 4.16 Effect of edible coating and essential oils on the weight loss (%) of soft cheese 77
Fig. 4.17 Effect of edible coating and essential oils on the DPPH activity (%)of soft cheese 80
Fig. 4.18 Effect of edible coating and essential oils on the POV (meq/kg) value of soft cheese 82
Fig. 4.19 Effect of edible coating and essential oils on the TBARS (mg MDA/kg) of soft cheese 85
Fig. 4.20 Standard casein 89Fig. 4.21 Break down of αS-1 casein during storage 89Fig. 4.22 Break down of αS-2 casein during storage 90Fig. 4.23 Break down of β casein during storage 90
Fig. 4.24 Effect of edible coating and essential oils on the TVC (cfu/g) of soft cheese 92
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Fig. 4.25 Effect of edible coating and essential oils on the flavor of soft cheese 95
Fig. 4.26 Effect of edible coating and essential oils on the appearance of soft cheese 98
Fig. 4.27 Effect of edible coating and essential oils on the body/texture of soft cheese 101
Fig. 4.28 Effect of edible coating and essential oils on the overall acceptability of soft cheese 104
Fig. 4.29 Effect of edible coating and essential oils on the moisture contents (%) of butter 108
Fig. 4.30 Effect of edible coating and essential oils on the pH of butter 111Fig. 4.31 Effect of edible coating and essential oils on the acidity (%) of butter 113
Fig. 4.32 Effect of edible coating and essential oils on the fat contents (%) of butter 116
Fig. 4.33 Effect of edible coating and essential oils on the ash contents (%) of butter 120
Fig. 4.34 Effect of edible coating and essential oils on the water activity (%) of butter 123
Fig. 4.35 Effect of edible coating and essential oils on the L* value of butter 126Fig. 4.36 Effect of edible coating and essential oils on the a* value of butter 127Fig. 4.37 Effect of edible coating and essential oils on the b* value of butter 127Fig. 4.38 Effect of edible coating and essential oils on the hardness (g) of butter 130Fig. 4.39 Effect of edible coating and essential oils on the cohesiveness of butter 132Fig. 4.40 Effect of edible coating and essential oils on the springiness of butter 135
Fig. 4.41 Effect of edible coating and essential oils on the gumminess (g) of butter 138
Fig. 4.42 Effect of edible coating and essential oils on the chewiness of butter 140
Fig. 4.43 Effect of edible coating and essential oils on the weight loss (%) of butter 143
Fig. 4.44 Effect of edible coating and essential oils on free fatty acid FFA (%) of butter 146
Fig. 4.45 Effect of edible coating and essential oils on the DPPH activity (%) of butter 149
Fig. 4.46 Effect of edible coating and essential oils on the POV (meq/kg) value of butter 152
Fig. 4.47 Effect of edible coating and essential oils on the TBARS (mg MDA/kg) of butter 155
Fig. 4.48 Effect of edible coating and essential oils on the TVC (cfu/g) of butter 158Fig. 4.49 Effect of edible coating and essential oils on the flavor of butter 161Fig. 4.50 Effect of edible coating and essential oils on the appearance of butter 164Fig. 4.51 Effect of edible coating and essential oils on the texture of butter 166Fig. 4.52 Effect of edible coating and essential oils on the overall acceptability
of butter 169
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LIST OF ABBREVIATION
Peppermint oil PMO
Ginger oil GO
Colove oil CO
Black cumin oil BCO
Corn Starch CS
Whey powder WP
Lactic acid bacteria LAB
Soft cheese SC
Figure Fig.
Essential oils EOs
Source of variation SOV
Mean Square MS
Sum of square SS
F value FV
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ABSTRACTThe current study was conducted to evaluate the impact of active edible coating and natural essential oils (EOs) on storage stability of butter and soft cheese (SC). The major issue in storage stability of butter is lipid oxidation due to high fat content while in SC is mold growth due to its high moisture content. EOs were used due to their multifarious role such as food flavoring, preservative (food safety), antioxidant and medicinal ingredient. Corn starch (CS) and whey powder (WP) based edible coating were developed for butter and SC respectively. Peppermint oil (PMO) and clove oil (CO) were used as active ingredient in SC coating while ginger oil (GO) and black cumin oil (BCO) in butter coating. Glycerol was added as plasticizer, xanthan gum to increase the viscosity, lecithin for emulsification of EOs in edible coating. Active edible coating was applied on SC and butter with brushing method. SC samples (T0= control; T1= edible coating of SC with WP; T2 = SC containing 0.5% CO; T3= SC containing 0.75% CO; T4= SC containing 1.0% CO; T5= WP based edible coating of SC containing 0.5% CO; T6= WP based edible coating of SC containing 0.75% CO; T7= WP based edible coating of SC containing 1.0% CO; T8= SC containing 1.5% PMO; T9= SC containing 2.0% PMO; T10= SC containing 2.5% PMO; T11= WP based edible coating of SC containing 1.5% PMO; T12= WP based edible coating of SC containing 2.0% PMO; T13= WP based edible coating of SC containing 2.5% PMO) were analyzed for physicochemical parameters, color value, water activity (aw), texture profile, antioxidant activity, total viable count (TVC), proteolysis and sensory attributes during 30 days of storage at 2-50C. Similarly butter samples (T0= control; T1= edible coating of butter with CS; T2 = butter containing 0.2% BCO; T3= butter containing 0.3% BCO; T4= butter containing 0.4% BCO; T5= CS based edible coating of butter containing 0.2% BCO; T6= CS based edible coating of butter containing 0.3% BCO; T7= CS based edible coating of butter containing 0.4% BCO; T8= butter containing 1.5% GO; T9= butter containing 2.0% GO; T10=butter containing 2.5% GO; T11=CS based edible coating of butter containing 1.5% GO; T12= CS based edible coating of butter containing 2.0% GO; T13= CS based edible coating of butter containing 2.5% GO) were also analyzed for physicochemical parameters, color value, water activity (aw), texture profile, free fatty acids (FFA), antioxidant activity, total viable count (TVC), and sensory attributes during 90 days of storage at 2-50C. SC and butter sample containing edible coating and natural EOs shows significant (p<0.05) results. Highest value of moisture (79.55 %) was noted in T9 SC sample while the lowest moisture (74.33%) was noted in control (T0) SC. Highest value of protein (13.55%) was observed in T12 while the minimum protein (13.30%) value was observed in control (T0) SC. Similarly highest value of moisture (16.53 %) was recorded in butter sample T10
while the lowest moisture (11.25%) was noted in control (T0) butter. Highest fat (79.80%) was recorded in control butter (T0) while the lowest fat (79.56%) was noted in T10. Best treatment of SC and butter was selected on the basis of sensory evaluation and storage stability. Texture profile of SC and butter also effected significantly due to edible
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coating and EOs. SC and butter with direct addition of EOs showed higher antioxidant and antimicrobial activity as compared to other samples. TVC (cfu/g) was minimum in all samples of SC and butter as compared to compare to their control (T0) due to the effect of EOs and edible coating. Incorporation of natural EOs and edible coating significantly improved all the sensory characteristics of SC and butter. Hence, it is concluded from the present study that storage stability of SC and butter can be enhanced by the judicious use of edible coating and natural EOs with better flavor and quality attributes.
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CHAPTER-1INTRODUCTION
Nowadays industries are facing problems against using chemical or synthetic
preservatives to enhance the shelf stability of dairy products due to consumer awareness
and their bad health effects. Cheese and butter are dairy products that consumed
worldwide directly and indirectly in a number of food products due to its therapeutic and
nutritional significance (Mallia et al., 2008). Cheese is very popular in the world due to
variety in functionality, flavor, texture and nutritional value. Soft cheese (SC) is
biologically and biochemically active due to its high level of moisture content and
subsequently undergoes changes in flavor, functionality, loss in moisture, mold growth
and texture that ultimately causes the deterioration (Fox et al., 2000). Similarly butter is
water-in-oil (w/o) emulsion composed of more than 80% fat and also water in the form of
tiny droplets, perhaps some milk solids not fat, with or without salt (sweet butter); and
the texture is a result of working/kneading during processing at appropriate temperatures
so as to establish a fat crystalline network that results in desired smoothness and the
butter is used as a spread, cooking fat, or a baking ingredient (USDA, 1989). Twenty one
pound of fresh milk is necessary to produce one pound of butter (Douma, 2004). Fat is a
major constituent in butter that plays a very vital role in flavor, nutritional value,
appearance, body/texture and shelf stability. Butter due to its high fat content is more
vulnerable to oxidative deterioration that leads to the reduction of nutritional quality and
also makes the food unacceptable for consumers. Oxidation of fats causes many human
diseases like cardiovascular diseases, membrane damage, cancer and ageing so that
antioxidants are added in foods to prevent or delay oxidation (Larick and Parker, 2001).
According to Mazzocchi et al, (2008), the big problem of the dairy industry is the
low storage stability of dairy products due to oxidation reactions such as fungal attack,
degradation oxidative rancidity and mold growth. At present to prevent these problems
synthetic or chemical preservatives are being used. Consumers have become more
conscious about the selection of food due to awareness about the use of synthetic or
chemical preservatives in the foods due to their toxicity and bad effects on health. So
natural antimicrobials and antioxidants had attracted great attention due to its harmless
health effect to enhance the shelf life and increase food stability and also considered to be
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an essential part of food. This has become a big challenge for industries to reduce the use
of conventional chemical preservatives for the safe end product (Campos et al., 2011).
One of the best approach to reduce food deterioration is to use edible films and
coatings (Soliva and Martin, 2003). In this perspective; the demand of edible coating is
increasing day by day. Edible coating is a thin layer of material that covers the surface of
the food and can be eaten as part of the whole product. Edible coatings have the
advantages that these are edible, give appealing appearance to food, non-polluting and
biodegradable (Kang et al., 2013). According to Lin and Zhao (2007) edible coatings are
formulated using different natural ingredients including lipids, proteins, polysaccharides
and by adding surfactants and plasticizers. The efficiency of edible coating also depends
on the method of application and adherence capacity of the coating. Spraying, dipping
and brushing methods are usually used to apply coatings on the surface of the food
products which performs a function as a semi-permeable membrane, regulates the
moisture loss and exchange of gases.
Quality of fresh and processed food can also be prolonged by using the new
technology of edible coatings that helps in enhancing the shelf life of food. Functional
properties of these edible coating can be enhance by the incorporation of active
ingredients (phenolic compounds, antioxidants and antimicrobials) into the emulsion that
is potentially beneficial for the prolong preservation (Sanchez et al., 2011). Active
ingredients are antioxidants and antimicrobials that can be synthetic or natural obtained
from natural sources like plant extracts, essential oils, Vitamin E (tocopherol), ascorbic
acid (vitamin C) and some other compounds. These ingredients can be used in
combination or individually as a replacer of synthetic ones due to their antioxidative and
antimicrobial activity (Perdones et al., 2014).
However, synthetic antioxidants needs legal approvals with recommended doses
for human consumption and some of synthetic one are BHT, BHA, PG, octyl gallate,
dodecyl gallate, and TBHQ (Andre et al., 2010). Synthetic antioxidants have some
drawbacks upon natural antioxidants as synthetic show hundred percent antioxidative
activity in comparison to the natural one because their activity affected by the presence of
other compounds that counteract the compounds activity. Natural antioxidants are not
present in maximum number but its usage is highly recommended by health authorities
2
that can be used in the food products as food flavorings, enhance the sensory properties
of food and preserve the food for longer storage period (Pokorny, 2007b) as they binds
the free radicles without defining the mechanism when food is oxidized (Pokorny, 2007).
Different natural antimicrobial agents are used to prevent the deterioration and
enhance the storage stability of SC and butter. Chemical antimicrobial compounds
include benzoic acid, sorbic acid, propionic acid and potassium sorbate may be
incorporated into edible coatings to prevent fungal and bacterial growth on the surface of
products but these have bad health effects (Cha and Chinnan, 2004). Natural active edible
coating is a carrier of active ingredients (antioxidants and antimicrobials) and can be
incorporated with enzymes, vitamins, minerals, colorants, flavors, probiotics and
neutraceuticals (Vasconez at el., 2009).
Major constituents of cheese are casein, water and lipids that make it a complex
product. Different packaging materials are being used to store the cheeses (such as SC,
cottage cheese, decorated cheese, cream cheese etc.) in modified atmosphere (MA) in
which gases like O2 are replaced with CO2 and N2 in the packaged form as they give long
shelf life (Mannheim and Soffer, 1996), there are yeasts and spoilage bacteria’s that
cause spoilage even in low O2 concentration in the package to avoid this smart packaging
material is in use as they give color indication on spoilage of product (Westall and
Filtenborg, 1998). During processing, handling and storage of cheese, the prevailing
uncontrolled condition affect the complex structure of cheese that further cause the
quality losses, unhygienic condition leads to the extensive bacterial and fungal
development on the surface of cheese. It disturbs its flavor and reduces its demand for
consumer and ecomically huge loss to the industry (De Oliveira et al., 2007). Water
losses in un- packaged cheese depends on its chemical nature that affect its texture,
proper packaging lowers the rate of water loss and suitable edible coating extended the
life of cheese (Pantaleao et al., 2007).
Edible packaging/coating has the potential to improve the food safety and quality
(i.e. improve stock management, increase shelf life expectancy and reduce waste), thus
focuses both retailers and consumers demands. These are the main reasons why edible
packaging systems (e.g. active packaging) are expected to play a key role in perishable
food sectors. However, technical limitations and high costs associated with these
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technologies (e.g. expensive bioactive agents) have raised constraints to a more extensive
implementation of edible packaging in the food industry (Kang et al., 2013).
So, the current issue in storage stability of butter is lipid oxidation due to high fat
content while in SC is mold growth due to its high moisture content. Keep in view the
bad health effects and consumer awareness about the use of synthetic antioxidants and
antimicrobial, problems related to the storage stability of SC and butter, consumer
demand about the use of natural essential oils as antioxidants and antimicrobial, unique
role of active edible coating, the research work was designed with the hypothesis to
produce SC and butter with improved quality and longer shelf life by direct combination
of natural essential oils; and application of active edible coating on SC and butter. To
check the above mentioned hypothesis, the current study was conducted with the
objectives (1) to formulate corn starch and whey powder based active edible coating for
butter and SC respectively (2) to explore the sensory and physicochemical impact of
active edible coating on the quality attributes of SC and butter during storage. Findings of
this study showed that active edible coating containing natural essential oils as active
ingredient improved the shelf stability of SC and butter with better flavor and quality
attributes as compared to products containing chemical or synthetic additives. Results of
current study could deliver new perception to dairy industry to fulfill increased demand
of consumer towards the use of natural additives/ preservatives. Hence edible coating
containing natural essential oils should be used at industrial scale to extend the shelf life
of SC and butter.
4
CHAPTER-2REVIEW OF LITERATURE
The current study was designed to evaluate the effect of essential oils and edible
coating on the quality attributes of soft cheese (SC) and butter from cream. Numerous
disciplines like nutrition, biotechnology, toxicology, enzymology, microbiology,
biochemistry and flavor chemistry involved in different characteristics of butter and SC,
attracted the scientists towards the study of butter and cheese. The literature relevant to
my research has been looked over to support the study under the following headings.
2.1 Edible Coating
Since past 10 years, on the demand of consumers for safe and hygiene food
products food engineers are in dire struggle to develop a technology for longer storage
and shelf life with environmental friendly packaging materials to protect the fresh food.
Edible coating or film is a thin layer of food material that is coated on a food surface with
generally recognized as safe (GRAS) ingredients. Edible packaging materials are used
differently for different food items as in case of meat, different meat casings are used for
cooking purpose that is not detached during cooking. These types of coating are required
to protect the food e.g. mechanically shield the food or avoid the food from
contamination or incorporation of microorganisms due to mass transfer. The coatings or
films certainly enhance the act, the film perform as vehicle for the integration of chemical
or natural antimicrobial, antioxidants, or functional probiotics, minerals, and vitamins are
components that can be incorporated into the edible coating. Different hydrocolloids
(proteins, polysaccharides and alginates) and lipids (constituted by the fatty acids,
glycerol or waxes) are the ingredients that can be used for the formation of edible
coatings. Edible coating is a thin layer of edible material that covers the surrounding of a
product and we can eat it with the product (Loura et al., 2011).
Starch
Glucose polymers joined with each other and make a polymeric anhydrous
structure known as starch. It has different types based on their structure or presence of
two types of glucose molecule in it. Presence of amylase that shows linear molecular
structure, second sugar polymer is amylopectin that have branched polymers of sugars
(Rodriguez et al., 2006). Being having strong mechanical properties starch is mostly used
5
in the products of food industries. Starches are used to generate a biodegradable
packaging material or coatings that partially or completely replace the cheap form of
plastic coating as these types of coatings are environmentally friendly (Xu et al., 2005).
Coatings are made up of different starch or by amylose polymer, the best example
is corn starch that can be used for any type of edible coatings, even aqueous gelatinized
solution is also used for the standing form of coatings after drying. Almost amylose 75%
and amylopectin 25% are present in corn starch a huge difference difference in starch is
because of starch amylose that is about 85% (Whistler and Daniel, 1985). Coatings that
are produced by using 71% of starch will not supports the 100% oxygen permeability
with lesser relative humidity than 100% studied by the Mark et al. (1966) and this
concept is for plasticized or un-plasticized products that having 16% glycerol in their
coating formula. Hydrophilic polymer in presence of plasticizers affect the absorption of
water molecules enhanced by the mobility of polymers as it normally increase the gas
permeability. High amylose starch etherified with propylene oxide that limitize the water
solubility and make hydroxyl-propylated derivative (Banker et al., 2000).
2.2 Introduction to natural antimicrobials and antioxidants
During recent decades, the consumer demands high quality and more safe food
which is free from chemical preservatives having extended shelf life (Brul and Coote,
1999). Food safety has drawn much attention due to increasing food borne illnesses. One
of the new techniques that is used for avoiding the chemical preservatives in foods is the
use of natural preservatives like herbs which contain active ingredients (Sánchez-
González et al., 2011). Natural antimicrobials and antioxidants have been focused due to
increased consumer demand. The government agencies have approved chemical
preservatives and many of them are still hazardous to health therefore scientists have
been focusing on natural products having potential antioxidant and antimicrobial
activities. On the other hand, food borne illnesses are increasing due to increased
resistance of some pathogens against certain chemical preservatives (Sofos, 2008;
Mathew et al., 2007).
Natural antimicrobials are more likely to be the solution of many food borne
illnesses which arise due to chemical preservatives. Natural preservatives can produce
better results as compared to other synthetic procedures and combinatorial chemistry.
6
Therefore, different types of compounds which are effective against variety of
microorganisms have been focused to meet consumer demand and for the wholesomeness
of food so as to protect the food from deterioration. A variety of secondary metabolites
and essential oils (Eos) can be obtained from edible, herbal and medicinal plants (Tiwari
et al., 2009). It has been scientifically confirmed that many natural antimicrobials have
been derived from plants and are in use for centuries (Gyawali and Ibrahim, 2012; Aires
et al., 2009). It has been observed that there is an increased trend of discovering new
antimicrobials of natural origin that can be used to increase the shelf life of certain foods
by preventing or inhibiting the growth of microorganisms (Fattouch et al., 2007;
Lanciotti et al., 2004). A single molecule having both antimicrobial and antioxidative
properties makes it more useful and suitable to be used as preservative in food industry.
The preservative potential of compounds in plants largely depends on the
concentration and structure of active components. A wide variety of chemical compounds
having antimicrobial activity have been found in plants which include thiosulfinates,
phenolics, saponin, glucosinates, flavonoids and different organic acids. The major plant
component having preservative activity include phenolic compounds such as acids,
ketones, aldehydes, terpenes, isoflavonoids, acids and aliphatic alcohols (Tiwari et al.,
2009). For example, the preservative potential of 46 different extracts of herbs and spices
was found to be associated with phenolic compounds (Shan et al., 2007). The authors
evaluated that the tested spices have significant antibacterial effect against Salmonella
anatum, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli and Bacillus
cereus. The red cabbage also had antimicrobial activity due to the presence of phenolic
compound i.e. anthocycanins (Hafidh et al., 2011). Essential oils contain phenolic
compounds such as olive oil (oleuropein), citrus oil, tea-tree oil (terpenoids) and
bergamot which have significant antimicrobial effects. However, it has been also reported
that non-phenolic oil compounds of cinnamon, coariander, parsley, muscadine, citral,
clove, rosemary, sage and lemongrass have strong effect against gram negative and gram
positive bacterial groups (Gutierrez et al., 2008; Lopes-Lutz et al., 2008; Holley and
Patel Improvement , 2005; Nguefack et al., 2007; Mandalari et al., 2007). There are
some non-phenolic components in essential oils for example garlic oil and isothiocyanate
are significantly effective against Gram- negative bacteria. Antimicrobials and
7
antioxidants from natural plants sources commonly includes herbs and spices (oregano,
rosemary, basil, thyme, sage, onion, cardamom and clove), vegetable and fruit (pepper,
garlic, guava, cabbage, xoconostle and onion) leaves and seeds such as caraway, nutgem,
olive leaves, grape seeds and parsley (Tajkarimi et al., 2010; Tiwari et al., 2009).
Antioxidant and antimicrobial activity of compounds derived from plants depends
on many factors which include the method of extracting essential oil, kind of solvent,
culture, volume of inoculum, postharvest condition, genetics, growth phase and extrinsic
or intrinsic properties of food such as water content, pH, antioxidants, fat, preservatives,
packaging procedure, incubation time/temperature, oxygen availability and concentration
as well as structure of food (Burt, 2004). The antibacterial effect of thyme and oregano
oils increases agains St. aureus and S. typhimurium at low oxygen levels. Therefore
vacuum packaging and essentials oil in combination seems to be an effective food
preservation technique (Solomakos et al., 2008).
2.3 Factors influencing the antimicrobial and antioxidant activity of natural
products
According to Tiwari et al. (2009) antimicrobial action of the natural compounds
influenced different factors like stage of development, botanical source, functional
groups, extraction method, structure composition and harvesting time of that natural
compound. In addition, food components and different kind of additives reacted with the
natural antimicrobial compound that’s play an imperative role in the food preservation.
For example, in food system negatively charged food particles reacted with cationic
antimicrobial peptides to may decrease the effect of antimicrobial peptide (Potter et al.,
2005). Most studies which were related to the antimicrobial effectiveness of natural
compounds have been showed in vitro using microbiological media (Tiwari et al., 2009;
Tajkarimi et al., 2010; Delaquis et al., 2002). Thus, there is very less understanding of
the efficiency of natural compounds when they were applied to different complex food
systems. Foods are not germ-free, so the results that are acquired by antagonistic assays
can be unfair by a diverse microbial population. Heat-treatments which are required to
destroy microbial flora in foods also alter the chemical composition of food and thus
disturb the natural efficiency of antimicrobial compounds. Thus the zones of
collaborations between natural antimicrobials and food components demand knowledge
8
to certify the efficacy of natural antimicrobials when applied on different food products.
The effect of food components, storage conditions and processing must be considered
when natural antimicrobials are mixed to the food products (Tiwari et al., 2009; Gutierrez
J et al., 2008; Singh et al., 2003; Potter et al., 2005).
Bacteriocin antimicrobials can be inclined by several factors containing the
emergence of bacteriocin-resistant bacteria, oxidation processes or conditions that
destabilize the biological activity of proteins such as proteases, poor solubility,
inactivation by food additives or binding to food components and irregular spreading in
the food matrix in addition to food pH ( Tiwari et al., 2009; Bastos, 2011). Settanni and
Corsetti, (2008) reported that when bacteriocins in mixture with other bacteriocins or
natural preservations were applied to target microorganism that reduced the resistance to
bacteriocins. For example, antimicrobial worth of bacteriocins LABP and LABB from
vegetable pickles Lactobacillus and batter isolates correspondingly, nisin separately and
in groupings were assessed. Bacteriocins LABP and LABB have been showed better
potent in blend with nisin or in combination with other bacteriocins shown than alone.
These bacteriocin combinations were also capable to hinder the growth of Pseudomonas
spp. and E. coli (Settanni and Corsetti, 2008).
2.4 Role of antimicrobial and antioxidant agents in edible coating
Natural antimicrobials extracted from organic sources like plant oils being used
preservation of food from ancient times. Precisely, different coatings that are made from
clove, mint, mustard, cinnamon, ginger, and garlic are all being used. Properties of EOs
such as antioxidants and antimicrobial have earlier been extensively studied. The
mechanism of EOs is not acknowledged entirely yet but it is anticipated that they have
similar mechanism. Demand of those foods which are natural and less treated is being
increased by consumers (Tajkarimi et al., 2010).
Glactomannan and chitosan were used to improve shelf life of cheese because it
has a great influence on the gaseous exchange. Chemical and microbial analysis was done
on it. Coatings showed reduction in oxygen and CO2 of the cheese. These coatings can be
used to reduce the water loss and the colour changes during the storage stability of
cheese. Edible coatings resist against temperature variation during storage of the product,
causes less hardness, improves the appearance and decrease post contamination of
9
cheese. According to Tajkarimi et al. (2010) many spices and herbs are added in edible
coating as natural preservative to save the dairy products against diverse pathogens,
Listeria monocytogenes and mostly plentiful E.Coli. Use of spices and herbs is
inadequate because of its high price and strong odour. A study showed in which ricotta
cheese was coated with a chitosan/whey protein edible film for increasing the shelf life of
ricotta cheese and it was stored under the improved atmosphere at 4oC, the coating had
35% lower oxygen and 21% carbon dioxide permeability and it had three times more
vapour permeability then the chitosan. After 30-days storage there was no change
between recorded pH of the control and coated ricotta cheese (Pierro et al., 2010).
Active edible coating treatments were studied on texture, lipid oxidation, sensory
analysis, color evaluation and moisture loss. Results shows that coated treatments
decreased the oil uptake, moisture loss and the reduction in lipid oxidation. There were
no significant differences in color, smell and taste among the treated and untreated
samples. The edible coated shows better results of chewiness, juiciness, texture and
overall acceptability (Zhong et al., 2014).
Natural essential oils as antimicrobial and antioxidant affect the functional,
textural and sensory properties of both the SC and butter. Natural essential oils increase
the moisture and protein retention; therefore cheeses and butter with greater shelf life can
be obtained. Different synthetic antimicrobial, antioxidant and natural plant extracts have
been used; and they behave differently in SC and butter. Numerous studies reported the
effect of synthetic antimicrobial and antioxidant in SC and butter but the impact of active
edible coating containing natural essential oils has not been studied so far. Keeping in
view this gap, current study was conducted to evaluate the effect of active edible coating
on the storage stability of SC and butter.
2.5 Historical use of essential oils
The word essential oil has been considered to be derived from the name termed in
16th century by the Swiss medicine reformer, Paracelsus von Hohenheim; who named the
active components of a drug Quinta essential. Approximately 3000 essential oils have
been discovered and about 300 of them are of commercial importance as they are used
for their fragrance and flavor (Burt, 2004). The application of essential oils have been in
use since ancient civilizations dating back to Middle east and East and then in Europe and
10
North Africa. In India more than 7000 years hydrosols were used as between 2000 and
3000 B.C, The Egyptians extensively used plants especially the aromatic plants for the
treatment of different diseases while the Persians are thought to have used the hydro-
distillation for the first time in 10000 B.C (Romane et al., 2012).
Even though herbs and spices have been in use since ancient times for their preservative
properties, flavor and aroma (Bauer et al., 2001), still only turpentine oil was stated in
Roman and Greek literature (Guenther, 1948). More than 2000 years ago the process of
distillation was first used in the east for the extraction of essential oil and it was further
developed in 9th century by the Arabs (Bauer et al., 2001). Villanova (ca. 1235–1311), a
Catalan physician is considered to be the first to write authentic information about the
distillation of essential oils. By the 13th century the production of essential oils was
started in pharmacies and their pharmacological properties were described in
pharmacopoeias (Bauer et al., 2001), but they were not widely used in Europe upto 16 th
century.
Distillation of essential oils were first accounted by the Villanova in (ca. 1235–
1311), who is a Catalan physician. Essential oils were first made by the pharmacist and
its effects were pharmacologically described in 13th century in different pharmacopoeias
but its uses are not properly defined or wide spread. The time since Europe traded to city
London was in 16th century (Crosthwaite, 1998). Use and distillation of essential oils
separately described by the two strass burgh physicians Brunschwig and Reiff, that told
about very little amount of compounds you can quantify or distillation in essential oils
that are turpentine, juniper, rosemary, clove, lavender, nutmeg and cinnamon. In 17th
century the use of essential oil was very well known in all pharmacies as many medical
stores have stock of 15 to 20 types of oils mentioned by the French physician (Guenther,
1948). But in 19th and 20th century, gradually its use for flavors and aromas become
secondary preference (Guenther, 1948).
2.6 Essential oils
Essential oils have been studied due to their preservative properties because the
presence of active ingredients in them, they were investigated due to their antioxidant,
anticarcinogenic, antimicrobial and antimutagenic properties (Sánchez et al., 2011).
Essential oils which are also called ethereal or volatile oils are oily liquids which are
11
obtained from the plant material (flowers, bark, leaves, twigs, buds, roots, wood, herbs
and fruit) and have radical scavenging properties (Sumonrat et al., 2008).
Different plant components have been found to possess antifungal properties and
several essential oils have have been reported to be involved in antifungal activities
without any health hazard (Sokmen et al., 1999). Findings of in vivo and in vitro studies
suggest that essential oils can be used as effective antifungal agents (Adam et al., 1998).
The plants were selected on the basis of literature due to their traditional use for curing
infectious dieseases (Panizzi et al., 1993; Crespo et al., 1990). However, there is only
limited information for the antifungal action of essential oils against plant and human
fungal pathogens. The major fungal pathogens of plants include Alternaria, Aspergillus
and Fusarium worldwide (Ghafoor and Khan, 1976). The methods for the extraction of
essential oils include expression, extraction or enfleurage, fermentation while the most
common method used for the commercial extraction of essential oils is steam distillation
(Burt, 2004). The compounds which are responsible for aroma and flavor exist in
different concentration in their natural sources.
The quantity and quality of the essential oils is influenced by the growth
conditions including certain part of plant, climate, soil characteristics and geographical
conditions. As essential oils are relatively volatile materials, the molecular weights are
generally less than 300 and also have common physical properties including high
refractive index, adequate solubility to add aroma, optical activity and solubility in
organic solvents like ether and alcohol (Mukhopadhyay, 2000). Eos as high value high
volume products has been found to be very favorable for the exploitation of the
supercritical fluid technology. Different types of foods processed include: spice extracts,
flavor, essential oils and fragrance from animals, plants and other sources; hop extraction
to obtain essential oils and alpha and beta acids, fractionation and purification of aroma
constituents (Khosravi, 2010).
Components of EOs
The composition of essential oils shows that they contain 85-99% lower
molecular weight compounds which include terpenes, terpenoids, aliphatic and aromatic
compounds (Smith-Palmer et al., 2001). The terpenes having molecular formula (C5H8)n
are naturally occurring hydrocarbons which are derived from isoprene (2-methyl
12
butadiene). Monoterpenes are formed by two isoprene units that represent almost 90% of
essential oils. Aromatic compounds are derived from phenylpropane e.g. phenols
(eugenol), aldehydes (cinnamaldehyde), alcohols (cinnamic alcohol), methylene dioxy
compounds (myristicine, apiole) and methoxy derivatives such as anethole and estragole
(Sánchez et al., 2011; Bakkali et al., 2008). Nitrogenous or sulphured components e.g.
isothiocyanate or glucosinolates derivatives of garlic and mustard oils, are secondary
metabolites of different plants, grilled or roasted commodities. Essential oils contains a
number or active compounds, mainly two of its spices giving a same flavor characteristic
belongs to one botanical category and same appearance that may be varied depending on
its growing region, harvesting time, extraction methods, and other environmental factors
(Bakkali et al., 2008). Different parts of one plant gives a variable composition of
essential oils that are chemical mixtures of terpenoids that are mainly monoterpenes,
sesquiterpens, and diterpens containing aliphatic hydrocarbons, alcohols, acids,
aldehydes, ketones, esters, and mostly C and n containing compounds (Benchaar et al.,
2008). Steam distillation is most frequently used method for extraction of essential oils
but other methods (expression, fermentation, effleurage) accounts for production of
essential oils (Burt, 2004). Terpenoids are made up of mixture r hydrogen and carbon, so
where terpenes also known as hydrocarbon. These are made up isoprene chain and some
terpenes have their links with oxygen. p-cymene, limonene, and pinene etc are examples
of terpenes (Jayasena and Jo, 2013).
Lipid peroxidation which shows the communication between oxygen molecule
and polyunsaturated fatty acids (PUFA), leads to food spoilage, cancer promotion and
aging organisms (Ashok and Ali, 1999). Radicals that take part in lipid peroxidation can
be scavenged by antioxidants due to health protective role of antioxidants (Takao et al.,
1994). They can be used as food additives.
Clove oil
Clove is a condiment that are also used in Asia for its naturally occurring
attributes mostly found in Indonesia Molucca Islands with botanical name Eugenia
Carypphyllata. Active ingredients are present in the clove one of them is eugenol that is
90 to 95% of total content of essential oil have potential for physiological disorders and
show antimicrobial activity (Moreira, 2005 and Briozzo et al., 1989). Oil contents from
13
cloves can be extracted by the distillation method from different portions of clove tree
like stems, leaves, flowers (Mylonasa et al., 2005). Clove essential oils considered GRAS
(Generally Regarded as Safe) for the consumption purposes up to the level of 1500 ppm
in all dairy products by the affiliation of United States of Food and Drug administration
(Kildeaa et al., 2004). World health organization generally recommended the tolerable
consumption level of clove essential oil according to the body weight that is 2.45 mg/kg
BW for human usage (Anderson et al., 1997). It is recorded that for intermediate
moisture food products content of clove essential oil found to resist the growth of
microorganisms up to safe level of product such as SC (Matan et al., 2006). Clove on the
whole is used as a flavoring additive in different food items as well as for dentistry
purposes to reduce the pain (Soto and Burhanuddin, 1995), antilisteric property also seen
in clove for SC and meat products (Menon and Garg, 2001). Gas chromatography is an
analytical technique that can be used for the determination of clove essential oils along
with other essential compounds are also quantified by GC as they also show potential of
antimicrobial and antioxidative properties (Dorman et al., 2000).
Significant reduced growth of Listeria at 70C and 300C were recorded in the
products in which 0.5 to 1% of clove oil concentration level was used. Essential oil of
clove have an active ingredient that if applied as edible coating resist the pathogenic
bacteria growth as a natural preservative. Essential oil of clove and mixture of other oils
when applied in combination on numerous cheese types with edible coatings shown
remarkable effect of antibacterial and antimicrobial as it has been seen a significant drop
off in growth of pathogen Listeria monocytogenes (Smith-Palmer et al., 1998). Having
potential against microbial load and bactericidal effect (Mourey and Canillac, 2002),
clove essential oil also possess other attributes that are antitoxic (Juglal et al., 2002),
antimycotic effect (Mari et al., 2003), antiparasitic (Pessoa et al., 2002)
Ginger
Flavonoids, polyphenols, phenolic acids and tannis are the active ingredients of
herbs and spices (Dawidowicz et al., 2006). Essential oils can be used as a food additive
to enhance the shelf life of products that can be used from plants of angiospermic
families (like Rutacea, asteracea, Lamiacea (Regnault-Roger et al., 2012). Spices and
condiments are the important ingredients of Asian foods being rich in antioxidants. The
14
common is ginger with botanical name Zingiber oficinale belongs to a family
Zingiberaece is being used worldwide (Basak et al., 2010). Ginger is known as medicinal
herb and its 24 family species are being used to treat the ailments related to human and
animal diseases (Bartley and Jacobs, 2000). It has potential to cop up the symptoms of
physiological disorders like asthma, nausea, respiratory tract infection, throat infections,
and other diarrheal conditions (Medoua et al., 2009). Ginger has a rich contents of
polyphenols and antioxidative components so it also exhibits a potential against cancer
(Karna et al., 2012), inflammation (Shim et al., 2011), act as antimicrobial and antifungal
(Alzoreky and Nakahara, 2003; Sharma, 2011). Essential oils extracted from plant
sources are catching attention of industries and other consumers that they can use the
essential oils for the coating of food product to enhance the shelf life of product to make
it more acceptable with value addition of oxidation (Ouattara et al., 2001). Ginger and
ginger rhizomes have phenolic compounds known as gingerols that exhibit the
antioxidative and anti-fungal activity. Being rich in antimicrobial and antioxidant
compounds, incorporating the essential oils in product is very beneficial (Yeh et al.,
2014).
2.7 Current use of essentials oils
The use of essential oils is compulsory declared by European union to apply all
these essential oils in products of food as antimicrobial, flavoring agents, antioxidents,
for perfumes essential oils can be used as (after shaves, fragrances),in pharmaceuticals to
incorporate their functional properties (Van Welie, 1997). Aroma therapy is a technique
in which abundant use of essential oils were found that is about 2% of the total market
depending upon the flavoring contents of essential oils. Different components in EOs
were extracted from plants or any synthetic sources could be used in food products
(Oosterhaven et al., 1995). Diversification in health promoting effect of essential oils like
antibacterial effects in dental caries or tooth aches (Manabe et al., 1987), different
commercial products are available for usage, feed supplements for weaning and lactation
of small babies (Ilsley et al., 2002), even essential oils are used as antiseptics purpose to
control ailments and have germicidal effect (Cox et al., 2000).
Food preservation is a technique used to preserve food for a long time by using
the preservatives that are commercially available e.g. essential oils can play role in the
15
preservation of food with their active ingredients that are available in market. Natural
food preservative that is manufactured by the company DOMCA originated by the S.A,
Alhendi in Spain. is “DMC base” generated the 505 of essential oils from variety of
plants like citrus, rosemary, and round about 50% from glycerol (Mendoza-Yepes et al.,
1997). Different herbs can be used from crops that have essential oils like “Protecta one”
and “Protecta two” blended produced by the breeding of Bavaria Apopka Florida, USA
and safe food additive to be used for consumption. Different solutions of sodium citrate
and sodium chloride are required for the extraction of essential oils as exact quantity of
essential oils cannot be determined, one plant must have two or more ingredients of
essential oils (Cutter, 2000). Essential oils are also used as an antimicrobial and sprout
suppressants in many vegetables especially in potatoes and insect repellents (Carson and
Riley, 1993).
Mechanism of action of essential oil
Antimicrobial activity of essential oils (EOs) is based on their active components
and their chemical compositions of compounds. These components are naturally present
in the plant either active or some enzymes are used for the generation of active
components when their reproductive organs are subjected to biotic or abiotic stress. There
is not a specific mechanism of active components to behave as an antimicrobial activity
but certain reactions happen in the entire bacterial cells that inhibit the growth of
microorganisms this quality is known as essential oil versatility. Essential oils have
different mode of actions and its effect on protecting the food in a number of ways that
are as follows, essential oils degrades the cell wall, it act on damaging cytoplasmic
membrane and its coagulation, it damages the protein membrane further leads from
amplification of permeability to the leakage substances of cell, causing the reduction of
ATP synthesis by reducing ATP pool and increase the hydrolysis by increasing the
membrane permeability (Nazzaro et al., 2013). An important property of essential oils
and their components is their hydrophobicity that separates the mitochondria and
bacterial cell membranes lipid’s, by tormenting the cell memberane and making them
more penetrable. Later on it leads to leakage of ions and other cell substances (Burt,
2004).
16
Hydrophobic molecules are less cooperative to get entry through by the gram
negative cell wall rather than gram positive cell walls therefore, essential oils being
hydrophobic are resistant to gram positive cell walls (Nazzaro et al., 2013). Organisms
that are gram negative bacteria more resistant to the entry of essential oils because of
their cell envelops that are containing complex inner double layer of membranes
contrast the organisms (bacteria and yeast) that have single membrane cell walls made up
of glycoproteins or beta glucan based structure. Capability of partitioning in lipid phase
of membrane, the rate and degree of separation of antimicrobial dissolution also the
factors that contributes to the resistance of essential oils. Hydrophobicity of these two
bacterial groups can be changed as gram negative cell walls have more hydrophobicity
can be replaced by the addition of porin proteins on the outer surface negative bacterial
cells that makes a channels for the passage of smaller molecules through the channel’s
likewise phenolic compounds present in essential oil, allow the entry of compounds into
periplasmic space that are cytoplasmic and glycoproteins layers (Holley and Patel, 2005).
2.8 Role of edible coating in dairy products
Cheese includes in most diverse group of dairy products encompasses diverse
qualities of aromas, textures and flavors resultant of specific ripening protocols and
manufacturer techniques. Extensive and un-controlled fungal and bacterial development
on surface of cheese may be due to the complex composition and prevailing
environmental conditions throughout processing line leads to the economic losses to
industry and health problems to consumers (Oliveira et al., 2007). Storage conditions and
chemical properties of unpackaged cheese affect water losses. Shelf life of cheese can be
enhanced by lowering the rate of water loss furthermore coating of cheese is a more
effective provision to control the prevailing issues (Pantaleao et al., 2007).
Commercially available coatings prepared by non-edible polymers like polyvinyl
acetate (PVA) are being used to control moisture content and microbial growth on final
products of cheese (Reps et al., 2002). Moreover, an increasing demand of consumers
and industries to augment the natural and biodegradable food packaging materials
without chemical preservatives by consumers and packaging industries (Cerqueira et al.,
2009). Being edible and biodegradable, coatings and whey protein films have gathered a
special attention because they offers an upgrade whey as a byproduct of cheese
17
manufacture industry. It is clear from previous review that whey protein products offers a
pure form of whey protein isolates (WPI) that possess mechanical features and retain
oxygen barrier properties (Mulvihill and Ennis, 2003) when compared to the best
polymers based films (Perez-Gago and Krochta, 2002; Khwaldia et al., 2004; Ramos et
al., 2012a). Whey protein isolates perform several functional qualities such as containing
microbial compounds that controls increased concentration of active ingredients
(Campos et al., 2011) further prolongs safety and shelf life of packaged (Ponc et al.,
2008). Moisture barrier, controlled oxidation of lipids, prevention in loss of aromas and
volatile compounds are some selective functional qualities of whey protein isolates
(Kester and Fennema, 1986). To extend the shelf life, applications of edible coatings on
different cheese products have been rarely explored. Kampf and Nussinovitch, (2000)
reported that the use of capa-carrageenan, gellan and alginate coatings in cheese products
whereas Cerqueira et al. (2010) described the use of chitosan and galactomannan
coatings on semi hard cheese products that showed the reduction of after coating
applications. Chitosan coatings with mixtures of lysozymes and natamycin also applied to
different cheese products packages (Mozzarella and Saloio cheese) that are helpful to
control post-packaging contamination and enhance shelf life by improving microbial
safety (Duan et al., 2007 and Fajardo et al., 2010).
To prevent the microbial growth on cheese surface, whey protein isolates can
serve as a carrier of lactic acid natamycin or chito - oligosaccharides but no work has
been done till date. Purposely, different combinations of antimicrobial coatings with base
matrix made up of WPI (10%, wt/wt), glycerol (5%, wt/wt), guar gum (0.7%, wt/wt),
sunflower oil (10%, wt/wt), and Tween 20 (0.2%, wt/wt) along with antimicrobial
compounds natamycin (0.25 g/L) and lactic acid (6 g/L), natamycin (0.25 g/L) and COS
(20 g/L), and natamycin (0.25 g/L), lactic acid (6 g/L),and COS (20 g/L). On the basis of
previous assumptions these formulation were selected coupled with struggle (Ramos et
al., 2012c, d, 2013), to optimize the antimicrobial coatings (Ramos, 2011). The efficiency
of these formulations were further analyzed against microbial, physicochemical and
microbiological features up to 60 days of storage and obtained results were compared
with the commercial coated and un-coated cheese. To check the effectiveness of these
formulations salio cheese was selected as it is consumed more and practiced with high
18
spoilage or pathogenic microflora on cheese surface leads to unfavorable mouth feel and
flavor. Besides all these issues, only found solution relied on use of PVA coatings
incorporated with antifungal agent (natamycin) onto its surface, hence our edible coating
was tested as a realistic alternative for actual shelf-life extension. Essential oils can also
be used as best source to develop coatings against spoilage bacterial or pathogenic
microflora. Another study conducted by Samy, (2011) who reported that addition of
clove oil with MIC of 2.0% in Feta cheese stored at 7°C for 14 days showed highly
significant antibacterial effect for E.coli and Vancomycin Enterococci.
2.9 Cytotoxicity and legal aspects for the use of EOs
Cytotoxicity
Hydrophobicity and due to diversity of different chemical compound groups
present in essential oils have numerous mechanisms mainly responsible for their
antibacterial activity (Skandamis and Nychas, 2002; Carson et al., 2002). Mainly
terpenes in essential oils have ability for the disruption of cell wall. Due to heavy leakage
of critical molecules cell death arises as a result of infiltration of terpenes to the lipid
structure of cell membrane that leads to the destruction of cell membrane and
denaturation of protein (Burt, 2004). Permeability of cytoplasmic membrane to ATP was
increased due to the releasing of lipopolysaccharides in high percentage (Sanchez-
Gonzalez et al., 2011). Nevertheless, volatile components of essential oils do not display
same mechanism of action all times according to Wendakoon and Sakaguchi (1995) with
the binding of proteins inhibition of amino acid decarboxylases occurred when
Cinnamon essential oil and its components against Enterobacter aerogenes was used.
Moreover, Origanum vulgare essential oil was also used in the suppression of
Staphylococcal enterotoxins (De Souza et al., 2010). In most of the microorganisms
cytotoxic effects of essential oils were witnessed. Smith-Palmer et al. (2001) studied that
due to absence of external membrane defense of cell wall in GP bacteria that prevents
dispersion of hydrophobic compounds so that Gram-positive bacteria are somewhat extra
sensitive to essential oils as compared to Gram-negative bacteria. In contrast, mint
essential oil has showed lesser reduction in the viable count of L. monocytogenes as
compared to S. enteritidis (Tassou et al., 1995). Dorman and Deans (2008) disclosed
diverse action of individual components which were present in essential oils against
19
Gram-positive and Gram-negative bacteria. This statement is consequence variation in
the chemical composition of essential oils due to their harvesting period and geographical
basis (Sanchez-Gonzalez et al., 2011). Amongst Gram-negative bacteria Pseudomonads
particularly P. aeruginosa was showed minimum sensitivity to the essential oils action
(Burt, 2004). The natural antimicrobial activity of essential oils is allocated to the
interactions, chemical configuration and components proportion (Dorman and Deans,
2008; Delaquis et al., 2002). In fact their biological properties are the effect of a
synergism of all components. It has been revealed that antimicrobial activity of essential
oils were better than the combination of the major components (Gill et al., 2002; Mourey
and Canillac, 2002). Even minor components have been showed their major role and
synergism phenomena occur. For instance, blend at concentration of eugenol 500 μg/ml
and cinnamaldehyde 250 μg/ml was used for growth’s complete distraction of Bacillus
sp., Micrococcus sp., Enterobacter sp and Staphylococcus sp for one month, but separate
substrates did not stoped the growth of microorganisms (Moleyar and Narasimham,
1992).
Legal aspects for the use of EOs
FDA and European Union (EU) are continually producing the most appreciated
laws and recommendations on the use of essential oils in several industries. Numbers of
essential oils have been registered by FDA and EU Commission and for their use in
agriculture and food industries to overcome the harmful effect of pathogenic
microorganisms. The EU Commission registered EOs, which means that FDA has
categorized these substances as generally recognized as safe (GRAS) (Bajpai et al.,
2011). For use as flavourings in food products several essential oils components have
been registered in the Commission of EU. The registered flavourings are believed to
existing no risk for the health of the consumer and comprise amongst others carvacrol,
citral, p-cymene, carvone, limonene, cinnamaldehyde, thymol, eugenol and menthol.
Later in 2001 methyl eugenol and estragole were deleted from the list due to their
genotoxic effect.
Spices are supposed as Class I preservative according to Food Safety and
Standards (Food Products Standards and Food Additives) regulations 2011 In India.
Adding of Class I additives in any food product is not prohibited, unless otherwise
20
provided in the regulations. Food contamination is massive public health problem all over
the world, but it could be overcome by the use of natural additives like essential oils
those are acquired from spices. In the recent years the statement that many essential oils
owns antimicrobial activity which has been demonstrated by quite investigations. The
optimal concentration and type of essential oils reliance on the food product used and
against which type of fungi or bacteria it is to be used. But if essential oils are commonly
applied as antifungals and antibacterial in any food product, then the organoleptical
influence should be considered as the use of naturally derived preservatives can be
increased acceptable flavor thresholds and vary the taste of food product. Therefore,
exploration in this area should be focused on the escalation of essential oils combinations
and utilization to attain effective antimicrobial activity at necessarily low concentrations
so as not to badly affect the organoleptic acceptability of the any food product.
21
CHAPTER-3
MATERIALS AND METHODSCurrent study was conducted to evaluate the effect of active edible coating on
shelf stability of soft cheese (SC) and butter; whey powder (WP) was used as base
material for SC and corn starch (CS) for butter coating. Ginger and black cumin essential
oil were used as active ingredient in butter coating while pepper mint and clove essential
oil in SC coating to prolong shelf stability. Analyses and trials for manufacturing of SC
and butter were performed in the D.T. Lab., NIFSAT, U.A.F. and Department of Food
Science, UMass, Amherst, U.S.A. Products were analyzed for different physicochemical,
microbiological, antioxidant parameters and sensory evaluation was also done during
storage at 2-50C temperature. Best treatment was selected on the basis of sensory
evaluation and shelf stability.
Table 3.1 Treatment plan for SC
Treatment Soft cheese coating Clove oil (CO) (%) Pepper mint (PMO) (%)
T0 (Control) _ _ _T1 + _ _T2 _ 0.5% _T3 _ 0.75% _T4 _ 1.0% _T5 + 0.5% _T6 + 0.75% _T7 + 1.0% _T8 _ _ 1.5%T9 _ _ 2.0%T10 _ _ 2.5%T11 + _ 1.5%T12 + _ 2.0%T13 + _ 2.5%
3.1 Procurement of raw materials
Raw buffalo milk was used for SC production obtained from the Dairy Farm,
U.A.F. Pakistan and cream for butter from local dairy farm of Faisalabad. Corn starch
(CS) from Rafhan Maize products Co. Ltd., xanthan gum and whey powder (WP) was
procured from local market of Faisalabad. All the chemicals and reagents used in the
22
study were purchased from local market. Lyophilized thermophilic starter cultures
(Lactobacillus Bulgaricus and Streptococcus Thermophilus) used for cheese
manufacturing from SACCO and rennet (chymosin) used for the coagulation of milk.
3.2 Manufacturing of active edible coating for SC
Edible coating for SC and butter was manufactured by following the method of
Yongling et al. (2011) with little bit modification. Four gram WP as base material of
coating, 2g xanthan gum to enhance the viscosity and 0.5 g lecithin as emulsifier was
taken in a 250 mL beaker. Eight ml glycerol as plasticizer was added in beaker and then
made the total vol. 100 mL by the addition of distill water. Continuous stirring was done
with sitter to homogenize the coating solution. Then put the beaker in water bath at 55 to
60 ºC for 8 min. to make the coating solution viscous and uniform. Cooling was done by
covering the beaker with aluminum foil at room temperature. After this three different
concentrations (as mentioned in Table 3.1) of clove and peppermint oil were added as
active ingredient in coating solution.
3.2.1 Analysis of active edible coating
3.2.1.1 pH
Method of Ong et al., (2007) was used to measure the pH of edible coating
solution. First of all take 10 milliliter (ml) of edible coating solution into the beaker then
placed it into the water bath and maintain at 50oC temperature to melt the coating, then
allowed it to cool at normal temperature. The buffer solutions were used for the
calibration of pH meter (Inolab WTW series 720) at 4 and 7 pH then measured the pH of
the edible coating solution at room temperature.
3.2.1.2 Acidity
According to the method of AOAC (2000), the acidity of coating was measured.
For this purpose 10 milliliter (mL) of edible coating solution was taken into 100 milliliter
(mL) of flask followed by the addition of 2-3 drops of phenolphthalein, titrated it against
0.1 normal solution of NaOH until pink color was appeared. The formula that mentioned
below was used to calculate the percentage of acidity
3.2.1.3 Viscosity
To determine the viscosity of edible coating solution, viscometer DV-I Brookfield
(LVDVE 230 model, Laboratories of Brookfield Engineering, Middleboro, MA) was
23
used according to the method that is followed by (Gassem and Frank, 1991). At 4-60°C
temperature, the viscosity of edible coating solution was determined. For this purpose, at
60 rpm the spindle number 1 was used as a measuring scale. The reading of viscometer
was calibrated in cp i-e centipoises as well as in torque percentage during 30th second.
3.2.1.4 Water Activity
For the determination of water activity, electronic hygropalm that is a water
activity meter (Aw-Win, Rotronic model, Karl-Fast probe well equipped) was used
followed by the method that described (El-Nimr et al. 2010).The water activity meter was
standardized to analyze the 3 milliliter (ml) of edible coating solution. After that the
reading was noted at room temperature from digital display.
3.3 Preparation of SC
a. Milk fat standardization
First of all de-creamed the raw buffalo milk and then by Gerber method determine
both the fat content of milk and cream. After that Pearson’s Square method was used to
standardize the milk at 3% fat for SC manufacturing.
b. Physicochemical analysis of milk
The milk samples were analyzed for physicochemical analysis following the
method of AOAC, (2000) as described in section 3.4.1.
Flow line of SC
Raw milk
Standardization of milk at 3% Fat
Pasteurization at (65ᵒC for 30 min.)
Addition of starter cultures @ 0.02% (31C)
Incubation at 31oC for 30-40 min.
24
Addition of rennet @ 1g/50L (31ᵒC 40-50 min)
Cutting the curd
Cooking the curd at 37-38 ºC for 20-25 min
Whey removal
Molding
Pressing (5Bar; 12-18 Hrs.)
Edible coating of cheese
Packaging
Storage at 2-50C
Fig. 3.1 Flow line for manufacturing of SC (SC)
3.4 Quality assessment of SC
3.4.1 Physico-chemical analysis of SC (SC)
3.4.1.1 Moisture content
The moisture of SC sample was computed by followed the standard method
(Method number 926.08: AOAC, 2000). For this purpose, the sample was placed in hot
air oven at about 100-106°C unless constant weight was recorded. For this purpose the
dish bottom was covered with sand and kept the dish in oven for one hour at 104ºC. Then
25
removed it from oven and kept into desiccator for cooling. The dish wt. was recorded and
label it W1. In next step 2 gram grated cheese sample was taken in a dish and label it as
W2 and then with the help of glass rod blend it properly with sand. Then put the china
dish in an oven for 5 hours at 104 ºC. After 5 hours, the dish was removed and left it in
desiccator for some times unless it is cooled, then measured the weight that is W3.
The moisture contents was computed by followed the below mentioned formula.
W2 (SC weight in grams) - W3 (dried cheese weight in grams)Moisture percentage = ----------------------------------------------------------------------------- x 100 W2 (SC weight in grams) 3.4.1.2 Fat content
Gerber method was used to determine the fat content in SC sample as described
by Marshal, (1993). In cellophane bag that is smear impervious enveloped about 3 gram
grated SC, take the SC sample in butyrometer followed by the addition of 10 milliliter
(mL) of H2SO4 and distilled water 3 milliliter (ml). At 60ºC temperature, 5 milliliter (ml)
distilled water was added alongside the butyrometer wall and at the end added 1ml of
amyl alcohol. The butyrometer was strongly shaken to completely dissolve the cheese
then placed it in water bath for 15 minutes at 85ºC for thorough digestion. After heating,
at 1300rpm butyrometer was centrifuged for 5 minutes, subsequently at 65 ºC kept in
water bath for 5 minutes and reading was noted for measuring the fat percentage.
3.4.1.3 Protein content
The protein in SC was determined through standard system of Kjeltech as
defined in AOAC (2000).Take SC sample about 1 gram in digestion flask that also
containing 25 milliliter (mL) of H2SO4 and two digestion Tablets. Sample was digested
unless solution became clear. After that distilled water was added to dilute the solution
and made the volume up to 250 milliliter. The NH4 trapped in H2SO4 during distillation
process that removed when added 40 percent NaOH solution and collected in the solution
of boric acid. Then, for the determination of N2 boric acid was titrated against 0.1 normal
solution of H2SO4.
Amount of N2 in percentage were computed followed by below mentioned formula;
0.0014 x H2SO4 Vol. in milliliter (as a blank sample) Nitrogen in percentage = -------------------------------------------------------------------- x 100 Weight of SC
26
Protein percentage in crude form = 6.38 x Nitrogen percentage 3.4.1.4 pH
Method of Ong et al, (2007) was used to determine the pH of SC sample. 20 gram
of SC blended with 12 milliliter (mL) of distilled water to prepare the slurry and used it
to measure the pH at 25oC with the help of pH meter (Hanna, HI-99161). Before use the
pH meter was calibrated with buffers of 4 and 7pH.
3.4.1.5 Acidity
According to the standard titration method of AOAC (2000) the method no. 920.124,
the acidity of SC samples was measured. For this purpose 1gram of cheese was blended with
slightly warm water and made the vol. up to 10 milliliter (ml), strongly blended or shaked
and filtered it. Then this filter solution was titrated against 0.1 normal solution of sodium
hydroxide, phenolphthalein used as indicator to obtained the pink color as an end point
Vol. of NaOH in mL × 0.009 Acidity percentage (as lactic acid) = ----------------------------------------------- × 100 SC weight in grams
3.4.1.6 Ash content
To determine the ash contents in SC, samples were burnt followed the method of
AOAC (2000) as described in method No. 935.42. For this purpose take 2 gram of cheese
sample, burnt it on flame till the smoke was disappeared. After this, it was kept in muffle
furnace for 4 hours at 550oC unless constant ash weight was obtained.
Ash weightAsh in percentage = ---------------------- × 100 Sample weight
3.4.2Color analysis
Method of Hunterlab, (2011) was used to determine the color of SC by using
Miniscan porTable colorimeter (Associates Laboratory of Hunter, Inc., Reston, VA). For
this purpose black and white plates kept in elastic bags for standardization of color
(QME355 3.5 mil, Vilutis & Co., Inc., Frankfurt, IL) that used for cheese. Determination
of color through ‘CIE (1978) *L (lightness) a & b values’ used by illuminant D 65. This
indicated the red (+) & green (-) and *b was indicated yellow (+) & blue (–).
3.4.3 Water Activity (aw)
aw of SC samples were determined according to the same method as described in 3.2.1.4.
27
3.4.4 Weight Loss
Method of Cerqueira et al. (2010) was used to determine the weight loss in SC
samples during storage.
Method
• Every cheese sample was measured in crystal tarred plates by means of weighing
balance.
• These plates were kept in fridge and after five days difference again measured.
• Loss of weight in grams or kilograms was computed by followed the below
mentioned formula:
Amount of Moisture in ‘g Kg-1’ = A−B ×1000A
Whereas,
A= initial wt. of sample
B= after dehydrating in drying oven at 70 °C, the final sample wt.
Loss of weight in percentage was computed followed the below mentioned formula:
Loss of weight in percentage= *DM / DM1 ×100 Whereas:*DM= wt. on 1st day
DM1 = wt. on subsequent days of examination
3.4.5 Assessment of proteolysis by RP-HPLC
Method of Verdini et al. (2004) was used to determine the freeze dried insoluble
part (pH 4.6) of SC samples during proteolysis. The freeze dried sample about 10
milligram was mixed in 0.5 milligram (mL) of urea that in solution form contained (urea
48 gram, Tris 2 gram, sodium citrate 1.3 gram & mercaptoethanol 300 microliter per 100
millilter) and TFA 1.5 milliliter per water solution. After mixing, this solution was
filtered through a filter of 0.2 micrometer (Fisher Scientific, Ireland) then inserted about
20 microliter in the system of HPLC (Shimadzu, Japan). A Shim-pack ‘CLC-ODS’ C-18
column, 250 centimeter × 4.6 millimeter, 5 micrometer col.(for chromatographic partition
at ambient temperature. At 214 nm wavelength, detection was carried out.
With solvent ‘A’ 0.1 percent trifluoro-acetic acid i-e TFA, the gradient elution
was used in water and for solvent ‘B’ 0.1 percent trifluoro-acetic acid in acetonitrile. The
gradient program was, original configuration 0 percent ‘B’ isocratic elution at 0 percent
28
‘B’ for 5 min., at linear step 25 percent ‘B’ in 5 min., at linear step to 35 percent ‘B’ for
30 min., at linear step to 50 percent ‘B’ in 10 minutes, at isocratic elution at 50 percent
‘B’ for 10 min. Stream rate was 1.0 mL per min.
3.5 Butter
3.5.1 Physicochemical analysis of cream
Cream was analyzed for pH (Ong et al., 2007), acidity (AOAC, 2000) as described in
section 3.4.1 and free fatty acids (FFA) according to the method of AOAC (1990).
Table 3.2 Treatment plan for butter
Treatment Butter coating Black cumin oil (BCO) (%)
Ginger oil (GO) (%)
T0 (Control) _ _ _T1 + _ _T2 _ 0.2% _T3 _ 0.3% _T4 _ 0.4% _T5 + 0.2% _T6 + 0.3% _T7 + 0.4% _T8 _ _ 1.5%T9 _ _ 2.0%T10 _ _ 2.5%T11 + + 1.5%T12 + + 2.0%T13 + + 2.5%
3.5.1.1 Free Fatty Acid (FFA)
According to the standard method of AOAC (1990) free fatty acids were
determined in cream. The percentage of free fatty acid was measured through titration, 95
percent ethanol was used to neutralize the cream against potassium hydroxide solution
(KOH). For this purpose, in a sterilized conical flask take the 10 gram cream sample with
the addition of 25 mL of 95 percent neutralized ethanol. It was appropriately mixed so
that ethanol was fully dissolved, then added phenolphthalein 1-2 drops as an indicator
and titrated it against 0.1 normal potassium hydroxide (KOH) solution, mixed it
continuously till the pink color became Table. The percentage of free fatty acids was
computed as follows:
% FFA = Vol. of alkali in mL ×28.2 × N ×100 Wt. of sample in gram
29
Whereas:
N = Normality of alkali (KOH) used
3.5.2 Preparation of active edible coating for butter
Method of Yongling et al. (2011) with slight modification was used for the
manufacturing of natural active edible coating solution for butter. Three gram CS, 2g
xanthan gum and 0.5 g lecithin was taken in a 250 mL beaker. Eight ml glycerol as
plasticizer was added in beaker and then made the total vol. 100 mL by the addition of
distill water. Continuous stirring was done with sitter to homogenize the coating solution.
Then put the beaker in water bath at 60-65 ºC for 8-10 min. to make the coating solution
viscous and uniform. Cooling was done by covering the beaker with aluminium foil at
room temperature. After that three different concentrations (as mentioned in Table 3.2) of
black cumin and ginger oil was added as active ingredient in coating solution and then
applied on butter.
3.5.3 Manufacturing of Butter Raw Cream
Pasteurization of cream at 63oC for 15 min
Aging of cream by keeping in incubator for a night
Churning of cream
Butter
Edible coating of butter
Packaging
Storage at 2-5 0C refrigeration temperature
30
Fig. 3.2 Flow line of manufacturing of Butter
3.5.4 Quality assessment of butter
3.5.4.1 Physico-chemical analysis of butter
Butter samples were analyzed for moisture, pH, acidity, fat, ash, water activity,
color analysis and weight loss according to the same methods as described in section 3.4.
3.5.5 Free Fatty Acid (FFA)
FFA of butter samples were determined according to the same method as
described in section 3.5.1.1.
3.6 Textural profile analysis of SC and butter
The texture profile analysis was conducted to check the texture of butter and SC.
For this purpose followed the methods that described by Zisu and Shah, (2007) by using
the compression plate test through Texture Analyzer (TA-XT plus) (Godalming,
STableMicro Systems, Surrey, UK). In air close-fitting plastic bags, butter and cheese
samples was placed and equilibrated for 18 hours at 8oC. Then, before analysis at 8ºC
made the cubes of all the samples with the help of stainless steel cutter. After this,
removed sample from incubator and instantly compressed about 30 percent of the initial
height in two successive cycles at the rate of one mL per second that is double
compression.
3.7 Total Viable Count in SC and butter sample
According to the method of APHA, (1992) total viable count (TVC) of butter and
SC was calculated.
Preparation of normal saline solution
The normal saline solution with the help of 8.9 gram per liter of sodium chloride
mixed in 1liter of distilled water was prepared and at 121oC autoclaved it for 15 minutes.
Media preparation
Nutrient agar was prepared by mixing the 2.8 gram nutrient ager in one liter of
distilled water, autoclaved it for 15 minutes at 121oC.
Sample preparation
Six to seven test tubes that are already sterilized were taken and categorized as 10 -
1 to 10-6 and took 9 mL of normal saline solution that was poured into all test tube one by
one. With the help of micropipette, transferred 1 mL of uniform sample into the 1 st tube
31
and properly mixed it. Then sample was moved from 1st tube to 2nd one. Sample was
inverted about five times to mix the ingredients properly. Then taken one mL of sample
and moved to the next tube and continued this process unless desired dilution was
obtained. This serial dilutions for every sample was done by the method described by
APHA (1992).
Pouring the plate
Transferred 0.5 mL of sample contents from every test tube on the surface of
nutrient agar and dispersed the sample properly through spreader and incubated these
samples for 24 hr. at 37°C.
Colony counting
The normal colony count from dilutions that presented the range of colonies from 30-300
cfu/g that measured with the help of colony counter.
TVC = Dilution factor x Ave. No. of colonies Vol. factor
3.8 Antioxidant analysis of SC and butter
3.8.1 2,2-Diphenyl-1-picrylhdrazyl (DPPH) scavenging activity
To check the free radical scavenging activity of butter and cheese samples
spectrophotometer was used at 517 nm wavelength. Method described by Ghafoor et al.
(2010) was used for the determination of 2, 2-Diphenyl-1-picrylhdrazyl (DPPH)
scavenging activity in butter and cheese samples. The solution of DPPH was prepared by
mixing it in methanol. The treatment sample in different concentration was taken about 5
to 25 milligram in test tube and introduce to the DPPH solution for reaction. After this
shake the test tube for complete mixing of DPPH solution, then placed the test tube in
dark room at about half an hour then checked the absorbance at 517 nm in
spectrophotometer. Butylated hydroxy toluene (BHT) was used as a standard in
spectrophotometer. The percentage of radical scavenging activity was checked according
to the equation mentioned below. The scavenging of free radical activity for every sample
can be indicated in percentage reduction in DPPH due to specified quantity of every
extract.
3.8.2 Peroxide value
For the determination of peroxide value (POV) in butter and cheese samples
followed the standard methods of AOAC (2005). For this purpose in 250 milliliter of
32
Erlenmeyer flask took the 5 gram of sample. Through continuous mixing homogenize the
sample after the addition of 3:2 vol.by vol. ratio of 30 % acetic acid-chloroform solution.
Then sample was filtered through whatman filter paper and added the 0.5 percent of 0.25
milliliter of saturated potassium iodide and properly mixed it and transferred to the
burette, unless yellow color was disappeared. Then titrate it against 0.02 Normal solution
of sodium thiosulfate 25 gram/liter concentration. After this starch solution was added,
and continue the titration unless blue color became vanished. In identical manner a blank
observation was recorded. Then POV was determined through the equation.
P0V (meq/kg) = ____(S-B) x N x1000 Wt. of sample (g)Where:
N = Normality of Na2S2O3
B = Volume of Na2S2O3 used for blank
S = Volume of Na2S2O3 used for sample
3.8.3 Measurement of Thiobarbituric acid reactive substance (TBARs) value
Method of Luo et al. (2011) was used to determine the (TBARS) value that is
thiobarbituric acid reactive substance in butter and cheese samples. For this purpose took
the 5gram sample and dispersed it in 20 milliliter of thiobarbituric acid solution
(0.37percent thiobarbituric acid, 15percent trichloro-acetic acid and 0.25mol/liter HCl).
Then for 10 minutes heated the sample in boiled water, cooled with water; then
centrifuged it for 20 minutes at 3600 rpm and ambient temperature. The absorbance was
checked at 532 nm in spectrophotometer. By the use of malondialdehyde (MDA), the
standard curve was checked & TBARS were stated as mg MDA/kg sample.
3.9 Sensory evaluation of SC and butter
According to the Ganesan et al. (2014) sensory evaluation of butter and SC was
done by a panel of nine inspectors drawn from faculty members and the students of post-
graduate of the institute. The inspectors were nominated on the basis of their interest in
sensory evaluation. The butter and SC samples were assessed for different parameters
flavor, odor, texture, overall acceptability and appearance. The samples were offered to
inspectors as 20 gram of butter and cheese. The Performa for sensory evaluation is given
in Appendix.
33
3.10 Statistical analysis
The data was statistically analyzed by using the SPSS 19 software at p < 0.05
confidence level explained by Steel et al. (1997) [18] to estimate the effect of active edible
coating and essential oils on acceptability and quality of SC and butter. Experiments were
recurring three times, standard deviations reported as a result of three times the samples
have been analyzed and inspection was done at least in duplicate. Tukey’s multiple range
test was used for the identification of significant differences between mean values.
34
CHAPTER-4
RESULTS ANDDISCUSSION
The current study was conducted in D.T. Lab., NIFSAT UAF, Faisalabad-
Pakistan and Department of Food science, UMass Amherst, USA. The study was done to
evaluate the impact of active edible coating on sensory attributes and physicochemical of
SC and butter. CO and PMO were used as active ingredient in whey protein based edible
coating for SC while black cumin and ginger oils were used as active ingredient in corn
starch based edible coating for butter. Three different levels of pepper mint (1.50, 2.0,
2.50%) and clove (0.5, 0.75, 1.0%) essential oil were used directly in SC and indirectly
by adding these concentrations in edible coating developed for SC. Similarly three
different levels of black cumin (0.2, 0.3, 0.4%) and ginger (1.5, 2.0, 2.5%) essential oil
were used directly in butter and indirectly by adding these concentrations in edible
coating developed for butter.
4.1 Physico-chemical analysis of milk
Raw milk used for SC manufacturing was evaluated for pH, ash, moisture,
acidity, protein, fat and total solids contents (Table 4.1).
4.2 Analysis of edible coating (EC)
Results of active EC are shown in in the Table 4.2.
35
Table 4.1 Compositional analysis of milk
Parameters Concentration
Fat contents 4.34±0.03%
Protein 3.59±0.02%
pH 6.8±0.02
Ash 0.78±0.02%
Moisture 86.75± 0.02%
Acidity 0.16±0.002%
Total solids (TS) 20.2±0.01%
Table 4.2 Analysis for edible coating of SC
Parameters Composition
Water Activity 0.65 ± 0.02
Viscosity 73 ±0.04cp
Acidity 0.17±0.03 %
pH 4.70±0.60
36
4.3 Physicochemical parameter of SC
4.3.1 Moisture
Moisture is a major constituent of SC plays an important role in imparting
different attributes like softness, and other quality characteristics that classify cheese into
different types. It also serves as major substrate for reactants and affects the textural or
sensory qualities of cheese (Lee et al., 2004).
The result of means squares for % moisture of SC samples presented in Table
4.3.1a. Results revealed that moisture contents of SC due to treatments, storage days and
their interactive effect showed a * (p<0.05), ** (p<0.01) and NS (p>0.05) effect
respectively. The mean values for moisture contents of SC are given in Table 4.3.1b.
Results revealed that active edible coating and essential oils have ** (p<0.05) effect on %
moisture of SC. Data shows that the highest moisture (79.55 %) was noted in SC sample
T9 (SC containing 2.0% peppermint oil) and T10 (SC formulated with 2.5% peppermint
oil) at 1 day while the lowest moisture (71.30%) was noted in control (T0) at 30days.
Results revealed that the SC samples which were prepared with whey protein based
active edible coating (containing clove and peppermint oil as active ingredient) and
cheese samples with direct incorporation of these essential oils show minimum moisture
loss during storage of 30 days as compared to control (T0) due to addition of EOs and
active edible coating.
Fig. 4.1 depicted that during storage of 30 days a regular decrease in moisture
content of SC was recorded in all samples which were formulated with active edible
coating and by direct addition of clove and peppermint essential oils. Mean SC values
shows that moisture was decreasing from 79.55% to 77.10%, 78.50% to 74.60% and
76.50% to 71.30% at 1, 15 and 30 days respectively.
Edible coating serve as a barrier against moisture loss by covering the surface of
SC samples so minimum moisture loss was noted in edible coated samples of SC.
Eugenol in CO and menthol in PMO have the antioxidant and antimicrobial potential so
binds the free radicals in SC samples that ultimately leads to minimum moisture loss as
compared to control (T0). The decline in moisture during storage of SC might be due to
the change of physical state of water in cheese because during storage the water gets
chemically bound which reduce the moisture
37
Table 4.3.1a MS for moisture contents of SC
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
351.428 104.022 17.829 303.594 776.873
175.714 8.002 0.686 3.614
48.62** 2.21* 0.19NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
66 68 70 72 74 76 78 8074.6
77.7577
77.677.75
7878.23
78.576.5
77.577.6
7878.4
78
71.376
74.275.2
75.575.75
7676.5
7475.275.475.5
75.7576.25
30 day 15 day 1 day
Cheese Moisture
Treatm
ent
Fig. 4.1 Effect of edible coating and EOs on the moisture (%) of soft cheese
38
Table 4.3.1b Effect of treated SC on the % moisture during storage
Treat Days Total
1 day 15 day 30 day
T0 77.10±0.514ab 74.60±0.912ab 71.30±0.779b 74.33±0.920B
T1 79.25±0.716a 77.75±1.241a 76.00±0.722ab 77.67±0.660A
T2 79.50±0.814a 77.00±1.438ab 74.20±0.797ab 76.90±0.931AB
T3 79.55±1.247a 77.60±1.311a 75.20±1.201ab 77.45±0.888A
T4 79.50±1.218a 77.75±1.207a 75.50±0.670ab 77.58±0.786A
T5 79.25±1.536a 78.00±0.953a 75.75±1.074ab 77.67±0.794A
T6 79.40±0.635a 78.23±0.918a 76.00±1.426ab 77.88±0.722A
T7 79.50±1.391a 78.50±0.791a 76.50±1.149ab 78.17±0.720A
T8 79.50±0.808a 76.50±1.253ab 74.00±1.449ab 76.67±0.996AB
T9 79.55±0.525a 77.50±1.005ab 75.20±0.918ab 77.42±0.756AB
T10 79.55±0.508a 77.60±1.484a 75.40±1.282ab 77.52±0.837A
T11 79.25±0.958a 78.00±0.589a 75.50±1.039ab 77.58±0.707A
T12 79.30±1.143a 78.40±1.426a 75.75±1.455ab 77.82±0.859A
T13 79.40±1.455a 78.00±1.507a 76.25±0.727ab 77.88±0.786A
Total 79.26±0.246A 77.53±0.300B 75.18±0.311CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
39
content in SC. The findings of this parameter match with the results of Tarakci and
Kucukoner (2006), they also noted that moisture in SC decrease during storage.
4.3.2 Fat contents
Flavor is mainly contributed by the presence of fat in cheese that improves
appearance, texture, structure and overall acceptability of cheese (Kucukoner and Haque,
2006). Stretching and melting properties are one of major attribute of fat that adds up to
cheese (Petersen et al., 2000).
The results of mean squares for % fat of SC samples are presented in Table
4.3.2a. Results revealed that storage days have ** (p<0.01) effect while treatments and
the interactive effect of treatments and storage days have NS (p>0.05) effect in % fat of
SC. The mean fat values for SC are given in Table 4.3.2b. Data showed that the highest
fat (3.34%) was noted in T1, T5, T6, T8 and T12 SC samples at 30 days while the lowest fat
(3.22%) was noted in T0 SC at 1day formulated with 3% milk fat. Results revealed that
there is NS difference among the SC samples which were formulated with WP based
active edible coating (containing clove and peppermint oil as active ingredient) and
cheese samples with direct incorporation of these essential oils have more fat as
compared to (T0) due to addition of EOs, active edible coating and fat retention property
of xanthan gum in coating.
Fig. 4.2 depicted that during storage of 30 days a minute increase in %fat was noted in
SC samples, which were formulated with active edible coating and by direct addition of
clove and peppermint essential oils. Mean value of SC samples shows that fat was
increasing from 3.23% to 3.27%, 3.25% to 3.28% and 3.32% to 3.34% at 1, 15 and 30
days respectively. Mean values of fat for all SC samples at 15 and 30 days of storage
showed a NS increase.
Results shows that minor increase in fat of SC that is might be due addition of PMO
and CO in the SC. Eugenol and menthol have the excellent antioxidant potential so binds the
free radicals in SC samples that ultimately leads to minimum lipolysis in SC samples as
compare to T0. The results of fat are in accordance with the Lopez et al. (2007), they also
reported that fat during storage remains almost same. During storage combination of
complex biochemical changes and microbiological changes occur such as proteolysis,
lipolysis and glycolysis (Farkye, 2004).
40
Table 4.3.2a MS for fat contents of SC
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.17091 0.00478 0.00813 1.25347 1.43729
0.085460.000370.000310.01492
5.73** 0.02NS
0.02NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
3.2 3.22 3.24 3.26 3.28 3.3 3.32 3.34 3.363.27
3.263.26
3.263.26
3.253.253.25
3.283.283.28
3.263.25
3.26
3.343.34
3.333.33
3.323.343.34
3.333.34
3.333.32
3.333.34
3.32
30 day 15 day 1 day
Cheese Fat
Treat
ment
Fig. 4.2 Effect of edible coating and EOs on the fat contents (%) of SC
41
Table 4.3.2b Effect of treated SC on the % fat during storage
Treat Days Total
1 day 15 day 30 day
T0 3.22±0.054 3.27±0.059 3.33±0.044 3.27±0.031A
T1 3.24±0.023 3.26±0.040 3.34±0.052 3.28±0.025A
T2 3.25±0.040 3.26±0.040 3.33±0.058 3.28±0.027A
T3 3.25±0.081 3.26±0.077 3.33±0.069 3.28±0.040A
T4 3.26±0.080 3.26±0.023 3.32±0.075 3.28±0.034A
T5 3.24±0.090 3.25±0.098 3.34±0.064 3.28±0.046A
T6 3.24±0.104 3.25±0.078 3.34±0.064 3.28±0.045A
T7 3.24±0.066 3.25±0.069 3.33±0.052 3.27±0.034A
T8 3.26±0.050 3.28±0.069 3.34±0.069 3.29±0.034A
T9 3.26±0.067 3.28±0.106 3.33±0.046 3.29±0.040A
T10 3.27±0.059 3.28±0.092 3.32±0.098 3.29±0.043A
T11 3.25±0.110 3.26±0.069 3.33±0.052 3.28±0.042A
T12 3.24±0.090 3.25±0.064 3.34±0.081 3.28±0.043A
T13 3.24±0.069 3.26±0.090 3.32±0.046 3.27±0.037A
Total 3.25±0.016B 3.26±0.016AB 3.33±0.014AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
42
4.3.3 Protein contentsProtein shows a very important role in contributing the flavor and taste to cheese
(Guinee et al., 2004). The results of mean squares for protein of SC samples are
presented in Table 4.3.3a. Results revealed that treatment, storage days and their
interaction have NS (p>0.05) effect on the protein of SC.
The mean values for protein contents of SC are given in Table 4.3.3b. It is evident
from data that the highest protein (13.55%) was recorded in T6 (WP based edible coating
of SC containing 0.75 % CO) and T12 (WP based edible coating of SC containing 2.0 %
PMO) at 1 day, while the lowest protein (13.30%) was observed in T0 SC at 30 days. The
results revealed that there is NS (p>0.05) difference among all the treatments, the SC
samples which were formulated with WP based active edible coating (containing clove
and peppermint oil as active ingredient) and cheese samples with direct incorporation of
these essential oils have more protein as compared to control (T0).
Fig. 4.3 depicted that during storage of 30 days a minute decrease in protein of SC
samples were recorded which were manufactured with active edible coating and by direct
incorporation of clove and peppermint essential oils in different concentration. It is
evident from the data that protein content was decreasing from 13.55% to 13.48%, 13.44% to
13.41% and 13.37% to 13.30% at 1, 15 and 30 days respectively. Mean protein values for SC
samples at 15 and 30 days of storage showed a NS reduction.
The ability to bind with water and making a colloidal suspension is a specific functional
quality of proteins. Casein micelles are the protein that makes colloidal suspension in
milk. Final quality and acceptance for cheese depends on the protein content and
biochemical changes that occur during ripening. Protein content contributes the melting
and stretching properties of different chees types (Guinee et al., 2007).
Menthol in PMO and eugenol in CO have the excellent antioxidant potential so
binds the free radicals in SC samples that ultimately leads to minimum proteolysis in SC
samples as compare to T0. So the protein contents of the SC samples remains almost the
same the minor increase or decrease might be due to proteolysis. Findings of this
experiment are not in line with the results of Foda et al. (2010) and O’Connor and
O’Brien, (2000) because they reported a minor increase in protein during storage.
Ashenafi and Busse, (1992) also reported 15± 1.5 content in SC during cold storage with
the addition of spearmint in dried form in the manufacturing of cheese.
43
Table 4.3.3a MS for protein contents of SC
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.6335 0.0384 0.0105 10.3656 11.0480
0.3167 0.0030 0.0004 0.1234
2.57NS
0.02NS
0.00NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
13.2 13.25 13.3 13.35 13.4 13.45 13.513.41
13.4413.41
13.4213.43
13.4413.44
13.4313.41
13.4213.42
13.4413.44
13.43
13.313.35
13.3213.33
13.3913.3613.36
13.3513.32
13.3313.34
13.3613.37
13.36
30 day 15 day 1 day
Cheese Protein
Treat
ment
Fig. 4.3 Effect of edible coating and EOs on the protein (%) of SC
44
Table 4.3.3b Effect of treated SC on the % protein during storage
Treat Days Total
1 day 15 day 30 day
T0 13.50±0.242 13.41±0.208 13.30±0.144 13.40±0.105A
T1 13.52±0.098 13.44±0.307 13.35±0.306 13.44±0.131A
T2 13.48±0.202 13.41±0.162 13.32±0.185 13.40±0.095A
T3 13.50±0.254 13.42±0.277 13.33±0.081 13.42±0.114A
T4 13.51±0.173 13.43±0.133 13.39±0.173 13.44±0.082A
T5 13.54±0.133 13.44±0.208 13.36±0.185 13.45±0.093A
T6 13.55±0.081 13.44±0.260 13.36±0.131 13.45±0.091A
T7 13.54±0.150 13.43±0.248 13.35±0.173 13.44±0.101A
T8 13.50±0.173 13.41±0.162 13.32±0.104 13.41±0.079A
T9 13.50±0.208 13.42±0.277 13.33±0.173 13.42±0.114A
T10 13.50±0.300 13.42±0.208 13.34±0.146 13.42±0.116A
T11 13.54±0.115 13.44±0.358 13.36±0.170 13.45±0.122A
T12 13.55±0.208 13.44±0.260 13.37±0.087 13.45±0.103A
T13 13.54±0.329 13.43±0.098 13.36±0.051 13.44±0.104A
Total 13.52±0.045A 13.43±0.052A 13.35±0.036AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
45
4.3.4 Ash contents Ash is a mixture of inorganic constituents and is first parameter used to analyze
the rough mineral content in any food stuff (El-Nasri et al., 2012). It is present in cheese that results from the burning of organic matter (Kirk and Sawyer, 1991).
The result of mean squares for % ash of SC samples presented in Table 4.3.4a revealed a significant (p<0.05) effect on ash contents of SC due to storage days, while treatments and their interactive effect have NS (p>0.05) effect for ash contents of SC. The mean values for ash of SC are given in Table 4.3.4b. It is evident from data that the highest ash (1.37%) was recorded in T11 (WP based edible coating of SC containing 1.5 % PMO) at 30 days while the lowest ash (1.30%) was noted in control (T0), T2 (SC containing 0.5% CO), T8 (SC containing 1.5% PMO) and T9 (SC containing 2.5% PMO) SC samples. Results showed that there is NS difference among the treatments, the SC samples which were prepared with whey protein based active edible coating (containing clove and peppermint oil as active ingredient) and cheese samples with direct incorporation of these essential oils have more ash as compared to control (T0).
Fig. 4.4 showed that during storage of 30 days minor increase in ash content was recorded in SC samples which were prepared with active edible coating and by direct addition of natural essential oils in different concentration. Mean value of all treatments of SC shows that ash content was increasing from 1.30% to 1.32%, 1.32% to 1.35% and 1.31% to 1.37% at 1, 15 and 30 days respectively. EOs and edible coating have NS effect on SC.
Results showed that active edible coating and essential oils have no effect on the ash contents of SC. During storage of SC different metabolic, microbiological and biochemical processes occur but it does not affect the ash contents in SC (Singh et al., 2003; Farkye, 2004). The results of this parameter match with the findings of Kassa, (2008) he also that ash in SC remains same during storage of 30 day.4.3.5 pH
pH is one of the most critical factor in manufacturing and for determining the quality of cheese like texture, taste, appearance, and flavor that can be affected by pH (McSweeney et al., 2004). Starter cultures are used for the production of LAB maintain the ultimate pH (Pandey et al., 2003) that further helps in rennet coagulation of milk (Odile et al., 2012; Jacob et al., 2011).
The results of mean squares for pH of SC samples are presented in Table 4.3.5a.
Results revealed a significant (p<0.05), ** (p<0.01) and NS (p>0.05) effect due to
treatments, storage
46
Table 4.3.4a MS for ash contents of SC
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.0096140.0126000.0057860.1174000.145400
0.004807 0.000969 0.000223 0.001398
3.44* 0.69NS
0.16NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
1.28 1.29 1.3 1.31 1.32 1.33 1.34 1.35 1.36 1.37 1.381.32
1.331.321.32
1.311.34
1.331.35
1.321.33
1.311.34
1.331.32
1.341.331.33
1.321.31
1.341.33
1.351.34
1.331.31
1.371.33
1.32
30 day 15 day 1 day
Cheese Ash
Treat
ment
Fig. 4.4 Effect of edible coating and EOs on the ash (%) of SC
Table 4.3.4b Effect of treated SC on the % ash during storage
47
Treat Days Total
1 day 15 day 30 day
T0 1.30±0.017 1.32±0.035 1.34±0.029 1.32±0.015A
T1 1.31±0.021 1.33±0.017 1.33±0.023 1.32±0.011A
T2 1.30±0.029 1.32±0.023 1.33±0.012 1.32±0.012A
T3 1.31±0.023 1.32±0.023 1.32±0.017 1.32±0.011A
T4 1.31±0.023 1.31±0.017 1.31±0.023 1.31±0.011A
T5 1.32±0.012 1.34±0.017 1.34±0.023 1.33±0.010A
T6 1.31±0.035 1.33±0.035 1.33±0.012 1.32±0.015A
T7 1.33±0.023 1.35±0.017 1.35±0.017 1.34±0.010A
T8 1.30±0.023 1.32±0.017 1.34±0.012 1.32±0.011A
T9 1.30±0.023 1.33±0.023 1.33±0.023 1.32±0.013A
T10 1.31±0.023 1.31±0.010 1.31±0.012 1.31±0.008A
T11 1.32±0.020 1.34±0.015 1.37±0.029 1.34±0.013A
T12 1.32±0.038 1.33±0.015 1.33±0.012 1.33±0.012A
T13 1.32±0.012 1.32±0.012 1.32±0.015 1.32±0.006A
Total 1.31±0.005 1.33±0.005 1.33±0.005In a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
Table 4.3.5a MS for pH of SC
48
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.6396140.2307710.0919860.7530001.715371
0.319807 0.017752 0.003538 0.008964
35.68** 1.98* 0.39NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
4.2 4.3 4.4 4.5 4.6 4.7 4.84.54
4.654.69
4.74.7
4.644.664.66
4.674.68
4.694.66
4.694.69
4.374.46
4.574.61
4.664.53
4.554.59
4.64.61
4.644.57
4.594.64
30 day 15 day 1 day
Cheese pH
Treat
ment
Fig. 4.5 Effect of edible coating and EOs on the pH of SC
49
Table 4.3.5b Effect of treated SC on the pH during storage
Treat Days Total
1 day 15 day 30 day
T0 4.66±0.064 4.54±0.058 4.37±0.029 4.52±0.050B
T1 4.77±0.040 4.65±0.040 4.46±0.052 4.63±0.050AB
T2 4.75±0.058 4.69±0.058 4.57±0.046 4.67±0.038AB
T3 4.76±0.069 4.70±0.046 4.61±0.046 4.69±0.035A
T4 4.74±0.052 4.70±0.081 4.66±0.046 4.70±0.033A
T5 4.76±0.058 4.64±0.040 4.53±0.052 4.64±0.042AB
T6 4.75±0.069 4.66±0.040 4.55±0.029 4.65±0.038AB
T7 4.74±0.035 4.66±0.040 4.59±0.098 4.66±0.039AB
T8 4.74±0.058 4.67±0.058 4.60±0.052 4.67±0.034AB
T9 4.75±0.046 4.68±0.046 4.61±0.029 4.68±0.029A
T10 4.74±0.046 4.69±0.040 4.64±0.029 4.69±0.024A
T11 4.76±0.035 4.66±0.092 4.57±0.064 4.66±0.044AB
T12 4.76±0.075 4.69±0.058 4.59±0.064 4.68±0.041A
T13 4.75±0.046 4.69±0.064 4.64±0.052 4.69±0.031A
Total 4.75±0.013A 4.67±0.014B 4.57±0.016CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
50
days and interactive effect of storage days and treatments respectively for pH of SC. The mean values for pH of SC are illustrated in Table 4.3.5b. Results showed that active edible coating and EOs have ** (p<0.01) effect on pH of SC. It is evident from data that the highest pH (4.77) was recorded in sample T1 (WP based edible coating of SC) at 1 day while the minimum pH (4.37) was noted in control (T0) SC at 30 days. Results showed that the SC samples which were prepared with whey protein based active edible coating (containing clove and peppermint oil as active ingredient) and cheese samples with direct incorporation of these essential oils shows minimum decrease in pH during storage of 30 days as compared to control (T0). The overall values for all the SC showed consistent decrease in pH during storage.
Fig. 4.5 showed that during storage of 30 days a consistent decrease in pH of SC was observed in all SC samples which were prepared with active edible coating and by direct addition of natural essential oils in different concentrations. Mean value of all SC samples (treatments) shows that pH was decreasing from 4.77 to 4.77, 4.70 to 4.54 and 4.66 to 4.37 at 1, 15 and 30 days respectively. Mean values of pH for all treatments at 15 days and 30 days of storage showed a ** increase.
Decrease in pH during storage of SC samples mainly due to the production of lactic acid by LAB (Souza et al., 2003) and it might be decrease due to the acidity of CO and PMO that is added in SC samples. The overall values for all treatments of SC shows that pH decrease during storage but the rate of reduction was slow in all samples as compare to control due to the effect of edible coating and EOs. The difference in pH during storage could be due to the variable activity of starter cultures and enzymes and during storage (Shakeel-ur-Rehman et al., 2003). The findings of this parameter match with the results of Ozkan et al. (2007) they also noted that pH in butter decrease during storage.4.3.6 Acidity
Acidity is an important factor for determining cheese texture, structure, functionality and also protects against microbiological spoilage (Ceylan et al., 2003). The result of mean squares for acidity of SC samples shown in Table 4.3.6a revealed a ** (p<0.01) effect on the acidity of SC due to treatments and storage days. However, interactive effect of storage days and treatments was found to have significant effect (p<0.05) for acidity of SC.The mean values for acidity of SC are illustrated in Table 4.3.6b. Results showed that active edible coating and essential oils have ** (p<0.01) effect on acidity of SC. It is evident from the
51
Table 4.3.6a MS for acidity of SC
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.5000570.0296930.0389430.0748000.643493
0.250029 0.002284 0.001498 0.000890
280.78** 2.56** 1.68*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.2 0.4 0.6 0.8 1 1.20.83
0.770.780.780.78
0.770.76
0.790.78
0.770.78
0.760.77
0.79
0.960.91
0.880.86
0.850.90.9
0.890.85
0.840.82
0.910.89
0.87
30 day 15 day 1 day
Cheese Acdity
Treat
ment
Fig. 4.6 Effect of edible coating and EOs on the acidity (%) of SC
52
Table 4.3.6b Effect of treated SC on the % acidity during storage
Treat Days Total
1 day 15 day 30 day
T0 0.750±0.015g-j 0.830±0.017b-h 0.960±0.015a 0.847±0.032A
T1 0.700±0.012j 0.770±0.012e-j 0.910±0.012ab 0.793±0.031B
T2 0.720±0.017j 0.780±0.023d-j 0.880±0.017abc 0.793±0.025B
T3 0.730±0.017ij 0.780±0.006d-j 0.860±0.012b-e 0.790±0.020B
T4 0.750±0.006g-j 0.780±0.017d-j 0.850±0.023b-f 0.793±0.017B
T5 0.720±0.006j 0.770±0.006e-j 0.900±0.012ab 0.797±0.027B
T6 0.720±0.012j 0.760±0.017f-j 0.900±0.017ab 0.793±0.028B
T7 0.730±0.017ij 0.790±0.012c-j 0.890±0.017ab 0.803±0.025AB
T8 0.730±0.017ij 0.780±0.017d-j 0.850±0.017b-f 0.787±0.019B
T9 0.730±0.017ij 0.770±0.006e-j 0.840±0.012b-g 0.780±0.017B
T10 0.740±0.017hij 0.780±0.012d-j 0.820±0.031b-i 0.780±0.016B
T11 0.720±0.012j 0.760±0.012f-j 0.910±0.012ab 0.797±0.029B
T12 0.730±0.006ij 0.770±0.012e-j 0.890±0.050ab 0.797±0.028B
T13 0.740±0.025hij 0.790±0.017c-j 0.870±0.020a-d 0.800±0.022AB
Total 0.729±0.004C 0.779±0.004B 0.881±0.007AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMO
53
T13= WP based edible coating of SC containing 2.5% PMOdata that the highest acidity (0.960%) was recorded in control SC sample T0 at 30 days
while the lowest acidity (0.700%) was noted in T1 (WP based edible coating of SC) at 1
day. It is obvious from the results that the SC samples which were formulated with whey
protein based active edible coating (containing clove and peppermint oil as active
ingredient) and cheese samples with direct incorporation of these essential oils shows
minimum increase in acidity during storage of 30 days as compared to control (T0). The
overall values for all the treatments showed gradual increase in acidity during storage.
Fig. 4.6 showed that during storage of 30 days a consistent increase in acidity
content of SC was observed in all samples which were prepared with active edible
coating and by direct addition of essential oils in different concentrations. Mean value of
all treatments shows that acidity was increasing from 0.70% to 0.75%, 0.72% to 0.83%
and 0.82% to 0.96% at 1, 15 and 30 days respectively. Mean values of acidity for all
treatments at 15 days and 30 days of storage showed a ** increase.
Increase in acidity during storage of SC was mainly due to the production of lactic
acid by LAB (Warsama et al., 2006; Hamid and Abdelrahman, 2012) and it might be
increase due to the acidity of CO and PMO that is added in SC samples. The overall
values of acidity for all SC samples increase during storage. The findings of this
parameter match with the results of (Ong et al., 2007a) they also noted that acidity in SC
samples increase during storage. The results match with Foda et al. (2009), they also
recorded that acidity increase during storage. Results also showed resemblance with
Eyassu, (2013) he reported 0.12 ±0.07 to 0.15± 0.05 in stored SC.
4.3.7 Water activity (aw)Water activity (aw) determine that how much amount of water in food is
available for the growth and activity of microorganisms or equilibrium amount of water
is available for hydration of materials that directly measures the shelf life of the product
(Koca and Metin, 2004).
Results of mean squares analysis for aw of SC samples are presented in Table
4.3.7a. It is obvious from the result that the treatments, storage days and their interactive
effect have ** (p<0.01) effect on the water activity of SC. The mean values for water
activity of SC are given in Table 4.3.7b. Results revealed that active edible coating and
essential oils have ** (p<0.01) effect on water activity of SC. It is evident from data that
the highest water activity (0.97) was recorded in SC sample T4 (SC containing 1.0%CO)
and T10 (whey protein based coating of SC
54
Table 4.3.7a MS for water activity of SC
SOV Df SS MS MS
DayTreatmentDay x TreatmentErrorTotal
2132684
125
2.2235570.1278360.0728430.0740002.498236
1.111779 0.009834 0.002802 0.000881
1262.02** 11.16** 3.18**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.90.7
0.750.77
0.80.81
0.750.77
0.80.79
0.810.83
0.770.790.8
0.490.53
0.620.65
0.690.59
0.610.64
0.660.7
0.720.63
0.660.67
30 day 15 day 1 day
Cheese Water Activity
Treat
ment
Fig. 4.7 Effect of edible coating and EOs on the water activity of SC
55
Table 4.3.7b Effect of treated SC on the water activity during storage
Treat Days Total
1 day 15 day 30 day
T0 0.960±0.015a 0.700±0.015d-h 0.490±0.017k 0.717±0.068E
T1 0.950±0.017a 0.750±0.012b-f 0.530±0.012jk 0.743±0.061DE
T2 0.960±0.023a 0.770±0.023b-e 0.620±0.012hij 0.783±0.050BCD
T3 0.960±0.029a 0.800±0.006bc 0.650±0.012ghi 0.803±0.046ABC
T4 0.970±0.012a 0.810±0.017bc 0.690±0.012e-h 0.823±0.041AB
T5 0.950±0.012a 0.750±0.006b-f 0.590±0.012ij 0.763±0.052CDE
T6 0.950±0.017a 0.770±0.012b-e 0.610±0.017hij 0.777±0.050BCD
T7 0.960±0.017a 0.800±0.012bc 0.640±0.017ghi 0.800±0.047ABC
T8 0.960±0.017a 0.790±0.012bcd 0.660±0.017f-i 0.803±0.044ABC
T9 0.960±0.029a 0.810±0.017bc 0.700±0.017d-h 0.823±0.039AB
T10 0.970±0.020a 0.830±0.029b 0.720±0.006c-g 0.840±0.038A
T11 0.950±0.023a 0.770±0.017b-e 0.630±0.012ghi 0.783±0.047BCD
T12 0.950±0.021a 0.790±0.006bcd 0.660±0.017f-i 0.800±0.043ABC
T13 0.960±0.023a 0.800±0.023bc 0.670±0.015f-i 0.810±0.043ABC
Total 0.958±0.005A 0.781±0.006B 0.633±0.010CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
56
containing 1.5%PMO) at 1st day while the lowest water activity (0.49) was noted in
control (T0) at 30 days. Results showed that the SC samples which were prepared with
whey protein based active edible coating (containing clove and peppermint oil as active
ingredient) and samples with direct incorporation of these essential oils have more water
activity as compared to control (T0) during storage of 30 days. The overall values for all
the treatments showed gradual decrease in water activity during storage.
Fig. 4.7 showed that during storage of 30 days a consistent decrease in water
activity in all SC samples which were prepared with active edible coating and by direct
incorporation of essential oils in different concentrations. Mean value of all treatments
shows that water activity was decreasing from 0.97 to 0.95, 0.83 to 0.70 and 0.72 to 0.49
at 1, 15 and 30 days respectively. Mean values of water activity for all treatments at 15
days and 30 days of storage showed a ** decrease.Edible coating serve as a barrier against moisture loss by covering the surface of SC
samples so less moisture loss was observed in edible coated samples of SC. PMO and CO
have excellent antioxidant and antimicrobial potential so binds the free radicals in SC
samples that ultimately results in higher aw as compared to control SC. The overall values of
aw in all SC samples decrease but the rate of reduction is slow in all SC samples as compare
to control SC. Results of aw match with
the findings of Koca and Metin, (2004), they also observed that aw decreases in SC
samples during storage.
4.3.8 Color Analysis The consumer’s first attractive tool towards food is its color that helps in either to
accept or reject the food (Gokmen and Sugut, 2007). Color is vital sensory factor in all
food systems that affect the acceptance, rejection and marketing of any food product. In
dairy products especially in processed food color act as a great sensual indicator of which
biochemical reactions are occurring in many food that changes the color (Yates and
Drake, 2007).
4.3.8.1 Lightness (L*) value of color The result of mean squares (ANOVA) for L* value of SC samples presented in
Table 4.3.8a revealed a ** (p<0.01) effect in L* value due to treatments of SC which
were manufactured with WP based active edible coating (containing clove and
peppermint oil as active ingredient) and samples with direct incorporation of clove and
peppermint oils in different concentrations. Table 4.3.8b presents the mean values of L*.
The difference is due to change in
57
Table 4.3.8a MS for color of SC
SOV df MS
Cheese L* Cheese a* Cheese b*
TreatmentErrorTotal
132841
29.692** 3.797
0.35039**0.02911
6.7341**0.1031
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 2 4 6 8 10 12Cheese L*
Treat
ment
Fig. 4.8Effect of edible coating and EOs on the L*value of SC
58
EOs concentration and active edible coating. Lowest value (77.41) was recorded in T13 (WP based edible coating of SC containing 2.5% PMO) while highest value (85.6) was attained in control (T0). All the SC samples are lighter in color than control (T0). 4.3.8.2 Redness (a*) value for color
The result of mean squares (ANOVA) for a* value of SC samples presented in Table 4.3.8a revealed a ** (p<0.01) effect in a* due to treatments of SC which were formulated with WP based active edible coating (containing clove and peppermint oil as active ingredient) and samples with direct incorporation of clove and peppermint oils in different concentration. Table 4.3.8b presents the mean values of a*. Highest value (-6.09) was recorded in T7 (WP based edible of SC coating containing 1.0%CO) while lowest value (-7.17) was attained in T10 (SC containing 2.5% PMO).The results of this parameter match with the findings of Foda et al. (2010) they also reported the almost same results.4.3.8.3 Yellowness (b*) value of color
The result of mean squares for b* value of SC presented in Table 4.3.8a revealed a ** (p<0.01) effect in b* due to SC samples which were formulated with WP based active edible coating (containing clove and peppermint oil as active ingredient) and samples with direct incorporation of clove and peppermint oils in different concentration. Table 4.3.8b presents the mean values of b*. Higher intensity (14.1) was recorded in T4
(SC containing 1.0% CO) lower value (9.3) was attained in T10 (SC containing 2.5% peppermint oil). All other SC samples have values in between these two treatments. Results of this parameter for yellowness (b*) values match with the findings of Cardenas et al. (2014) they also reported almost the same results that yellowness (b*) values within range of 10.76± 0.16 to 10.93±0.36 in SC.4.4 Texture analysis
Texture of food is a combination of rheological and structural attributes of a product defined by international organization for standardization (ISO) that can be analyzed by the tactile, mechanical, and appropriately where needed then visionary, auditory are used (Fox et al., 2000). Texture of cheese is a basic signal that is given by customer for the appearance and texture of cheese because customer’s acceptance and rejection strongly affect the marketing of cheese or any food product (Hicsasmaz et al., 2000). Texture plays an important role in determing the quality and characteristics of cheese that is mostly influenced by the compositional and processing parameters (Wium et al., 2003).
59
T0
T2
T4
T6
T8
T10
T12
5 6 7 8 9 10 11Cheese a* (-ve)
Trea
tmen
t
Fig. 4.9 Effect of edible coating and EOs on the a*value of SC
T0
T2
T4
T6
T8
T10
T12
7 7.5 8 8.5 9 9.5 10 10.5Cheese b*
Trea
tmen
t
Fig. 4.10 Effect of edible coating and EOs on the b*value of SC
60
Table 4.3.8b Effect of treated SC on the L*, a*and b* value
Treatment Cheese L* Cheese a* Cheese b*
T0 85.60±1.276A -6.45±0.052A-D 12.90±0.237A
T1 84.70±1.547AB -6.19±0.104AB 13.10±0.092AB
T2 85.10±0.889A -6.37±0.133ABC 13.40±0.156AB
T3 84.90±1.085AB -6.33±0.110ABC 13.50±0.121AB
T4 84.80±1.120AB -6.26±0.115AB 14.10±0.139AB
T5 81.90±0.693ABC -6.21±0.104AB 13.20±0.150AB
T6 80.15±1.449ABC -6.17±0.121AB 13.25±0.306B
T7 80.00±1.386ABC -6.09±0.052A 13.40±0.271BC
T8 79.80±0.525ABC -6.90±0.058DEF 10.20±0.069BC
T9 79.15±1.051BC -7.00±0.081EF 10.00±0.144CD
T10 78.30±0.906C -7.17±0.081F 9.30±0.069D
T11 78.00±0.745C -6.57±0.052A-E 11.60±0.312E
T12 77.60±1.403C -6.64±0.167B-E 12.10±0.162E
T13 77.40±1.117C -6.77±0.064C-F 12.70±0.121EIn a column or row means having similar letters are statistically NS (P>0.05). T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
61
4.4.1 Hardness
The amount of force required to bite the sample entirely when placed in the
middle of molar teeth is called Hardness (Meullenet and Gross, 1999). Chemical,
physicochemical constituents composition of cheese give its hardness that is solid to fat
ratio. Existence of heterogeneities in the cheese composition like granules connection in
curd, fissures and cracks also expressed as cheese hardness (Gunasekaran and Ak, 2003).
The results of mean squares for hardness of SC samples are shown in Table
4.4.1a. Results showed that treatments and storage days have ** (p<0.01) effect while
their interactive effect have NS (p>0.05) variation for hardness of SC.
The mean hardness values for SC are given in Table 4.4.1b. It is evident from data
that the highest value of hardness (1192.33) was recorded in control (T0) at 30 days while
the lowest value of hardness (1035) was noted in T10 (SC containing 2.5% PMO) SC
sample at 1 day. Results showed that there is a ** difference among all the SC samples
which were prepared with WP based active edible coating (containing CO and PMO as
active ingredient) and samples with direct incorporation of these natural essential oils.
Control cheese sample (T0) have more hardness as compared to all other SC samples due
to the effect of active edible coating and addition of natural essential oils.
Fig. 4.11 depicted that during storage of 30 days a continuous decrease in
hardness of SC was noted in SC samples which were formulated with active edible
coating and natural essential oils in different concentrations. Mean SC values shows that
hardness was decreasing from 1,045 to 1,035, 1,130.34 to 1,062 and 1,192.34 to 1,095 at
1, 15 and 30 days respectively. Mean hardness values for SC at 15 and 30 days of storage
showed a ** decrease.
Decrease in hardness of SC samples containing active edible coating and natural
EOs was might be due to the proteolysis that causes changes in protein structure that
leads to compactness (Bryant et al, 1995). Results of the present study match with
Sipahioglu et al. (1999), who reported that hardness of feta cheese decrease during
storage. Zisu and Shah (2005) also found out the similar results in Mozzarella cheese.
The results of decline in cheese hardness are also in accordance with the findings of
Karaman and Akalin (2013) they reported decrease hardness in Turkish white cheeses.
62
Table 4.4.1a MS for hardness of SC
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
114489.1 30301.7 6662.9 63504.0214957.7
57244.6 2330.9 256.3 756.0
75.72** 3.08** 0.34NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
950 1000 1050 1100 1150 1200 12501130.34
107010751073
1070106710661064
10731071
1068106610641062
1192.341110
11251122
111711071105
11021122
11181115
110010971095
30 day 15 day 1 day
Cheese Hardness
Treat
ment
Fig. 4.11 Effect of edible coating and EOs on the hardness (g) of SC
63
Table 4.4.1b Effect of treated SC on the hardness (g) during storage
Treat Days Total
1 day 15 day 30 day
T0 1072.33±11.795 1130.33±21.310 1192.33±5.239 1131.67±18.757A
T1 1038.00±10.970 1070.00±9.815 1110.00±24.249 1072.67±13.248B
T2 1040.00±21.362 1075.00±24.249 1125.00±14.434 1080.00±16.015B
T3 1038.00±20.207 1073.00±13.856 1122.00±9.815 1077.67±14.367B
T4 1038.00±12.702 1070.00±16.166 1117.00±17.898 1075.00±13.910B
T5 1045.00±17.321 1067.00±12.124 1107.00±12.702 1073.00±11.534B
T6 1044.00±6.928 1066.00±8.660 1105.00±12.702 1071.67±10.160B
T7 1044.00±7.506 1064.00±30.022 1102.00±19.053 1070.00±13.505B
T8 1037.00±21.362 1073.00±8.660 1122.00±23.094 1077.33±15.505B
T9 1037.00±6.928 1071.00±13.856 1118.00±19.053 1075.33±13.715B
T10 1035.00±13.279 1068.00±13.279 1115.00±9.815 1072.67±13.119B
T11 1045.00±8.083 1066.00±19.053 1100.00±14.434 1070.33±10.828B
T12 1043.00±5.774 1064.00±17.321 1097.00±20.207 1068.00±11.116B
T13 1043.00±17.898 1062.00±16.166 1095.00±13.279 1066.67±10.994B
Total 1042.81±3.409C 1072.81±4.557B 1116.24±5.095AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
64
4.4.2 CohesivenessCohesiveness is the ability of internal bond strength that allows a product to
crumble (Bourne, 1978) and maintains the body of that product (Zoon, 1991).
The results of mean squares for cohesiveness of SC samples are shown in Table
4.4.2a. Results revealed that treatments and storage days have ** (p<0.01) effect while
their interactive effect have NS (p>0.05) variation for cohesiveness of SC.
The mean cohesiveness values for SC are given in Table 4.4.2b. It is evident from
data that the highest value of cohesiveness (0.616) was recorded in T1 (WP based edible
coating of SC), T7 (WP based edible coating of SC containing 1.0%CO) and T13 (WP
based edible coating of SC containing 2.5% PMO) at 1 day while the lowest value of
cohesiveness (0.534) was noted in control SC sample T0 at 30 days. Results revealed that
there is a ** difference among all the SC samples which were prepared with WP based
active edible coating (containing clove and peppermint oil as active ingredient) and
samples with direct incorporation of these natural essential oils. T0 SC sample have more
cohesiveness as compared to all other SC samples due to active edible coating and
addition of natural essentials oils.
Fig. 4.12 depicted that during storage of 30 days a significant decrease in
cohesiveness of SC was observed in SC samples which were formulated with edible
coating and different concentrations of natural EOs. Mean cohesiveness value shows that
cohesiveness was decreasing from 0.611 to 0.595, 0.598 to 0.565 and 0.578 to 0.535 at 1,
15 and 30 days respectively. Mean cohesiveness values for SC samples showed **
decrease at 15 and 30 days of storage.
Overall cohesiveness value of SC samples decrease during storage, outcomes of
cohesiveness match with Bhaskaracharya, (2004). According to his results cohesiveness
also decreases during storage. Cheese cohesiveness is associated with proteolysis.
4.4.3 SpringinessThe bouncing property of sample and returning to its original shape after
consective bites is known as the springiness of that sample (Civille and Szczesniak,
1973).
Results of mean squares for springiness of SC samples are shown in Table 4.4.3a.
Results revealed that treatments and storage have** (p<0.01) effect while their
interactive effect have * (p<0.05) variation in springiness of SC.
65
Table 4.4.2a MS for cohesiveness of SC
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.0311535 0.0066279 0.0019410 0.0169260 0.0566484
0.0155767 0.0005098 0.0000747 0.0002015
77.30** 2.53** 0.37NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0.5 0.52 0.54 0.56 0.58 0.6 0.620.565
0.5890.5950.5960.597
0.5920.594
0.5970.5970.598
0.580.591
0.5930.596
0.5350.565
0.5740.575
0.5770.567
0.5690.571
0.575333333333333
0.5780.58
0.5690.569
0.572
30 day 15 day 1 day
Cheese Cohesiveness
Treat
ment
Fig. 4.12 Effect of edible coating and EOs on the cohesiveness of SC
66
Table 4.4.2b Effect of treated SC on the cohesiveness during storage
Treat Days Total
1 day 15 day 30 day
T0 0.594±0.007 0.565±0.005 0.534±0.004 0.565±0.009B
T1 0.611±0.012 0.589±0.013 0.565±0.004 0.588±0.008A
T2 0.608±0.004 0.595±0.006 0.574±0.010 0.592±0.006A
T3 0.608±0.005 0.596±0.005 0.575±0.010 0.593±0.006A
T4 0.609±0.014 0.597±0.009 0.577±0.004 0.594±0.007A
T5 0.610±0.005 0.592±0.012 0.567±0.011 0.590±0.008A
T6 0.610±0.003 0.594±0.007 0.569±0.009 0.591±0.007A
T7 0.611±0.010 0.597±0.003 0.571±0.005 0.593±0.007A
T8 0.608±0.005 0.597±0.005 0.575±0.007 0.593±0.006A
T9 0.608±0.013 0.598±0.013 0.578±0.005 0.595±0.007A
T10 0.607±0.006 0.580±0.008 0.580±0.010 0.589±0.006A
T11 0.609±0.008 0.591±0.005 0.569±0.006 0.590±0.007A
T12 0.609±0.005 0.593±0.007 0.569±0.006 0.590±0.007A
T13 0.611±0.011 0.596±0.009 0.572±0.013 0.593±0.008A
Total 0.608±0.002A 0.591±0.002B 0.570±0.002CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
67
Table 4.4.3a MS for springiness of SC
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.0310047 0.0659195 0.0101753 0.0170200 0.1241195
0.0155024 0.0050707 0.0003914 0.0002026
76.51** 25.03** 1.93*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.1 0.2 0.3 0.4 0.5 0.60.439
0.4570.4690.478
0.5090.4760.4820.488
0.4810.520.521
0.4710.476
0.507
0.4070.4310.439
0.4530.496
0.4310.437
0.4740.4930.499
0.5110.4810.4860.495
30 day 15 day 1 day
Cheese Springiness
Treat
ment
Fig. 4.13 Effect of edible coating and EOs on the springiness of SC
68
Table 4.4.3b Effect of treated SC on the springiness during storage
Treat Days Total
1 day 15 day 30 day
T0 0.467±0.004e-i 0.439±0.004g-j 0.407±0.008j 0.438±0.009H
T1 0.482±0.013b-h 0.457±0.011f-i 0.431±0.008ij 0.457±0.009GH
T2 0.494±0.012a-f 0.469±0.008e-i 0.439±0.005g-j 0.467±0.009FG
T3 0.512±0.008a-e 0.478±0.011b-i 0.453±0.006f-j 0.481±0.010DEF
T4 0.519±0.009abc 0.509±0.008a-e 0.496±0.003a-f 0.508±0.005ABC
T5 0.489±0.004a-f 0.476±0.007c-i 0.431±0.008ij 0.465±0.009FG
T6 0.492±0.005a-f 0.482±0.007b-h 0.437±0.008hij 0.470±0.009EFG
T7 0.499±0.006a-f 0.488±0.003a-f 0.474±0.007c-i 0.487±0.005C-F
T8 0.519±0.009abc 0.481±0.007b-h 0.493±0.013a-f 0.498±0.007A-D
T9 0.524±0.009ab 0.520±0.010abc 0.499±0.005a-f 0.514±0.006AB
T10 0.530±0.010a 0.521±0.003abc 0.511±0.009a-e 0.521±0.005A
T11 0.511±0.009a-e 0.471±0.007d-i 0.481±0.009b-h 0.488±0.007C-F
T12 0.514±0.016a-e 0.476±0.009c-i 0.486±0.003a-g 0.492±0.008B-E
T13 0.518±0.006a-d 0.507±0.008a-e 0.495±0.010a-f 0.507±0.005ABC
Total 0.505±0.003A 0.484±0.004B 0.467±0.005CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
69
The mean values for springiness of SC are given in Table 4.4.3b. It is evident from data that the highest value of springiness (0.530) was recorded in T10 (SC containing 2.5% peppermint oil) at 1 day while the lowest value of springiness (0.407) was noted in control SC sample T0 at 30 days. Results revealed that there is a ** difference among all the SC samples which were prepared with WP based active edible coating (containing clove and peppermint oil as active ingredient) and samples with direct incorporation of these natural essential oils. T0 SC sample have less springiness as compared to all other SC samples due to addition of essentials oils and active edible coating.
Fig. 4.13 showed that during storage of 30 days a continuous decrease in springiness of SC was observed in all samples which were prepared with active edible coating and natural essential oils in different concentrations. Mean SC values shows that springiness was decreasing from 0.97 to 0.95, 0.83 to 0.70 and 0.72 to 0.49 at 1, 15 and 30 days respectively. Mean springiness values for SC samples at 15 days and 30 days of storage showed a ** decrease. Elasticity of cheese decreases with the progress of ripening because proteolysis breaks down the casein network (Hort and Le Grys, 2001). Bhaskarcharya (2004) also reported that there is a significant decrease in the springiness of cheese during first 30 days of ripening.4.4.4 Gumminess
Cheese hardness is because of gumminess and chewiness (Awad et al., 2013). Confrontation or resistance that is offered by cheese cube cuddling in between fore finger and thumb or during mastication is called gumminess (Wium et al., 1997). For cheese and semisolid foods gumminess is better term to be used it is also called an energy that is required for splitting of the semisolid to make a bolus for proper swallowing. Gumminess is the product of hardness and cohesiveness (Lee et al., 1978).
The results of mean squares for gumminess of SC samples are shown in Table 4.4.4a. Results revealed that treatments have NS (p>0.05) effect while storage days and their interactive effect have* (p<0.05) variation on the gumminess of SC.The mean gumminess values for SC are given in Table 4.4.4b. Data showed that the maximum value of gumminess (666.30) was noted in T0 SC sample at 30 days while the minimum value of gumminess (623.87) was noted in SC sample T1 (WP based edible coating of SC) also at 30 days. Results revealed that there is a ** difference among the SC samples which were formulated with WP based active edible coating (containing CO and PMO as active ingredient)
70
Table 4.4.4a MS for gumminess of SC
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
247.7 3760.9 4067.1 12828.8 20904.5
123.8 289.3 156.4 152.7
0.81NS
1.89* 1.02NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
600 610 620 630 640 650 660 670640.07
626.87639.6639.5
638.7631.7
633.2635.2
640.6640.5
638.7628.34
631633
666.3623.87
645.8645.2
644.5627.7628.7629.3
645.2646.2646.7
624.24624.2
626.3
30 day 15 day 1 day
Cheese Gumminess
Treat
ment
Fig. 4.14 Effect of edible coating and EOs on the gumminess (g) of SC
71
Table 4.4.4b Effect of treated SC on the gumminess (g) during storage
Treat Days Total
1 day 15 day 30 day
T0 641.27±2.282 640.07±2.205 666.30±6.083 649.21±4.712A
T1 630.87±1.729 626.87±10.012 623.87±2.205 627.20±3.168B
T2 632.30±5.774 639.60±8.141 645.80±6.466 639.23±3.948AB
T3 631.10±10.508 639.50±11.085 645.20±6.813 638.60±5.244AB
T4 632.20±4.099 638.70±7.275 644.50±10.335 638.47±4.227AB
T5 637.40±8.603 631.70±1.155 627.70±4.907 632.27±3.204AB
T6 636.80±2.117 633.20±8.025 628.70±4.330 632.90±2.945AB
T7 637.80±8.487 635.20±4.734 629.30±6.409 634.10±3.588AB
T8 630.50±10.681 640.60±7.679 645.20±8.776 638.77±5.055AB
T9 630.50±5.831 640.50±5.023 646.20±3.984 639.07±3.394AB
T10 628.30±10.046 638.70±9.757 646.70±5.600 637.90±5.104AB
T11 634.73±10.483 628.33±10.829 624.23±5.263 629.10±4.855AB
T12 635.20±5.369 631.00±8.083 624.20±8.487 630.13±4.052AB
T13 637.30±2.021 633.00±8.256 626.30±4.099 632.20±3.159AB
Total 634.02±1.671A 635.50±1.880A 637.44±2.381AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
72
and samples with direct addition of these natural essential oils. T0 SC sample have more gumminess as compared to all other SC samples due to active edible coating and addition of natural essentials oils.
Fig. 4.14 depicted that during storage of 30 days a decline in gumminess of SC was noted in SC samples which were formulated with edible coating and different concentrations of essential oils. Mean SC value shows that gumminess was decreasing from 641.27 to 628.3, 640.5 to 626.87 and 666.3 to 623.87 at 1, 15 and 30 days respectively. Mean gumminess values for SC at 15 days and 30 days of storage showed a ** decrease.
Decrease in gumminess during storage is due to the proteolysis in SC that causes the compactness in protein matrix (Karaman and Akalin, 2013). The results of the present study match with the Drake et al, (1996) who showed decrease in gumminess of cheese during storage. 4.4.5 Chewiness
During swallowing the energy required to chew the food is called chewiness (Zoon, 1991). There is a correlation between hardness and chewing because more harder cheese results in more force to apply and chew the food stuff as chewiness defined as the number of mastication that are applied on a certain amount of sample to enhance the satisfactory swallowing (Beal and Mittal, 2000).
The results of mean squares for chewiness of SC samples are shown in Table 4.4.5a. Results revealed that treatments and storage days have ** (p<0.01) effect on springiness of SC while their interactive effect have * (p<0.05) impact for chewiness of SC.
The mean chewiness values for SC are given in Table 4.4.5b. Data showed that highest value of chewiness (382.10) was recorded in T4 (WP based edible coating of SC containing 0.5% clove oil) at 1 day while the lowest value of chewiness (264.70) was noted in T0 SC sample at 30 days. Results revealed that there is a ** difference among all the SC samples which were prepared with WP based active edible coating (containing clove and peppermint oil as active ingredient) and samples with direct incorporation of these natural essential oils. T0 SC sample have less chewiness as compared to all other SC samples due to active edible coating and addition of natural essentials oils.
Fig. 4.15 depicted that during storage of 30 days a decrease in chewiness of SC was noted in SC samples which were formulated with active edible coating and different concentrations of natural essential oils. Mean SC values shows that chewiness was decreasing
73
Table 4.4.5a MS for chewiness of SCSOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
15286.1 36445.5 7649.4 12950.1 72331.1
7643.1 2803.5 294.2 154.2
49.58** 18.18** 1.91*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 50 100 150 200 250 300 350276.7
288300
305.7325.1
300.7305.2310308.2
333.1332.8
296.7300.4
320.9
264.7270.3
283.5292.3
319.7270.5274.7
298.3318.1322.5
330.5301.1303.4
310.1
30 day 15 day 1 day
Cheese Chewiness
Treatm
ent
Fig. 4.15 Effect of edible coating and EOs on the chewiness (g) of SC
74
Table 4.4.5b Effect of treated SC on the chewiness (g) during storage
Treat Days Total
1 day 15 day 30 day
T0 294.80±7.54b-g 276.70±5.66efg 264.70±4.00g 278.73±5.28H
T1 305.68±8.30b-g 288.00±11.75c-g 270.30±8.15fg 287.99±6.99GH
T2 312.40±6.50b-e 300.00±9.91b-g 283.50±6.61d-g 298.63±5.73E-H
T3 323.20±8.50bcd 305.70±4.15b-g 292.30±7.71b-g 307.07±5.69D-G
T4 382.10±10.59a 325.10±4.71bc 319.70±8.95bcd 342.30±10.84A
T5 311.70±6.04b-e 300.70±4.43b-g 270.50±3.35fg 294.30±6.60FGH
T6 313.30±4.85b-e 305.20±7.93b-g 274.70±2.59efg 297.73±6.50E-H
T7 318.30±3.99bcd 310.00±5.83b-f 298.30±6.15b-g 308.87±3.97C-F
T8 327.30±5.03bc 308.20±8.27b-f 318.10±5.97bcd 317.87±4.29B-E
T9 330.40±8.24b 333.10±8.06b 322.50±10.28bcd 328.67±4.73ABC
T10 333.00±9.17b 332.80±5.01b 330.50±4.39b 332.10±3.30AB
T11 325.20±6.73bc 296.70±7.08b-g 301.10±4.10b-g 307.67±5.38D-G
T12 326.50±6.32bc 300.40±9.96b-g 303.40±6.67b-g 310.10±5.68C-F
T13 330.20±7.05b 320.90±8.15bcd 310.10±8.74b-f 320.40±4.95BCD
Total 323.86±3.41A 307.39±2.95B 297.12±3.58CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
75
from 382.1 to 294.8, 333.1 to 276.7 and 333.5 to 264.7 at 1, 15 and 30 days respectively.
Mean chewiness values for SC samples at 15 and 30 days of storage showed a **
decrease. The results
of chewiness are not similar with the Awad et al. (2005) who showed an increase in
chewiness of cheese during ripening.
4.5 Weight loss The results of mean squares (ANOVA) for weight loss of SC samples are
presented in Table 4.5a. It was evident from the data that treatments, storage days and
their interactive effect shows a ** (p<0.01) effect on the weight loss of SC.
The mean values for weight loss of SC are given in Table 4.5b. Results revealed
that active edible coating and essential oils have ** (p<0.01) effect on weight loss of SC.
Data showed that the highest value of weight loss (11.87%) was recorded in T0 SC
sample at 30 days while the lowest value of weight loss (2.99%) was noted in T4 (SC
containing 1.0% CO) at 15 days. Results revealed that there is a ** difference among all
the SC samples which were prepared with WP based active edible coating (containing
clove and peppermint oil as active ingredient) and samples with direct incorporation of
these natural EOs. T0 SC sample have more
weight loss as compared to all other SC samples due to active edible coating and addition
of natural essentials oils. Mean weight loss values shows that all the butter samples have
0% weight loss at 1 day.
Fig. 4.16 depicted that during storage of 30 days increase in weight loss of SC
samples which were formulated with active edible coating and different concentrations of
clove and peppermint oil. Mean SC values shows that weight loss was increasing from
2.99% to 5.28% and 7.93% to 11.87% at 15 and 30 days respectively. Mean weight loss
values for SC samples at 15 days and 30 days of storage showed a ** increase. Findings
of this parameter match with the results of Tarakci and Kucukoner (2006), they also
noted that moisture in SC decrease during storage.Edible coating serve as a barrier against moisture loss by covering the surface of SC
samples so less moisture loss was observed in edible coated samples of SC. PMO and CO
have excellent antioxidant and antimicrobial potential so binds the free radicals in SC
samples that ultimately leads to minimum weight loss as compared to control (T0) SC. The
findings of this parameter match with the results of Tarakci and Kucukoner (2006), they
also noted that moisture in SC decrease during storage.
76
Table 4.5a MS for weight loss of SC
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
1 13 13 56 83
597.067 46.229 3.805 2.649
649.749
597.067 3.556 0.293 0.047
12624.20** 75.19** 6.19**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 2 4 6 8 10 12 14
11.8710.07
8.958.4
7.939.15
9.018.7
9.058.65
8.19.3
9.18.8
30 day 15 day
Cheese Weight Loss
Treat
ment
Fig. 4.16 Effect of edible coating and EOs on weight loss (%) of SC
77
Table 4.5b Effect of treated SC on the % weight loss during storage
Treat Days Total
1 day 15 day 30 day
T0 - 5.28±0.114g 11.87±0.214a 8.58±1.478A
T1 - 4.25±0.093h 10.07±0.104b 7.16±1.303B
T2 - 3.41±0.108j-m 8.95±0.127cd 6.18±1.241D-G
T3 - 3.20±0.055lm 8.40±0.064def 5.80±1.163GHI
T4 - 2.99±0.101m 7.93±0.161f 5.46±1.108I
T5 - 4.03±0.150hij 9.15±0.163c 6.59±1.149CD
T6 - 3.91±0.058h-k 9.00±0.158cd 6.46±1.141CDE
T7 - 3.60±0.110h-m 8.70±0.231cde 6.15±1.146EFG
T8 - 3.46±0.064i-m 9.05±0.104cd 6.26±1.251DEF
T9 - 3.30±0.025klm 8.65±0.218cde 5.98±1.200FGH
T10 - 3.06±0.053m 8.10±0.069ef 5.58±1.128HI
T11 - 4.12±0.150hi 9.30±0.064c 6.71±1.161C
T12 - 4.01±0.061hij 9.10±0.196c 6.56±1.142CDE
T13 - 3.80±0.075h-l 8.80±0.069cd 6.30±1.119C-F
Total - 3.74±0.093B 9.08±0.148AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
78
4.6 Antioxidant analysis4.6.1 Diphenyl-2-picrylhdrazyl (DPPH) activity
Results of mean squares for DPPH of SC samples are presented in Table 4.6.1a.
Results showed that mean square value for treatments and storage days for DPPH values
have a ** (p<0.01) variation while their interaction have NS (p>0.05) effect on the
DPPH value of SC.
The mean DPPH activity values for SC are illustrated in Table 4.6.1b. It is
obvious from the results that active edible coating and essential oils have ** (p<0.01)
effect on DPPH activity of SC. Data revealed that the highest value of DPPH activity
(90.27%) was recorded in T4 (SC containing 1.0% CO) at 1st day while the lowest value
of DPPH activity (51.76%) was noted in control SC (T0) at 30 days. Results revealed that
there is a ** difference among all the SC samples which were prepared with WP based
active edible coating (containing clove and peppermint oil as active ingredient) and
samples with direct incorporation of these natural essential oils. Control SC (T0) have
less DPPH activity as compared to all other SC samples due to active edible coating and
addition of natural essentials oils.
Fig. 4.17 showed that during storage of 30 days decrease in DPPH activity of SC was
noted in all SC samples which were manufactured with active edible coating and
different concentrations of clove and peppermint oil. Mean value of all SC samples
shows that DPPH activity was decreasing from 90.27% to 76.68%, 81.7% to 63.34% and
70.6% to 51.76% at 1, 15 and 30 days respectively. Mean values of DPPH activity for all
treatments at 15 days and 30 days of storage showed a ** decrease.
Antioxidant potential of EOs are mainly due to acting as a substrate for radicals
(like superoxide), free radical-scavenging, hydrogen-donation, singlet oxygen quenching
and metal ion chelation (Al-Mamar et al., 2002). Eugenol in CO and menthol in PMO are
very effective in shelf life extension of soft cheese due to their antioxidant potential by
scavenging the radicals that are involved in lipid oxidation. The results of DPPH activity
are in agreement with the findings of (Hala et al., 2010), they also noted that DPPH
activity decrease during storage period.
4.6.2 Peroxide value (POV)Oxidation of food results in production of free radicles that interact with other
molecules to produce aldehydes and ketones and yields bad flavor in food that can be
calculated by using the peroxide value (Ozkan et al., 2007).
79
Table 4.6.1a MS for DPPH activity of SC
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
7796.7 2624.6 126.1 399.5
10947.0
3898.36 201.89 4.85 4.76
819.70** 42.45** 1.02
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
40 45 50 55 60 65 70 75 80 8563.34
65.277.2
79.381.7
76.0776.8277.2576.9
78.2580.35
75.976.3576.9
51.7653.32
67.2468.81
70.665.3165.97
66.767.0367.267.89
65.0265.4166.1
30 day 15 day 1 day
Cheese DPPH
Treat
ment
Fig. 4.17 Effect of edible coating and EOs on the DPPH activity (%) of SC
80
Table 4.6.1b Effect of treated SC on the % DPPH activity during storage
Treat Days Total
1 day 15 day 30 day
T0 76.68±1.34 63.34±0.95 51.76±0.70 63.93±3.64D
T1 76.80±1.05 65.20±0.47 53.32±0.34 65.11±3.41D
T2 84.40±1.67 77.20±0.79 67.24±0.96 76.28±2.56BC
T3 86.23±1.14 79.30±1.71 68.81±0.26 78.11±2.60ABC
T4 90.27±0.61 81.70±0.88 70.60±0.79 80.86±2.87A
T5 83.10±1.28 76.07±1.68 65.31±1.08 74.83±2.68C
T6 84.18±1.82 76.82±0.58 65.97±1.45 75.66±2.73BC
T7 84.96±0.66 77.25±1.55 66.70±1.38 76.30±2.72BC
T8 84.10±1.90 76.90±1.06 67.03±1.03 76.01±2.57BC
T9 86.02±0.99 78.25±1.25 67.20±0.77 77.16±2.78BC
T10 89.07±1.22 80.35±0.50 67.89±1.98 79.10±3.15AB
T11 83.01±2.24 75.90±0.87 65.02±1.62 74.64±2.75C
T12 83.89±1.78 76.35±1.70 65.41±1.52 75.22±2.81C
T13 84.54±0.76 76.90±1.52 66.10±1.07 75.85±2.74BC
Total 84.09±0.64A 75.82±0.82B 64.88±0.86CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
81
Table 4.6.2a MS for POV value of SC
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
47.0562 0.0618 0.0869 0.1656 47.3705
23.5281 0.0048 0.0033 0.0020
11764.05** 2.41** 1.70*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.5 1 1.5 2 2.50.72530.7210.7140.7060.6940.7190.7150.710.7150.7090.7050.7240.7150.71
1.931.83
1.721.711.711.731.721.721.721.721.711.7211.721.72
30 day 15 day 1 day
Cheese POV
Treat
ment
Fig. 4.18 Effect of edible coating and EOs on the POV (meq/kg) of SC
Table 4.6.3b Effect of treated SC on the POV (meq/kg) during storage
82
Treat Days Total
1 day 15 day 30 day
T0 0.289±0.006d 0.725±0.020c 1.924±0.049a 0.980±0.245A
T1 0.285±0.009d 0.720±0.017c 1.826±0.024ab 0.944±0.229AB
T2 0.280±0.006d 0.714±0.014c 1.719±0.034b 0.904±0.213B
T3 0.275±0.003d 0.706±0.014c 1.710±0.051b 0.897±0.213B
T4 0.274±0.003d 0.694±0.009c 1.705±0.052b 0.891±0.213B
T5 0.281±0.006d 0.720±0.015c 1.725±0.014b 0.909±0.214AB
T6 0.280±0.006d 0.715±0.020c 1.720±0.047b 0.905±0.214B
T7 0.280±0.015d 0.710±0.006c 1.715±0.023b 0.902±0.213B
T8 0.284±0.006d 0.715±0.020c 1.720±0.044b 0.906±0.213B
T9 0.284±0.007d 0.710±0.012c 1.714±0.043b 0.903±0.212B
T10 0.279±0.006d 0.705±0.013c 1.710±0.040b 0.898±0.212B
T11 0.285±0.013d 0.724±0.014c 1.721±0.035b 0.910±0.213AB
T12 0.284±0.007d 0.715±0.014c 1.720±0.040b 0.907±0.213B
T13 0.284±0.007d 0.710±0.020c 1.715±0.052b 0.903±0.213B
Total 0.282±0.002C 0.713±0.004B 1.739±0.013AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
83
The result of mean squares for POV of SC samples are shown in Table 4.6.2a. Results revealed that treatments and storage days have ** (p<0.01) effect while their interactive effect have * (p<0.01) variation on the peroxide value of SC. The mean POV values for SC are given in Table 4.6.2b and results showed that active edible coating and essential oils have ** (p<0.01) effect on POV of SC. Data revealed that the maximum POV (1.924) was noted in (T0) at 30 days while the minimum POV (0.274) was noted in T4 (SC containing 1.0% CO) at 1 day. All the SC samples which were prepared with WP based active edible coating (containing clove and peppermint oil as active ingredient) and samples with direct incorporation of these natural essential oils show consistent increase in POV during storage of 30 days as compared to control (T0).
Fig. 4.18 showed that during storage of 30 days a continuous increase in POV of SC was noted in SC samples which were manufactured with edible coating and different concentrations of essential oils. Mean value of all treatments shows that POV was increasing from 0.274 to 0.289, 0.694 to 0.726 and 1.705 to 1.925 at 1, 15 and 30 days respectively. Mean peroxide value for all SC samples at 15 and 30 days of storage showed a ** increase.
There is inverse relationship between EOs concentration and POV as the concentration of EOs increase in coating and cheese POV decrease and vice versa but overall values of all the samples decrease during storage. Free radicals are produced during storage; leads to the formation of aldehydes and ketones results in the production of off flavor/ bad smell and rancidity that ultimately causes the spoilage of cheese. PMO and CO binds the free radicals that ultimately prevent the spoilage of SC. Results of POV are in line with the findings of (Mervat et al., 2010; O'Connor and O'Brien, 2006), they also observed that POV decrease during storage.4.6.3 Measurement of TBARS value
TBARS is routinely a parameter or index point for lipid oxidation or oxidative deterioration that leads to spoilage of the food product (Luo et al., 2011).
The results of mean squares for TBARS of SC samples are shown in Table 4.6.3a. It was evident from the data that treatment, storage days and their interactive effect shows a ** (p<0.01) effect on the TBARs value of SC. The mean values for TBARS of SC are given in Table 4.6.3b. Results showed that active edible coating and essential oils have ** (p<0.01) effect on TBARS of SC. Data revealed that the maximum value of TBARS (0.393) was noted in T0 at 30 days while the lowest TBARS (0.139) was noted in T4 (SC containing 1.0% clove
84
Table 4.6.3a MS for TBARS of SC
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.6150300.0432720.0127200.0113480.682370
0.307515 0.003329 0.000489 0.000135
2276.28** 24.64** 3.62**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.450.307
0.2910.226
0.2210.217
0.230.2280.2250.230.227
0.2210.2340.231
0.226
0.3930.376
0.3170.312
0.3060.3210.318
0.3120.3210.318
0.3140.326
0.3220.312
30 day 15 day 1 day
Cheese TBARS
Treatm
ent
85
Fig. 4.19 Effect of edible coating and EOs on the TBARS (mg MDA/kg) of SC
Table 4.6.3b Effect of treated SC on the TBARS value (mg MDA/kg) during storage
Treat Days Total
1 day 15 day 30 day
T0 0.163±0.003d 0.307±0.007b 0.393±0.008a 0.288±0.034A
T1 0.167±0.004d 0.291±0.003b 0.376±0.008a 0.278±0.030A
T2 0.151±0.005d 0.226±0.008c 0.317±0.009b 0.231±0.024BC
T3 0.147±0.005d 0.221±0.008c 0.312±0.009b 0.227±0.024BC
T4 0.139±0.003d 0.217±0.004c 0.306±0.009b 0.221±0.024C
T5 0.163±0.007d 0.230±0.005c 0.321±0.004b 0.238±0.023BC
T6 0.157±0.003d 0.228±0.011c 0.318±0.008b 0.234±0.024BC
T7 0.153±0.003d 0.225±0.008c 0.312±0.005b 0.230±0.023BC
T8 0.155±0.002d 0.230±0.008c 0.321±0.015b 0.235±0.025BC
T9 0.151±0.002d 0.227±0.006c 0.318±0.009b 0.232±0.024BC
T10 0.146±0.007d 0.221±0.002c 0.314±0.003b 0.227±0.024BC
T11 0.165±0.003d 0.234±0.005c 0.326±0.006b 0.242±0.023B
T12 0.159±0.005d 0.231±0.009c 0.322±0.002b 0.237±0.024BC
T13 0.157±0.008d 0.226±0.010c 0.312±0.008b 0.232±0.023BC
Total 0.155±0.002C 0.237±0.004B 0.326±0.004AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PM
86
oil) at 1 day. All the SC samples which were prepared with WP based active edible
coating (containing clove and peppermint oil as active ingredient) and samples with
direct incorporation of these natural essential oils show consistent increase in TBARS
during storage of 30 days as compared toT0 SC. The overall values for the SC samples
showed gradual increase in TBARS during storage.
Fig. 4.19 depicted that during storage of 30 days continuous increase in TBARs
values of SC samples were noted which were formulated with edible coating and
different concentrations of natural essential oils. Mean SC values shows that TBARs was
increasing from 0.139 to 0167, 0.217 to 0.307 and 0.306 to 0.393 at 1, 15 and 30 days
respectively. Mean TBARS values for SC samples at 15 and 30 days of storage showed a
** increase.
Results indicate that there is a negative correlation between natural essential oil
concentration and TBARS value as the concentration of CO and PMO increases in edible
coating TBARS value decreases and vice versa but higher concentration affects the
sensory
attributes. Mean TBARS values of samples at 15 and 30 days of storage showed a
significant increase. Oxidation reactions of lipids produce secondary metabolites
malonaldehyde that upon reaction with TBA produce pink color. Eugenol in CO and
menthol in PMO due to their antioxidant activities inhibit the production of TBARS.
Results of TBARS match with the findings of Simsek, (2011) he also observed that
TBARS value decrease during storage that might be due to storage condition.
4.7Proteolysis
Structural changes along with flavor and color development are the significant
effects of proteolysis in SC (Orlyuk and Stepanishchev, 2014). Formation of
plasminogen to plasmin, milk microflora, industrial process conditions and ripening
interval conditions are the factors that that contribute in proteolysis of SC. Proteolysis
can be increased by the temperature required for enzyme activity, lower salt
concentration, amount of rennet required, water activity (aw), and encouraging pH
activity (Kalit et al., 2005). Soft texture of SC is also because of breaking down of
complex proteins into simpler poly peptides and small WSP that are not presents in
protein (Upreti et al., 2006).
87
4.7.1 RP-HPLCAlpha s-1, Alpha s-2 and beta casein
Proteolysis is a biochemical process that happens during ripening of cheese
causes the breakdown of complex proteins e.g. casein broken down into the other smaller
fractions by the process of hydrolysis that is the addition of water. Alpha s-1, alpha s-2
and β-caseins undergo by the hydrolysis of casein degradation in SC. Casein was used as
a standard to check the percentage of hydrolysis, so the amount of casein hydrolysis
during ripening was monitored. During proteolysis, water insoluble fractions were used
to study the proteolysis in which alpha s-1, alpha s-2, and β-caseins are known of casein
standard. Alpha s-1, alpha s-2 and β-casein of the casein standard are shown in Fig.20.
Fig. 4.20, 4.21 and 4.22 illustrated the peak areas of alpha s-1, alpha s-2 and β-
caseins respectively. On quantification of SC, three different peaks will be attained on
their specific retention times along with standard curve. Moisture to protein (M/P) ratio
makes a significant difference in the peak areas of the casein fractions. Results revealed
that more breakdown of casein was observed in all the SC samples as compared to
control (T0) SC due to the effect of EOs and edible coating. Treatment (T4) containing
more moisture so more proteolysis was seen and lowest breakdown was seen in control
(T0) SC sample especially in case of alpha s-1, alpha s-2, and beta-caseins. Because of the
proteolysis, less intact casein is available and reduced peak areas were recorded in alpha
s-1, alpha s-2 and beta casein during the storage of SC samples.
Results obtained were studied and analyzed that growth in ripening of SC causes
the reduction of peak area of casein fractions. The reduced peak area of casein is because
of degradation of proteins either by coagulant or any protease enzyme or bacterial starter
culture that enhances the proportion of hydrolyzed proteins or decline in the level of
intact casein during storage of cheese (McSweeney et al., 1993). Alpha s-1 casein
hydrolysis results in the production of alpha s-1 fraction-I that results in case of alpha s-1
casein (Ceruti et al., 2012). A study conducted on SC by Michaelidou et al. (1998) for
the generation of peptides that are produced during the hydrolysis of casein and
concluded that peptide profile vary depending upon the hydrolysis as alpha s-1 have
potential of some peptide and some peptides also originate from C-Terminal of beta-
casein. There is a decrease in beta casein during hydrolysis of casein because some
fraction of beta casein converted into capa-casein by the action of plasmin is reported by
Fox and McSweeney, (1996)
88
Fig. 4.20 Standard casein
T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T130
500
1000
1500
2000
2500
3000
Fig. 4.21 Break down of αS1 casein during storage
89
T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T130
200
400
600
800
1000
1200
1400
1600
1800
Fig. 4.22 Break down of αS2 casein during storage
T0 T1 T2 T3 T4 T5 T6 T7 T8 T9 T10 T11 T12 T130
500
1000
1500
2000
2500
Fig. 4.23 Break down of β casein during storage
4.8 Microbiological analysis
90
4.8.1 Total Viable count (TVC)The results of mean squares for TVC of SC samples are shown in Table 4.8.1a. It
was evident from the data that treatments, storage days and their interactive effect shows
a ** (p<0.01) effect on the total viable count (TVC) of SC.
The mean values for total viable count of SC are given in Table 4.8.1b. Results
showed that active edible coating and essential oils have ** (p<0.01) effect on total
viable count of SC.
Data showed that the highest value of total viable count (92.0x104) was recorded in
control SC sample T0 at 30 days while the lowest value of total viable count (0.21x104)
was noted in T4 (SC containing 1.0% CO) at 1 day. All the SC samples which were
prepared with WP based active edible coating (containing clove and peppermint oil as
active ingredient) and samples with direct incorporation of these natural essential oils
show increase in total viable count during storage of 30 days as compared to control (T0)
SC.
Fig. 4.24 depicted that during storage of 30 days increase in total viable count of
SC was noted in SC samples which were formulated with edible coating and different
concentrations of natural essential oils. Mean value of SC samples shows that total viable
count was increasing from 0.21x104 to 4.4x104, 0.59x104 to 69.01x104 and 5.3x104 to
92.0x104 at 1, 15 and 30 days respectively. Mean total viable count values for SC
samples at 15 and 30 days of storage showed a ** increase.
Eugenol in clove oil and menthol in pepper mint oil due to their antimicrobial
activity either control/prevent micro-organisms growth (including pathogenic
microorganisms) or prolong the storage stability of cheese by controlling the natural
spoilage process (food preservation). Results indicate that growth and multiplication rate
of TVC was slow in samples prepared with WP based active edible coating (containing
clove and peppermint oil as active ingredient) as compared to T0. Conclusively, the most
important factor for growth of microorganisms is concentration of essential oil. Findings
of TVC match with the results of Gokçe et al. (2010), they also reported that TVC
consistently increase during storage. In another study clove, thyme, cinnamon and bay
EOs were tested against Salmonella enteritidis and L. monocytogenes. Results indicate
that clove oil was more effective against Salmonella enteritidis (Burt, 2004).
Table 4.8.1a MS for total viable count of SC
91
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
85856 24475 23035 973
134340
42928.2 1882.7 886.0 11.6
3705.38** 162.51** 76.47**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 10 20 30 40 50 60 70 80 90 10069.01
656.35
0.630.59
5.825.75.576.46.365.935.785.725.6
9285
786.5
5.369.34
6156
8780.5
597069.5
67.6
30 day 15 day 1 day
Cheese TVC
Treat
ment
Fig. 4.24 Effect of edible coating and EOs on the total viable count (cfu/g) of SC
Table 4.8.1b Effect of treated SC on the TVC (X 104 cfu/g) during storage
92
Treat Days Total
1 day 15 day 30 day
T0 4.40±0.098g 69.00±3.783de 92.00±3.251a 55.13±13.190A
T1 4.30±0.040g 65.00±2.575ef 85.00±2.568ab 51.43±12.177A
T2 4.20±0.069g 6.35±0.110g 78.00±4.290bcd 29.52±12.188BC
T3 0.32±0.003g 0.63±0.006g 6.50±0.104g 2.48±1.006E
T4 0.21±0.003g 0.59±0.012g 5.30±0.092g 2.03±0.819E
T5 4.05±0.087g 5.82±0.075g 69.33±2.995cde 26.40±10.771CD
T6 3.98±0.052g 5.70±0.040g 61.00±4.820ef 23.56±9.466D
T7 3.80±0.064g 5.57±0.069g 56.00±2.601f 21.79±8.589D
T8 4.26±0.040g 6.40±0.052g 87.00±2.480ab 32.55±13.634B
T9 4.04±0.064g 6.36±0.098g 80.50±3.470bc 30.30±12.594BC
T10 3.70±0.069g 5.93±0.115g 59.00±2.886ef 22.88±9.075D
T11 4.02±0.069g 5.78±0.075g 70.00±4.387cde 26.60±10.927CD
T12 3.95±0.087g 5.72±0.075g 69.50±4.078cde 26.39±10.845CD
T13 3.88±0.029g 5.60±0.104g 67.60±2.045de 25.69±10.496CD
Total 3.51±0.209C 13.89±3.411B 63.34±4.059AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
4.9 Sensory Evaluation
93
Sensory properties of food play an important role on the overall acceptance of
food based on sensory evaluation is the human response on evaluating food composition
by following the principles and methods towards products acceptability (Braghieri et al.,
2014).Consumer likes and dislikes towards food on the basis of his/her perception is
called sensory perception (Drake et al., 2006). Cheeses are mostly categorized on the
basis of their sensory qualities (SISS, 2012). Sensory evaluation is done by the
experienced panel of judges to measure the sensory food stuff (Stone et al., 2012).
4.9.1 Flavor
Selection, acceptance and ingestion of food are three sensory stuffs that add up
the flavor (Hassan et al., 2013). The flavoring compounds are produced by the
breaking of milk constituents in which lactose and proteins are major contributing
factor to flavor along with citrate, and milk lipids (Singh et al., 2003). Flavor is the
most appealing character and quality parameter for SC. The development of flavor in
cheese quite complex as it is the resultant element of ripend cheese during which
complex enzymatic digestion, physico-chemical and chemical reactions occur (Zishu
and Shah, 2007).
The result of mean squares for flavor of SC samples presented in Table 4.9.1a
revealed a ** (p<0.01) effect on flavor of SC due to treatments and storage days while
their interactive effect was also found to have a ** effect (p<0.01).
The mean values for flavor of SC are given in Table 4.9.1a and results showed
that active edible coating and essential oils have ** (p<0.01) effect on flavor of SC.
Data showed that the maximum flavor score (9.52) was recorded in SC sample T4 (SC
containing 1.0% CO) at 1 day while the lowest value of flavor (3.52) was noted in
control (T0) SC sample at 30 days. All the SC samples which were prepared with WP
based active edible coating (containing clove and peppermint oil as active ingredient)
and samples with direct incorporation of these natural essential oils show continuous
decrease in flavor during storage of 30 days as compared to control (T0).
Fig. 4.25 depicted that during storage of 30 days a continuous decrease in flavor
of SC was observed in all SC samples which were prepared with active edible coating
and different concentrations of natural essential oils. Mean SC samples values shows that
flavor was
Table 4.9.1a MS for flavor of SC
94
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
21326
210251
626.265 99.929 41.270 45.243812.707
313.132 7.687 1.587 0.215
1453.42** 35.68** 7.37**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 1 2 3 4 5 6 7 8 96.02
6.777.25
8.028.52
7.026.98
7.57
7.257.5
77.25
7.02
3.524.02
4.254.27
6.544
4.254.52
5.027
4.274.25
4.5
30 day 15 day 1 day
Cheese Flavor
Treat
ment
Fig. 4.25 Effect of edible coating and EOs on the flavor of SC
Table 4.9.1b Effect of treated SC on the flavor during storage
95
Treat Days Total
1 day 15 day 30 day
T0 7.00±0.317def 6.02±0.367fg 3.52±0.135i 5.51±0.389E
T1 7.48±0.187cde 6.77±0.206ef 4.02±0.135hi 6.09±0.375D
T2 8.00±0.304bcd 7.25±0.171de 4.25±0.076hi 6.50±0.409CD
T3 9.00±0.165ab 8.02±0.371bcd 4.27±0.033hi 7.09±0.511B
T4 9.52±0.125a 8.52±0.145abc 6.50±0.097ef 8.18±0.312A
T5 8.02±0.174bcd 7.02±0.274def 4.00±0.026hi 6.34±0.426CD
T6 8.52±0.194abc 6.98±0.130def 4.00±0.026hi 6.50±0.461CD
T7 9.50±0.134a 7.50±0.141cde 4.25±0.143hi 7.08±0.530B
T8 8.00±0.274bcd 7.00±0.157def 4.52±0.060hi 6.51±0.369CD
T9 8.00±0.134bcd 7.25±0.112de 5.02±0.135gh 6.76±0.315BC
T10 8.52±0.145abc 7.50±0.229cde 7.00±0.237def 7.67±0.190A
T11 8.00±0.153bcd 7.00±0.285def 4.27±0.095hi 6.42±0.397CD
T12 9.00±0.093ab 7.25±0.226de 4.25±0.163hi 6.83±0.485BC
T13 8.52±0.178abc 7.02±0.227def 4.50±0.184hi 6.68±0.416BC
Total 8.36±0.090A 7.22±0.084B 4.60±0.108CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
96
decreasing from 9.52 to 7.0, 8.52 to 6.02 and 6.5 to 3.52 at 1, 15 and 30 days respectively. Mean flavor values for SC samples at 15 and 30 days of storage showed a ** decrease.
Edible coating and incorporation of EOs in SC samples significantly affects the sensory properties of SC. Results revealed that flavor score decrease during storage but the reduction rate in flavor score was less as compare to control SC that might be due to proteolysis and lipolysis. The overall score of all the SC samples decrease during storage. Similar results were found by Songul et al, (2014) they also reported that flavor score decrease during storage. 4.9.2 Appearance
To judge the visual quality of cheese, the appearance of cheese is the important factor that gives the best impression to the judge. Shape of cheese shreds, surface consistency and color are some visual characteristics of cheese (Clark et al., 2009).
Results of mean squares for appearance of SC samples are presented in Table 4.9.2a. Results revealed that treatments and storage days have ** (p<0.01) effect while their interactive effect have * (p<0.05) variation on the appearance of SC. The mean appearance values for SC are given in Table 4.9.2b. Results showed that active edible coating and essential oils have ** (p<0.01) effect on appearance of SC. Data showed that the highest appearance value (9.50) was recorded in SC sample T3 (SC containing 0.75.0% CO) and T4 (SC containing 1.0% CO) at 1 day while the lowest value of appearance (2.98) was noted in control (T0) at 30 days. The SC samples which were formulated with WP based edible coating (containing clove and peppermint oil as active ingredient) and samples with direct incorporation of these natural essential oils show consistent decrease in appearance during storage of 30 days as compared to control (T0) SC. The overall values for all the treatments showed gradual decrease in appearance during storage.
Fig. 4.26 depicted that during storage of 30 days a continuous decrease in appearance of SC was observed in SC samples which were formulated with edible coating and different concentrations of natural essential oils. Mean value of SC samples shows that appearance was decreasing from 9.0 to 7.0, 7.47 to 5.98 and 3.52 to 2.52 at 1, 15 and 30 days respectively. Mean appearance values for SC at 15 days and 30 days of storage showed a ** decrease.
Edible coating and incorporation of EOs in SC samples significantly affects the sensory properties of SC. Results revealed that appearance/color value decrease during storage but the reduction rate in color value was less as compare to control SC that might be due to proteolysis
97
Table 4.9.2a MS for appearance of SC
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
21326
210251
1400.871 19.511 10.539 53.640
1484.561
700.435 1.501 0.405 0.255
2742.20** 5.88** 1.59*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 1 2 3 4 5 6 7 85.98
6.57.02
7.257.47
6.526.746.75
77
7.256.756.84
7
2.523
3.273.25
3.5232.98
3.53.25
3.483.52
33
3.52
30 day 15 day 1 day
Cheese Appearance
Treat
ment
Fig. 4.26 Effect of edible coating and EOs on the appearance of SC
98
Table 4.9.2b Effect of treated SC on the appearance during storage
Treat Days Total
1 day 15 day 30 day
T0 9.00±0.183a 5.98±0.366d 2.52±0.183e 5.83±0.658E
T1 9.00±0.237a 6.50±0.224cd 3.00±0.129e 6.17±0.607B-E
T2 9.50±0.124a 7.02±0.366cd 3.27±0.092e 6.59±0.634ABC
T3 9.50±0.153a 7.25±0.171c 3.25±0.154e 6.67±0.633AB
T4 9.50±0.124a 7.47±0.163bc 3.52±0.130e 6.83±0.607A
T5 8.52±0.187ab 6.52±0.187cd 3.00±0.258e 6.01±0.565DE
T6 8.50±0.171ab 6.73±0.167cd 2.98±0.259e 6.07±0.569CDE
T7 8.50±0.198ab 6.75±0.092cd 3.50±0.139e 6.25±0.509B-E
T8 9.02±0.187a 7.00±0.373cd 3.25±0.092e 6.42±0.595A-D
T9 9.00±0.183a 7.00±0.366cd 3.48±0.130e 6.49±0.569A-D
T10 9.00±0.139a 7.25±0.112c 3.52±0.130e 6.59±0.559ABC
T11 8.52±0.194ab 6.75±0.208cd 3.00±0.129e 6.09±0.566CDE
T12 8.52±0.217ab 6.83±0.158cd 3.00±0.263e 6.12±0.572B-E
T13 8.52±0.130ab 7.00±0.373cd 3.52±0.183e 6.34±0.526A-E
Total 8.90±0.060A 6.86±0.075B 3.20±0.053CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
99
and lipolysis. It is also by pH and acidity because during storage pH decrease and acidity
increase results in the production of LAB that leads to variation in the appearance. The
overall score of all the SC samples decrease during storage. Similar results were found
by Songul et al, (2014) they also reported that color value decrease during storage.
4.6.3Texture/BodyMastication is a process that involves chewing with saliva, a practice for sensory
evaluation of cheese texture while force is applied to deform the body of cheese.
Mastication process gives the perception about the sensory texture of cheese that cannot
be measured by any sensitive equipment (Foegeding and Drake, 2007).
The results of mean squares for body/texture of SC samples are shown in Table
4.9.3a. Results revealed that treatments and storage days have ** (p<0.01) effect while
their interactive effect have * (p<0.05) variation for the body/texture of SC. The mean
texture values for SC are illustrated in Table 4.9.3b. Results showed that active edible
coating and essential oils have ** (p<0.01) effect on texture of SC. Data showed that the
maximum texture score (9.50) was recorded in SC sample T7 (WP based edible coating of
SC containing 1.0% CO) at 1 day while the lowest value of texture (2.23) was noted in
control (T0) at 30 days. The SC samples which were formulated with WP based edible
coating (containing clove and peppermint oil as active ingredient) and samples with
direct addition of these natural essential oils show regular decrease in texture during
storage of 30 days as compared to control (T0).
Fig. 4.27 depicted that during storage of 30 days a continuous decrease in
texture/body of SC was observed in SC samples which were formulated with edible
coating and different concentrations of natural essential oils. Mean SC samples shows
that body/texture was decreasing from 9.5 to 8.0, 6.52 to 5.0 and 4.25 to 2.24 at 1, 15 and
30 days respectively. Mean texture/body value for SC samples at 15 and 30 days of
storage showed a ** decrease.
Edible coating and incorporation of EOs in SC samples significantly affects the
sensory properties of SC. Results revealed that texture value decrease during storage but
the reduction rate in texture value was less as compare to control SC that might be due to
proteolysis and lipolysis. The overall score of all the SC samples decrease during storage.
Similar results were found by Songul et al. (2014) they also reported that texture value
decrease during storage.
100
Table 4.9.3a MS for texture of SC
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
21326
210251
1066.190 48.499 3.649 18.087
1136.424
533.095 3.731 0.140 0.086
6189.64** 43.32** 1.63*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 1 2 3 4 5 6 75
5.55.25
5.756.02
6.276.27
6.525.5
5.7566.02
6.286.28
2.243.5
3.253.5
3.754
4.244.25
3.523.5
3.743.74
4.024.25
30 day 15 day 1 day
Cheese Texture
Treat
ment
Fig. 4.27 Effect of edible coating and EOs on the texture/body of SC
101
Table 4.9.3b Effect of treated SC on the texture/body during storage
Treat Days Total
1 day 15 day 30 day
T0 8.00±0.365e 5.00±0.263j 2.23±0.021n 5.08±0.588J
T1 8.25±0.056de 5.50±0.097hij 3.50±0.058lm 5.75±0.474HI
T2 8.27±0.109de 5.25±0.043ij 3.25±0.043m 5.59±0.502I
T3 8.52±0.098cde 5.75±0.076ghi 3.50±0.058lm 5.92±0.499E-H
T4 8.75±0.085bcd 6.02±0.040fgh 3.75±0.043klm 6.17±0.497DEF
T5 9.00±0.106abc 6.27±0.115fg 4.00±0.026kl 6.42±0.498BCD
T6 9.27±0.152ab 6.27±0.080fg 4.23±0.049k 6.59±0.505AB
T7 9.50±0.077a 6.52±0.087f 4.25±0.043k 6.76±0.523A
T8 8.27±0.092de 5.50±0.097hij 3.52±0.031lm 5.76±0.474GHI
T9 8.50±0.077cde 5.75±0.076ghi 3.50±0.058lm 5.92±0.497FGH
T10 9.00±0.139abc 6.00±0.052fgh 3.73±0.182klm 6.24±0.528CDE
T11 8.48±0.190cde 6.02±0.266fgh 3.73±0.084klm 6.08±0.482EFG
T12 9.02±0.091abc 6.28±0.087fg 4.02±0.017kl 6.44±0.497A-D
T13 9.02±0.145abc 6.28±0.105fg 4.25±0.123k 6.52±0.479ABC
Total 8.70±0.059A 5.89±0.056B 3.68±0.058CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
102
4.9.4 Overall acceptability
Performa for sensory evaluation was used to assess the score or value for overall
acceptability of SC.
The results of mean squares for overall acceptability of SC samples are shown in
Table 4.9.4a. Results showed that treatments and storage days have ** (p<0.01) effect
while their interactive effect * (p<0.05) variation for the overall acceptability of SC.
The mean overall acceptability values for SC are given in Table 4.9.4b. Results
showed that edible coating and essential oils have ** (p<0.01) variation on overall
acceptability of SC. Data showed that the highest overall acceptability value (9.73) was
recorded in SC sample T9 (SC containing 2.5% peppermint oil) while the lowest value of
overall acceptability (2.50) score was found in control. All the SC samples which were
prepared with WP based active edible coating (containing clove and peppermint oil as
active ingredient) and samples with direct incorporation of these natural EOs show
continuous decline in overall acceptability during storage of 30 days as compared to
control (T0).
Fig. 4.28 depicted that during storage of 30 days a continuous decrease in overall
acceptability of SC was recorded in SC samples which were formulated with edible
coating and different concentrations of clove and peppermint oils. Mean value of all
treatments shows that body/texture was decreasing from 9.74 to 8.0, 7.0 to 5.25 and 4.0
to 2.5 at 1, 15 and 30 days respectively. Mean overall acceptability score for SC samples
at 15 and 30 days of storage showed a ** decrease.
Edible coating and incorporation of EOs in SC samples significantly affects the
sensory properties of SC. Results revealed that overall acceptability score was decrease
during storage but the reduction rate in overall acceptability was less as compare to
control SC that might be due to proteolysis and lipolysis. The overall score of all the SC
samples decrease during storage. Similar results were found by Songul et al, (2014), they
also reported that overall acceptability score decrease during storage.
103
Table 4.9.4a MS for overall acceptability of SC
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
21326
210251
1182.815 51.162 14.167 62.872
1311.016
591.407 3.936 0.545 0.299
1975.38** 13.15** 1.82*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 1 2 3 4 5 6 7 86
6.276.27
6.527
5.255.52
5.256
76.75
5.55.75
5.5
2.53
3.53.5
43
3.253.25
3.523.5
42.983
3.27
30 day 15 day 1 day
Cheese Overall Acceptability
Treat
ment
Fig. 4.28 Effect of edible coating and EOs on the overall acceptability of SC
104
Table 4.9.4b Effect of treated SC on the overall acceptability during storage
Treat Days Total
1 day 15 day 30 day
T0 8.02±0.135cd 6.00±0.373e-h 2.50±0.129j 5.51±0.568D
T1 8.52±0.259abc 6.27±0.120e-h 3.00±0.129ij 5.93±0.558BCD
T2 8.73±0.167abc 6.27±0.120e-h 3.50±0.190ij 6.17±0.526ABC
T3 9.50±0.124ab 6.52±0.217efg 3.50±0.183ij 6.51±0.602AB
T4 9.00±0.246abc 7.00±0.259de 4.00±0.144i 6.67±0.513A
T5 8.00±0.447cd 5.25±0.112h 3.00±0.263ij 5.42±0.523D
T6 8.27±0.120bc 5.52±0.194fgh 3.25±0.154ij 5.68±0.505CD
T7 8.27±0.285bc 5.25±0.092h 3.25±0.214ij 5.59±0.513CD
T8 9.00±0.225abc 6.00±0.366e-h 3.52±0.183ij 6.17±0.563ABC
T9 9.73±0.076a 7.00±0.365de 3.50±0.026ij 6.74±0.630A
T10 8.75±0.214abc 6.75±0.281ef 4.00±0.026i 6.50±0.485AB
T11 8.02±0.371cd 5.50±0.224gh 2.98±0.031ij 5.50±0.517D
T12 8.25±0.310c 5.75±0.089fgh 3.00±0.259ij 5.67±0.536CD
T13 8.50±0.316abc 5.50±0.224gh 3.27±0.120ij 5.76±0.535CD
Total 8.61±0.085A 6.04±0.087B 3.30±0.059CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of SC with WP T2 = SC containing 0.5% COT3= SC containing 0.75% COT4= SC containing 1.0% COT5= WP based edible coating of SC containing 0.5% COT6= WP based edible coating of SC containing 0.75% COT7= WP based edible coating of SC containing 1.0% COT8= SC containing 1.5% PMOT9= SC containing 2.0% PMOT10= SC containing 2.5% PMOT11= WP based edible coating of SC containing 1.5% PMOT12= WP based edible coating of SC containing 2.0% PMOT13= WP based edible coating of SC containing 2.5% PMO
105
Butter
4.10 Physico-chemical analysis of cream
Fresh cream used for manufacturing of butter was analyzed for different
parameters such as fat, protein, pH, acidity, total solids, ash and free fatty acids (Table
4.10). The results of cream match to the findings of Ozkan et al. (2007) they had reported
almost the same results of cream.
4.11 Analysis for edible coating of butter
Table 4.10 Physico-chemical analysis of cream
Parameters Mean
Fat 28.5±0.01 %
Protein 0.36± 0.01%
Ph 6.64±0.01
Acidity 0.15±0.01 %
Total solids 54.5± 0.02%
Ash 0.75± 0.02 %
Free fatty acids 0.23± 0.01%
Table 4.11 Physico-chemical analysis of edible coating of butter
Parameters Mean
pH 5.65±0.01
Acidity (%) 0.15±0.02
Viscosity (cP) 7568±0.05
Water activity 0.04±0.01
106
4.12 Physicochemical Analysis of Butter4.12.1 Moisture
Moisture is tightly bound in butter and present as water in oil (w/o) emulsion
form. It affects the quality of butter because as the quantity of moisture in butter increases
rate of firmness decreases (Olson and Price, 1999).
The result of mean squares (ANOVA) for moisture content of butter samples
shown in Table 4.12.1a revealed a ** (p<0.01) variation in moisture contents of butter
due to treatments, storage days and also the interactive effect of treatments and storage
days.
The mean moisture values for butter are given in Table 4.12.1b. Results showed
that edible coating and essential oils have ** (p<0.01) effect on % moisture of butter.
Data revealed that the highest moisture (16.53 %) was recorded in butter sample T10
(butter containing 2.5% GO) at 1 day while the lowest moisture (11.25%) was noted in
control (T0) at 90 days. Butter samples which were formulated with CS based active
edible coating (containing BCO and GO as active ingredient) and samples with direct
addition of these natural essential oils shows minimum moisture loss during storage of 90
days as compared to control (T0) butter. The overall values for all the butter treatments
showed gradual decrease in moisture during storage.
Fig. 4.29 depicted that during storage of 90 days a continuous decrease in
moisture content of butter was observed in all butter samples which were prepared with
active edible coating and natural essential oils in different concentrations. Mean value of
all butter treatments shows that moisture was decreasing from 16.52% to 16.30%,
15.40% to 13.50% and 14.0% to 11.25% at 1, 45 and 90 days respectively. Mean
moisture values for butter treatments at 45 and 90 days of storage showed a ** decrease.
Edible coating serve as a barrier against moisture loss by covering the surface of
butter samples so minimum moisture loss was noted in edible coated samples of butter.
Thymol in BCO and gingerol in GO have the antioxidant and antimicrobial potential so
binds the free radicals in butter samples that ultimately leads to minimum moisture loss
as compared to control (T0) but the overall values for all treatments of butter showed
gradual decrease in moisture during storage. The findings of this parameter match with
the results of Foda et al. (2010), they also noted that pH in butter decrease during
storage.
107
Table 4.12.1a MS for moisture contents of butter
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
233.3399 20.0511 12.3670 16.6523282.4102
116.6699 1.5424 0.4757 0.1982
588.52** 7.78** 2.40**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
10 11 12 13 14 15 1613.5
1415
15.115.25
1515.1
15.2515.00
15.1515.25
15.215.3515.4
11.2512
12.7512.9
13.2513
13.113.5
13.2513.3513.4
13.7513.8
14
90 day 45 day 1 day
Butter Moisture
Treat
ment
Fig. 4.29 Effect of edible coating and EOs on the moisture contents (%) of butter
108
Table 4.12.1b Effect of treated butter on the % moisture during storage
Treat Days Total
1 day 45 day 90 day
T0 16.50±0.185a 13.50±0.231f 11.25±0.092h 13.75±0.766C
T1 16.30±0.127ab 14.00±0.162c-f 12.00±0.335gh 14.10±0.631BC
T2 16.50±0.208a 15.00±0.225b-e 12.75±0.191fg 14.75±0.555AB
T3 16.52±0.323a 15.10±0.127a-e 12.90±0.248fg 14.84±0.541A
T4 16.52±0.416a 15.25±0.289a-d 13.25±0.202fg 15.01±0.501A
T5 16.30±0.294ab 15.00±0.318b-e 13.00±0.283fg 14.77±0.503AB
T6 16.30±0.260ab 15.10±0.225a-e 13.10±0.248fg 14.83±0.482A
T7 16.35±0.231ab 15.25±0.248a-d 13.50±0.306f 15.03±0.435A
T8 16.50±0.294a 15.00±0.234b-e 13.25±0.300fg 14.92±0.490A
T9 16.52±0.197a 15.15±0.260a-e 13.35±0.127fg 15.01±0.470A
T10 16.53±0.416a 15.25±0.167a-d 13.40±0.179fg 15.06±0.475A
T11 16.30±0.252ab 15.20±0.133a-e 13.75±0.191ef 15.08±0.382A
T12 16.35±0.445ab 15.35±0.150abc 13.80±0.087def 15.17±0.396A
T13 16.35±0.358ab 15.40±0.433abc 14.00±0.110c-f 15.25±0.379A
Total 16.42±0.068A 14.97±0.097B 13.09±0.120CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
109
4.12.2 pHpH of butter plays a very important role in quality assessment of butter. Flavor
perception of butter is directly affected by pH (Naveena et al., 2004).
The result of mean squares for pH of butter samples presented in Table 4.12.2a
revealed a significant (p<0.05), ** (p<0.01) and NS (p>0.05) variation in moisture
contents of butter samples due to treatments, storage days and their interactive effect of
treatments and storage days respectively for pH of butter.
The mean pH values for butter are given in Table 4.12.2b. Results showed that edible
coating and essential oils have ** (p<0.01) effect on pH of butter. Data revealed that the
highest pH (5.31) was recorded in butter sample T11 (CS based edible coating of butter
containing 1.5% GO) at 1 day while the minimum pH (4.85) was noted in control (T0) at
90 days. Butter samples which were formulated with CS based active edible coating
(containing black cumin and ginger oil as active ingredient) and samples with direct
incorporation of these natural essential oils shows minimum decrease in pH during
storage of 90 days as compared to control (T0) butter sample. The overall values for all
the butter treatments showed gradual decrease in pH during storage.
Fig. 4.30 depicted that during storage of 90 days a continuous decrease in pH of
butter was observed in butter samples which were formulated with edible coating and
natural EOs in different concentrations. Mean values for treatments of butter shows that
pH was decreasing from 5.32 to 5.23, 5.26 to 5.04 and 5.23 to 4.85 at 1, 15 and 30 days
respectively. Mean values of pH for butter samples at 15 days and 30 days of storage
showed a ** decrease.
pH reduction during storage of butter mainly due to the production of lactic acid
by LAB and it might be decrease due to the acidity of GO and BCO that is added in
butter samples. The overall values for all treatments of butter shows that pH decrease
during storage but the rate of reduction was slow in all samples as compare to control due
to the effect of edible coating and EOs. The findings of this parameter match with the
results of Ozkan et al. (2007) they also noted that pH in butter decrease during storage.
4.12.3 AcidityThe result of mean squares (ANOVA) for acidity of butter samples presented in
Table 4.12.3a revealed a ** (p<0.01) variation in acidity of butter samples due to
treatments, storage days and also the interactive effect of treatments and storage days.
110
Table 4.12.2a MS for pH of butter
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.43163 0.40951 0.19766 1.36300 2.40180
0.215810.031500.007600.01623
13.30** 1.94* 0.47NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
4.6 4.7 4.8 4.9 5 5.1 5.2 5.35.05
5.135.255.25
5.245.24
5.265.23
5.225.23
5.255.255.26
5.23
4.854.96
5.175.18
5.235.16
5.185.19
5.175.2
5.225.17
5.195.18
90 day 45 day 1 day
Butter pH
Treat
ment
Fig. 4.30 Effect of edible coating and EOs on the pH of butter
111
Table 4.12.2b Effect of treated butter on the pH during storage
Treat Days Total
1 day 45 day 90 day
T0 5.22±0.058 5.05±0.079 4.85±0.067 5.04±0.064B
T1 5.32±0.075 5.13±0.087 4.96±0.069 5.14±0.065AB
T2 5.30±0.087 5.25±0.075 5.17±0.052 5.24±0.041AB
T3 5.29±0.081 5.25±0.081 5.18±0.075 5.24±0.043AB
T4 5.28±0.098 5.24±0.040 5.23±0.075 5.25±0.038A
T5 5.31±0.035 5.24±0.035 5.16±0.064 5.24±0.032AB
T6 5.31±0.064 5.26±0.035 5.18±0.029 5.25±0.029A
T7 5.29±0.098 5.23±0.069 5.19±0.081 5.24±0.044AB
T8 5.28±0.046 5.22±0.058 5.17±0.087 5.22±0.037AB
T9 5.28±0.069 5.23±0.075 5.20±0.046 5.24±0.034AB
T10 5.27±0.069 5.25±0.081 5.22±0.110 5.25±0.045A
T11 5.31±0.115 5.25±0.064 5.17±0.092 5.24±0.051AB
T12 5.30±0.052 5.26±0.087 5.19±0.069 5.25±0.039A
T13 5.29±0.064 5.23±0.121 5.18±0.040 5.23±0.044AB
Total 5.29±0.017A 5.22±0.019B 5.15±0.023CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
112
Table 4.12.3a MS for acidity of butter
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
1.6546290.0700930.0895710.0434001.857693
0.827314 0.005392 0.003445 0.000517
1601.25** 10.44** 6.67**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.90.6
0.550.490.5
0.480.490.5
0.490.51
0.50.51
0.490.50.5
0.820.76
0.70.68
0.660.7
0.690.68
0.650.62
0.60.7
0.670.66
90 day 45 day 1 day
Butter Acidity
Treat
ment
Fig. 4.31 Effect of edible coating and EOs on the acidity (%) of butter
113
Table 4.12.3b Effect of treated butter on the % acidity during storage
Treat Days Total
1 day 45 day 90 day
T0 0.400±0.010k 0.600±0.015ef 0.820±0.031a 0.607±0.062A
T1 0.390±0.012k 0.550±0.015fg 0.760±0.012ab 0.567±0.054B
T2 0.400±0.006k 0.490±0.012ghi 0.700±0.012bc 0.530±0.045BC
T3 0.410±0.000jk 0.500±0.015gh 0.680±0.010cd 0.530±0.040BC
T4 0.420±0.012ijk 0.480±0.010ghij 0.660±0.010cde 0.520±0.036C
T5 0.390±0.015k 0.490±0.006ghi 0.700±0.015bc 0.527±0.046C
T6 0.400±0.006k 0.500±0.020gh 0.690±0.021bcd 0.530±0.043BC
T7 0.410±0.006jk 0.490±0.015ghi 0.680±0.010cd 0.527±0.040C
T8 0.420±0.006ijk 0.510±0.015g 0.650±0.012cde 0.527±0.034C
T9 0.420±0.006ijk 0.500±0.015gh 0.620±0.015def 0.513±0.030C
T10 0.430±0.010h-k 0.510±0.010g 0.600±0.021ef 0.513±0.026C
T11 0.400±0.012k 0.490±0.006ghi 0.700±0.006bc 0.530±0.045BC
T12 0.410±0.010jk 0.500±0.023gh 0.670±0.010cde 0.527±0.039C
T13 0.410±0.006jk 0.500±0.006gh 0.660±0.015cde 0.523±0.037C
Total 0.408±0.003C 0.508±0.006B 0.685±0.009AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
114
The mean acidity values for butter are given in Table 4.12.3b. Results showed
that edible coating and essential oils have ** (p<0.01) effect on acidity of butter. Data
shows that the high value of acidity (0.82%) was noted in control (T0) butter at 90 days
while the lower value of acidity (0.43%) was noted in T10 (butter containing 2.5% ginger
oil) at 1 day. The butter samples which were formulated with CS based active edible
coating (containing BCO and GO as active ingredient) and samples with direct
incorporation of these natural essential oils shows minimum increase in acidity during
storage of 90 days as compared to T0 butter. The overall values for all the treatments
showed gradual increase in acidity during storage.
Fig. 4.31 depicted that during storage of 90 days a continuous increase in acidity
of butter was observed in butter samples which were formulated with active edible
coating and natural essential oils in different concentrations. Mean acidity values for all
SC samples at 15 days and 30 days of storage showed a highly significant increase.
Increase in acidity during storage of butter mainly due to the production of lactic
acid by LAB and it might be increase due to the acidity of GO and BCO that is added in
butter samples. The findings of this parameter match with the results of Povolo and
Contarini (2003) they also noted that acidity in butter increase during storage.
4.12.4 Fat content
Fat is a major constituent and quality assessment parameter in butter. It plays a
very important role to improve the appearance, texture/body, flavor and overall
acceptability of butter (Shukla et al., 1994).
The result of mean squares (ANOVA) for fat content of butter samples are shown
in Table 4.12.4a. Results revealed that treatments, storage days and their interactive
effect have NS (p>0.05) variation in the % fat of butter.
The mean fat values far butter are shown in Table 4.12.4b. Results showed that
edible coating and EOs have NS (p>0.05) effect on fat of butter. Data showed that the
highest fat (79.80%) was recorded in control butter sample T0 at 90 days while the lowest
fat (79.48%) was noted in T1 (CS based edible coating of butter) at 1 day. Finding of this
experiment shows that there is NS difference among butter samples which were
formulated with CS based edible coating (containing BCO and GO as active ingredient)
and samples with direct addition of these natural EOs.
115
Table 4.12.4a MS for fat content of butter
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.932 0.048 0.044 85.166 86.190
0.466 0.004 0.002 1.014
0.46NS
0.00NS
0.00NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
70 72 74 76 78 80 8279.679.5579.5179.5279.5279.5279.579.579.5579.5479.5679.5279.5179.52
79.879.7579.779.779.779.6879.6779.6779.779.779.6879.779.6979.7
90 day 45 day 1 day
Butter Fat
Treat
ment
Fig. 4.32 Effect of edible coating and EOs on the fat content (%) of butter
116
Table 4.12.4b Effect of treated butter on the % fat during storage
Treat Days Total
1 day 45 day 90 day
T0 79.50±0.638 79.60±0.572 79.80±0.150 79.63±0.255A
T1 79.48±0.612 79.55±0.687 79.75±0.595 79.59±0.319A
T2 79.51±0.064 79.51±0.624 79.70±0.115 79.57±0.187A
T3 79.51±0.121 79.52±0.456 79.70±0.693 79.58±0.244A
T4 79.52±0.531 79.52±0.098 79.70±0.127 79.58±0.163A
T5 79.49±0.572 79.52±0.266 79.68±0.456 79.56±0.227A
T6 79.50±0.375 79.50±0.641 79.67±0.804 79.56±0.317A
T7 79.50±0.566 79.50±0.577 79.67±0.341 79.56±0.255A
T8 79.53±0.375 79.55±0.421 79.70±0.468 79.59±0.213A
T9 79.54±0.803 79.54±0.577 79.70±0.341 79.59±0.303A
T10 79.56±0.462 79.56±1.010 79.68±0.795 79.60±0.395A
T11 79.50±0.843 79.52±0.843 79.70±1.057 79.57±0.461A
T12 79.51±0.254 79.51±0.803 79.69±0.323 79.57±0.262A
T13 79.52±0.722 79.52±0.774 79.70±0.531 79.58±0.343A
Total 79.51±0.121A 79.53±0.141A 79.70±0.123AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
117
Fig. 4.32 depicted that during storage of 90 days a minute increase in % fat of
butter was observed butter samples which were formulated with edible coating and
natural EOs in different concentrations. Mean fat values of butter shows that fat was
increasing from 79.48% to 79.56%, 79.50% to 79.60% and 79.67% to 79.80% at 1, 45
and 90 days respectively. Mean fat values for all butter samples at 45 and 90 days of
storage showed a ** increase.
Results shows that minor increase in fat of butter that is might be due addition of
BCO and GO in the butter. Thymol in BCO and gingerol in GO have the excellent
antioxidant potential so binds the free radicals in butter that ultimately leads to minimum
lipolysis in butter samples as compare to T0. The slight increase in % fat of butter might be
due to loss of moisture. The findings of this parameter match with the results of Sagd et
al. (2004),Simsek, (2011) and Ahmad et al. (2016), they also recorded the slight increase
in fat during storage of butter.
4.12.5 Ash content
Ash is a mixture of inorganic constituents and is first parameter used to analyze
the rough mineral content in any food stuff (El-Nasri et al., 2012). It is present in cheese
that results from the burning of organic matter (Kirk and Sawyer, 1991).
The result of mean squares (ANOVA) for ash content of butter samples are
shown in Table 4.12.5a.Results revealed that treatments, storage days and their
interactive effect have NS (p>0.05) variation in % ash of butter. The mean ash values for
butter are given in Table 4.12.5b. Results showed that edible coating and essential oils
have a NS (p>0.05) effect on %ash of butter. Data revealed that the highest ash content
(0.83%) was recorded in butter sample T9 (butter containing 2.0%GO) and T10 (butter
containing 2.5%GO) at 90 days while the lowest ash (0.79%) was noted in control (T0) at
1 day. Results showed that there is a NS difference among treatments which were
formulated with CS based active edible coating (containing BCO and GO as active
ingredient) and samples with direct addition of these natural essential oils.
Fig. 4.33 depicted that during storage of 90 days a minute increase in % ash of
butter was observed butter samples which were formulated with active edible coating and
different concentrations of essential oils. Mean ash values for butter samples shows that
ash was increasing from 0.79% to 0.82%, 0.80% to 0.82% and 0.81% to 0.83% at 1, 45
118
and 90 days respectively. Mean ash values for treatments of butter at 45 and 90 days of
storage showed a NS increase. Results of ash in butter match with the findings of Ewe
and Loo (2016), they also observed that ash content in butter remains almost same during
storage of butter.
119
Table 4.12.5a MS for ash contents of butter
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.0048143 0.0057714 0.0013857 0.0658000 0.0777714
0.0024071 0.0004440 0.0000533 0.0007833
3.07NS
0.57NS
0.07NS
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0.7 0.72 0.74 0.76 0.78 0.8 0.82 0.840.8
0.810.810.81
0.80.8
0.820.82
0.80.820.82
0.80.810.81
0.810.820.82
0.810.82
0.810.820.820.82
0.830.83
0.810.820.82
90 day 45 day 1 day
Butter Ash
Treat
ment
Fig. 4.33 Effect of edible coating and EOs on the ash (%) of butter
120
Table 4.12.5b Effect of treated butter on the % ash during storage
Treat Days Total
1 day 45 day 90 day
T0 0.80±0.015 0.80±0.010 0.81±0.017 0.80±0.007A
T1 0.81±0.017 0.81±0.017 0.82±0.012 0.81±0.008A
T2 0.80±0.017 0.81±0.023 0.82±0.012 0.81±0.009A
T3 0.80±0.023 0.81±0.023 0.81±0.006 0.81±0.010A
T4 0.79±0.012 0.80±0.012 0.82±0.023 0.80±0.009A
T5 0.79±0.012 0.80±0.017 0.81±0.017 0.80±0.008A
T6 0.81±0.006 0.82±0.012 0.82±0.012 0.82±0.005A
T7 0.81±0.012 0.82±0.017 0.82±0.023 0.82±0.009A
T8 0.80±0.012 0.80±0.006 0.82±0.023 0.81±0.008A
T9 0.81±0.015 0.82±0.015 0.83±0.017 0.82±0.008A
T10 0.82±0.026 0.82±0.017 0.83±0.012 0.82±0.010A
T11 0.80±0.012 0.80±0.010 0.81±0.012 0.80±0.006A
T12 0.80±0.023 0.81±0.015 0.82±0.017 0.81±0.010A
T13 0.81±0.021 0.81±0.012 0.82±0.012 0.81±0.008A
Total 0.80±0.004A 0.81±0.004A 0.82±0.004AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
121
4.12.6 Water activity (aw)
Water activity (aw) is a very important parameter to assess the storage stability of
a food product. It gives us the estimation how much the water is active the product is
more susceptible to spoilage if the aw is higher (Zamfir et al., 2006).
Results of mean square for aw of butter samples are shown in Table 4.12.6a.
Results revealed that treatments, storage days and their interaction have highly **
(p<0.01) effect on aw of butter. The mean water activity values butter are given in Table
4.12.6b. Results showed that edible coating and essential oils have ** (p<0.01) variation
on water activity of butter. Data showed that the highest water activity (0.95) was
recorded in butter sample T4 (butter containing 0.4%BCO), T9 (butter containing 2.0%
GO) and T10 (butter containing 2.5% GO) at 1 day while the lowest water activity (0.51)
was noted in control (T0) butter at 90 days. All the butter samples which were formulated
with CS based edible coating (containing BCO and GO as active ingredient) and samples
with direct addition of these natural essential oils have more water activity as compared
to T0 butter during storage of 90 days. Fig. 4.34 depicted that during storage of 90 days a
continuous decrease in aw of butter was noted in butter samples which were formulated
with edible coating and different concentrations of EOs. Mean value of butter shows that
water activity was decreasing from 0.95 to 0.93, 0.82 to 0.68 and 0.71 to 0.51 at 1, 45 and
90 days respectively. Mean values of water activity for butter samples at 45 and 90 days
of storage showed a ** decline.
Edible coating serve as a barrier against moisture loss by covering the surface of
butter samples so less moisture loss was observed in edible coated samples of butter. Thymol
in BCO and gingerol in GO due to their excellent antioxidant and antimicrobial potential
binds the free radicals in butter samples that ultimately results in higher aw as compared to
control. The overall values of aw in butter samples decrease but the rate of reduction is slow in
all butter samples as compare to control. Results of aw match with the findings of Koca and
Metin, (2004), they also observed that aw decreases in butter during storage.
122
Table 4.12.6a MS for aw contents of butter
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
2.0983430.1074930.0546570.0592002.319693
1.049171 0.008269 0.002102 0.000705
1488.69** 11.73** 2.98**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.90.68
0.730.75
0.780.81
0.740.770.78
0.760.79
0.820.76
0.780.8
0.510.56
0.60.62
0.660.590.590.6
0.650.68
0.710.63
0.660.67
90 day 45 day 1 day
Butter Water Activity
Treat
ment
Fig. 4.34 Effect of edible coating and EOs on the aw of butter
123
Table 4.12.6b Effect of treated butter on the aw during storage
Treat Days Total
1 day 45 day 90 day
T0 0.940±0.015a 0.680±0.015e-j 0.510±0.020m 0.710±0.063F
T1 0.930±0.017a 0.730±0.006c-h 0.560±0.006lm 0.740±0.054EF
T2 0.940±0.029a 0.750±0.017b-f 0.600±0.012jkl 0.763±0.050CDE
T3 0.940±0.012a 0.780±0.012bcd 0.620±0.012jkl 0.780±0.047B-E
T4 0.950±0.017a 0.810±0.012bc 0.660±0.017g-k 0.807±0.043AB
T5 0.930±0.017a 0.740±0.006b-g 0.590±0.006klm 0.753±0.049DE
T6 0.930±0.012a 0.770±0.012bcd 0.590±0.006klm 0.763±0.049CDE
T7 0.940±0.023a 0.780±0.017bcd 0.600±0.012jkl 0.773±0.050B-E
T8 0.940±0.023a 0.760±0.006b-e 0.650±0.012h-k 0.783±0.043BCD
T9 0.950±0.017a 0.790±0.017bcd 0.680±0.017e-j 0.807±0.040AB
T10 0.950±0.025a 0.820±0.017b 0.710±0.012d-i 0.827±0.036A
T11 0.940±0.015a 0.760±0.017b-e 0.630±0.006i-l 0.777±0.045B-E
T12 0.930±0.021a 0.780±0.015bcd 0.660±0.006g-k 0.790±0.040A-D
T13 0.940±0.017a 0.800±0.015bc 0.670±0.012f-k 0.803±0.040ABC
Total 0.939±0.004A 0.768±0.006B 0.624±0.008CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
124
4.12.7 Color
The consumer’s first attractive tool towards food is its color that helps in
either to accept or reject the food (Gokmen and Sugut, 2007). Color is vital sensory
factor in all food systems that affect the acceptance, rejection and marketing of any
food product. In dairy products especially in processed food color act as a great
sensual indicator of which biochemical reactions are occurring in many food that
changes the color (Yates and Drake, 2007).
4.12.7.1 L* valueThe result of mean squares (ANOVA) for L* value of butter samples are shown in
Table 4.12.7.1a. Results revealed that treatments have * (p<0.05) effect in L* value
which were formulated with CS based edible coating (containing BCO and GO as active
ingredient) and samples with direct addition of black cumin and ginger oils in different
concentration. Tab 4.12.7.1b represents the mean values of L*. The difference is due to
change in EOs concentration and active edible coating. Lowest value (82.77%) was
recorded in T13 (CS based active edible coating of butter containing 2.5% GO) while
highest value (86.04%) was attained in T1 (CS based edible coating of butter). Color
variation in butter fat might be due to feed, size of fat globule, season of the year and
addition of essential oils. Results of the L* value match with the findings of Lee and Min
(2014) who reported that L* value of butter samples remain unchanged for 2 months as
compare to control butter.
4.12.7.2 a* valueThe result of mean squares (ANOVA) for a* value of butter samples presented in
Table 4.12.7.2a revealed a ** (p<0.01) effect in a* value due to treatments which were
manufactured with CS based active edible coating (containing BCO and GO as active
ingredient) and samples with direct incorporation of BCO and GO in different
concentration. Table 4.12.7b represents the mean values of a*. Lowest value (-4.95) was
recorded in T10 (butter containing 2.5% GO) while highest value (-2.77) was attained in
T7 (CS based edible coating of butter containing 1.0% CO).
4.12.7.3 b* valueThe result of mean squares (ANOVA) for b* value of butter samples are shown in
Table 4.12.7.3a. Results showed that treatments have ** (p<0.01) effect on b* value
which were formulated with CS based edible coating (containing BCO and GO as active
ingredient) and
125
Table 4.12.7a MS for color parameters of butter
SOV Df MS
Butter L* Butter a* Butter b*
TreatmentErrorTotal
132841
2.489*1.049
1.6963** 0.0091
2.3278** 0.1136
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 2 4 6 8 10 12Butter L*
Treat
ment
Fig. 4.35 Effect of edible coating and EOs on the L* value of butter
126
T0
T2
T4
T6
T8
T10
T12
2 2.5 3 3.5 4 4.5 5Butter a* (-ve)
Trea
tmen
t
Fig. 4.36 Effect of edible coating and EOs on the a* value of butter
T0
T2
T4
T6
T8
T10
T12
10 11 12 13 14 15 16 17 18 19 20Butter b*
Trea
tmen
t
Fig. 4.37 Effect of edible coating and EOs on the b* value of butter
127
Table 4.12.7b Effect of treated butter on the L*, a*and b* value
Treatment Cheese L* Cheese a* Cheese b*
T0 84.35±0.525A -3.07±0.064A 11.60±0.150A
T1 86.03±0.274AB -3.01±0.035AB 11.90±0.156AB
T2 84.30±0.803AB -3.20±0.023AB 12.30±0.346ABC
T3 84.30±0.895AB -3.70±0.035ABC 12.90±0.191BCD
T4 84.25±0.629AB -3.90±0.046BCD 13.10±0.237BCD
T5 85.60±0.502AB -2.83±0.046CDE 12.05±0.144CDE
T6 85.50±0.745AB -2.80±0.052CDE 11.90±0.260DE
T7 85.20±0.548AB -2.77±0.029DE 11.90±0.231DEF
T8 84.05±0.583AB -4.65±0.115EF 13.90±0.092DEF
T9 83.70±0.485AB -4.78±0.058FG 14.10±0.104EF
T10 83.40±0.277AB -4.95±0.029G 14.30±0.167EF
T11 85.10±0.636AB -3.20±0.064H 12.45±0.092EF
T12 84.10±0.681AB -3.31±0.058HI 12.80±0.225EF
T13 82.77±0.272B -3.45±0.055I 13.10±0.150F
In a column or row means having similar letters are statistically NS (P>0.05). T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
128
samples with direct addition of black cumin and ginger oils in different concentration.
Table 4.12.7b represents the mean values of b*. Higher intensity (14.3) was recorded in
T10 (butter containing 2.5% ginger oil) while the lowest value (11.6) was attained inT0
butter, the remaining butter samples have values in between these two treatments.
4.13 Texture analysis4.13.1 HardnessThe amount of force required to bite the sample entirely when placed in the middle of
molar teeth is called Hardness (Meullenet and Gross, 1999).
The result of mean squares for hardness of butter samples are presented in Table
4.13.1a. Results revealed that treatments and storage days have ** (p<0.01) effect on the
hardness of butter while their interactive effect have NS (p>0.05) for hardness of butter.
The mean hardness values for butter are given in Table 4.13.1b. Data show that
the highest value of hardness (34.85) was recorded in T0 butter at 90 days while the
lowest value of hardness was noted in T10 (butter containing 2.5% ginger oil) at 1 day.
Results showed that there is a ** difference among all the butter samples which were
prepared with corn starch based active edible coating (containing black cumin and ginger
oil as active ingredient) and samples with direct incorporation of black cumin and ginger
oils in different concentration. Control butter sample (T0) have more hardness as
compared to other butter samples due to the effect of active edible coating and addition
of black cumin and ginger oil in different concentrations.
Fig. 4.38 depicted that during storage of 90 days a continuous decrease in
hardness of butter was observed in butter samples which were formulated with edible
coating and natural essential oils. Mean values of butter shows that hardness was
decreasing from 24.45 to 23.20, 29.65 to 25.9 and 34.85 to 32.25 at 1, 45 and 90 days
respectively. Mean hardness value for butter samples at 1, 45 and 90 days of storage
showed a ** decrease.
4.13.2 CohesivenessCohesiveness is the ability of internal bond strength that allows a product to
crumble (Bourne, 1978) and maintains the body of that product (Zoon, 1991).
The result of mean squares for cohesiveness of butter samples presented in Table
4.13.2a revealed a ** (p<0.01) effect on the cohesiveness of butter due to treatments and
storage days. However, interactive effect of storage days and treatments was found to
have NS (p>0.05) effect for cohesiveness of butter.
129
Table 4.13.1a MS for hardness of butter
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
1823.12 46.15 10.08 61.60
1940.95
911.561 3.550 0.388 0.733
1243.09** 4.84** 0.53
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
20 22 24 26 28 30 32 34 3629.65
28.327.9
27.126.55
2828
27.727.2
26.325.9
27.727.527.4
34.8534.1
33.2533.05
32.3533.95
33.633.2
3332.7
32.2533.5
33.333.1
90 day 45 day 1 day
Butter Hardness
Treat
ment
Fig. 4.38 Effect of edible coating and EOs on the hardness (g) of butter
130
Table 4.13.1b Effect of treated butter on the hardness (g) during storage
Treat Days Total
1 day 45 day 90 day
T0 24.35±0.50 29.65±0.58 34.85±0.33 29.62±1.53A
T1 24.45±0.39 28.30±0.53 34.10±0.76 28.95±1.43AB
T2 24.20±0.36 27.90±0.47 33.25±0.34 28.45±1.33ABC
T3 24.00±0.49 27.10±0.16 33.05±0.38 28.05±1.34BC
T4 23.90±0.31 26.55±0.63 32.35±0.83 27.60±1.29BC
T5 24.40±0.23 28.00±0.65 33.95±0.64 28.78±1.42AB
T6 24.30±0.43 28.00±0.24 33.60±0.51 28.63±1.37AB
T7 24.30±0.19 27.70±0.46 33.20±0.46 28.40±1.31ABC
T8 23.95±0.23 27.20±0.35 33.00±0.40 28.05±1.33BC
T9 23.70±0.33 26.30±0.50 32.70±0.51 27.57±1.36BC
T10 23.20±0.49 25.90±0.44 32.25±0.62 27.12±1.37C
T11 24.20±0.32 27.70±0.33 33.50±0.95 28.47±1.39ABC
T12 24.15±0.44 27.50±0.53 33.30±0.87 28.32±1.37ABC
T13 24.05±0.57 27.40±0.30 33.10±0.33 28.18±1.34BC
Total 24.08±0.10C 27.51±0.17B 33.30±0.17AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
131
Table 4.13.2a MS for cohesiveness of butter
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
4.25144 0.03574 0.00930 0.02867 4.32515
2.12572 0.00275 0.00036 0.00034
6227.98** 8.05** 1.05
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1
0.6370.596
0.5720.563
0.5470.5810.574
0.560.561
0.550.539
0.5780.562
0.552
0.8740.8620.857
0.8320.814
0.8580.8510.8450.842
0.8160.793
0.8490.836
0.825
90 day 45 day 1 day
Butter Cohesiveness
Treatm
ent
Fig. 4.39 Effect of edible coating and EOs on the cohesiveness of butter
132
Table 4.13.2b Effect of treated butter on the cohesiveness during storage
Treat Days Total
1 day 45 day 90 day
T0 0.402±0.006 0.637±0.011 0.874±0.021 0.638±0.068A
T1 0.404±0.006 0.596±0.008 0.862±0.009 0.621±0.067AB
T2 0.396±0.006 0.572±0.009 0.857±0.013 0.608±0.067A-D
T3 0.388±0.008 0.563±0.005 0.832±0.015 0.594±0.065B-E
T4 0.387±0.002 0.547±0.003 0.814±0.010 0.583±0.062DE
T5 0.401±0.005 0.581±0.003 0.858±0.019 0.613±0.067ABC
T6 0.400±0.008 0.574±0.008 0.851±0.013 0.608±0.066A-D
T7 0.397±0.003 0.560±0.006 0.845±0.014 0.601±0.066BCD
T8 0.390±0.005 0.561±0.008 0.842±0.012 0.598±0.066B-E
T9 0.382±0.005 0.550±0.008 0.816±0.018 0.583±0.063DE
T10 0.375±0.008 0.539±0.006 0.793±0.010 0.569±0.061E
T11 0.398±0.003 0.578±0.009 0.849±0.023 0.608±0.066A-D
T12 0.392±0.005 0.562±0.013 0.836±0.015 0.597±0.065B-E
T13 0.388±0.005 0.552±0.009 0.825±0.019 0.588±0.064CDE
Total 0.393±0.002C 0.569±0.004B 0.840±0.005AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
133
The mean values for cohesiveness of butter are given in Table 4.13.2b. It is
evident from data that the highest value of cohesiveness (0.874) was recorded in control
butter sample (T0) at 90 days while the lowest value of cohesiveness (0.375) was noted in
T10 (butter containing 2.5% ginger oil) at 1 day. Results revealed that there is a **
difference among all the butter samples which were prepared with CS based active edible
coating (containing BCO and GO as active ingredient) and samples with direct
incorporation of BCO and GO in different concentration. Control butter sample (T0) have
more cohesiveness as compared to all other butter samples due to the effect of active
edible coating and addition of black cumin and ginger oil in different concentrations.
Fig. 4.39 showed that during storage of 90 days a consistent increase in
cohesiveness of butter was observed in all samples which were prepared with active
edible coating and natural essential oils. Mean value of all butter samples shows that
cohesiveness was increasing from 0.375 to 0.404, 0.539 to 0.637 and 0.793 to 0.874 at 1,
45 and 90 days respectively. Mean values of cohesiveness for all treatments of butter
showed a ** increase at 45 days and 90 days of storage.
4.13.3 Springiness
The bouncing property of sample and returning to its original shape after
consective bites is known as the springiness of that sample (Civille and Szczesniak,
1973).
The result of mean squares for springiness of butter samples presented in Table
4.13.3a revealed a ** (p<0.01), *(p<0.05) and NS (p>0.05) effects on springiness of
butter due to treatments, storage days and their interactive effect respectively.
The mean springiness values for butter are given in Table 4.13.3b. Data revealed
that the highest value of springiness (0.509) was recorded in T10 (butter containing 2.5%
ginger oil) at 1day while the lowest value of springiness (0.455) was noted in T0 butter at
90 days. Results revealed that there is a ** difference among all the butter samples which
were prepared with CS based active edible coating (containing BCO and GO as active
ingredient) and samples with direct incorporation of BCO and GO in different
concentration. T0 butter sample have less springiness as compared to other butter samples
due to the effect of active edible coating and addition of BCO and GO in different
concentrations.
134
Table 4.13.3a MS for Springiness of butter
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.01454 0.00525 0.00041 0.01711 0.03731
0.00727 0.00040 0.00002 0.00020
35.71** 1.98* 0.08
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0.43 0.44 0.45 0.46 0.47 0.48 0.49 0.5
0.470.475
0.4790.486
0.490.472
0.4760.4820.483
0.4880.493
0.4760.48
0.483
0.4550.463
0.4680.475
0.4820.464
0.4670.4720.473
0.480.488
0.4680.474
0.477
90 day 45 day 1 day
Butter Springiness
Treat
ment
Fig. 4.40 Effect of edible coating and EOs on the springiness of butter
Table 4.13.3b Effect of treated butter on the springiness during storage
135
Treat Days Total
1 day 45 day 90 day
T0 0.493±0.005 0.470±0.010 0.455±0.006 0.473±0.007B
T1 0.491±0.007 0.475±0.009 0.463±0.008 0.476±0.006AB
T2 0.495±0.003 0.479±0.011 0.468±0.005 0.481±0.005AB
T3 0.499±0.011 0.486±0.007 0.475±0.006 0.487±0.005AB
T4 0.505±0.012 0.490±0.005 0.482±0.003 0.492±0.005AB
T5 0.492±0.009 0.472±0.009 0.464±0.005 0.476±0.006AB
T6 0.495±0.006 0.476±0.005 0.467±0.005 0.479±0.005AB
T7 0.499±0.008 0.482±0.012 0.472±0.010 0.484±0.006AB
T8 0.497±0.005 0.483±0.008 0.473±0.003 0.484±0.004AB
T9 0.502±0.010 0.488±0.005 0.480±0.009 0.490±0.005AB
T10 0.509±0.012 0.493±0.009 0.488±0.014 0.497±0.007A
T11 0.494±0.008 0.476±0.010 0.468±0.005 0.479±0.006AB
T12 0.498±0.005 0.480±0.008 0.474±0.014 0.484±0.006AB
T13 0.500±0.007 0.483±0.012 0.477±0.005 0.487±0.005AB
Total 0.498±0.002A 0.481±0.002B 0.472±0.002CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
Fig. 4.40 depicted that during storage of 90 days a continuous decrease in
springiness was noted in butter samples which were formulated with active edible
136
coating and natural essential oils in different concentrations. Mean value of butter
samples shows that springiness was decreasing from 0.509 to 0.491, 0.493 to 0.470 and
0.488 to 0.455 at 1, 45 and 90 days respectively. Mean springiness values for all the
butter samples at 45and 90 days of storage showed a ** decrease.
4.13.4 GumminessConfrontation or resistance that is offered by food product during cuddling in
between fore finger and thumb or during mastication is called gumminess (Wium et al.,
1997).
The result of mean squares for gumminess of butter samples are presented in
Table 4.13.4a. Results revealed that treatments and storage days have ** (p<0.01) effect
while their interactive effect have NS (p>0.05) variation on the gumminess of butter.
The mean values for gumminess of butter are given in Table 4.13.4b. It is evident
from the results that the highest value of gumminess (30.46) was recorded in control
butter sample (T0) at 90 days while the lowest value of gumminess (8.70) was noted in
butter sample T10 (butter containing 2.5% GO) at 1 day. Results revealed that there is a **
difference among all the butter samples which were formulated with CS based active
edible coating (containing BCO and GO as active ingredient) and samples with direct
incorporation of BCO and GO in different concentration. Control butter sample (T0) have
more gumminess as compared to all other butter samples due to the effect of active
edible coating and addition of BCO and GO in different concentrations.
Fig. 4.41 showed that during storage of 90 days a increase in gumminess of butter
was observed in all samples which were prepared with active edible coating and different
concentrations of essential oils. Mean value of all the butter samples shows that
gumminess was increasing from 8.7 to 9.88,13.96 to 18.89 and 25.57 to 30.46 at 1, 15
and 30 days respectively. Mean values of gumminess for all treatments at 45 days and 90
days of storage showed a ** decrease.
4.13.5 Chewiness During swallowing the energy required to chew the food is called chewiness
(Zoon, 1991). There is a correlation between hardness and chewing because harder butter
results in more force to apply and chew the food stuff as chewiness defined as the
number of
Table 4.3.4a MS for gumminess of butterSOV Df Sum of squares Mean squares F-value
137
DayTreatmentDay x TreatmentErrorTotal
2132684
125
7455.05 98.63 26.92 54.48
7635.07
3727.52 7.59 1.04 0.65
5747.77** 11.70** 1.60
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 5 10 15 20 25 30 3518.89
16.8715.96
15.2614.53
16.2716.07
15.5215.26
14.4613.96
16.0115.46
15.13
30.4629.4
28.527.49
26.3329.23
28.628.05
27.7826.68
25.5728.44
27.8427.31
90 day 45 day 1 day
Butter Gumminess
Treat
ment
Fig. 4.41 Effect of edible coating and EOs on the gumminess (g) of butter
Table 4.13.4b Effect of treated butter on the gumminess (g) during storage
Treat Days Total
138
1 day 45 day 90 day
T0 9.790±0.101 18.890±0.219 30.460±0.854 19.713±3.002A
T1 9.880±0.124 16.870±0.254 29.400±0.433 18.717±2.859AB
T2 9.580±0.185 15.960±0.139 28.500±1.000 18.013±2.794B-E
T3 9.320±0.237 15.260±0.289 27.490±0.416 17.357±2.679C-F
T4 9.250±0.167 14.530±0.306 26.330±0.566 16.703±2.532EF
T5 9.780±0.156 16.270±0.242 29.230±0.985 18.427±2.874ABC
T6 9.720±0.139 16.070±0.260 28.600±0.647 18.130±2.781BC
T7 9.650±0.173 15.520±0.242 28.050±0.635 17.740±2.721B-E
T8 9.340±0.092 15.260±0.260 27.780±0.501 17.460±2.723B-E
T9 9.050±0.231 14.460±0.162 26.680±0.393 16.730±2.611DEF
T10 8.700±0.064 13.960±0.121 25.570±0.618 16.077±2.498F
T11 9.630±0.260 16.010±0.505 28.440±0.848 18.027±2.777BCD
T12 9.470±0.208 15.460±0.433 27.840±1.004 17.590±2.723B-E
T13 9.330±0.156 15.130±0.127 27.310±0.941 17.257±2.664C-F
Total 9.464±0.062C 15.689±0.191B 27.977±0.253AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
Table 4.13.5a MS for chewiness of butterSOV df SS MS FV
139
DayTreatmentDay x TreatmentErrorTotal
2132684
125
1566.46 11.50 3.52 8.26
1589.75
783.232 0.885 0.135 0.098
7964.68** 9.00** 1.38
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 2 4 6 8 10 12 14 168.87
8.027.65
7.427.12
7.687.65
7.487.37
7.056.88
7.627.427.31
13.8613.62
13.3413.06
12.6913.56
13.3613.2413.14
12.8112.47
13.3113.19
13.03
90 day 45 day 1 day
Butter Chewiness
Treat
ment
Fig. 4.42 Effect of edible coating and EOs on the chewiness (g) of butter
Table 4.13.5b Effect of treated butter on the chewiness (g) during storage
Treat Days Total
140
1 day 45 day 90 day
T0 4.850±0.055 8.870±0.075 13.860±0.306 9.193±1.306A
T1 4.850±0.064 8.020±0.179 13.620±0.179 8.830±1.284AB
T2 4.740±0.075 7.650±0.098 13.340±0.115 8.577±1.264BC
T3 4.650±0.098 7.420±0.046 13.060±0.208 8.377±1.239BCD
T4 4.670±0.035 7.120±0.081 12.690±0.075 8.160±1.187CD
T5 4.810±0.092 7.680±0.064 13.560±0.236 8.683±1.290AB
T6 4.820±0.069 7.650±0.110 13.360±0.285 8.610±1.259BC
T7 4.820±0.092 7.480±0.202 13.240±0.344 8.513±1.248BC
T8 4.640±0.081 7.370±0.052 13.140±0.178 8.383±1.254BCD
T9 4.540±0.029 7.050±0.092 12.810±0.329 8.133±1.228CD
T10 4.430±0.110 6.880±0.121 12.470±0.219 7.927±1.192D
T11 4.750±0.121 7.620±0.133 13.310±0.495 8.560±1.267BC
T12 4.710±0.040 7.420±0.069 13.190±0.202 8.440±1.252BC
T13 4.660±0.069 7.310±0.173 13.030±0.427 8.333±1.242BCD
Total 4.710±0.025C 7.539±0.077B 13.191±0.083AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GOmastication that are applied on a certain amount of sample to enhance the satisfactory
swallowing (Beal and Mittal, 2000).
141
The result of mean squares for chewiness of butter samples presented in Table
4.13.5a revealed a ** (p<0.01) effect due to treatments and storage days while their
interaction effect have NS (p>0.05) effect for chewiness of butter.
The mean chewiness values for butter are given in Table 4.13.5b. Data showed
that the highest value of chewiness (13.86) was recorded in control butter sample (T0) at
90 days while the lowest value of chewiness (4.43) was noted in T10 (butter containing
2.5% GO) at 1 day. Results revealed that there is a ** difference among the butter
samples which were formulated with CS based active edible coating (containing BCO
and GO as active ingredient) and samples with direct incorporation of BCO and GO in
different concentration. T0 butter have more chewiness as compared to other butter
samples due to the effect of active edible coating and addition of BCO and GO in
different concentrations.
Fig. 4.42 depicted that during storage of 90 days a increase in chewiness of butter
was observed in all samples which were prepared with active edible coating and different
concentrations of natural essential oils. Mean value of all butter samples shows that
chewiness was increasing from 4.43 to 4.85, 6.88 to 8.87 and 4.43 to 4.85 at 1, 45 and 90
days respectively. Mean values of chewiness for all treatments at 45 and 90 days of
storage showed a ** increase. The results of the present study match with the Awad et al.
(2005) who showed an increase in chewiness of cheese during ripening.
4.14 Weight lossThe results of mean squares (ANOVA) for weight loss of butter samples are
shown in Table 4.14a. Data showed that treatments and storage days shows a ** (p<0.01)
effect while their interactive effect have * (p<0.05) variation on the weight loss of butter.
The mean weight loss values for butter samples are given in Table 4.14b. Results
showed that edible coating and essential oils have ** (p<0.01) effect on weight loss of
butter. Data showed that the highest value of weight loss (9.05%) was recorded in T0
butter at 90 days while the lowest value of weight loss (2.50%) was noted in T4 (butter
containing 0.4% BCO) at 15days. Results revealed that there is a ** difference among
the butter samples which were formulated with CS based active edible coating
(containing BCO and GO as active ingredient) and butter samples with direct
incorporation of BCO and GO. T0 butter sample have more
Table 4.14a MS for weight loss of butter
SOV Df SS MS FV
142
DayTreatmentDay x TreatmentErrorTotal
1 13 13 56 83
646.020 6.276 1.020 1.877
655.193
646.020 0.483 0.078 0.034
19273.90** 14.40** 2.34*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
6.5 7 7.5 8 8.5 9 9.5
9.058.85
8.378.05
7.558.7
8.488.1
8.558.42
8.058.78
8.558.2
90 day 45 day
Butter Weight Loss
Treat
ment
Fig. 4.43 Effect of edible coating and EOs on weight loss (%) of butter
Table 4.14b Effect of treated butter on the % weight loss during storage
143
Treat Days Total
1 day 45 day 90 day
T0 - 3.210±0.040f 9.050±0.139a 6.130±1.307A
T1 - 3.010±0.035fg 8.850±0.104ab 5.930±1.307AB
T2 - 2.850±0.058fg 8.370±0.075bcd 5.610±1.235B-F
T3 - 2.670±0.017fg 8.050±0.185de 5.360±1.206EFG
T4 - 2.500±0.064g 7.550±0.064e 5.025±1.130G
T5 - 3.000±0.023fg 8.700±0.069abc 5.850±1.275ABC
T6 - 2.900±0.058fg 8.480±0.139a-d 5.690±1.250B-F
T7 - 2.830±0.058fg 8.100±0.156de 5.465±1.181DEF
T8 - 2.900±0.017fg 8.550±0.098a-d 5.725±1.264B-E
T9 - 2.780±0.035fg 8.420±0.156bcd 5.600±1.263B-F
T10 - 2.620±0.052g 8.050±0.069de 5.335±1.215FG
T11 - 3.000±0.046fg 8.780±0.214abc 5.890±1.296ABC
T12 - 2.930±0.029fg 8.550±0.202a-d 5.740±1.260BCD
T13 - 2.850±0.046fg 8.200±0.196cd 5.525±1.200C-F
Total - 2.861±0.029B 8.407±0.067AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
144
weight loss as compared to all other butter samples due to edible coating and addition of
natural essentials oils. Mean weight loss values shows that all the butter samples have
0% weight loss at 1 day.
Fig. 4.43 depicted that during storage of 90 days increase in weight loss of butter
was observed in all samples which were prepared with active edible coating and different
concentrations of BCO and GO. Mean value of butter samples shows that weight loss
was increasing from 2.50% to 3.21% and 7.55% to 9.05% at 45 and 90 days respectively.
Mean weight loss values for butter samples at 45 and 90 days of storage showed a **
increase in weight loss.
Edible coating serve as a barrier against moisture loss by covering the surface of
butter samples so minimum weight loss was noted in edible coated samples of butter.
BCO and GO have the antioxidant and antimicrobial potential so binds the free radicals
in butter samples that ultimately leads to minimum weight loss as compared to control
(T0) control. Findings of this parameter match with the results of Tarakci and Kucukoner
(2006), they also noted that moisture in SC decrease during storage.
4.15 Free fatty acids The result of mean squares for free fatty acids of butter samples are shown in
Table 4.15a. Results revealed that treatments and storage days have ** (p<0.01) effect in
free fatty acids of butter while their interactive effect have * (p<0.05) variation for FFA
of butter. The mean free fatty acids values for butter are given in Table 4.15b. Results
showed that edible coating and essential oils have ** (p<0.01) effect on free fatty acids
of butter. Data showed that the maximum free fatty acids (0.627%) value was noted in T0
butter at 90 days while the minimum FFA (0.216%) value was recorded in T1 (edible
coating of butter with CS) at 1 day. Butter samples which were formulated with CS based
active edible coating (containing BCO and GO as active ingredient) and samples with
direct incorporation of these natural essential oils shows consistent increase in free fatty
acids during storage of 90 days as compared to T0 butter. The overall values for butter
showed gradual increase in free fatty acids during storage.
Fig. 4.44 depicted that during storage of 90 days a continuous increase in free
fatty acids of butter was noted in butter samples which were formulated with edible
coating and different concentrations of essential oils. Mean values of butter shows that
FFA was increasing from 0.216% to 0.224%, 0.317% to 0.409% and 0.525% to 0.627%
at 1, 45 and 90 days respectively.
145
Table 4.15a MS for free fatty acid (FFA) of butter
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
2.4202080.0313050.0192280.0324522.503193
1.210104 0.002408 0.000740 0.000386
3132.28** 6.23** 1.91*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.1 0.2 0.3 0.4 0.5 0.6 0.70.409
0.3760.3420.3390.336
0.3490.3450.342
0.3250.3210.317
0.3490.3460.342
0.6270.603
0.5520.546
0.5390.5590.5560.552
0.5360.531
0.5250.5560.551
0.545
90 day 45 day 1 day
Butter Free Fatty Acid
Treat
ment
Fig. 4.44 Effect of edible coating and EOs on free fatty acid (FFA) (%) of butter
146
Table 4.15b Effect of treated butter on the % FFA during storage
Treat Days Total
1 day 45 day 90 day
T0 0.218±0.008f 0.409±0.012d 0.627±0.025a 0.418±0.060A
T1 0.216±0.005f 0.376±0.007de 0.603±0.020ab 0.398±0.056AB
T2 0.219±0.006f 0.342±0.009e 0.552±0.009bc 0.371±0.049BC
T3 0.220±0.006f 0.339±0.006e 0.546±0.016bc 0.368±0.048BC
T4 0.220±0.003f 0.336±0.014e 0.539±0.015bc 0.365±0.047C
T5 0.218±0.010f 0.349±0.008de 0.559±0.022bc 0.375±0.050BC
T6 0.219±0.004f 0.345±0.006de 0.556±0.008bc 0.373±0.049BC
T7 0.219±0.007f 0.342±0.004e 0.552±0.009bc 0.371±0.049BC
T8 0.220±0.003f 0.325±0.018e 0.536±0.014c 0.360±0.047C
T9 0.222±0.008f 0.321±0.012e 0.531±0.008c 0.358±0.046C
T10 0.224±0.005f 0.317±0.008e 0.525±0.009c 0.355±0.045C
T11 0.219±0.006f 0.349±0.006de 0.556±0.015bc 0.375±0.049BC
T12 0.220±0.006f 0.346±0.005de 0.551±0.026bc 0.372±0.049BC
T13 0.220±0.005f 0.342±0.006e 0.545±0.014bc 0.369±0.048BC
Total 0.220±0.001C 0.346±0.004B 0.556±0.005AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
147
Mean values of free fatty acids for butter samples at 45 and 90 days of storage showed a
** increase.
FFA increase during storage in butter samples mainly due to the oxidation of fat
and lipolysis but the rate of FFA production was slow in the coated butter samples as
compare to control due to effect of active edible coating and EOs. EOs due to their
excellent antioxidant potential prevents lipid oxidation so less FFA production was
observed. The findings of the FFA are similar with the results of Samet et al., (2009),
they also observed that FFA increase in butter during storage.
4.16 Anti-oxidative analysis
4.16.1 Diphenyl-2-picrylhdrazyl (DPPH) activity
The results of mean squares (ANOVA) for DPPH activity of butter samples are
shown in Table 4.16.1a. It was evident from the data that treatments and storage days
shows a ** (p<0.01) effect while the interaction of treatments and storage days was found
to have non-significant effect (p>0.05) on the DPPH activity of butter.
Mean DPPH values for butter samples are given in Table 4.16.1b. Results
revealed that active edible coating and essential oils have ** (p<0.01) variation on DPPH
activity of butter. Data showed that the highest value of DPPH activity (84.10%) was
recorded in T10 (butter sample containing 2.5% ginger oil) at 1 day while the lowest value
of DPPH activity (48.95%) was noted in control (T0) butter sample at 90 days. Results
revealed that there is a ** difference among all the butter samples which were prepared
with CS based active edible coating (containing BCO and GO as active ingredient) and
butter samples with direct incorporation of BCO and GO. Control butter sample (T0)
have minimum DPPH activity as compared to all other butter samples due to active
edible coating and addition of natural essentials oils in different concentration.
Fig. 4.45 depicted that during storage of 90 days decrease in DPPH activity of
butter was observed in butter samples which were formulated with active edible coating
and different concentrations of BCO and GO. Mean value of all treatments shows that
DPPH activity was decreasing from 84.1% to 74.35%, 78.9% to 61.9% and 65.6% to
48.95% at 1, 45 and 90 days respectively. Mean DPPH values for all butter samples at 45
and 90 days of storage showed a ** decrease in DPPH activity.
148
Table 4.6.1a MS for DPPH activity of butter
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
8414.5 1958.1 100.2 425.6
10898.3
4207.25 150.62 3.85
5.07
830.46** 29.73** 0.76
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
40 45 50 55 60 65 70 75 80 8561.9
63.271.32
73.476.75
68.2370.4
72.174.65
75.678.9
70.3471.3
73.64
48.9551.75
60.5561.3
64.657.45
59.560.2
61.963.1
65.658.9
60.162.3
90 day 45 day 1 day
Butter DPPH
Treat
ment
Fig. 4.45 Effect of edible coating and |EOs on the DPPH activity (%) of butter
149
Table 4.16.1b Effect of treated butter on the % DPPH activity during storage
Treat Days Total
1 day 45 day 90 day
T0 74.35±0.62 61.90±1.47 48.95±1.13 61.73±3.71F
T1 74.42±1.31 63.20±0.74 51.75±0.71 63.12±3.31F
T2 79.38±1.19 71.32±1.47 60.55±1.41 70.42±2.81CDE
T3 81.20±1.06 73.40±0.53 61.30±0.79 71.97±2.92BCD
T4 83.95±1.24 76.75±2.11 64.60±0.53 75.10±2.91AB
T5 76.42±1.78 68.23±1.32 57.45±1.11 67.37±2.84E
T6 78.20±0.81 70.40±1.29 59.50±0.69 69.37±2.75DE
T7 80.31±1.39 72.10±1.78 60.20±0.99 70.87±3.00CDE
T8 80.42±0.83 74.65±1.56 61.90±1.71 72.32±2.83BCD
T9 82.60±1.82 75.60±1.15 63.10±1.21 73.77±2.94ABC
T10 84.10±1.66 78.90±2.10 65.60±1.05 76.20±2.88A
T11 78.35±0.56 70.34±0.95 58.90±1.36 69.20±2.87DE
T12 79.50±1.92 71.30±1.78 60.10±0.82 70.30±2.92CDE
T13 81.60±1.40 73.64±0.79 62.30±1.35 72.51±2.86BCD
Total 79.63±0.55A 71.55±0.78B 59.73±0.73CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
150
Thymol and thymoquonone in BCO and gingerol in GO as active components are very effective to prolong the shelf life of edible coated butter due to their antioxidant potential to scavenge free radicals involved in lipid oxidation in butter that ultimately prevent the spoilage of butter samples. Antioxidant potential of EOs are mainly due to acting as a substrate for radicals (like superoxide), free radical-scavenging, hydrogen-donation, singlet oxygen quenching and metal ion chelation (Al-Mamar et al., 2002). The results of DPPH activity are in agreement with the findings of Sultan et al. (2009), they also noted that DPPH activity decrease during storage period. Hala, (2010) also showed the same results.4.16.2 Peroxide value
Oxidation of food results in production of free radicles that interact with other molecules to produce aldehydes and ketones and yields bad flavor in food that can be calculated by using the peroxide value (Ozkan et al., 2007).
The result of mean squares (ANOVA) for POV of butter samples are presented in Table 4.16.2a. Results revealed that treatments, storage days and their interactive effect have ** (p<0.01) effect in peroxide value of butter samples. Mean peroxide value of butter are given in Table 4.16.2b and results showed that edible coating and essential oils have ** (p<0.01) effect on peroxide value of butter. Data showed that the highest peroxide value (0.890) was recorded in control butter sample T0 at 90 days while the lowest peroxide value (0.030) was noted in T10 (butter containing 2.5% ginger oil) at 1 day. The butter samples which were formulated with CS based active edible coating (containing BCO and GO as active ingredient) and samples with direct incorporation of these natural essential oils show consistent increase in POV during storage of 90 days as compared to control (T0). The overall values for all the butter samples showed gradual increase in POV during storage. Fig. 4.46 depicted that during storage of 90 days a regular increase in POV of butter was noted in butter samples which were formulated with edible coating and different concentrations of EOs. Mean value of all the butter samples shows that POV was increasing from 0.05 to 0.15, 0.26 to 0.71 and 1.13 to 1.81 at 1, 45 and 90 days respectively. Mean POV for all butter samples at 45 days and 90 days of storage showed a ** increase.
EOs concentration and POV showed the negative correlation between them in butter samples as the concentration of EOs increase POV decrease and vice versa but overall values of the samples decrease during storage. Free radicals are produced during the storage of butter
151
Table 4.16.2a MS for peroxide value (POV) of butter
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
39.06750 1.21554 0.34690 0.0954840.72543
19.53375 0.09350 0.01334 0.00114
17134.87**82.26** 11.74**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 20.71
0.550.4
0.380.35
0.470.44
0.420.35
0.310.26
0.410.38
0.36
1.811.61
1.41.37
1.321.45
1.421.4
1.31.21
1.131.42
1.371.33
90 day 45 day 1 day
Butter POV
Treat
ment
Fig. 4.46 Effect of edible coating and EOs on the POV (meq/kg) of butter
152
Table 4.16.2b Effect of treated butter on the POV (meq/kg) during storage
Treat Days Total
1 day 45 day 90 day
T0 0.150±0.003no 0.710±0.008h 1.810±0.035a 0.890±0.244A
T1 0.140±0.003op 0.550±0.007i 1.610±0.023b 0.767±0.219B
T2 0.090±0.003op 0.400±0.012jkl 1.400±0.040cde 0.630±0.198CDE
T3 0.070±0.003op 0.380±0.006jkl 1.370±0.035cde 0.607±0.196DEF
T4 0.050±0.003op 0.350±0.006klm 1.320±0.029def 0.573±0.192FG
T5 0.110±0.002op 0.470±0.006ij 1.450±0.046c 0.677±0.201C
T6 0.100±0.006op 0.440±0.012ijk 1.420±0.035cd 0.653±0.198CD
T7 0.100±0.012op 0.420±0.015jkl 1.400±0.029cde 0.640±0.196CDE
T8 0.070±0.002op 0.350±0.006klm 1.300±0.035ef 0.573±0.186FG
T9 0.040±0.002op 0.310±0.012lm 1.210±0.017fg 0.520±0.177GH
T10 0.030±0.002p 0.260±0.007mn 1.130±0.017g 0.473±0.168H
T11 0.090±0.001op 0.410±0.006jkl 1.420±0.035cd 0.640±0.201CDE
T12 0.080±0.002op 0.380±0.006jkl 1.370±0.035cde 0.610±0.195DEF
T13 0.070±0.003op 0.360±0.007j-m 1.330±0.029de 0.587±0.191EF
Total 0.085±0.005C 0.414±0.017B 1.396±0.026AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
153
leads to the formation of aldehydes and ketones results in the production of off flavor/ bad smell and rancidity that ultimately causes the spoilage of butter. Thymol in BCO and gingerol in GO binds the free radicals; ultimately prevent the spoilage of butter samples (as described in SC due to the same mode of action of EOs). Results of POV are in line with the findings of Ozkan et al, (2007) and Simsek, (2011), they also observed that POV decrease during storage.4.16.3 Measurement of TBARs valueTBARs value is routinely a parameter or index point for lipid oxidation or oxidative deterioration that leads to spoilage of the food product (Luo et al., 2011).
The result of mean squares (ANOVA) for TBARS value of butter samples presented in Table 4.16.3a revealed a ** (p<0.01) effect in TBARs value of butter samples due to treatments, storage days and also their interactive effect. The mean Table 4.16.3b for TBARS of butter showed that edible coating and EOs have ** (p<0.01) effect on TBARS of butter. Data showed that the maximum value of TBARS (0.370) was noted in T0 butter at 90 days while the minimum TBARS (0.040) value was recorded in T10
(butter containing 2.5 ginger oil) at 1 day. The butter samples which were formulated with CS based edible coating (containing BCO and GO as active ingredient) and samples with direct incorporation of these natural essential oils show consistent increase in TBARS during storage of 90 days as compared to control (T0). The overall values for all the butter samples showed gradual increase in TBARS during storage. Fig. 4.47 depicted that during storage of 90 days a regular increase in TBARS value of butter was noted in butter samples which were formulated with edible coating and different concentrations of essential oils. Mean value of the butter samples shows that TBARS was increasing from 0.04 to 0.08, 0.06 to 0.26 and 0.20 to 0.37 at 1, 45 and 90 days respectively. TBARS value for all the butter samples at 45 and 90 days of storage showed a ** increase.
Results revealed that butter samples containing active edible coating and essential oils have better antioxidant potential as compare to control butter sample. There is a negative correlation between natural essential oil concentration and TBARS as the concentration of BCO and GO increases in edible coating of butter samples TBARS value decreases and vice versa but higher concentration affects the sensory attributes (as mentioned in soft cheese). Oxidation reactions of lipids produce secondary metabolites malonaldehyde that upon reaction with TBA produce pink color. Thymol in BCO and gingerol in GO due to their antioxidant activities inhibit the production of TBARS. Results of TBARS match with the findings of Simsek, (2011) and
154
Table 4.16.3a MS for TBARS of butter
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
0.7132430.1350860.0353570.0050880.888774
0.356621 0.010391 0.001360 0.000061
5887.62** 171.55** 22.45**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.40.26
0.210.16
0.150.14
0.170.17
0.150.14
0.110.09
0.170.160.16
0.370.32
0.260.24
0.210.27
0.250.24
0.230.2
0.160.27
0.250.24
90 day 45 day 1 day
Butter TBARS
Treat
ment
Fig. 4.47 Effect of edible coating and EOs on the TBARS value (mg MDA/kg) of butter
155
Table 4.16.3b Effect of treated butter on the TBARS value (mg MDA/kg) during storage
Treat Days Total
1 day 45 day 90 day
T0 0.080±0.001kl 0.260±0.006cd 0.370±0.012a 0.237±0.042A
T1 0.080±0.000kl 0.210±0.006fg 0.320±0.006b 0.203±0.035B
T2 0.070±0.000klm 0.160±0.006hi 0.260±0.006cd 0.163±0.028DE
T3 0.060±0.000lmn 0.150±0.000hi 0.240±0.006de 0.150±0.026FG
T4 0.050±0.000mn 0.140±0.000i 0.210±0.000fg 0.133±0.023H
T5 0.080±0.000kl 0.170±0.000h 0.270±0.006c 0.173±0.027CD
T6 0.070±0.000klm 0.170±0.000h 0.250±0.000cde 0.163±0.026DE
T7 0.070±0.000klm 0.150±0.000hi 0.240±0.006de 0.153±0.025EFG
T8 0.060±0.000lmn 0.140±0.002i 0.230±0.008ef 0.143±0.025GH
T9 0.050±0.000mn 0.110±0.002j 0.200±0.006g 0.120±0.022I
T10 0.040±0.000n 0.090±0.004jk 0.160±0.003hi 0.097±0.017J
T11 0.090±0.000jk 0.170±0.008h 0.270±0.006c 0.177±0.026C
T12 0.070±0.003klm 0.160±0.009hi 0.250±0.006cde 0.160±0.026EF
T13 0.060±0.003lmn 0.160±0.006hi 0.240±0.008de 0.153±0.026EFG
Total 0.066±0.002C 0.160±0.006B 0.251±0.008AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
156
Dagdemir et al, (2009), they also observed that TBARS value decrease during storage
that might be due to storage condition.
4.17 Microbiological analysis4.17.1 Total viable count (TVC)
The results of mean squares (ANOVA) for total viable count (TVC) of butter
samples are presented in Table 4.17.1a. Results revealed that treatments, storage days
and their interactive effect shows a ** (p<0.01) effect on the total viable count (TVC) of
butter.
The mean Table 4.17.1b for total viable count of butter showed that the highest
value of total viable count (79.0x104) was recorded in control butter sample (T0) at 90
days while the lowest value of total viable count (0.07x104) was noted in butter sample
T4 (butter containing 0.4% BCO) at 1 day. Results revealed that there is a ** difference
among all the butter samples which were prepared with CS based active edible coating
(containing BCO and GO as active ingredient) and samples with direct incorporation of
black cumin and ginger oils in different concentration. Control butter sample (T0) have
more total viable count as compared to all other butter samples due to the effect of active
edible coating and addition of BCO and GO in different concentrations.
Fig. 4.48 depicted that during storage of 90 days increase in total viable count of
butter was noted in butter samples which were manufactured with edible coating and
different concentrations of essential oils. Mean values of butter shows that total viable
count was increasing from 0.07x104 to 0.45x104,0.52x104 to 5.1x104 and 6.2x104 to
79.0x104 at 1, 45 and 90 days respectively. Mean values of total viable count for butter
samples at 45 and 90 days of storage showed a ** increase.
Data showed that growth and multiplication rate of TVC was slow in samples
prepared with CS based active edible coating (containing black cumin and ginger oil as
active ingredient) as compared to T0 due to the effect of edible coating that create
hindrance against microorganisms by covering the surface of butter samples.
Conclusively, the most important factor for growth of microorganisms is concentration of
essential oil. Findings of TVC match with the results of Gokçe et al. (2010), they also
reported that TVC consistently increase during storage. In another study clove, thyme,
cinnamon and bay EOs were tested against Salmonella enteritidis and L. monocytogenes.
Results indicate that clove oil was more effective against Salmonella enteritidis (Burt,
2004).
157
Table 4.17.1a MS for total viable count (TVC) of butter
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
2132684
125
47763.5 7594.9 13768.1 381.0
69507.5
23881.7 584.2 529.5 4.5
5264.97** 128.80** 116.74**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 10 20 30 40 50 60 70 80 905.1
0.4440.430.760.52
4.184.03
3.794.383.943.724.284.093.85
7965
518.4
6.252
3928
7144
1367
4533.01
90 day 45 day 1 day
Butter TVC
Trea
tmen
t
Fig. 4.48 Effect of edible coating and EOs on the TVC (cfu/g) of butter
158
Table 4.17.1b Effect of treated butter on the TVC (X 104 cfu/g) during storage
Treat Days Total
1 day 45 day 90 day
T0 0.45±0.006j 5.10±0.139ij 79.00±5.089a 28.18±12.806A
T1 0.45±0.009j 0.44±0.009j 65.00±1.518b 21.96±10.768BC
T2 0.44±0.009j 0.43±0.006j 51.00±2.672cd 17.29±8.463DE
T3 0.10±0.003j 0.76±0.012j 8.40±0.237hi 3.09±1.334H
T4 0.07±0.000j 0.52±0.006j 6.20±0.075hij 2.26±0.987H
T5 0.43±0.006j 4.18±0.110ij 52.00±1.463c 18.87±8.311CD
T6 0.42±0.006j 4.03±0.075ij 39.00±1.704ef 14.48±6.171EF
T7 0.40±0.006j 3.79±0.104ij 28.00±0.525g 10.73±4.348G
T8 0.44±0.006j 4.38±0.104ij 71.00±3.995b 25.27±11.504AB
T9 0.39±0.012j 3.94±0.029ij 44.00±1.127de 16.11±6.999DE
T10 0.30±0.003j 3.72±0.087ij 13.00±0.202h 5.67±1.898H
T11 0.44±0.009j 4.28±0.081ij 67.00±1.990b 23.91±10.803B
T12 0.42±0.009j 4.09±0.104ij 45.00±0.936cde 16.50±7.149DE
T13 0.41±0.006j 3.85±0.035ij 33.00±0.763fg 12.42±5.174FG
Total 0.37±0.019C 3.11±0.260B 42.97±3.544AIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
159
4.18 Sensory Evaluation
4.18.1 Flavor
The flavor and smell are the amalgamation of senses interaction. Taste, smell, and
chemical irritation give a combined effect as flavor (Keast and Breslin 2003). Selection,
acceptance and ingestion of food are three sensory stuffs that add up the flavor (Hassan et
al., 2013).
The result of mean squares (ANOVA) for flavor of butter samples presented in
Table 4.18.1a revealed a ** (p<0.01) effect in flavor of butter samples due to treatments,
storage days and also their interactive effect.
The mean Table 4.18.1b for flavor of butter showed that active edible coating and
essential oils have ** (p<0.01) effect on flavor of butter. Results revealed that the
maximum flavor score (9.25) was recorded in butter sample T4 (butter containing 0.4%
BCO) and T7 (CS based edible coating of butter containing 0.4% BCO) at 1 day while the
lowest value of flavor (4.85) was noted in control (T0) at 90 days. All the butter samples
which were prepared with corn starch based active edible coating (containing BCO and
GO as active ingredient) and samples with direct incorporation of these natural essential
oils show consistent decrease in flavor during storage of 90 days as compared to control
(T0).
Fig. 4.49 depicted that during storage of 90 days a continuous decrease in flavor
of butter was noted in all butter samples which were manufactured with edible coating
and different concentrations of essential oils. Mean value of all butter samples shows that
flavor value was decreasing from 9.25 to 6.75, 8.25 to 5.75 and 4.77 to 3.27 at 1, 45 and
90 days respectively. Mean values of flavor for all butter samples at 45 and 90 days of
storage showed a ** decrease.
Edible coating and incorporation of EOs in butter samples significantly affects
the sensory properties of butter. Results revealed that flavor score decrease during
storage but the reduction rate in flavor score was less as compare to control butter that
might be due to proteolysis and lipolysis. The overall score of all butter samples decrease
during storage. Similar results were found by Songul et al, (2014) they also reported that
flavor score decrease during storage.
160
Table 4.18.1a MS for flavor of butter
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
21326
210251
634.396 97.825 41.207 79.860853.289
317.198 7.525 1.585 0.380
834.10** 19.79** 4.17**
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 1 2 3 4 5 6 7 8 95.75
6.487
7.778.25
6.756.77
7.276.77
77.27
6.777
6.74
3.273.82
44
6.273.753.75
4.024.25
4.776.75
43.98
4.24
90 day 45 day 1 day
Butter Flavor
Treat
ment
Fig. 4.49 Effect of edible coating and EOs on the flavor of butter
161
Table 4.18.1b Effect of treated butter on the flavor during storage
Treat Days Total
1 day 45 day 90 day
T0 6.75±0.310e-h 5.75±0.214hi 3.27±0.071k 5.26±0.375E
T1 7.25±0.099c-g 6.48±0.194fgh 3.82±0.273jk 5.85±0.373DE
T2 7.75±0.324b-f 7.00±0.365d-h 4.00±0.259jk 6.25±0.429CD
T3 8.77±0.167ab 7.77±0.320b-f 4.00±0.263jk 6.84±0.517BC
T4 9.25±0.128a 8.25±0.324a-d 6.27±0.167gh 7.92±0.324A
T5 8.00±0.373a-e 6.75±0.208e-h 3.75±0.099jk 6.17±0.454CD
T6 8.23±0.204a-d 6.77±0.109e-h 3.75±0.099jk 6.25±0.459CD
T7 9.25±0.099a 7.27±0.095c-g 4.02±0.261jk 6.84±0.531BC
T8 7.75±0.281b-f 6.77±0.320e-h 4.25±0.163jk 6.26±0.385CD
T9 7.75±0.310b-f 7.00±0.369d-h 4.77±0.206ij 6.51±0.348CD
T10 8.25±0.324a-d 7.27±0.320c-g 6.75±0.205e-h 7.42±0.217AB
T11 8.00±0.369a-e 6.77±0.095e-h 4.00±0.365jk 6.26±0.438CD
T12 8.50±0.224abc 7.00±0.447d-h 3.98±0.261jk 6.49±0.489CD
T13 8.48±0.217abc 6.73±0.105e-h 4.23±0.095jk 6.48±0.431CD
Total 8.14±0.098A 6.97±0.091B 4.35±0.116CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
162
4.18.2 Appearance / colorFor the color of butter carotenes are the main constituents that contribute in light
yellow or bright golden color (Cross and Overby, 1988).The result of mean squares (ANOVA) for appearance of butter samples presented
in Table 4.18.2a revealed a ** (p<0.01) variation in appearance of butter samples due to treatments and storage days while their interactive effect have *(p<0.05) variation in appearance of butter samples.
The mean Table 4.18.2b for appearance of butter showed that active edible coating and essential oils have ** (p<0.01) effect on appearance of butter. Data showed that the highest appearance value (9.52) was recorded in butter sample T4 (butter containing 0.4% BCO) at 1 day while the lowest value of appearance (2.52) was noted in control (T0) butter at 90 days. All the butter samples which were prepared with CS based active edible coating (containing BCO and GO as active ingredient) and samples with direct incorporation of these natural essential oils show consistent decrease in appearance during storage of 90 days as compared to T0 butter.
Fig. 4.50 depicted that during storage of 90 days a continuous decline in appearance score of butter was notes in all butter samples which were manufactured with edible coating and different concentrations of EOs. Mean value of all butter samples shows that appearance was decreasing from 9.52 to 8.25, 7.27 to 5.75 and 3.27 to 2.52 at 1, 45 and 90 days respectively. Mean values of appearance for all butter samples at 45 and 90 days of storage showed a ** increase.Edible coating and incorporation of EOs in butter samples significantly affects the
sensory properties of SC. Results revealed that appearance/color value decrease during
storage but the reduction rate in color value was less as compare to control butter that
might be due to lipolysis. It is also by pH and acidity because during storage pH decrease
and acidity increase results in the production of LAB that leads to variation in the
appearance. The overall score of all butter samples decrease during storage. Similar
results were found by Songul et al, (2014) they also reported that color value decrease
during storage.
4.18.3Texture/Body
The result of mean squares (ANOVA) for texture/body of butter samples
presented in Table 4.18.3a revealed a ** (p<0.01) effect in texture/body of butter samples
due to treatments
163
Table 4.18.2a MS for appearance of butter
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
21326
210251
1379.669 18.252 11.026 53.053
1462.000
689.834 1.404 0.424 0.253
2730.56** 5.56** 1.68*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 1 2 3 4 5 6 7 85.75
6.256.5
77.27
6.246.56.5
6.776.77
76.56.52
6.78
2.522.75
33.02
3.272.75
3.023.25
33.273.27
2.922.98
3.25
90 day 45 day 1 day
Butter Apperance
Treat
ment
Fig. 4.50 Effect of edible coating and EOs on the appearance of butter
164
Table 4.18.2b Effect of treated butter on the appearance during storage
Treat Days Total
1 day 45 day 90 day
T0 8.77±0.229ab 5.75±0.205e 2.52±0.174f 5.68±0.629D
T1 8.75±0.223ab 6.25±0.136de 2.75±0.112f 5.92±0.603BCD
T2 9.25±0.191ab 6.50±0.184de 3.00±0.365f 6.25±0.636ABC
T3 9.25±0.148ab 7.00±0.366d 3.02±0.259f 6.42±0.642AB
T4 9.52±0.130a 7.27±0.161cd 3.27±0.071f 6.68±0.631A
T5 8.27±0.105bc 6.23±0.161de 2.75±0.208f 5.75±0.560CD
T6 8.27±0.390bc 6.50±0.184de 3.02±0.130f 5.93±0.547BCD
T7 8.25±0.099bc 6.50±0.190de 3.25±0.154f 6.00±0.509BCD
T8 8.75±0.173ab 6.77±0.206de 3.00±0.263f 6.17±0.590A-D
T9 8.75±0.235ab 6.77±0.307de 3.27±0.154f 6.26±0.565ABC
T10 8.75±0.148ab 7.00±0.282d 3.27±0.095f 6.34±0.564AB
T11 8.25±0.112bc 6.50±0.141de 2.92±0.194f 5.89±0.545BCD
T12 8.25±0.198bc 6.52±0.147de 2.98±0.259f 5.92±0.543BCD
T13 8.52±0.194ab 6.78±0.215de 3.25±0.171f 6.18±0.542A-D
Total 8.68±0.066A 6.60±0.067B 3.02±0.055C
In a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GOTable 4.18.3a MS for texture of butter
165
SOV Df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
21326
210251
1052.497 34.214 3.133 16.133
1105.977
526.248 2.632 0.121 0.077
6849.93** 34.26** 1.57*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 1 2 3 4 5 6 74.77
5.255
5.485.75
66.02
6.255.25
5.55.775.75
6.026.02
3.253.5
3.023.25
3.53.74
44
3.273.25
3.53.5
3.754.02
90 day 45 day 1 day
Butter Texture
Treat
ment
Fig. 4.51 Effect of edible coating and EOs on the texture of butter
Table 4.18.3b Effect of treated butter on the texture during storage
166
Treat Days Total
1 day 45 day 90 day
T0 7.75±0.214e 4.77±0.206j 3.25±0.092lm 5.26±0.464H
T1 8.02±0.296de 5.25±0.076hij 3.50±0.026klm 5.59±0.461FG
T2 8.02±0.130de 5.00±0.063ij 3.02±0.031m 5.34±0.501GH
T3 8.25±0.118cde 5.48±0.070ghi 3.25±0.043lm 5.66±0.498EF
T4 8.52±0.060bcd 5.75±0.043fgh 3.50±0.026klm 5.92±0.498CDE
T5 8.75±0.085abc 6.00±0.063fg 3.73±0.033kl 6.16±0.499BC
T6 9.02±0.130ab 6.02±0.070fg 4.00±0.058k 6.34±0.502AB
T7 9.23±0.152a 6.25±0.072f 4.00±0.058k 6.49±0.523A
T8 8.25±0.056cde 5.25±0.076hij 3.27±0.033lm 5.59±0.498FG
T9 8.50±0.106bcd 5.50±0.063ghi 3.25±0.043lm 5.75±0.523DEF
T10 8.77±0.109abc 5.77±0.084fgh 3.50±0.026klm 6.01±0.525CD
T11 8.50±0.077bcd 5.75±0.076fgh 3.50±0.058klm 5.92±0.497CDE
T12 8.75±0.118abc 6.02±0.263fg 3.75±0.043kl 6.17±0.504BC
T13 9.00±0.089ab 6.02±0.087fg 4.02±0.259k 6.34±0.505AB
Total 8.52±0.057A 5.63±0.054B 3.54±0.040CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
167
and storage days while their interactive effect have * (p<0.05) effect in texture/body of
butter samples.
Mean Table 4.18.3b for texture of butter showed that edible coating and EOs have
** (p<0.01) effect on texture of butter. Data shows that the maximum texture value (9.00)
was recorded in butter sample T13 (CS based edible coating of butter containing 2.5%
GO) at 1 day while the lowest value of texture appearance (2.52) was noted in control
(T0) at 90 days. All the butter samples which were prepared with corn starch based active
edible coating (containing BCO and GO as active ingredient) and samples with direct
incorporation of these natural essential oils show consistent decrease in texture during
storage of 90 days as compared to control (T0). The overall values for all the butter
samples showed gradual decrease in texture during storage.
Fig. 4.51 showed that during storage of 90 days a consistent decrease in
texture/body of butter was recorded in all butter samples which were prepared with
active edible coating and different concentrations of essential oils. Mean value of all
treatments shows that texture/body was decreasing from 9.24 to 7.75, 6.02 to 4.77 and
4.02 to 3.02 at 1, 45 and 90 days respectively. Mean values of texture/body for all butter
samples at 45 days and 90 days of storage showed a ** decrease.
Edible coating and incorporation of EOs in butter samples significantly affects
the sensory properties of butter. Results revealed that texture value decrease during
storage but the reduction rate in texture value was less as compare to control butter that
might be due to proteolysis and lipolysis. The overall score of all butter samples decrease
during storage. Similar results were found by Songul et al, (2014) they also reported that
texture value decrease during storage.
4.18.4 Overall acceptability
Performa for sensory evaluation was used to assess the score or value for overall
acceptability of butter.
The result of mean squares for overall acceptability of butter samples presented in
Table 4.18.4a revealed a ** (p<0.01) variation in overall acceptability of butter samples
due to
treatments and storage days while their interactive effect have * (p<0.05) variation in
overall acceptability of butter samples.
168
Table 4.18.4a MS for overall acceptability of butter
SOV df SS MS FV
DayTreatmentDay x TreatmentErrorTotal
21326
210251
1151.216 51.375 14.699 67.743
1285.033
575.608 3.952 0.565 0.323
1784.35** 12.25** 1.75*
NS = (P>0.05); * = (P<0.05); ** = (P<0.01)
T0
T2
T4
T6
T8
T10
T12
0 1 2 3 4 5 6 7 85.75
66.02
6.276.75
55.25
55.77
6.776.5
5.275.5
5.25
2.52.75
3.243.25
3.752.75
33
3.243.27
3.752.75
33.02
90 day 45 day 1 day
Butter Overall Acceptability
Treat
ment
Fig. 4.52 Effect of edible coating and EOs on the overall acceptability of butter
169
Table 4.18.4b Effect of treated butter on the overall acceptability during storage
Treat Days Total
1 day 45 day 90 day
T0 7.50±0.224c-f 5.75±0.214ghi 2.50±0.129k 5.25±0.513C
T1 8.00±0.246bcd 6.00±0.259ghi 2.75±0.277k 5.58±0.543BC
T2 8.50±0.224abc 6.02±0.261ghi 3.23±0.095k 5.92±0.534AB
T3 9.25±0.099ab 6.27±0.095f-i 3.25±0.099k 6.26±0.596A
T4 8.75±0.214abc 6.75±0.112d-g 3.75±0.281jk 6.42±0.512A
T5 7.73±0.340cde 5.00±0.447ij 2.75±0.112k 5.16±0.526C
T6 8.02±0.371bcd 5.25±0.171hi 3.00±0.263k 5.42±0.520BC
T7 8.00±0.365bcd 5.00±0.373ij 3.00±0.258k 5.33±0.531BC
T8 8.75±0.297abc 5.77±0.211ghi 3.23±0.095k 5.92±0.559AB
T9 9.50±0.124a 6.77±0.105d-g 3.27±0.154k 6.51±0.623A
T10 8.52±0.194abc 6.50±0.190e-h 3.75±0.198jk 6.26±0.485A
T11 7.77±0.120cde 5.27±0.154hi 2.75±0.198k 5.26±0.504C
T12 8.00±0.274bcd 5.50±0.190ghi 3.00±0.263k 5.50±0.513BC
T13 8.25±0.324abc 5.25±0.092hi 3.02±0.259k 5.51±0.537BC
Total 8.32±0.089A 5.79±0.086B 3.09±0.063CIn a column or row means having similar letters are statistically NS (P>0.05). Interaction means comparison is represented by (a-z) letters in above Table while overall means are denoted by (A-Z) letters.T0= CT1= Edible coating of butter with CST2 = Butter containing 0.2% BCOT3= Butter containing 0.3% BCOT4= Butter containing 0.4% BCOT5= CS based edible coating of butter containing 0.2% BCOT6= CS based edible coating of butter containing 0.3% BCOT7= CS based edible coating of butter containing 0.4% BCOT8= Butter containing 1.5% GOT9= Butter containing 2.0% GOT10=Butter containing 2.5% GOT11=CS based edible coating of butter containing 1.5% GOT12=CS based edible coating of butter containing 2.0% GOT13=CS based edible coating of butter containing 2.5% GO
170
The mean Table 4.18.4b for overall acceptability of butter showed that edible
coating and essential oils have ** (p<0.01) variation on overall acceptability of butter.
Results revealed that the highest overall acceptability value (9.50) was recorded in butter
sample T9 (butter containing 2.5% GO) at 1 day while the lowest value of overall
acceptability (2.50) was noted in T0 butter at 90 days. All the butter samples which were
prepared with CS based active edible coating (containing BCO and GO as active
ingredient) and samples with direct incorporation of these natural essential oils show
consistent decline in overall acceptability during storage of 90 days as compared to T0
butter.
Fig. 4.52 depicted that during storage of 90 days a continuous decline in overall
acceptability of butter was noted in all butter samples which were manufactured with
edible coating and different concentrations of essential oils. Mean value of all treatments
shows that overall acceptability was decreasing from 9.50 to 7.50, 6.77 to 5.0 and 3.75 to
2.50 at 1, 45 and 90 days respectively. Mean values of overall acceptability for all the
butter samples at 45 days and 90 days of storage showed a ** (p<0.01) decrease.
Edible coating and incorporation of EOs in butter samples significantly affects
the sensory properties of butter. Results revealed that overall acceptability score was
decrease during storage but the reduction rate in overall acceptability was less as
compare to control butter that might be due to lipolysis. The overall score of all butter
samples decrease during storage. Similar results were found by Songul et al, (2014), they
also reported that overall acceptability score decrease during storage. The score of the
overall acceptability of butter match with the findings of Foda et al. (2010).
171
CONCLUDING REMARKS
Storage stability of SC (SC) and butter can be enhanced by using active edible
coating and natural essential oils (EOs)
Direct addition of natural EOs was more effective as compared to their use in
edible coating
As the concentration of EOs increase storage stability of SC and butter increases
and vice versa but higher concentration of EOs effects the sensory characteristics
Antioxidant activity (DPPH, POV) of SC and butter consistently decreases during
storage
Texture profile of SC and butter also decreases during storage
Total viable count (TVC) of butter and SC increases as the storage period increase
Rate of free fatty acids (FFA) production in butter continuously increases during
storage
Rate of proteolysis was higher in SC samples containing high moisture to protein
(M/P) ratio during storage of 30 days due to the effect of EOs and edible coating
172
CHAPTER-5SUMMARY
Nowadays industries are facing problems against using chemical or synthetic
additives to prolong the shelf stability of food due to consumer awareness and their bad
health effects. Cheese and butter are dairy products that consumed worldwide directly
and indirectly in a number of food products due to its therapeutic and nutritional
significance. Cheese is very popular in the world due to variety in functionality, flavor,
texture and nutritional value. SC is biologically and biochemically active due to its high
level of percent moisture and subsequently undergoes changes in flavor, functionality,
loss in moisture, mold growth, texture, and causes oxidation during storage that
ultimately causes the deterioration. Similarly fat is a major constituent in butter and
contains more than 78% fat and present in emulsion form i.e. water in oil (w/o). Butter
also contain (milk solid not fat) MSNF and water in the form of small droplets. Fat in
butter plays a very vital role in flavor, nutritional value, appearance, body/texture and
shelf stability. Butter due to its high fat content is more vulnerable to oxidative
deterioration that leads to the reduction of nutritional quality and also makes the food
unacceptable for consumers. Oxidation of fats causes many human diseases like
cardiovascular diseases, membrane damage, cancer and ageing so that antioxidants are
added in foods to prevent or delay oxidation.
Problems related to storage stability of SC and butter can be tackled through
numerous strategies including the application of natural EOs and edible coating. Natural
edible coating and use of natural essential oils is the strategy that is more beneficial to
handle the problems related to storage stability along with health benefits as compared to
chemical preservatives. The current study was conducted to evaluate the effect of edible
coating and natural essential oils (EOs) on storage stability of SC and butter. Corn starch
(CS) and whey powder (WP) based edible coating were developed for butter and SC
respectively. Peppermint oil (PMO) and clove oil (CO) were used as active ingredient in
SC coating while ginger oil (GO) and black cumin oil (BCO) in butter coating. Glycerol
was added as plasticizer, xanthan gum to increase the viscosity, lecithin for
emulsification of EOs in edible coating. Edible coating was applied on SC and butter
with brushing method. SC (T0= control; T1= edible coating of SC with WP; T2 = SC
173
containing 0.5% CO; T3= SC containing 0.75% CO; T4= SC containing 1.0% CO; T5=
WP based edible coating of SC containing 0.5% CO; T6= WP based edible coating of SC
containing 0.75% CO; T7= WP based edible coating of SC containing 1.0% CO; T8= SC
containing 1.5% PMO; T9= SC containing 2.0% PMO; T10= SC containing 2.5% PMO;
T11= WP based edible coating of SC containing 1.5% PMO; T12= WP based edible
coating of SC containing 2.0% PMO; T13= WP based edible coating of SC containing
2.5% PMO) was analyzed for physicochemical parameters, color value, water activity
(aw), texture profile, antioxidant activity, total viable count (TVC), proteolysis and
sensory attributes during 30 days of storage at 2-50C. Similarly butter (T0= control; T1=
edible coating of butter with CS; T2 = butter containing 0.2% BCO; T3= butter containing
0.3% BCO; T4= butter containing 0.4% BCO; T5= CS based edible coating of butter
containing 0.2% BCO; T6= CS based edible coating of butter containing 0.3% BCO; T7=
CS based edible coating of butter containing 0.4% BCO; T8= butter containing 1.5% GO;
T9= butter containing 2.0% GO; T10=butter containing 2.5% GO; T11=CS based edible
coating of butter containing 1.5% GO; T12= CS based edible coating of butter containing
2.0% GO; T13= CS based edible coating of butter containing 2.5% GO) was also analyzed
for physicochemical parameters, color value, water activity (aw), texture profile, free fatty
acids (FFA), antioxidant activity, total viable count (TVC), and sensory attributes during
90 days of storage at 2-50C.
Mean values of physicochemical analysis of SC and butter showed that highest
value of moisture (79.55 %) was noted in T9 SC sample while the lowest moisture
(74.33%) was noted in control (T0) SC. Highest value of protein (13.55%) was observed
in T12 while the minimum protein (13.30%) value was observed in control (T0) SC.
Similarly highest value of moisture (16.53 %) was recorded in butter sample T10 while the
lowest moisture (11.25%) was noted in control (T0) butter. Highest fat (79.80%) was
recorded in control butter (T0) while the lowest fat (79.56%) was noted in T10. Reduction
rate of pH in EOs and edible coated sample of SC and butter was lower as compare to
their control. Similarly the acidity increase slowly in EOs and edible coated sample of SC
and butter due to development of lactic acid but the trend to increase in acidity was slow
as compared to their control. Slight increase in fat was observed in butter and SC
174
samples. Similarly the treatments have NS effect on the protein of SC samples during
storage.
Texture profile of SC and butter effected significantly due to edible coating and
EOs. SC and butter with direct addition of EOs showed higher antioxidant activity as
compared to other samples. TVC (cfu/g) was minimum in all samples of SC and butter as
compared to compare to their control (T0) samples due to the effect of EOs and edible
coating. Higher rate of proteolysis was observed in T4 SC sample due to high moisture to
protein ratio (M/P) while minimum proteolysis was observed in control due to greater
moisture during storage. EOs and edible coating have ** effect on the FFA therefore
minimum FFA production was observed in all butter samples during storage as compared
to control butter. Addition of natural EOs and edible coating significantly improved all
the sensory characteristics of SC and butter. Hence, it is concluded from the present study
that storage stability of SC and butter can be enhanced by using natural edible coating
and natural EOs with better flavor and quality attributes as compared to SC and butter
containing chemical or synthetic additives.
175
RECOMMENDATIONSEdible coating should be used by dairy industry to prolong the storage stability of
SC and butter. It can be recommended from our research findings that essential oils
should be used to prevent microbial spoilage in SC while oxidative rancidity in butter.
Potential of natural EOs as antimicrobial and antioxidants should be explored for their
use as replacer of synthetic antimicrobial and antioxidants in SC and butter.
LIMITATIONS AND FUTURE PROSPECTSThe concentration of EOs in the edible coating and storage stability of soft cheese
and butter have positive correlation as the concentration of EOs increases storage stability
increases and vice versa but higher concentration of EOs in the active edible coating
effects the sensory characteristics. Results of current study could deliver new perception
to dairy industry to fulfill increased demand of consumer towards the use of natural
additives/ preservatives. In further work, different additives should be used to overcome
the sensory effect of EOs. Different combinations and concentrations of these and some
other natural essential oils should also be explored for better results.
176
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