LOW TEMPERATURE EXTRACTION OF PLANTS BY LIQUIFICATE GASES. 10. WASTE OF CHOKEBERRY FRUITS (Aronia melanocarpa (Michx) Elliott.)

Journal of

EcoAgriTourismJournal of

EcoAgriTourismBulletin of Agri-ecology Agri-food Bioengineering and Agritourism

Vo

l 9

(20

13)

Nr

1 (

26)

Biodiversity

CalitaTerra

Sustainable Tourism and Hospitality

Ideas and Concepts

Sanogeneous Food

NewsJournal of EcoAgriTourism

ISSN 1844-8577

Journal of EcoAgriTourism is a follow up by translation in English of

ldquoRevista de EcoAgroTurismrdquo ISSN 1841-642X first issued in 2005

Journal of EcoAgriTourism CalitaTerra Vol 9 no1 2013

23

LOW TEMPERATURE EXTRACTION OF PLANTS BY LIQUIFICATEGASES WASTE OF CHOKEBERRY FRUITS

(Aronia melanocarpa (Michx) Elliott)

P MERDZHANOV ANGELOV-ROMOVA N NENOV MZLATANOV G ANTOV T ATANASOV P DENEV A STOYANOVA

Abstract The lipid composition from waste of chokeberry (Aronia melanocarpa (Michx)Elliott) extraction with freon134a (1112-tetrafluorethane) and with freon 134a+acetonewas analyzed using GC and HPLC The main components in the triacylglycerol fractionswere linoleic (478 ndash 572) oleic (264 ndash 284) and palmitic (110 ndash 155) acids -Tocopherol (593 ndash 614) predominated in the tocopherol fractions and -sitosterol(743) ndash in the sterol fraction

Keywords waste of aronia fruits lipid composition

1 Introduction

Aronia (Aronia melanocarpa (Michx) Elliott)family Rosaceae originates from the eastern partsof North America It is industrially cultivated inmany European countries as an industrial cropThe chokeberries are good raw material for theproduction of various beverages including juicesnectars syrups jellies jams tea wines andliquors [11] The berries contain anthocyanins flavonolsvaluable minerals vitamins and dietary fibres[11 12 15 19 20 22]Chokeberries have a long traditional use in folkmedicine Among the health benefits ofchokeberries are inhibition of cancer cellproliferation antimutagenic effecthepatoprotective effect cardioprotective effectand antidiabetes effect [21] Data about the chemical composition of thelipid fraction of chokeberries fruits are scarceIn the lipid fraction of the fruits from thechokeberries growing in Bulgaria Antova [2] andZlatanov [24] have been identified leic (214)linoleic (711) and palmitic acid (51) In thelipid fraction incorporates phospholipids (28)mainly phosphatidylinositol (298) sterols(12) mainly -sitosterol (898) andtocopherols (555 mgkg) mainly -tocopherol(706) In our previous paper we established inthe lipid fraction from the waste the many acidsbut in different quality - linoleic (397) oleic(307) and palmitic (198) acids The maincomponents in the tocopherol fraction were -tocopherol (597) and -sitosterol (826) andcampesterol (64) ndash in the sterol fraction [1]

In the lipid fraction investigated by Anwar etal [3] the prevailing fatty components werelinoleic (279) oleic (6190) and palmiticacids (32) as well as sterols mainly -sitosterol (660) 5-avenasterol (180) andcampesterol (50) Currently in many countries medical plants areprocessed by extraction with liquefied gases

2 air freon 134a and other) The producedextracts are harmless thatrsquos why they can bewidely used in food and flavour industrycosmetics and medicine The usage of liquefiedgases overcomes the drawbacks of installationsworking with volatile polar and apolar solvents[13] The aim of present study is producing of wasteof chokeberry by using freon 134a and mix offreon 134a and acetone in laboratory installationand determination of their chemical compositionand characteristics for possible application innatural food product

2 Materials and Methods

Samples The plant material was aronia fruitwaste which was dried at room temperature The moisture content (79) was determinedby drying up to constant weight at 105o

(Russian Pharmacopoeia 1990) The used solvent 1112 tetrafluorethane (freon134a) is apolar with relative permittivity at 20o

and 100 kHz 1013 dipole moment 2060 Debayand polarization 138 cm3mol Its dynamicviscosity and surface tension at 20o are small198 as and 8 mNm respectively whichallows easier penetration into plant cells and


24

benefits extraction of cells associatedcompounds Its pressure at 20o is 057 MPawhich allows the extraction process to be carriedout at acceptable pressures from 02 to 07 MPaIts specific heat of vaporization at the appliedthermal regiment is low (about 200 kJkg) whichdetermines small energy consumption forextraction process The solvent is also chemicallyinert and it is well compatible with copper andcarbon steel The influence of freon134a on theGreen House Effect with coefficient HGWP =0285 and its higher price are its majordisadvantagesThe other solvent is acetone Extraction The air-waste are ground separatelyin an attrition mill to a size of 015 ndash 025 mmand extracted in laboratory scale 1 l volumeextractor using subcritical liquefied gas co-extraction The used solvent was mixture ofliquefied gas tetrafluoroethane and acetone inproportion (tetrafluoroethane acetone 101) fortotal amount of acetone of 49 g The weight ofsample was 280 g The extraction temperaturewas 25 ndash 30oC and absolute extraction pressurewas 070 ndash 080 MPa The extraction processfeatured triple exchange of solvent with durationof each stage of 30 min Total duration ofextraction process was 90 min The solvents was partly removed in a rotaryvacuum evaporator the residue was transferred toa pre-weighed glass vessel and the rest of thesolvent was removed under stream of nitrogen toa constant weight in order to determine the oilcontent [6] Fatty acids The total fatty acid composition ofthe oil was determined by GC aftertransmethylation of the respective sample with2N methanolic KOH at 50oC according toChristie [4] Fatty acid methyl esters (FAME)were purified by TLC on 20 cm x 20 cm platescovered with 02 mm Silica gel 60 G layer(Merck Darmstadt Germany) with mobile phasen-hexaneacetone 100 8 (by volume) Determination was performed on a gaschromatograph equipped with a 30 m x 025 mmx 25 m (ID) capillary EC 30-Wax column(Hewlett Packard GmbH Vienna Austria) and aflame ionization detector The columntemperature was programmed from 130oC (hold1 min) at 65oCmin to 170 OC at 3oCmin to215oC (hold 9 min) at 40oCmin to 230oC (hold 1min) injector were 270oC and detectortemperatures were 280oC Hydrogen was thecarrier gas at a flow rate 08 mlmin split was501

Identification was performed by comparison ofretention times with those of a standard mixtureof FAME subjected to GC under identicalexperimental conditions [5 7]

Sterols Unsaponifiables were determined byweight after saponification of the lipid fractionand extraction with hexane [10] Theunsaponifiable matters (100 mg preciselymeasured) was applied on 20 cm x 20 cm glassplates (ca 1 mm thick Silica gel G layer) anddeveloped with n-hexane acetone 100 8 (byvolume) Free sterols (Rf = 04) were detectedunder UV light by spraying the edges of eachplate with 2acute7acute-dichlorofluorescein they werethen scraped transferred to small glass columnsand eluted with diethyl ether The solvent wasevaporated under a stream of nitrogen and theresidue was weighed in small glass containers toa constant weight Sterol composition wasdetermined by GC using HP 5890 gaschromatograph (Hewlett Packard GmbH ViennaAustria) equipped with a 25 m x 025 mm DB ndash 5capillary column (Agilent Technologies SantaClara CA USA) and a flame ionization detector Temperature gradient was from 90oC (hold 2min) up to 290oC at a rate 15oCmin and then upto 310oC at a rate of 4oCmin (hold 10 min) theinjector temperature was 300oC and the detectortemperature was 320oC Hydrogen was used ascarrier gas at a flow rate 08 mlmin split 50 1 Identification was confirmed by comparison ofretention times with those of a standard mixtureof sterols [9]

Tocopherols Tocopherols were determineddirectly in the oil by high performance liquidchromatography (HPLC) by a Merck-Hitachi(Merck Darmstadt Germany) unit equipped witha 250 mm x 4 mm Nucleosil Si 50-5 column(Merck Darmstadt Germany) and a fluorescentdetector Merck-Hitachi F 1000 The operatingconditions were as follows mobile phase n-hexane dioxan 96 4 (by volume) flow rate 10mlmin excitation 295 nm emission 330 nm 20

kl 1 solution of crude oil were injected Tocopherols were identified by comparing theretention times to those of authentic individualpure tocopherols The tocopherol content wascalculated on the base of tocopherol peak areas inthe sample vs tocopherol peak area of thestandard tocopherol solution [8]


25

2 Results and Discussions

The lipid fractions were 007 in the extractwith freon 134a and 027 in extract withmixture freon 134a+acetone As seen the resultsfor the yields of extracts are comparable withliterature data are almost very small with thesefor the lipid fraction obtained by traditionalsolvents The extracts are odorless viscous products withamber color

The fatty acid composition is presented inTable 1 Data show that 11 fatty acids weredetected constituting 100 of the total oilcontent The main fatty acids in lipid fraction wereoleic linoleic and palmitic acids In the extractsaccording to data from the literature are identifiedas predominating the same three fatty acids butin different quantities which could be explainedby the influence of the origin of the raw

Table 1 Fatty acid composition (ww)Fatty acids Extract with

freon 134aExtract with

freon 134a+acetone1 120 Lauric 16 092 140 Myristic 11 073 160 Palmitic 155 1104 161 Palmitoleic 04 025 170 Margaric 02 026 180 Stearic 28 127 181 Oleic 284 2648 182 Linoleic 478 5729 183 Linolenic 11 0910 200 Arachidic 09 1111 201 Gadoleic 02 02

The correlation unsaturated saturated fattyacids was 779 221 in the extract with freon134a and 849 151 in extract with freon 134a+acetone

Their distribution profiles are presented onFigures 1 and 2 Palmitic acid predominated inthe fraction of saturated fatty acids and oleic andlinoleic were predominant among the unsaturatedacids

123

123

Fig 1 Distribution of fatty acids in extractwith freon 134a

1 - Saturated acids (221)2 - Monounsaturated acids (290)3 - Polyunsaturated acids (489)

Fig 2 Distribution of fatty acids in extract withfreon 134a+acetone


Regarding the individual presence of oleic andlinoleic acid the oil from waste of chokeberrieswas similar to the oils from other nontraditional

materials such as grape seeds watermelontobacco and poppy seeds (Popov and Ilinov1986 Shterbakov 1963) In lipid fractions formwaste of aronia was found to contain very high


26

amounts of the saturated palmitic acid (110 ndash155) which comes close to the levels in otheroils like olive oil (75 - 200) and corn oil (80ndash 190) [14] Sterols are present in the so called non-saponificated part from the lipid fraction and inthe extract with freon 134a its not determined By extraction with freon 134a+acetone theircontent in non- saponificated part is 137 andthe total content in the extract was found to be29 The individual sterol composition ispresented in Table 2 -sitosterol predominated in

the sterol fraction It is obvious from the datathat regarding its sterol content and compositionextract with freon 134a from waste ofchokeberries was similar to the findings for lipidfraction with hexane from waste of aronia fruits[1] lipid fraction from the fruits of the Apiaceaefamily [23]

Table 2 Sterol composition (ww)Sterols Extract with freon 134a+acetone

1 Cholesterol -2 Campesterol 313 Stigmasterol 384 - Sitosterol 7435 5- Avenasterol 836 7- Stigmasterol 337 7- Avenasterol 72

not detected

The total content of tocopherols in the lipidfraction was comparatively higher ndash 1530 mgkgfor extract with freon 134a and 1287 mgkg forextract with freon 134a+acetone The tocopheroland tocotrienol composition are presented inTable 3 The -tocopherol predominated in theextracts followed by -tocopherol and -tocopherol Regarding the low content of -

tocopherol the examined extracts proved superiorto a number of common food oils for example ndashcorn oil (500 ndash 620) and soya oil (600 ndash850) thus showing similarity to some non-traditional oils such as pyrene oils of morello(931) and apricot (960) [16 17]

Table 3 Tocopherol composition (ww) Tocopherols and tocotrienol Extract with


freon 134a+acetone1 -Tocopherol 191 1022 -Tocotrienol 07 -3 -Tocopherol 614 5934 -Tocotienol - 435 -Tocopherol 188 2146 -Tocopherol - 48

not detected

Conclusion

For the first time in Bulgaria new extracts fromwaste of chokeberries (Aronia melanocarpa(Michx) Elliott) were produced by extractionwith liquefied gases - freon 134a and freon134a+ cetone The yield of produced extractswas small but their chemical composition wasinteresting The extracts from waste of

chokeberries can be used as a non-traditionalmaterial rich in biologically active substances asoleic and linoleic acids -sitosterol and -tocopherol for an additive in fodder mixtures inorder to enrich them with valuable nutrients

References

1 Angelova-Romova M P Merdzhanov PDenev M Zlatanov A Stoyanova MKratchanova Lipid composition of


27

chokeberry fruits (Belarus) 12 2012 1

76 ndash 802 Antova G Lipid composition of chokeberry

black currant and rose seed oils SciencesWorks of the University of FoodTechnologies v 43 1998 2 361 ndash 365

3 Anwar F R Przybylski M Rudzinska EGruczynska J Bain Fatty acid tocopheroland sterol composition of Canadian prairiefruit seed lipids Journal of American OilChemistry and Society v 85 2008 953 ndash959

4 Christie W Lipid Analysis The Oily PressBridgwater (3rd Edition) England 2003

5 Codex Stan 210 ndash 1999 Codex standard fornamed vegetable oils Revisions 2001 20032009 Amendment 2005 2011

6 ISO 659 Oilseeds Determination of oilcontent (Reference method) 2009

7 ISO 5508 Animal and vegetable fat and oilsAnalysis by gas chromatography of methylesters of fatty acids 1990

8 ISO 9936 Animal and vegetable fat and oilsDetermination of tocopherols and tocotrienolcontents (HPLC method) 2006

9 ISO 12228 Animal and vegetable fat and oilsDetermination of individual and total sterolscontents (Gas chromatographic method)1999

10 ISO 18609 Animal and vegetable fat andoils Determination of unsaponifiable matter(Method using hexane extraction) 2000

11 Kulling SE HM Rawel Chokeberry(Aronia melanocarpa) A review on thecharacteristic components and potential healtheffects Planta Medica v 74 2008 1625-1634

12 Lehmann H Die Aroniabeere und ihreverarbeitung Fluumlssiges Obst v 57 1990746-752

13 Nenov N Low temperature extraction ofessential oil bearing plants by liquificategases 1 Laboratory installation Sciences

Works of the University of FoodTechnologies v 53 2006 2 195-200

14 OBrien RD WE Farr PJ WanIntroduction to fats and oils technology (2rd

Edition) AOCS Press Champaign IL 200015 Oszmianski J A Wojdylo Aronia

melanocarpa phenolics and their antioxidantactivity European Food Research andTechnology v 221 2005 809-813

16 Popov A P Ilinov Chemistry of lipidsNauka i Iskustvo Sofia 1986

17 Russian Pharmacopoeia (11th Edition)Moscow 1990

18 Shterbakov V Biochemistry of oil plantsMoscow 1963

19 Strigl AW E Leitner W Pfannhauser Dieschwarze Apfelbeere (Aronia melanocarpa)als natuumlrliche Farbstoffquelle DeutscheLebensmittel Rundschau v 91 1995 177-180

20 Tanaka T A Tanaka Chemical componentsand characteristics of black chokeberryJournal of the Japanese Society for FoodScience and Technology v 48 2001 606-610

21 Valcheva-Kuzmanova S A BelchevaCurrent knowledge of Aronia melanocarpa asa medicinal plant Folia Medica v 48 200611-17

22 Wu X L Gu R Prior S McKayCharacterization of anthocyanins andproanthocyanidins in some cultivars of RibesAronia and Sambucus and their antioxidantcapacity Journal of Agricultural and FoodChemistry v 52 2004 7846-7856

23 Zlatanov M S Ivanov Untersuchungen uumlberdie Sterinzusammensetzung der Samenoumlleeiniger Vertreter der Familie Apiaceae FeetWissenschaft Technologie v 97 1995

10381-38324 Zlatanov M Lipid composition of Bulgarian

chokeberry black currant and rose seed oilsJournal of the Science of Food andAgriculture v 79 1999 12 1620-1624


23

LOW TEMPERATURE EXTRACTION OF PLANTS BY LIQUIFICATEGASES WASTE OF CHOKEBERRY FRUITS

(Aronia melanocarpa (Michx) Elliott)

P MERDZHANOV ANGELOV-ROMOVA N NENOV MZLATANOV G ANTOV T ATANASOV P DENEV A STOYANOVA

Abstract The lipid composition from waste of chokeberry (Aronia melanocarpa (Michx)Elliott) extraction with freon134a (1112-tetrafluorethane) and with freon 134a+acetonewas analyzed using GC and HPLC The main components in the triacylglycerol fractionswere linoleic (478 ndash 572) oleic (264 ndash 284) and palmitic (110 ndash 155) acids -Tocopherol (593 ndash 614) predominated in the tocopherol fractions and -sitosterol(743) ndash in the sterol fraction

Keywords waste of aronia fruits lipid composition

1 Introduction

Aronia (Aronia melanocarpa (Michx) Elliott)family Rosaceae originates from the eastern partsof North America It is industrially cultivated inmany European countries as an industrial cropThe chokeberries are good raw material for theproduction of various beverages including juicesnectars syrups jellies jams tea wines andliquors [11] The berries contain anthocyanins flavonolsvaluable minerals vitamins and dietary fibres[11 12 15 19 20 22]Chokeberries have a long traditional use in folkmedicine Among the health benefits ofchokeberries are inhibition of cancer cellproliferation antimutagenic effecthepatoprotective effect cardioprotective effectand antidiabetes effect [21] Data about the chemical composition of thelipid fraction of chokeberries fruits are scarceIn the lipid fraction of the fruits from thechokeberries growing in Bulgaria Antova [2] andZlatanov [24] have been identified leic (214)linoleic (711) and palmitic acid (51) In thelipid fraction incorporates phospholipids (28)mainly phosphatidylinositol (298) sterols(12) mainly -sitosterol (898) andtocopherols (555 mgkg) mainly -tocopherol(706) In our previous paper we established inthe lipid fraction from the waste the many acidsbut in different quality - linoleic (397) oleic(307) and palmitic (198) acids The maincomponents in the tocopherol fraction were -tocopherol (597) and -sitosterol (826) andcampesterol (64) ndash in the sterol fraction [1]

In the lipid fraction investigated by Anwar etal [3] the prevailing fatty components werelinoleic (279) oleic (6190) and palmiticacids (32) as well as sterols mainly -sitosterol (660) 5-avenasterol (180) andcampesterol (50) Currently in many countries medical plants areprocessed by extraction with liquefied gases

2 air freon 134a and other) The producedextracts are harmless thatrsquos why they can bewidely used in food and flavour industrycosmetics and medicine The usage of liquefiedgases overcomes the drawbacks of installationsworking with volatile polar and apolar solvents[13] The aim of present study is producing of wasteof chokeberry by using freon 134a and mix offreon 134a and acetone in laboratory installationand determination of their chemical compositionand characteristics for possible application innatural food product

2 Materials and Methods

Samples The plant material was aronia fruitwaste which was dried at room temperature The moisture content (79) was determinedby drying up to constant weight at 105o

(Russian Pharmacopoeia 1990) The used solvent 1112 tetrafluorethane (freon134a) is apolar with relative permittivity at 20o

and 100 kHz 1013 dipole moment 2060 Debayand polarization 138 cm3mol Its dynamicviscosity and surface tension at 20o are small198 as and 8 mNm respectively whichallows easier penetration into plant cells and


24







25









123

123








26





not detected






not detected

Conclusion



References



27






























24







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123

123








26





not detected






not detected

Conclusion



References



27






























25









123

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26





not detected






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Conclusion



References



27






























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not detected






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Conclusion



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27






























27





























LOW TEMPERATURE EXTRACTION OF PLANTS BY LIQUIFICATE GASES. 10. WASTE OF CHOKEBERRY FRUITS (Aronia melanocarpa (Michx) Elliott.)

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