Anthocyanin v nhng nguyn liu cha anthocyanin - phn 2 Hnh 3: S chuyn ha cu trc ca phn t Anthocyanin trong mi trng lng

Dng quinonoidal base c to thnh thng thng pH > 3. Cation flavylium tng i bn mi trng acid (nh pH < 3). Trong mi trng trung tnh hay acid yu, Anthocyanin tn ti ch yu cc dng khng mu. V vy, s n nh ca cc dng mang mu, c bit l dng quinonoidal base, mt phn l do s xut hin ca cc gc acid gn vo phn t ng, hay cn gi l s co-pigment ha ni phn t. S d s co-pigment ha ni phn t gip tng bn ca phn t anthocyanin l do s xp chng ca vng thm trong gc acid vi vng pyrylium ca phn t anthocyanin, gim bt kh nng b cc phn t nc tn cng hnh thnh cc hp cht carbinol, chalcone khng mu. Ngoi ra, hiu ng copiment ha ngoi phn t cng c th gip lm bn cc cu trc quinoidal base hay ion flavylium do s c mt ca nhng phc cht khng mu, nh cc flavone, flavonol. Trong qu trnh x l nhit v nh sng, qu trnh copigment ha ngoi phn t gi vai tr quan trng, ngn chn s mt mu c th xy ra. C ch phn hy do nhit ni chung do s chuyn ha thun nghch ca cc ion flavylium thnh cc dng chalcone. Chalcone l cht trung gian trong c hai qu trnh phn hy do nhit v nh sng ca anthocyanin. Theo cc nghin cu gn y (Davis v cng s, 1983; Mazza v Brouillard,1990) hiu ng co pigment ha chnh l c ch lm bn mu chnh ca anthocyanin trong thc vt do n lm gim t l gia dng khng mang mu (chalcone) v dng mang mu thng qua vic to phc vi dng mang mu, ngn chn qu trnh chuyn ha t dng mang mu sang dng khng mang mu. Trong dung dch, Anthocyanin dng khng acyl ha hay acyl ha mt n v ng vai tr nh mt cht ch th pH, tn ti dng acid hay base ty thuc vo gi tr pH. pH acid (pH < 3),

dung dch anthocyanin cho mu m. Khi pH tng, mu sc ca dung dch s nht dn v chuyn sang khng mu v cui cng c mu tm hay xanh dng pH cao (pH > 6). 7. Mt s yu t nh hng n mu ca Anthocyanin: So vi a s cc cht mu thin nhin, anthocyanin l cht mu c bn km hn, n ch th hin tnh bn trong mi trng acid. Ngoi ra, n c th phn hy to thnh dng khng mu v sn phm cui cng ca s phn hy c dng mu nu cng vi nhng sn phm khng tan. S phn hy anthocyanin c th xy ra trong qu trnh trch v tinh ch chng, ng thi s phn hy ny cn xy ra trong qu trnh x l v bo qun cc sn phm thc phm. bn ca cc anthocyanin ph thuc vo nhiu yu t nh: cu trc ha hc ca anthocyanin, pH, nhit , s c mt ca copigment, ion kim loi, oxy, acid ascorbic, SO 2 , nh sng, enzyme, ng v cc sn phm bin tnh ca chng. 7.1. Cu trc: 7.1.1.Cu trc chuyn ha: Trong mi trng nc cc anthocyanin t nhin ging nh cht ch th pH. pH thp, xanh pH trung gian, v khng mu pH cao. Ti pH cho, tn ti mt cn bng gia 4 cu trc ca anthocyanin v aglycone: quinonoidal (anhydro) bazo (A) mu xanh, cation flavylium (AH + ) mu , carbinol pseudobase (B), v chalcone (C) khng mu. Hnh 4: Cu trc chuyn ho ca anthocyanin trong nc Khi pH < 2.0, cc anthocyanin tn ti ch yu dng cation flavylium mu (R 3 =0-ng) hoc mu vng (R 3 =H). Khi pH tng, s mt prton xy ra nhanh thnh dng quinononidal mu

xanh dng hoc mu . Dng quinonoidal thng tn ti nh mt hn hp v pK a ca nhm OH v tr 4,7 v 5 l tng t. Khi yn, s chuyn ha xy ra : c s tch nc ca cation flavylium cho ra dng pseudobase (hemiacetal), Dng ny cn bng vi dng chalcone vng m khng mu. Hai dng Cis (C E ) v transchalcone(C Z ) c to thnh t carbinol pseudobase bi phn ng m vng nhanh v isomer ha chm. C 2 chalcone ny khc nhau so vi chalcone bnh thng l bi chng c nhm chc carbonyl cnh vng B, trong chalcone bnh thng c nhm carbonyl k cn vng A. i. Cu trc ha hc: bn mu v cng mu ca cc anthocyanin ph thuc vo v tr v s lng ca cc nhm hydroxyl, methoxyl, ng v cc ng c acyl ha. Khi s nhm hydroxyl trong vng B tng,cc i hp thu vng thy c dch chuyn v pha c bc sng di hn v mu thay i t cam n xanh dng. V d: Bc sng hp thu cc i trong dung dch HCL 0,01% MeOH i vi: pelgonidin: max = 520 nm (cam) cyaniding: max = 535 nm ( cam) dephinidin: max = 545 nm ( xanh) Khi nhm methoxyl thay th nhm hydroxyl th ta thu c kt qu ngc li. Nhm hydroxyl ti v tr C-3 c ngha quan trng v dung dch Anthocyanin chuyn t mu cam vng n mu ,

iu ny gii thch s khc nhau gia Anthocyanin c mu trong khi 3 deoxyanthocyanin : apigenidin, luteolinidin v tricetinidin c mu vng nhng 3 deoxyanthocyanidin bn hn cc anthocyanidin khc (Mazza v Brouillard, 1987; Iacobucci v Sny, 1983). S c mt ca nhm hydroxyl ti v tr C-5 v nhm th v tr C-4, c 2 bin ha dng c mu thng qua s ngn cn cc phn ng hydrat ha n n s to thnh dng khng mu. Khi mc hydroxyl ha cc aglycone tng, tnh bn ca cc anthocyanin s gim. Tuy nhin khi tng s methoxyl ha, s thu c kt qu ngc li. V d: s c mt ca nhm OH v tr 4 v 7 trong phn t lm bn ha ng k cc pigment, trong khi , s methoxyl ha c nhm hydroxyl ny lm gim bn. Cc anthocyanin c glycosyl ha v acyl ha s cho dng mu xanh, S glycol ha nhng nhm OH t do lm tng tnh bn ca anthocyanin. V vy, cc diglucoside bn hn cc monogluside ca cng mt anthocyanin. Anthocnin c cha 2 hay nhiu nhm acyl (nh ternatin, platyconin, cinerarrin, gntiodenphin v zebrrinin) l bn trong mi trng trung tnh hoc acid yu do lin kt hydro gia cc nhm hydroxyl ca cc nhn phenolic trong anthocyanin v acid vng thm. Brouillard (19811982) v Goto cng vi cng vin (1982-1983) kho st rng cc anthocnin diacylate ha c bn ha bi s lin kt cht kiu sandwich nh s tng tc gia vng anthocyanin v 2 nhm acyl vng thm. 7.2. PH: Trong mi trng nc, pH c nh hng ng k ln mu sc ca anthocyanin (Brouillard 1984,Mzza v Brouillard 1987). Cu trc, bn mu, mu sc ca anthocyanin thay i theo s thay i ca ph. S thay i cu trc ca anthocyanin khi pH thay i c cp trong phn cu trc chuyn ha. Nm 1956 Lukton nghin cu thy rng: ph = 2 4.5 th vn tc phn hy pelargonidin 3- glucoside trong dch p ca qu du khng xc nh c khi khng c O

2 . Khi c mt O 2 ph tng nhanh, s bin tnh tng ln mnh lit. Hnh 5: nh hng ca pH ln tc thy phn ca qu du c un nng ti 45 0 C trong trng hp c Oxygen hoc Nitrogen n nm 1972, Adams nghin cu thy rng ph = 2 4 t nh hng n s phn hy ca Anthocyanin trong sut qu trnh gia nhit khi khng c O 2 . Khi c mt O 2 th Anthocyanin b bin tnh mnh ph trn cu trc ca chng cng vi bn mu thay i vi s thay i ph. 7.3. Nhit : Trong qu trnh ch bin v bo qun thc phm, Anthocyanin d dng b bin tnh di tc dng ca nhit (Markakis 1974). Hnh 6: S bin tnh ca Anthocyanin 3,5 di Glucoside ti pH 3,7 Khi mt cu trc ca anthocyanin bn vi s gia tng ca pH th n cng bn vi s gia tng ca nhit . S hydroxyl ha cc aglycone lm gim tnh bn ca anthocyanin, trong khi s methxyl ha, glycosyl ha, acyl ha s cho kt qu ngc li. V d: Anthocyanin 3- glycoside c bn nhit ln nht ti pH = 1.8 - 2.0 vi s c mt ca oxy trong khi anthocyanidin 3,5-diglycoside c bn nhit ln nht ti pH = 4.0 5.0 C ch ca s phn hy nhit xy ra khng nhng ph thuc vo nhit m cn ph thuc vo bn thn anthocyanin . Drazdina cho rng coumarin glycoside l sn phm phn hy ph bin ca anthocyanin 3,5 diglycoside v c ch ca s phn hy l: u tin, cation flavylium chuyn

thnh dng quinnidal base, tip theo l hnh thnh nhiu dng sn phm trung gian v cui cng thu c dn xut coumarin v thnh phn tng ng vi vng B ca anthocyanidin, S bin tnh ny khng nhng c xc tin bi nhit m cn b nh hng bi oxy. Anthocyanidin 3 glycoside th khng hnh thnh nhng dn xut coumrin m bc u tin ca s phn hy bao gm s chuyn ha ca dng carbinol pseudobase khng mu, sau l s m vng pyrylium hnh thnh dng chalcone trc khi thy phn lin kt glycoside. Adams cho rng, cc anthocyanidin 3-glycoside khi c nung nng pH = 2.0 4.0 u tin s b thy phn lin kt glycoside ( 100 o C) sau l s bin i aglycone thnh chalcone. S bin tnh hn na dn n hnh thnh dng sn phm mu nu c bit l vi s c mt ca oxy. Khi un nng lu di cc anthocyanin c th b phn hu v mt mu, c bit l cc anthocyanin ca dy ty, anh o, c ci. Ngc li cc anthocyanin ca phc bn t en cng trong iu kin li khng b thay i.Nhn chung khi gia nhit, cc cht mu d dng b phn hu, cn cc cht mu vng th kh hn. 7.4. Oxy: Oxy v nhit c xem l nhng tc nhn c trng xc tin s phn hy ca anthocyanin, do m sinh ra nhng dng sn phm khng mu hoc mu nu. Chnh s oxy ha trc tip dng carbinol pseudobase gy ra s kt ta v ng vng trong nc tri cy. Trong qu trnh un nng cc anthocyanin b oxy ha mnh lit. Oxy v nhit l nhng tc nhn xc tin c bit nht trong nc p ca blueberry, cherry (anh o), currant, nho, raspberry v du. Lng anthocyanin cn li ca du s ln hn khi ng chai di iu kin chn khng

hoc nitrogen (Baravingas v Cain, 1965). bn ca cc pigment ca nho cn c s dng nh l cht mu nc gii kht c tng ln khi ng hp vi nitrogen. 7.5. Enzyme Nhiu enzyme ni sinh trong t bo ca cy c kh nng lm mt mu anthocyanin. Nhng enzyme ny c gi chung l anthocyanase. Da vo c tnh ca cc enzyme m ngi ta phn lm 2 nhm; Glycosidase v polyphenol oxidase (PPO). Cc enzyme ny thu c t nm (fugal). Glycosidase: l enzyme thy phn lin kt glycoside ca anthocyanin to ra ng t do v aglycone ny km bn hn rt nhiu v mt mu rt nhanh khi c mt ca catecol(Huang 1955). Polyphenol oxidase (PPO): tc dng ln anthocyanin vi s c mt ca Odiphenol thng qua c ch oxy ha kt hp. Theo Gromeck v Markakis, s thm vo glycosidase v PPO xc tc cho qu trnh peroxide ha phn hy anthocyanin. Hnh 7: S bin tnh anthocyanin vi phn ng oxy ho catechol Peng v Markakis ngh mt c ch m trong O-quinine c to thnh bi s oxy ha anthocyanin. Blom phn lp c enzyme anthocyanin-glycosidase t Aspergillus Niger v ch ra nh hng ca n trong qu trnh thy phn lin kt glycoside ca anthocyanin. PPO l enzyme c nh hng yu ln anthocyanin dng quinonoidal base d b thy phn bi PPO hn dng cation flavylium. Tc phn hy bi PPO ph thuc vo s thay th m hnh ca vng B v mc glycosyl ha. 7.6. nh sng Cc anthocyanin thng khng bn khi tip xc vi tia t ngoi, nh sng thy c, v cc ngun php x khc. nh sng c 2 nh hng n anthocyanin: Tng cng cho qu trnh sinh tng hp. Xc tin s bin tnh ca chng. Nm 1964, Siegenman cho rng nhng qu to ging s chuyn sang mu xanh

khi trong bng ti. Nm 1968 Vanburen v cc cng s tng trnh rng cc diglucoside c acyl ha v methyl ha th cc Anthocyanin trong ru bn nht khi ngoi nh sng, cc diglucoside khng b acyl ha l t bn hn v monoglucoside l km bn nht, nm 1975 Palamidis v Markakis tm thy rng nh sng thc y qu trnh phn hy anthocyanin trong nc gii kht c CO 2 c phi mu vi anthocyanin t xc nho. Macccarone v cng s nghin cu s quang ha ca anthocyanin v ch ra rng , anthocyanin diglycosyl ha ti v tr C3 v C-5 l bn hn cc anthocyanin mono glycoyl ha ti v tr C-3 ng thi chng bn hn so vi cc aglycone tng ng. 7.7. ng v cc sn phm bin tnh ca chng: nng 100 ppm, ng v cc sn phm phn hy ca chng c tc dng thc y s phn hy cc anthocyanin, trong fructose, arabinose, lactose v sorbose c kh nng phn hy anthocyanin mnh hn glucose, sucrose, v maltose. Tc phn hy ca anthocyanin lin quan n tc phn hy ca ng. Cc sn phm phn hy ca ng gm c: furfura l,5 hydroxymethyl furfural v acctaldehyde thu c t phn ng Mailard hoc t s oxy ha ca acid ascorbic, polyuronic hoc bn thn cc anthocyanin. Nhng sn phm phn hy ny d dng ngng t vi cc anthocyanin hnh thnh nhng hp cht phc tp c mu nu sm. S phn hy anthocyanin vi s c mt ca furfural v HMF trc tip ph thuc vo nhit v c thy r nht l trong nc tri cy s c mt ca n lm tng thm hiu qu phn hy ca tt c cc loi ung v cc dn xut ca chng. Hnh 8: Phn ng ngng t ca a) Cyanidin v furfural

b) Cyanidin ketobase v furfural 7.8. Cc ion kim loi: Mt s ion kim loi a ha tr c th tng tc vi cc anthocyanin c nhm OH v tr ortho gy ra hiu ng su mu (bathocromic). Hin tng ny xy ra khi kim loi tip xc vi anthocyanin trong qu trnh ch bin rau qu hoc s cho thm cc mui kim loi vo trong thc phm. Sistrunk v Cash chng minh rng c th bn ha mu ca dch trch du bng cch thm vo mui thic. Francis (1977) cng b rng, cc ion Ca, Fe, Al to thm s bo v cho anthocyanin ca nc p tri mn vit qut (cranberry), nhng s bin i mu xy ra l do s to phc gia ion kim loi v tannin, kt qu sau cng l khng c li. Trong cng nghip hp, s mt mu ca nhng tri cy c cha anthocyanin l do c phn ng vi thic ca hp (Culpepper v caldwell, 1972). Trong phn ng vi thic, anthocyanin ng vai tr nh cht kh cc catod hoc anod. Cht kh cc catod c th b kh bi hydro mi