Đề tài anthocyanin và những nguyên liệu chứa anthocyanin

Truong Dai hoc Ky Thuat Cong NgM TP.HCM Khoa Cong nghe Thvc Ph~m BO MON Cong Nghe Thuc Ph~m DO ANMON HOC • Cong Ngh~ Thvc Ph~m (MSMH: 20471014) DE TAl: ANTHOCY ANIN vA NHONG NGUYEN LIBu CHUA ANTHOCYANIN GVHD: Co Tdn Thj Cue Phuong SVTH: 1. Huynh Thj Thanh Tuy~n_0851100344 2. Tr~n Thi Xuan Hong , 0851100091 Nganh: Cong Nghe Thuc Ph~m Narn hoc: 2011 Mt}clt}c I. T6ng quan v~ Anthocyanin 3 1. Gioi thieu 4 2. C~u true hoa h9C cua Anthocyanin 4 3. Tinh chat cua Anthocyanin 5 Cong dung 7 4. Su phan b6 cua Anthocyanin 8 5. Qua trinh sinh tong hop Anthocyanin 8 5.1. Con duong sinh t6ng hop Anthocyanin trong t~ bao thuc v~t 9 5.2. Cac y~u t6 anh huong d€n qua trinh sinh t6ng hop 9 6. Sir chuyen d6i C&U true cua Anthocyanin trong mo i truong long 9 7. MQt s6 y~u t6 anh huang d€n dQ mau cua Anthocyanin 11 7.1 . C~u true 12 7.1.1 C~u true chuyen hoa 13 7.1.2. C~u true hoa h9C 14 7.2. PH 15 7.3. Nhiet dQ 16 7.4. Oxy 17 7.5. Enzyme 19 7.6. Anh sang 20 7.7. Duong va cac san pharn bien tinh cua chung 22 7.8. Cac ion kim loai 25 7.9. Sunphurdioxide/So-) 28 7.10. Acid Ascorbic 30 8. Kha nang irng dung va cac nguon nguyen lieu trong san xuat Anthocyanin hien nay .. 34 9. Vai tro cua cac hop chat Anthocyanin 37 9.1. D6i voi thuc v~t 40 9.2. D6i voi sire khoe con nguoi 12 9.2.1. Heat tinh chong oxy hoa 13 9.2.2. Heat tinh chong ung thu 14 9.2.3. Heat tinh chong cac benh tim mach 15 K~t luan 16 II. Cac nguyen li~u co chua Anthocyanin 17 1. Blip cai tim 19 1.1. T6ng quan 20 1.2. Thanh ph~n hoa h9C 22 Gioi thieu v~ phuong phap trich ly 25 Cac y~u t6 anh huang d~n qua. trlnh trich ly 28 1.3. Quy trinh cong nghe chiet tach Anthocyanin tir blip cai tim 30 Ti~n hanh thi nghiem 14 Phuong phap thi nghiem 15 Muc dich thi nghiem 16 Chu§n bi thi nghiem 17 B6 tri thi nghiem Phuong phap nghien cuu Ket qua nghien CUD 0 ••••••••••••••••••• 0 ••••••••••••••••••• 19 20 22 1.4. K~t luan 25 2. Gao nep than 19 2.1. Tcng quan 20 2.2. Thanh phan hoa h9C 22 2.3. Qua trinh va tinh che Anthocyanin tt'l' gao nep than 30 2.4. Nhan xet va k~t luan 15 3. Dau tay 19 3.1. T6ng quan 20 3.2. Thanh phan hoa h9C 22 3.3. Phuong phap nghien ciru 30 3.3.1. Nguyen lieu 14 3.3.2. Phuong phap 15 3.3.3. Ti~n hanh thi nghiem 16 3.3.4. K~t qua nghien ciru v~ cac di;ictinh cua Anthocyanin 17 Nhan xet 16 3.4. K~t luan 17 III. Cac ch~t Anthocyanin thuong mai 17 IV. Phuong phap nghien ciru, bao v~ mau do cua Anthocyanin 19 l.Phuong phap nghien ciru 20 So d6 nghien ciru 22 2. Bao v~ man do cua Anthocyanin 22 Ml}c Ll}c_Bang: Bang l.Cac Anthocyanin trong mot s6 nguon thuc vat.. Bang 2. Thanh phan hoa h9C cua bap cai Bang 3. Anh huang cua dung moi den ham luong va do mau cua Anthocyanin 17 19 20 Bang 4. Ty i~ dung moi va bam hrong Anthocyanin 17 Bang 5. Thanh phan hoa h9C cua gao nep than 19 Bang 6. Thanh phan dinh duong cua qua dau tay 20 Bang 7. K~t qua ham hrong Anthocyanin thea dO chin cua qua dau 17 Bang 8. Ph§n tram man con lai thea thai gian a 9Soc tai cac PH khac nhau cua Anthocyanin tho chi~t trong dung moi mroc Sulfite va etanol/Hjf) 19 Ml}.CLl}.c_Hinh: Hinh 1. cAu true cua mot s6 Anthocyanin tv nhien 17 Hinh 2. Con duong sinh t6ng hop Anthocyanin 19 Hinh 3. SI! chuyen hoa cAutrue cua phan tIl' Anthocyanin trong moi truong long 20 Hinh 4. cAu true chuyen hoa cua Anthocyanin trong nuoc 17 Hinh S. Anh huang cua PH len t6c do thuy phan cua qua dau duoc dun nong tai 4SoC trong truong hQP co Oxygen hoac Nitrogen 19 Hlnh 6. Sir bien tinh cua Anthocyanin 3,S_diglucoside tai PH 3,7 20 Hinh 7. Sir bi~n tinh cua Anthocyanin voi phan ung oxy h6a Catechol. 20 Hinh 8. Phan irng ngung tu cua a) Cyanidin va furfural va b) cyanidin ketobase va furfural 17 Hinh 9. So d6 Jurd d6i voi phan irng thuan nghich giira S02 va Anthocyanin 19 Hinh 10. Sir chuyen hoa Malvin thanh Malvone boi H202 tao thanh S\I' oxy hoa vitamin C 20 Hinh 11. Fla-2-ene duoc tao thanh nr phan ling gitta vitamin C va A 17 Hinh 12. Sir tao phirc gitra Cyanidin va DNA 19 Hinh 13. B~p cai tim 20 Hinh 14. Cong tlnrc cAutao cua Cyanidin 3,S_diglucoside 20 Hlnh IS. Qua trinh trich Iy mot doan 20 Hinh 16. Qua trinh trich ba dean giao dong 17 Hlnh 17. Qua trinh trich nhieu doan nghich dong 19 Hinh 18. So d6 quy trinh cong nghe chiet tach Anthocyanin tir b~p cai tim 20 Hinh 19. 1)6 thi bi~u di~n anh huang cua cac h~ dung mo i. 20 Hinh 20. Anh huang cua ty I~dung moi (Etanol-mroc ) dSn ham IUQ'ngAnthocyanin 19 Hinh 21. Hat nSp than 20 Hinh 22. Qua dau tay 20 Hinh 23. KSt qua ham luong Anthocyanin a cac nong d9 S02 khac nhau 20 Hinh 24. KSt qua ham luong Anthocyanin a cac nhiet d9 khac nhau 20 I. Tang quan v~ Anthocyanin: 1. Gioi tbi~u: Cac Anthocyanin hien thuoc nh6m cac chat man tu nhien tan trong mroc Ian nhat trong th€ gioi thuc vat. Thuat ngii' anthocyanin b~t nguon nr tieng Hy Lap, trong do Anthocyanin la S\T ket h9]J giira Anthos - nghia la hoa va Kysanesos - nghia la mau xanh. Tuy nhien, khong chi co mau xanh, anthocyanin con mang d€n cho thuc v~t nhieu mau s~c me ro' khac nhir hong, do, cam va cac gam mau trung gian. Cac anthocyanin anthocyanidin hay aglycon. M6i anthocyanidin khi mAt h€t nhom dirong duoc goi la co th~ bi glycosyl h6a acylate boi cac loai duong va cac acid khac tai cac vi tri khac nhau. Vi th€ hrong anthocyanin anthocyanidin 2. nr Ian han 15-20 IAn. C§u true boa hoc cua Anthocyanin: Anthocyanin thuoc nhom cac h9]J chitjlavonoid, co kha nang hoa tan trong mroc va chua trong cac khong bao, V~ ban chit, cac Anthocyanin la nhtrng hop chit glycoside cua cac d~n xuit polyhydroxy va polymethoxy cua 2-phenylbenzopyrylium hoac mu6i flavylium. Cho d€n nay, nguoi ta da xac dinh duoc 18 loai aglycon khac nhau, trong do 6 Joai ph6 bien nhit Ia pelargonidin, cyanidin, delphinidin, peonidin, petunidin Trong W nhien, Anthocyanin rAt hiem khi hydroxy W do a vi tri C-3 va maldivin. a trang thai tu do (khong bi glycosyl h6a). Nhom lam cho phan tir anthocyanidin tro nen khoug 6n dinh va lam giarn kha nang hoa tan cua no so voi anthocyanin tuong (mg. Vi vay, su glycosyl h6a luon di6n ra, d~u tien 0 vi tri nhorn 3-hydroxy. Neu co them mot phan nr duong sua, vi tri ti€p thea bi glycosyl h6a thuong g~p nhit la a C-5. Ngoai ra, su glycosyl h6a con co th~ g~p a cac vi tri C- 7, C-3', C-5 '. Loai duong ph6 bien nhit la glucose, ngoai ra ciing co mot vai loai monosaccharide (nhu galactose, rammose, arabinose), cac loai disaccharide (chu y€u la rutinose, sambubiose hay sophorose) hoac trisaccharide tham gia vao qua. trinh glycosyl h6a. Su methoxyl h6a cac anthocyanin va cac glucoside tuong zing di6n ra thong thuoug nhat Ia a vi tri C-3' va C-5', ciing co the g~p a vi tri C-7 va C-5. Tuy nhien, cho d€n nay, nguoi ta v~n chua tim thay mot hop chit nao bi glycosyl h6a hay bi methoxyl h6a tren tit cit cac vi tri C-3, 5, -7 va -4' do dn thiet phai con it nhAt mot nhom hydroxyl tu do a C-5, -7 bay -4' d~ hinh thanh dang du true quinonoidal base (dang du true cua anthocyanin tlnrong t6n tai trong khong bao thuc v~t co pH ill 2,5 -7,5). ORs Hinh 1: C6.u true cua mot s6 Anthocyanidin tu nhien. Cac Anthocyanin tuong ung luon dutrc glycosyl hoa 0' nhom hydroxy C-3 Su acyl hoa ciing co th~ xay ra 6 vi tri C-3 cua phan nr duong hay ester hoa 6 nhorn hydroxy C-6. Cac nhom acyl hoa ehinh la cac phenolic acid nhu p-coumeric, caffeic, ferulic hay sinapic acid va mot loat cac acid nhu acetic, malic, malonic, axalic va succinic. 3. Tinh chit cua Anthocyanin: Anthocyanin tinh khidt 0' dang tinh th~ hoac vo dinh hinh la hO'P eh~t kha phan cue nen tan t6t trong dung moi phan eve. Anthocyanin hoa tan t6t trong mroc va trong dich bao hoa. Khi k€t hop voi duong la cho phan nr anthocyanin hoa tan han. Mau s~c anthocyanin thay d6i phu thuoc vao nhiet d(j, cac ch~t mau va nhi~u y€u t6 khac, ... Khi tang s6 luong nhorn OH trong vong benzene thi mau cang xanh d~m. Mire d(j methyl hoa cac nhorn OH 6 vong benzene cang eao thi mau cang do. Neu nhorn OH 6 vi tri thir ba k€t hop voi cac g6e duong thi mau s~e ding se thay d6i thea s6 luong cac g6e duong duoc dinh vao nhi~u hay it. Cac Anthocyanin ciing phu thuoc nit manh vao pH cua moi tnrong: )io> Khi pH> 7 cac Anthocyanin co mau xanh va khi pH < 7 cac Anthocyanin co mau do. )io> o pH = 1 cac Anthocyanin thuong a dang mu6i oxonium man cam d~n do. )io> o pH = 4+5 chung co th~ chuyen v~ dang baza Cacbinol hay baza Chalcon khong mau. )io> o pH = 7+8 lai v~ dang bazo Quinoidal Anhydro mau xanh. Mau sitc cua anthocyanin can co the thay d6i do hfrp thu a tren polysaccharide. Khi dun nong lau dai cac anthocyanin co the pha huy va mfrt mau, Anthocyanin co buoc song hfrp thu trong mien nhin thay, kha nang hfrp thu cue dai tai buoc song 510+540nm. D:1:. hung tinh (helltim) o - .~. Qulnold.11>:1:•• nlon(hellxanh) Hinh 3: S1!-'chuyen hoa cdu true cua phdn tu'Anthocyanin trong moi truong long Dang quinonoidal base duoc tao thanh thong thuong b~n (y (y pH> 3, Cationflavylium tuong d6i moi tnrong acid (rum pH < 3), Trong moi truong trung tinb hay aeid y~u, Anthocyanin t6n tai chu y~u (y cac dang khong mau, Vi vay, S\I' 6n dinh ella cac dang mang mau, d~e biet lit dang quinonoidal base, mot phan lit do su xuat hien cua cac g6c acid g~n vito phan tll' duong, hay eon goi la su co-pigment hoa noi phan ill', So' di su co-pigment hoa noi phan ill' giup tang do b~n ella phan nr anthocyanin la do 5\{ x~p chong ella vong thorn trong g6c aeid voi vong pyrylium ella phan nr anthocyanin, giam bot kha nang bi cac phan tll' nuoc tAn cong hinh thanh cac hop ch~t carbinol, chalcone khong mau. Ngoai ra, hieu irng copiment hoa ngoai phan tir ciing eo th~ giup lam ben cac du true quinoidal base hay ionflavylium do 5\1' eo mat ella nhirng plnrc eh~t khong mau, nhu cac flavone, flavonol, Trong qua trinh xu' Iy nhiet va anh sang, qua trinh copigment hoa ngoai phdn ttc gilt vai tro quan trong, ngan chan su mit man co th~ xay ra. Co ch~ phan huy do nhiet noi chung do sir chuyen hoa thuan nghich cua cac ionjlavylium thanh cac dang chalcone. Chalcone lit chat trung gian trong ca hai qua trinh phan huy do nhiet va anh sang cua anthocyanin. Theo cac nghien ciru g~n day (Davis va cting su; 1983; Mazza va Brouillard,1990) hieu irng co-pigment hoa chinh la CO" che lam b~n mau chinh cua anthocyanin trong thuc vat do no lam giam ti l~ gitla dang kh6ng mang mau (chalcone) va dang mang mau thong qua viec tao plnrc voi dang mang mau, ngan chan qua trinh chuyen hoa nr dang mang mau sang dang khong mang mau. Trong dung dich, Anthocyanin dang kh6ng acyl hoa hay acyl hoa mot dan vi dong vai tro nhu m(>tch~t chi thi pH, t6n tai o' dang acid hay base tuy thuoc vao gia tri pH. d pH acid (pH < 3), dung dich anthocyanin cho man do darn, Khi pH tang, mau s~c cua dung dich se nhat d~n va chuyen sang kh6ng mau va cuo: cung co mau tim hay xanh duong 6 pH cao (pH> 6). 7. M9t s~ y~u t~ anh huong d~n d9 mau cua Anthocyanin: So voi da s6 cac chat mau thien nhien, anthocyanin la ch~t mau co d(>ben kern hon, no chi thS hien tinh ben trong moi truong acid, Ngoai ra, no co th~ phan My tao thanh dang kh6ng mau va san pham cu6i cung cua S\I'phan huy co dang mau nan cong voi nhirng san pharn kh6ng tan. • S\I'phan My anthocyanin co thS xay ra trong qua trlnh trich va tinh ch~ chung, d6ng thai su phan huy nay con xay ra trong qua trinh xu • ly va bao quan cac san pharn thuc pharn, D(> b~n cua cac anthocyanin phu thuoc vao nhieu y~u t6 nhu: cfru true hoa h9C cua anthocyanin, pH, nhiet d(>, su co m?t cua copigment, ion kim loai, oxy, acid ascorbic, S02, anh sang, enzyme, duong va cac san pham bi~n tinh cua chung. 7.1. ciu true: 7.1.1.CAu true chuyen hoa: Trong m6i tnrong mroc cac anthocyanin tu nhien gieng nhu ch~t chi thi pH. Do 6 pH thap, do xanh 6 pH trung gian, va khong man 6 pH cao. Tai pH da cho, t6n tai mot din b~ng gitta 4 cfru true cua anthocyanin va aglycone: quinonoidal (anhydro) bazo (A) mau xanh, cation flavylium (AH+) mau do, carbinolpseudo base (B), va chalcone (C) kh6ng mau. R ~ OH -----=:.. R OH AH A 1~ OH OH -----=:.. ...,....-- + R OH OGI OH OH B C Hinh 4: Cdu true chuyen hOGcua anthocyanin trong nutrc Khi pH < 2,0, cac anthocyanin t6n tai chu y~u dang cation flavylium mau do (Rr=Osdtrcng] hoac mau vang (R3=H), Khi pH tang, sir m§t proton xay ra nhanh thanh dang quinononidal mau xanh duong hoac mau do, Dang quinonoidal thirong t6n tai nhu mot h6n hQP vi pKa cua nhom OH a vi tri 4',7 va 5 la tuong tu. Khi d€ yen, su chuyen hoa xay ra : co su tach mroc cua cation flavylium cho ra dang pseudo base (hemiacetal), Dang nay din bang voi dang chalcone vong Hai dang Cis (CE) va trans-chalcone-(Cz) phan img rna rna khong mau, duoc tao thanh ur carbinol pseudo base boi vong nhanh va isomer hoa cham. Cit 2 chalcone nay khac nhau so vci chalcone binh thuong la boi chung co nhorn chirc carbonyl 0' canh vong B, trong do chalcone binh thuong co nhorn carbonyl i. ciu o' k~ can vong A. true hoa hec: £)9 b~n mau va ctrong d9 rnau cua cac anthocyanin phu thuoc vao vi tri va s6 hrong cua cac uhorn hydroxyl, methoxyl, duong va cac duong duoc acyl hoa. Khi s6 nhorn hydroxyl trong vong B tang.cue dai h§p thu a vung th§y duoc dich chuyen v~ phia co buoc song dai hon va mau thay d6i tir cam d~n xanh duong. Vi du: Buoc song h§p thu cue dai trong dung dich HCL 0,01 % MeOH d6i voi: • pelgonidin: A,max = 520 nrn (cam) • cyaniding: Amax • dephinidin: Amax = 535 run (do cam) = 545 nrn (do xanh) Khi nhorn methoxyl thay th~ nhorn hydroxyl thi ta thu duoc k~t qua nguoc lai. Nhorn hydroxyl tai vi tri C-3 co y nghia quan trong vi dung dich Anthocyanin chuyen tir mau cam vang d~n mau do, f)i~u nay giai thich sir khac nhau giita Anthocyanin co mau do trong khi do 3 deoxyanthocyanin : apigenidin, luteolinidin va tricetinidin co mau vang nhung 3 - deoxyanthocyanidin b~n hon cac anthocyanidin khac (Mazza va Brouillard, 1987; Iacobucci va Sueny, 1983). Su co m~t cua nhorn hydroxyl tai vi tri C-5 va nhom th~ co mau thong qua S\1" ngan a vi tri C-4, ca 2 bien hoa dang can cac phan ung hydrat hoa d~n d~n su tao thanh dang khong man. Khi rmrc do hydroxyl hoa cac aglycone tang, tinh ben cua cac anthocyanin se giam. Tuy nhien khi tang S\I" methoxyl hoa, se thu duoc k~t qua nguoc lai. Vi du: su co m~t cua nhom OH 6 vi tri 4' va 7 trong phan tu lam ben h6a dang k~ cac pigment, trong khi do, S\T methoxyl h6a co nhom hydroxyl nay lam giam do ben. Cac anthocyanin duoc glycosyl hoa va acyl hoa se cho dang mau xanh, Su glycol hoa nhfrng nhorn OH 1\1" do lam tang tinh ben cua anthocyanin. Vi vay, cac diglucoside b~n hon cac rnonogluside cua cung mot anthocyanin. Anthocnin co chua 2 hay nhieu nhorn acyl (nhu ternatin, platyconin, cmerarnn, gntiodenphin va zebrrinin) la ben trong moi truong trung tinh boac acid y~u do lien k~t hydro gina cac nhorn hydroxyl cua cac nhan phenolic trong anthocyanin va acid vong thorn. Brouillard (1981-1982) va Goto cung voi cong vien (1982-1983) khao sat rang cac anthocnin diacylate hoa duoc b~n hoa boi sir lien k~t ch~t kieu sandwich nho S1,l' tuong tac giua vong anthocyanin va 2 nhom acyl vong thorn. 7.2. PH: Trong moi tnrong mroc, pH co anh huong dang k~ len mau sac cua anthocyanin (Brouillard 1984,Mzza va Brouillard 1987). C~u true, dO ben mau, mau s~c cua anthocyanin thay d6i theo su thay d6i cua ph. S\1"thay d6i du true cua anthocyanin khi pH thay d6i da: duoc d~ cap trong phan du true chuyen hoa. Nam 1956 Lukton nghien ciru thfry rang: a ph = 2 - 4.5 thi van toe phan huy pelargonidin 3- glucoside trong dich ep cua qua dau khong xac dinh duoc khi khong co O2. Khi co mat O2 ph tang nhanh, su bien tinh tang len manh liet, +~ 5 ....E PH=3,5 02 200 PH=4.0 400 600 800 KOIO -4 hour-I Hinh 5: Anh huang cua pH len t6c do thuy phan cua qua dau duoc dun nong tai 45°C trong truong hop co Oxygen hoac Nitrogen D~n narn 1972, Adams nghien ciru thfry rang 0- ph = 2 - 4 it anh huong d~n su phan My cua Anthocyanin trong suot qua trinh gia nhiet khi khong co O2. Khi co mat O2 thi Anthocyanin bi bi~n tinh rnanh 0- ph tren du true cua chung cung voi dO b~n mau thay d6i voi sir thay d6i ph. 7.3. Nhi~t de.>: Trong qua trinh ch~ bien va bao quan thuc pham, Anthocyanin d~ dang bi bien tinh duoi tac dung cua nhiet do (Markakis 1974). OH'(JO:::::-O /. OGI OGr OR / I R -.Q, OGI OR Hinh 6: S\f bien tinh cua Anthocyanin 3,5 - di Glucoside tai pH 3,7 Khi mot du true cua anthocyanin ben voi su gia tang cua pH thi no ciing b6n voi su gia tang cua nhiet dQ. S1,l· hydroxyl hoa cac aglycone lam giam tinh b6n cua anthocyanin, trong khi sir methxyl hoa, glycosyl hoa, acyl hoa se cho k~t qua nguoc lai, Vi du: Anthocyanin 3- glycoside co dQ b6n nhiet Ian nhl1t tai pH = 1.8 - 2.0 voi sir co m~t cua oxy trong khi anthocyanidin 3,5-diglycoside co dQ b6n nhiet Ian nhat tai pH = 4.0 - 5.0 Co ch~ cua su phan huy nhiet xay ra khong nhfrng phu thuoc vao nhiet dQ rna con phu thuoc vao ban than anthocyanin do. Drazdina cho r~ng coumarin glycoside la san pham phan huy phi> bi~n cua anthocyanin 3,5 - diglycoside va co ch~ cua S1,l· phan My la: d~u tien, cation flavylium chuyen thanh dang quinonidal base, tiep theo la hinh thanh nhieu dang san pham trung gian va cu6i cung thu duoc dftn xuit coumarin va thanh phan tuong ung voi vong B cua anthocyanidin, S\I' bien tinh nay khong nhung duoc xuc ti~n boi nhiet dQ rna con bi anh huang boi oxy. Anthocyanidin 3 - glycoside thi khong hinh thanh nhtrng dftn xuit coumrin rna buoc d~u tien cua sau do la glycoside. S1,l· S1,l· phan huy bao gem sir chuyen hoa cua dang carbinol pseudobase khong mau, rna vong pyrylium d€ hinh thanh dang chalcone tnroc khi thuy phan lien k~t Adams cho rang, cac anthocyanidin 3-glycoside khi duoc nung nong 6 pH = 2.0 - 4.0 dAu tien se bi thuy phan lien kSt glycoside (6 100°C) sau do la su biSn d6i aglycone thanh chalcone. Sir biSn tinh han nira d§n dSn hinh thanh dang san pham mau nau d~c biet ill voi su co m~t cua oxy. Khi dun nong lau dai cac anthocyanin co th~ bi phan huy va m~t mau, d~c biet la cac anthocyanin cua day tay, anh dao, cu cai. Nguoc lai cac anthocyanin cua phuc ben nr den cling trong di~u kien do lai khong bi thay d6i.Nhin chung khi gia nhiet, cac ch~t man do d~ dang bi phan huy, con cac ch~t mau vang thi kho hun. 7.4.0xy: Oxy va nhiet do duoc xem la nhfrng tac nhan d~c trung XlIC tien su phan huy cua anthocyanin, do do rna sinh ra nhirng dang san pham khong mau hoac man nau. Chinh SI,1' oxy hoa true tiSp dang carbinol pseudobase da gay ra sir ket tua va dong vang trong mroc trai cay. Trong qua trinh dun nong cac anthocyanin bi oxy hoa manh li~t. Oxy va nhiet do la nhirng tac nhan xuc tien d~c biet nh~t trong mroc ep cua blueberry, cherry (anh dao), currant, nho, raspberry va dau. LUQ'nganthocyanin con lai cua dau se 16n hun khi dong chai duoi di~u kien chan khong hoac nitrogen (Baravingas va Cain, 1965). DO b~n cua cac pigment cua nho con duoc slf dung nhir la ch~t rnau 6 nuoc giai khat duoc tang len khi dong hOp voi nitrogen. 7.5. Enzyme Nhi~u enzyme noi sinh trong tS bao cua cay co kha nang lam m~t mau anthocyanin. Nhirng enzyme nay duoc goi chung la anthocyanase. Dua vao dac tinh cua cac enzyme rna nguci ta phan lam 2 nhom; Glycosidase va polyphenol oxidase (PPO). Cac enzyme nay thu duoc ttl' n~m (fugal). • Glycosidase: la enzyme thuy phan lien kSt glycoside cua anthocyanin tao ra duong nr do va aglycone nay kern b~n han r~t nhieu va m~t mall r~t nhanh khi co mat cua catecol(Huang 1955). • Polyphenol oxidase (PPO): tac dung len anthocyanin voi su co mat cua O-diphenol thong qua co ch~ oxy hoa ket hQP. Theo Gromeck va Markakis, su them vao glycosidase va PPO xuc tac cho qua trinh peroxide hoa phan huy anthocyanin. Minh 7: S1,1' bi~n tinh anthocyanin voi phan ling oxy hoa catechol Peng va Markakis da: d~ nghi mot co' ch~ rna trong do O-quinine duoc tao thanh boi su oxy hoa anthocyanin. Blom phan l?p duoc enzyme anthocyanin-glycosidase nr Aspergillus Niger va chi ra anh huang cua no trong qua trinh thuy phan lien k~t glycoside cua anthocyanin. PPO la enzyme co iinh huang y~u len anthocyanin dang quinonoidal base d~ bi thuy phan boi PPO han dang cation flavylium. T6c d9 phan huy boi PPO phu thuoc vao str thay th~ mo hlnh cua vong B va rmrc d9 glycosyl boa. 7.6. Anh sang Cac anthocyanin thuong khong ben khi ti~p xuc voi tia ttl' ngoai, anh sang th~y dUQ'C,va cac nguon phap xa khac, Anh sang co 2 anh huang d~n anthocyanin: • Tang cuong cho qua trinh sinh t6ng hop. • Xuc ti~n su bien tinh cua chung. Narn 1964, Siegenrnan eho r~ng nhttng qua tao giong do se chuyen sang rnau xanh khi d~ trong b6ng t6i. Nam 1968 Vanburen va cac cong S1,1' tuong trinh r~ng cac diglueoside duoc acyl hoa va methyl hoa thi cac Anthocyanin trong nrou ben nheit khi d€ ngoai anh sang, diglucoside khong bi acyl hoa la it ben han va monoglucoside cac la kern ben nhat, nam 1975 Palamidis va Markakis d1:ltim thay r~ng anh sang thuc dAy qua trinh phan huy anthocyanin trong mroc giai khat co CO2 duoc phdi mau voi anthocyanin tir xac nho. Macccarone va cong S\T d1:l nghien cuu sir quang hoa cua anthocyanin va chi ra rang , anthocyanin diglycosyl hoa tai vi tri C3 va C-5 la ben han cac anthocyanin mono glycoyl hoa tai vi tri C-3 d6ng thai chung ben han so voi cac aglycone nrong irng, 7.7. Dlfb'ng 6 n6ng va cac san phiim bi~n tinh ella chang: de>100 ppm, duong va cac san pharn phan huy cua chung co tac dung thuc dAy su phan huy cac anthocyanin, trong do fructose, arabinose, lactose va sorbose co kha nang phan huy anthocyanin manh han glucose, sucrose, va maltose. T6c de>phan huy cua anthocyanin lien quan d@nt6c de> phan huy cua duong, Cac san pharn phan huy cua duong g6m co: furfura 1,5hydroxymethyl furfural va acetaldehyde thu duoc ttl" phan irng Mailard hoac til"S\!' oxy hoa cua acid ascorbic, poJyuronic boac a ban than cac anthocyanin. Nhirng san pham phan huy nay d~ dang ngung tu voi cac anthocyanin hinh thanh nhirng hop chfrt phirc tap co man nau s~m. S\!' phan huy anthocyanin voi SI,!" co m~t cua furfural va HMF true ti~p phu thuoc vao nhiet de>va duoc thay ro nhfrt la trong mroc trai cay S\I' co m~t cua no lam tang them hieu qua phan huy cua teitca cac loai duong va cac d~n xufrt cua chung. OH OH + OH 11110 OH OH OH '-oAc~ ~ .....-: ........ H OH Jk_Q Hinh 8: Phan ling ngung tu cua a) Cyanidin va furfural b) Cyanidin keto base va furfural 7.8. Cac ion kim loai: Mot s6 ion kim loai da hoa tri co th~ nrong UtC voi cac anthocyanin co nhorn OH (; vi tri ortho gay ra hieu irng sau mau (bathocromic). Hien tuong nay xay ra khi kim loai ti~p xuc vo i anthocyanin trong qua trlnh che bi~n rau qua hoac S\1' cho them cac mudi kim loai vao trong thuc pham. Sistrunk va Cash dii chung minh dng co th~ ben hoa mau cua dich trich dau bang each them vao do muoi thiec. Francis (1977) cong b6 rang, cac ion Ca, Fe, Al tao them su bao v~ cho anthocyanin cua nuoc ep trai man viet qu~t (cranberry), nhung su bien d6i mau xay ra la do S\1' tao plnrc giita ion kim loai va tannin, k~t qua sau cung la khong co 19'i. Trong cong nghiep d6 hop, sir mAt mau cua nhirng trai cay co chua anthocyanin la do co phan ling voi thiec cua d6 hOp (Culpepper va caldwell, 1972). Trang phan ung voi thiec, anthocyanin dong vai tro nhu cb~t khir cue catod boac anod. Ch~t klnr cue catod co th~ bi khu boi hydro moi sinh til" phan irng gi"(1'akim loai va acid, con ch~t khu CI!C anod thuong la cac anthocyanin co it nh~t 2 nhorn hydroxylo vi tri ortho. 7.9. Sunphurdioxide (S02) Cac anthocyanin thuong bi m~t mall khi co mat cua S02. Hien tuong nay thuong xay ra trong qua trinh xu' Iy cac san pham thuc pham co chira anthocyanin bang S02. Qua trinh khir nay co th~ la thuan nghich hoac b~t thuan nghich, Trai cay va mroc qua duoc xir Iy bang mot IUQ'ng trung blnh S02 (500 - 2000 ppm), lam m~t rnau cac anthocynin cua chung tnroc khi ch~ bien, han nira, chung duoc desunfit hoa va mau anthocyanin duoc phuc h6i. Jurd dii d~ nghi sa d6 phan ung thuan nghich gina S02 va anthocyanin trong do, dang co mau flavylium phan irng voi ion bisulphate tao thanh chrornene-Z{hoac 4)- sulphonic acid. Hinh 9: Sa d6 Jurd d6i voi phan irng thuan nghich giu'a S02 va anthocyanin S02 (y nong d9 r~t th~p (khoang 30 ppm) co th~ ire che su bien tinh do enzyme cua anthocyanin trong qua anh dao nhung khong lam mit mau chung (Goodman va Markakis, 1965 ). Su dy mau b~t thuan nghich xay ra trong qua trlnh t~y qua. voi luong Ian S02 (0.8 - 1.5%) va soda (004 1.0%) duoc dung trong qua trlnh t~y qua. Phan ung b~t thuan nghich nay chua duoc biet hoan toano 7.10. Acid ascorbic Nbi~u nha khao sat (Beatie va cong 1953; Markakis va cong sir sir 1943; Pederson va cong su 1947; Kertesz 1952; Mescbter 1957; Starr va Francis 1968) quan sat S\T bi6n mit d6ng thoi cua acid ascorbic va anthocyanin trong nuoc trai cay t6n trti: va d€ nghi mot tuong tac co th~ co gitta 2 hop ch~t nay. Acid ascorbic hien dien trong h am cua nguyen lieu, Nhiet dO co tac dung tang t6c d(> khuech tan vii giarn do nhot, phan nr chat hoa tan chuyen dong d6 dang khi khuech tan giifa cac phan tir dung moi, tuy nhien nhiet dO Iii yeu t6 gioi han, vi khi nhiet d(> cao qua co th~ xay ra cac phan ung khac khong cAn thiet gay kh6 khan cho cac qua trinh cdng nghe ... Thai. gian trich Iy cling anh huang d6n hieu sufrt trfch Iy. Khi thai gian tang len IUQ'ngchfrt khuech tan tang nhung thai gian phai co gioi han, khi da: dat duoc rmrc do trich ly cao nhfrt neu keo dai thai gian se khong mang lai hieu qua kinh t6. 1.3.Quy trinh cong ngh~ chi~t tach Anthocyanin to: b~p cai tim: Nguyen li~u ~ Rea sach bfug mroc, de rao ~ Can (20g/g6i) ~ Nghien nho ~ Xuly ~ N gam trong dung m6i ~ Trich lv ~ Dich trich . ~ Phan tich Hinh 18: Sa db quy trinh cong nghe chi~t tach Anthocyanin ttl' b~p cai tim. Tj~n hanh thi nghiem: • Nguyen lieu - H6a chftt sir dung: B~p cai tim Ethanol HCll% KCI CH3COONa • Dung cu - thi~t bi: May do quang pho pHk~ Tu lanh dong Thi~t bi sfty chan khong Phuong phap thi nghiem: Thi nghiem 1: Nghien ciru anh huong cua nhiet dO b6c ~m va do ~m nguyen lieu d~n kha nang chiet tach va do ben man anthocyanin M1}C dich thi nghiern: Tim ra nhiet dO b6c ~m va do ~m thfch hop d~ cho hieu su§t trich ly cao nhftt va do b€n mau nhftt. Chu§n b] thi nghi~m: Nguyen lieu b~p cai tim duoc nra sach Dung moi Ethanol- mroc - HCl (voi 1'y l~ 1:1 va I%HCl) Be} tri thi nghiem: Thi nghiem duoc b6 tri hoan toan ng~u nhien voi 2 nhan t6 va 3 l~n l~p lai. Nhan t6 A la nhiet dhlip thu cue dai cua anthocyanin 6' buoc song 515 nm va 420 nm va ap dung cong thirc (2.2), ta tinh duoc de:>mau cua anthocyanin co duoc nr cac dung moi khac nhau. K~t qua de:>mau va mau s~c cam nhan duoc bing mat cling duoc th~ hien 6' bang 3 va hinh 19. Bang 3: Anh huang cua dung moi d~n ham IUQ'ngva de:>mall anthocyanin Dung Methanol moi Ethanol - mroc - mroc - Nuoc - hydrochloric - NuO'c - NuO'c - formic acid acetic acid 0,492 0,452 0,107 3,134 3,032 2,825 2,059 do den do den vang do vang do xanh xanh xanh xanh HCl HCl acid Ham Iuong, % 0,949 1,046 De:>mau 3,267 Chi s6 Mau cam nh~n vang bang m~t Moi tnrong kiern nhat xanh do ._S l.2 S ro 1 >-.. (.) 0 ..t:: ...... ~ ~ 0 3.5~.,.... 3 § >-.. 2.5g 2 ~ ro l.5;:::l ,,.... 0.8 0.6 0.4 0.2 1 S 0.5.~ o [...]... dung len anthocyanin voi su co mat cua O-diphenol thong qua co ch~ oxy hoa ket hQP Theo Gromeck va Markakis, su them vao glycosidase va PPO xuc tac cho qua trinh peroxide hoa phan huy anthocyanin Minh 7: S1,1' bi~n tinh anthocyanin voi phan ling oxy hoa catechol Peng va Markakis da: d~ nghi mot co' ch~ rna trong do O-quinine duoc tao thanh boi su oxy hoa anthocyanin Blom phan l?p duoc enzyme anthocyanin- glycosidase... cac Anthocyanin trong nrou ben nheit khi d€ ngoai anh sang, diglucoside khong bi acyl hoa la it ben han va monoglucoside cac la kern ben nhat, nam 1975 Palamidis va Markakis d1:ltim thay r~ng anh sang thuc dAy qua trinh phan huy anthocyanin trong mroc giai khat co CO2 duoc phdi mau voi anthocyanin tir xac nho Macccarone va cong S\T d1:l nghien cuu sir quang hoa cua anthocyanin va chi ra rang , anthocyanin. .. My cua ca hai acid ascorbic va anthocyanin Nhung san pham khong mau, Acid dehyroascorbic ciing c6 th~ lam mat mau anthocyanin nhung tai t6c dQ th~p han acid ascorbic o on -+-_/ phan huy cua duong, Cac san pharn phan huy cua duong g6m co: furfura 1,5hydroxymethyl furfural va acetaldehyde thu duoc ttl" phan irng Mailard hoac til"S\!' oxy hoa cua acid ascorbic, poJyuronic boac a ban than cac anthocyanin Nhirng san pham phan huy nay d~ dang ngung tu voi cac anthocyanin hinh thanh nhirng hop chfrt phirc tap co man nau s~m S\!' phan huy anthocyanin. .. desunfit hoa va mau anthocyanin duoc phuc h6i Jurd dii d~ nghi sa d6 phan ung thuan nghich gina S02 va anthocyanin trong do, dang co mau flavylium phan irng voi ion bisulphate tao thanh chrornene-Z{hoac 4)- sulphonic acid Hinh 9: Sa d6 Jurd d6i voi phan irng thuan nghich giu'a S02 va anthocyanin S02 (y nong d9 r~t th~p (khoang 30 ppm) co th~ ire che su bien tinh do enzyme cua anthocyanin trong qua ... hanh Cy-3-glucoside, 3-galactoside, 3diglucoside va 3-laminarriobioside, Pn-3-g1ucoside Brassica oleraea B~p cai tim (red cabbage) Cy-3-sophoroside-5-g1ucoside cacyl h6a voi malonoyl, p-coumaroyl,... c::~ , , c v ~ " - 'u