Brigham Young University BYU ScholarsArchive All eses and Dissertations 2018-04-01 Effects of Acid Whey Marination on Tenderness, Sensory and Other Quality Parameters of Beef Eye of Round Jason Kim Brigham Young University Follow this and additional works at: hps://scholarsarchive.byu.edu/etd Part of the Nutrition Commons is esis is brought to you for free and open access by BYU ScholarsArchive. It has been accepted for inclusion in All eses and Dissertations by an authorized administrator of BYU ScholarsArchive. For more information, please contact [email protected], [email protected]. BYU ScholarsArchive Citation Kim, Jason, "Effects of Acid Whey Marination on Tenderness, Sensory and Other Quality Parameters of Beef Eye of Round" (2018). All eses and Dissertations. 6758. hps://scholarsarchive.byu.edu/etd/6758
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Brigham Young UniversityBYU ScholarsArchive
All Theses and Dissertations
2018-04-01
Effects of Acid Whey Marination on Tenderness,Sensory and Other Quality Parameters of Beef Eyeof RoundJason KimBrigham Young University
Follow this and additional works at: https://scholarsarchive.byu.edu/etd
Part of the Nutrition Commons
This Thesis is brought to you for free and open access by BYU ScholarsArchive. It has been accepted for inclusion in All Theses and Dissertations by anauthorized administrator of BYU ScholarsArchive. For more information, please contact [email protected], [email protected].
BYU ScholarsArchive CitationKim, Jason, "Effects of Acid Whey Marination on Tenderness, Sensory and Other Quality Parameters of Beef Eye of Round" (2018).All Theses and Dissertations. 6758.https://scholarsarchive.byu.edu/etd/6758
Effects of Acid Whey Marination on Tenderness, Sensory and Other Quality Parameters of Beef Eye of Round
Jason Kim Department of Nutrition, Dietetics, and Food Science, BYU
Master of Science
The growth of the Greek-style yogurt market is causing many problems for dairy companies who are trying to handle the voluminous whey by-product. Acid whey, unlike sweet whey, has a low amount of protein and high amounts of lactic acid, calcium, and other minerals. Therefore, it has limited commercial value to the food industry and often requires additional processing for disposal. Lactic acid and calcium solutions have shown efficacy in increasing the tenderness of beef and other types of meat. The purpose of this project is to investigate the use of acid whey, with its high amounts of lactic acid and calcium, to tenderize beef (eye of round, IMP 171C) during marination.
This study evaluated the effects of marination of utilizing acid whey in improving quality
parameters of beef. 13 roasts (Top round steaks from USDA Select steers) were randomly assigned to one of six marination treatments: (1) calcium chloride, (2) lactic acid, (3) phosphate (4) acid whey (lot 1), (5) acid whey (lot 2), and (6) control. Steaks were marinated in vacuum pouches, aged for 48 hours, cooked to 70º C and evaluated by a consumer sensory panel and other quantitative tests (texture analyzer, colorimeter, collagen, cook loss, and pH). Marination with acid whey increased the tenderness and juiciness ratings without decreasing hedonic liking scores for the overall flavor or aftertaste of the beef samples. Keywords: acid whey, beef, eye of round, Greek yogurt whey, hedonic scale, colorimeter, liking, marination, tenderness, ta.xt2, sensory
ACKNOWLEDGMENTS
I would like to express my gratitude to the faculty and staff of the department of
Nutrition, Dietetics, and Food Science for their time in teaching and mentoring me throughout
my undergraduate and graduate degrees at BYU. I am grateful to Dr. Steele, Dr. Jefferies and Dr.
Dunn for their help on my committee, and Dr. Dunn for serving as my committee chair. I could
not have produced as fine of research without their expert guidance along the way. Thanks are
also in order for the students of the food research lab and my fellow graduate students who
helped me think through many problems.
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TABLE OF CONTENTS
TITLE PAGE ................................................................................................................................... i
ABSTRACT .................................................................................................................................... ii
ACKNOWLEDGMENTS ............................................................................................................. iii
TABLE OF CONTENTS ............................................................................................................... iv
LIST OF TABLES ......................................................................................................................... vi
LIST OF FIGURES ...................................................................................................................... vii
Table 1 Proximate composition (%) and mineral content (ppm) of acid whey ............................ 11
Table 2 Consumer acceptance and hedonic ratings for beef eye of round steaks marinated in
different solutions ......................................................................................................................... 12
Table 3 Means for soluble collagen, insoluble collagen, cook loss percentage, pH, and shear
force .............................................................................................................................................. 13
Table 4 Effect of acid whey and other marinade treatments on color of beef eye of round. ........ 15
Table 5 Proximate Composition (%) and Mineral Content (ppm) of Acid Whey ........................ 27
Table 6 Mineral Analysis (ppm) ................................................................................................... 27
vii
LIST OF FIGURES
Figure 1 Scanning electron micrograph (SEM) of raw beef muscles with different treatments
aMeans for sensory panel rating for overall acceptability, appearance, flavor, tenderness, juiciness and aftertaste (n=110). Acceptability, appearance, flavor, and aftertaste were calculated based on a 9-point hedonic scale,1=dislike extremely, 9=like extremely. Tenderness was calculated based on a 5-point JAR scale, 5=definitely too tender, 3=just-about-right, 1=definitely not tender enough. Juiciness was calculated based on a 5-point JAR scale, 5=definitely too juicy, 3=just-about-right, 1=definitely not juicy enough. Like super-scripts within a column represent no significant difference (p>0.05). Compositional and Textural Properties
The pH for the acid whey (5.18) treated beef was significantly lower than the control
sample and all other treatments. This result was unexpected because the lactic acid and whey
marinades were initially at the same pH. This difference in pH could possibly be explained by
the inherent microbial load of the acid whey, that may still have continued to produce lactic acid
13
while the meat was marinated; or alternatively the pH difference may have resulted from
presence of natural buffers in the whey samples.
The statistical differences for tenderness, exhibited in the consumer panel, were not
picked up using the Warner-Bratzler attachment on the texture analyzer. There was no significant
difference in peak force detected between any of the treatments (see Table 3). This is clearly a
case where the human sensory organ is more accurate than the instrument. We specifically used
the semitendinosus muscle in this study because it is a tougher cut of beef, which could show
potential enhancements from marination pretreatment. However, DeYonge-Freeman and others
(2000) reported no improvement in semitendinosus tenderness after calcium chloride injection.
Aktas and Kaya (2001), reported that longissimus dorsi decreased in peak force when treated
with lactic acid. However, this tenderization effect was attributed to the change in pH to below
4.0. Similarly, Ertbjerg and others (1995) showed that lactic acid injected at low concentration
(0.3 M), leading to a pH of 5.2 -- which is close to the isoelectric point of the major myofibrillar
protein, did not improve beef texture, while injection at 1.0 M resulted
Table 3 Meansa for soluble collagen, insoluble collagen, cook loss percentage, pH, and shear force. Treatment
aPercent cook loss was calculated using weights taken immediately prior to and following cooking. pH was determined immediately after the 48 hour marination period using a surface pH probe. Shear force was calculated by recording the peak force applied to the sample core. Like super-scripts within a column represent no significant difference (p>0.05).
14
in a meat pH of 4.6 and decreased meat toughness. Since the meat pH for all the treatments
evaluated in our study did not fall below pH 5.0 (acid whey treatment being the lowest, at pH
5.18), it is possible that lower pH marinade treatments may have resulted in greater differences
in tenderness as manifested by texture analyzer peak force. Lower pH treatments were not
evaluated, due to our interest in evaluating the efficacy of untreated acid whey, at its native pH.
The only significant increase in soluble collagen content was observed in the phosphate
treatment, which showed a nearly 1.8-fold increase (see Table 3) in collagen solubility compared
to the control. There was no significant difference in the amount of insoluble collagen for any of
the treatments (see Table 3). Collagen has swelling properties under acidic or alkali conditions,
and swollen collagen can be more readily converted into gelatin at high temperatures such as
those encountered during cooking. In a number of previous studies, increased collagen solubility
under acidic or alkali conditions resulted in improvements of meat tenderness. For example,
Naveena and others (2011) reported that collagen solubility of buffalo meat increased with
increasing ammonium hydroxide concentrations. Oreskovich and others (1992) noted an increase
in soluble collagen values in beef marinated with 0.7M acetic acid (pH 2.50), compared to those
in control (non-buffer) and 0.1M NaCl (pH 6.50) marinated beef. Chang and others (2010)
suggested that marination with weak organic acids causes the denaturation of intramuscular heat-
soluble collagen. Even though the lactic acid and acid whey contained weak organic acids, the
concentration at which the treatments were applied to each sample possibly did not lower the pH
enough to result in a change in the soluble collagen content. While the acid whey and phosphate
pH levels were significantly different from control (the phosphate sample was near neutral, the
acid whey sample remained above pH 5), only the phosphate had an effect on the soluble
collagen.
15
As expected, the phosphate treatment had the lowest cook loss percentage (19.72%) and
was significantly lower than all the other treatments (p>0.05) (see Table 3). Phosphates are well
known for increasing water holding capacity and reducing cook loss in meat products (Roldan
and others 2014). However, it is interesting to note that, of all marinade treatments, the acid
whey treated sample had the next lowest percentage cook loss at 34.78% which was only about
5% higher than the control and significantly lower than the calcium chloride and lactic acid
treatments. Gault (1984, 1985), and Rao and Gault (1990), and Offer and others (1989) found
that meat treated with acidic marinades with a pH below 5.0 suffered less cooking loss. This may
help explain the lower cook loss for the acid whey treated sample, compared to the other non-
phosphate marinades.
Color
Before the samples were cooked, the acid whey treatment was significantly darker than
the control sample, but not significantly different from control on the red-green (a*) or blue-
yellow
Table 4 Effect of acid whey and other marinade treatments on colora of beef eye of round. Before Cooking After Cooking
aThe three CIE color coordinates are defined as: L*, where 0 = black and 100 = white; a*, where negative values indicate green, while positive values indicate red; and b*, where negative values indicate blue and positive values indicate yellow. Like super-scripts within a column represent no significant difference (p>0.05).
16
(b*) scales (see Table 4). After cooking, the acid whey marinated meat was no longer
significantly darker than the control, but was significantly less red and less yellow. Compared to
the other treatments, the uncooked acid whey samples were also significantly darker than lactic
acid, but lighter than phosphate and calcium treatments, both of which were similar to the
control. The calcium chloride treatment was significantly darker than all other treatments and
control, though not significantly darker than the acid whey.
The color effects reported in Table 4, for the uncooked samples, differ somewhat from
color changes resulting from the natural decline in pH of beef as it ages. Generally, the natural
pH drop in beef during aging, leads to a lighter color (Wojciak 2014), whereas in our study, the
lowest pH product (acid whey marination) resulted in a darker colored product, compared to
control. The minerals and sugars present in the whey may have played a role in the darkening
effect, possibly by altering the oxidation state of the myoglobin.
Considering all the differences in cooked beef color across the sample spectrum, acid
whey treated beef was the only marinated sample that was at statistical parity with the control
sample for consumer acceptance of appearance, with the two most preferred samples being the
acid whey and control sample.
Electron Microscopy
Figure 1 shows an electron micrograph of uncooked muscle and connective tissue from
the bovine semitendinosus muscles 48 h after each treatment (control-no treatment, acid whey,
calcium chloride, lactic acid, and phosphate). Each image examines the intramuscular collagen
matrix; and the effects of each treatment on the collage structure can be seen by comparison to
the untreated control.
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The unmarinated, raw control sample shows bundles of regular muscle fibers with an associated,
crisscrossed network of fairly tight collagenous fibers. The acid whey sample, by comparison,
shows a much looser, more open collagen structure, especially compared to the calcium chloride
sample. The lactic acid treated sample is quite similar to the control, while the phosphate treated
sample shows very loose, widely separated and disordered collagen fibers, very much unlike any
of the other samples.
The electron micrographs support the findings of the collagen extraction data in that there
are no dramatic differences in the appearance of the collagen in any of the samples, with the
exception of the phosphate treated sample. The control-no treatment, lactic acid and calcium do
not have any physical sign of collagen breakdown; however, the acid whey sample seems to
show signs that some of the collagen fibers are starting to breakdown.
18
Control Acid Whey
Calcium Chloride Lactic Acid
Phosphate
Figure 1 Scanning electron micrograph (SEM) of raw beef muscles with different treatments (control-no treatment, acid whey, calcium chloride, lactic acid, and phosphate)
19
Conclusion
Untreated acid whey appears to be a suitable base for beef marinade. Consumer
acceptance testing showed that marination of beef in acid whey resulted in the highest overall
flavor acceptance scores, and significantly improved the tenderness and juiciness of samples
relative to the control, without any indication of negative off-flavors. However, the increase in
tenderness and juiciness observed in consumer testing was not large enough for analytical
instrumentation to detect with statistical significance, and was not manifested in the
soluble/insoluble collagen results. Acid whey resulted in more significant cook loss than
control, though significantly less than the non-phosphate marinades evaluated. The color of acid
whey treated beef was different than control, but did not significantly affect consumer
acceptance. The effects of lactic acid and calcium were individually evaluated in other
treatments, and no apparent synergistic effects on meat tenderization with the combination of
calcium and lactic acid present in acid whey were observed. The main driver in tenderization
seems to be the change in pH from the marination.
20
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Appendix
Texture Analyzer Test Settings
Each day before it was used the texture analyzer was calibrated for height and force of 50 mm
with a return speed of 10 mm/sec and a contact force of 1 g. The following below are the settings
for the TAXT2 with the Warner Bratzler attachment.
Test Mode – Compression
Pretest Speed – 3.3 mm/sec
Test Speed – 3.3 mm/sec
Post Test Speed – 5 mm/sec
Target Mode - Distance
Distance - 30 mm
Trigger Type - Auto (Force)
Tigger Force - 20 g
Advanced Options – Off
Collagen Method - Detailed
The cooked steaks after the texture analyzer test were ground into a homogenous mixture
using a food processor for 1 min. Ground samples (4g) were placed into 50mL polycarbonate
centrifuge tubes in duplicate. Ringer’s solution (22mL; NaCl, CaCl, and KCl) was added to each
tube and the samples were homogenized in a Tissue Master Homogenizer 125 at 20,000 rpm for
1 min. The samples were then heated in a water bath at 50C for 15 min and subsequently
centrifuged (HN-SII Benchtop Centrifuge) at 2500 G at room temperature for 5 min. The
supernatant was filtered through #5 filter paper (Thermo Fisher Scientific) into an Erlenmeyer
26
flask labeled as soluble collagen. The pellet was rinsed with ¼ strength Ringer’s Solution
(10mL) and centrifuged as previously described. The supernatant was filtered into the soluble
collagen flask and the pellet and filter paper were placed in a flask labeled as insoluble collagen.
Sulfuric acid (3.5M/30 mL) was added to both the soluble and insoluble portions. The flasks
were covered and placed in an oven (Lab Line Imperial II Radiant Heat Oven) at 105C for at
least 20 h. The hot hydrolysate labeled insoluble was diluted to 100mL with water, mixed and
filtered through Fisherbrand #5 filter paper. The hot hydrolysate labeled soluble was taken out
and boiled until the volume was less than 25 mL and then it was diluted to 25mL with water,
mixed and filtered through Fisherbrand #5 filter paper. The samples were pipetted (insoluble;
0.36 mL diluted hydrolysate and 5.64 mL of water; soluble; 6 mL of diluted hydrolysate) into
borosilicate glass 13 x 100mm test tubes. One milliliter oxidant (citric acid monohydrate, sodium
hydroxide, sodium acetate trihydrate, 1-propanol, and chloramine-t, pH=6) was added and the
samples were mixed and allowed to stand for 20 min. One mL color reagent (perchloric acid, 4-
dimehtylaminobenzaldehyde, and 2-propanol) was added and mixed. The resulting mixture was
heated to 60 C in a water bath for 15 min and then allowed to cool in cold tap water bath for 3
min. Absorbance of samples was read at 558nm using a VWR UV-1600PC Spectrophotometer.
Hydroxyproline standards were used to generate a calibration curve and collagenous connective
tissue content was multiplied by 7.52 and 7.25 to determine soluble and insoluble collagen,
respectively (Cross, Carpenter, and Smith, 1973).
27
Table 5 Complete Proximate Composition (%) and Mineral Content (ppm) of Acid Wheys Acid
Sensory Comments P r o d u c t s 1 - 935 935 No Treatment 2 - 146 146 Acid Whey 3 - 213 213 Calcium 4 - 527 527 Lactic Acid 5 - 798 798 Phosphate R e s u l t s 1 17 1-935 The taste was absent while the texture seemed unsettling. This was my least favorite
of the selections today.
1 19 1-935 tasted very good
1 40 1-935 It was really hard to answer all of these questions with the amount of sample that we
had. I didn't know how many more questions I was going to have to answer and two bites was
not enough to remember everything. If I could answer all of the questions at the same time, then
I could have gone through and rated everything about one sample, and then move on to the next
sample. ...I didn't know I was going to have to rank them, I think that is close to my opinon, but
there were so many samples and so many questions.
1 45 1-935 This sample is slightly over cooked. But still the best.
1 48 1-935 GReat. Could maybe use a touch of tenerizer or pounding.
1 51 1-935 this sample is the only one that really smelled like a steak to me.
1 55 1-935 This was the best of the five...
1 66 1-935 This sample was the best tasting as far as flavor, tenderness, and juiciness.
1 76 1-935 935 had the most/best flavor of the samples that I enjoyed.
1 90 1-935 I liked the doneness and flavor of 935 the best, but in all cases, l was surprised by
how watery all the steak tasted.
85
1 101 1-935 While 935 appeared the darkest, I thought it had the best flavor and about the right
level of juicyness.
1 107 1-935 This might've tasted the best by just a tiny bit, but the overall presentation brought it
down to 2nd for me.
1 109 1-935 I like it! good presentation and flavor, needed to be juicy to reach perfection.
1 14 2-146 Sample 146 was near perfect! It was juicier and tasted better - but wasn't as raw as
sample 798
1 19 2-146 tasted very good
1 33 2-146 very tough and difficult to chew
1 48 2-146 REALLY chewey
1 57 2-146 I like the outter sear.
1 71 2-146 Very well prepared and had an excellent taste.
1 80 2-146 This one felt like the clear winner to me. The others were either too dry or too bland.
1 102 2-146 Best overall sample, but all samples could have used a little salt.
1 109 2-146 Is correct the point of cooking.
1 18 3-213 A little on the rare side.
1 19 3-213 too much grizzel
1 38 3-213 too pink for me to eat :(
1 39 3-213 Looked too raw to me.
1 48 3-213 NExt best. Not as chewey as the other 3.
1 49 3-213 A little to red for me - didn't seem well done.
1 57 3-213 Very pale.
1 65 3-213 #213 wasn't cooked very well, but it had the best flavor.
86
1 109 3-213 Needs to bleed to be juicy. If it is 'rare'
1 19 4-527 tasted very good
1 37 4-527 Seems too rare.
1 38 4-527 Too pink to eat, and you can see a small vessel :(
1 48 4-527 chewy
1 49 4-527 Way too red, color turned me off a bit.
1 57 4-527 appearance is just right.
1 71 4-527 While I wouldn't have considered it as ''well done,'' this sample was exceptionally
good. The taste and moistness were very good.
1 89 4-527 Best one for juciness and flavor
1 107 4-527 This had hardly any taste and was way too rare for me.
1 109 4-527 It is assumed that needs to be juicy. It was not tender.
1 15 5-798 Looks scary. I think I always expect more color.
1 16 5-798 Sample 798 one looks under cooked, I had to really talk myself into eating it. There's
a strange sheen to the meat. The seared edges add flavor, this one didn't offer that seared flavor.
1 18 5-798 Does not appear cooked enough. Appears rare and too much juicy liquid.
1 19 5-798 Little bloodier but very good
1 48 5-798 chewy, but not quite as band as the last 2
1 49 5-798 Closer to well done, but still a little too pink.
1 53 5-798 This is the only sample I deliberately did not finish.
1 57 5-798 Very pale.
1 58 5-798 This one was my favorite
1 83 5-798 I felt this one was over cooked.
87
1 89 5-798 Dry
1 100 5-798 it was an off flavor not exactly meat off flavor
1 102 5-798 Toughest and driest of the lot.
1 107 5-798 This was my second favorite in taste by just a tiny bit, but the overall presentation
(appearance, juiciness, aroma, flavor, etc) made it the best for me.