Tap chi Khoa hpc va Phat trign 2010: Tap 8, sd 6: 994 - 1003 T R U I N G DAI HOC NONG NGHIEP HA N Q I
MO HINH HOA QUA TRINH CHIET POLYPHENOL TUT VO VAI
Modelling the Extraction of Phenolics from Litchi Fruit Pericarp Lai Thi Ngoc Ha', Nguyen Thj Thu Hirong, Phan Thj Hang^
'Khoa Cong nghe thuc phdm, Tru&ng Dgi hgc Nong nghiep Ha Ngi
^L&p Bdo qudn che bien A K51, Tru&ng Dgi hgc Nong nghiep Hd Ngi Dja chi email tac gia lien lac: [email protected] / i j Z u n v > . . . u . , S ; = T 6 M T A T
Ham lu'O'ng polyphenol tdng s6, anthocyanin tdng so va kha nang khang oxi hoa cua vo vai dirac xac djnh. K4t qua cho t h i y ham lu'p'ng polyphenol trong vo vai cao (96,25 mgGAE/g CK), do vay co th6 la ngudn polyphenol ti§m nang irng dung du'O'c trong cdng nghiep t h y c p h i m . Kdt qua cho t h i y ham lu'O'ng polyphenol trong vo vai cao (96,25 mgGAE/g CK), do vay co thd la ngudn polyphenol tiem nang irng dung dirac trong cdng nghiep thirc p h i m . Anh hu'ang ciia mot s d ydu td cdng nghe nhu":
ndng do ethanol, nhiet do, pH va tho'i gian den hieu suat thu hdi polyphenol dup'c nghien ciru. Md hinh md ta qua trinh tach chidt polyphenol tir vd vai du'O'c xay dirng Y= 62745,17 + 3946,87X^1 + 6643,00X^ - 9949,96X^ - 1392,63X^,X^ + 2718,14X^iX^ + 9880,41X^X^.
Tir khda: Chidt, kha nang khang oxi hda, md hinh, nhiet dp, pH, polyphenol, vo vai.
.: . .•vy-''. :r..\ .••;.;^: v.ij ^r^ii; hh - ; n S U M M A R Y
The total phenolics, total monomeric anthocyanin and the antioxidant capacity of the litchi pericarp were determined. The results showed that the litchi pericarp is rich in polyphenol (96.25 mgGAE/g DW), and it is an important source of phenolic which could be used in food processing technologies. Several influencing factors on the phenolic extraction from litchi fruit pericarp were studied such as ethanol concentration, temperature,pH and time. The simulation model of the extraction process was determined: Y= 62,745.17 + 3,946.87X^1 + 6,643.00X^2 - 9,949.96X% - 1,392.63X^X^ + 2,718.14X^X^ + 9,880.41X^X%
Key words: Antioxidant capacity, extraction, litchi pericarp, model, pH, polyphenol, temperature.
I . D A T V A N D E „ , ...^ ...•.-..:.. pha buy eae dai phan tii sinh hpc (ADN, Stress oxi hda dae trflng bdi sfl ma't can protein, lipid) va gay benh cho cd the.
bang gifla san xua't cac gd'c tfl do va hoat Trong s6'cac cha't ch6'ng oxi hda tfl nhien ddng cua cae cha't chd'ng oxi hda trong ed the cd trong rau qua, polyphenol la nhdm cha't dflde eoi la nguyen nhan eiia ra't nhieu benh ra't dflde quan tam bdi le polyphenol the hien trong do cd ung thu, tim mach, suy giam he nhflng dac tinh sinh hpc quy dae biet la kha than kinh (Alzheimer, Parkinson) va lao hda nang chdng oxi hda, chd'ng viem, chd'ng di sdm (Pineemail va Defraigne, 2004; Edeas, iing va kha nang khang khuan (Tapiero va es., 2006). Ke't qua nhieu nghien cflu cho tha'y ed 2002; Alberto va cs., 2006). Cae polyphenol ed mpt mo'i lien he nghich gifla kha nang xua't tinh khii gi6ng vitamine C, khii cac g6'c tfl do hien cac can benh tren va che dp an giau rau va vd hoat chflng. Ngoai ra, polyphenol lam qua. Giai thich hdp ly cho mo'i lien he nghich tang hieu qua chd'ng oxi hda ciia nhieu chat nay la sfl cd mat eua cac cha't chd'ng oxi hda chdng oxi hda tfl nhien khac nhfl vitamine tfl nhien ed trong rau qua. Cac cha't chd'ng C, E va cac earotenoid. Nhieu ke't qua thii oxi hda tfl nhien trong rau qua se vd boat cac nghiem cho tha'y che dp an giau polyphenol gd'c tfl do khie'n chung khdng con kha nang (qua, rau, ngfl co'c nguyen dang, rUdu vang
Mo hinh hoa qua trinh chiet polyphenol tfl' vo vai dd, tra) cho phep han ehe sfl xua't hien
stress oxi hda va cac benh lien quan (Hung va cs., 2004; Haliwell, 1994). Polyphenol ddi dao trong la che giup ngfldi dan chau A tre lau, khde va sd'ng thp. Polyphenol trong khoai tay mau tim, dd trdng tren cae cao nguyen Nam My da giup ngfldi dan Peru, Bolivia eo sfle khde t6't, t r a n h dflde nhieu loai benh nguy hiem.
Viet Nam la nude nhiet ddi gid mfla, hoa trai ed quanh nam. Ra't nhieu loai cay an qua, cay rau, cay thud'e ed ham Ifldng polyphenol cao bfla hen la ddi tfldng nghien eflu xae dinh boat tinh sinh hpe, nghien cflu che bie'n cae san pham cd gia tri kinh te', sinh hpc cao nhfl cay di, cay vai, cay Iflu, eay tao meo, cac loai che dang, cay rau die'p ca...
Trong so' cac loai rau qua giau polyphenol, qua vai cd ham Ifldng polyphenol cao (eao hdn 15% so vdi nho) trong dd phan vd qua chfla ham Ifldng polyphenol kha ldn (80-120 mg GAE/g CK, Ruenroengkhn va es., 2008).
Dich chiet polyphenol tfl vd vai cd tac dung kim ham sfl tdng hdp ADN eua te' bao ung thu, do dd kim ham sU phat trien cua khdi u (Wang va es., 2006). Phan vd chiem 15%
khdi Ifldng qua vai tfldi (Ruenroengkhn va es., 2008) va la phan phe pham ciia cdng nghe che bie'n vai. Day bfla hen la ngudn nguyen lieu ddi dao cho san xua't dich giau polyphenol flng dung trong cdng nghiep thfle pham va dflde pham.
San xua't dich giau polyphenol tfl thfle vat ludn bat dau bang e6ng doan chiet polyphenol va phUdng phap chiet bang dung moi la phfldng phap thfldng dflde sii dung nha't. Hieu qua chiet polyphenol theo phfldng phap nay phu thupe rat nhieu yeu t6': loai dung m6i sii dung, ndng dp dung m6i, nhiet dp chiet, thdi gian chiet, pH eiia dung dich chiet... (Chirinos va es., 2007; Silva va cs., 2007; Todaro va es., 2009; Pompeu va es., 2009). Kh6ng ed quy trinh tach chiet chung cho ta't ca eae loai thfle vat. Mdi loai nguyen lieu thfle vat khac nhau vdi dac diem eau tao khac nhau va thanh phan polyphenol khac
nhau se chiu anh hfldng khac nhau eiia cae yeu td' ke tren va cd dieu kien tdi flu de chiet polyphenol. Muc dich cua nghien eflu nay la dflng phan mem toan hpe md ta anh hfldng cua mdt so' yeu td' c6ng nghe de'n hieu qua chiet polyphenol tfl vd vai.
2. VAT LIEU VA PHUONG PHAP NGHIEN Ctru
2.1. Vat lieu v a hoa chat "' "
Vd vai dupe thu tai Trfldng Dai hpe Nong nghiep Ha Npi thang 6 nam 2009. Vd cdn tfldi, ed mau dd tfldi, dflde riia sach, de rao nflde, ddng khd d nhiet dp -50°C bang may d6ng kh6 trong hai ngay. Sau dd, vd khd dflde nghien nhd va bao quan d nhiet dp - 23''C dfldi nito khi.
Acid gallic, DPPH (2,2 - diphenyl - 1 - picrylhydrazyl), thud'e thii Folin - Cioealteu, Trolox (6-hydroxy-2, 5, 7, 8-tetramethyl-2- carbocylie acid) eiia Sigma (Dfle). Cae dung moi ethanol, methanol, aceton; hoa chat NaaCO,,, KCI, CHjCOONa.SHaO, acid acetic, acid chlohydric ciia Trung Qu6c.
2.2. Cac thi nghiem khao s a t
Trflde khi tien hanh thi nghiem mo hinh hda, mpt so' thi nghiem khao sat dflde thflc hien de xac dinh sO bp khoang anh hfldng cua eae yeu to' de'n hieu qua chiet polyphenol tfl vd vai.
2.2.1. Thi nghiem khdo sdt 1: Anh hudng cua logi dung moi •" '^ -! •; ; - 'n-,::
0,2 g vd vai dang hot -I- 20 ml dung mdi nghien eflu, thdi gian lae 3 gid vdi van td'e lie 200 vdng/phut, nhiet dp 30°C, h tam trong 30 phflt vdi 6000 v/ph. Ca't dudi dung mdi va hda tan trd lai len 10 ml bang nflde eat thu dflde dich chiet polyphenol. Xae dinh ham Ifldng polyphenol tdng sd'ciia dich chiet.
2.2.2. Thi nghiem khdo sdt 2: Anh hudng nhiet do
0,2 g vd vai dang hot -i- 20 ml dung mdi rut ra tfl thi nghiem khao sat 1, thdi gian
Lai Thi Nggc Ha, Nguyin Thi Thu Huong, Phan Thi HSng lac 3 gid vdi van tdc lac 200 vdng/phut, li
tam trong 30 phut vdi 6000 vdng/phut. Ca't dudi dung m6i va hda tan trd lai len 10 ml bang nflde ca't thu dflde dich chiet polyphenol. Xac dinh h a m Ifldng polyphenol tdng sd'ciia dich chiet.
2.2.3. Thi nghiem khdo sdt 3: Anh hudng thai gian
0,2 g vd vai dang bdt -i- 20 ml dung mdi rut ra tfl thi nghiem khao sat 1, van t6'e lae 200 vdng/phflt, nhiet dp 30°C, h tam trong 30 phut vdi 6000 vdng/phut. Cat dudi dung m6i va hda tan trd lai len 10 ml bang nflde ca't thu dflde dich chiet polyphenol. Xac dinh ham Ifldng polyphenol tdng sd' cua dich chiet.
2.3. Thi n g h i e m m o h i n h h o a va toi xiu h o a Thi nghiem m6 hinh hda dflde tie'n hanh vdi 3 yeu to' anh hfldng sau: Xj- Ndng dp ethanol (% v/v); X^ - Nhiet dp xii ly ("C); X3 - pH dung dich chiet. Khoang bien ddi eua cac yeu t6' anh hfldng dflde rut ra tfl cae thi nghiem khao sat va dfla tren cac tai lieu tham khao. Ham mue tieu Y la ham lUdng polyphenol tdng sd' thu dflde tfl lg cha't khd vd vai (ng GAE/g CK).
Dang cua md hinh:
Y = bo + b j X , + b2X2 + b3X3 -1- bi2XiX2 + 013X1X3 -*- b23X2X3
2.4. Cac phifdng p h a p p h a n tich h o a sinh 2.4.1. Ham lugng polyphenol tong so
Ham Ifldng polyphenol tdng so' dflde xac dinh bang phfldng phap Folin-Ciocalteu (Singleton va Rossi, 1965). Dfla 500 \i\ dich chiet vao o'ng nghiem, them 250 ^il thud'e thii Folin - Cioealteu IN, lac deu sau dd them 1250 pi Na2C03 7,5%. Sau 30 phut, do dp ha'p thu ciia hdn hdp tai 755 nm. Gallic acid dflde dflng lam cha't chuan (y = 0,0299x -t- 0,0095; R2 = 0,9989). Ham lfldng polyphenol dflde bieu dien theo dfldng Ifldng acid gallic trong 1 g cha't kh6 vd vai (|ig GAE/g CK - |ig Gallic Acid Equivalent/g cha't kh6).
2.4.2. Ham lugng anthocyanin tong so Ham Ifldng anthocyanin tdng s6' dflde xae dinh bang phfldng phap pH vi sai fAGAC
Method 2005.02). Cho 0,2 g vd vai vao 10 ml methanol 80% chfla HCI ndng dp 0,1%. Hdn hdp dflde trpn deu va chiet b 4°C trong 24 gid. H6n hdp sau dd dflde ly tam, dich trong thu dflde la dich chiet anthocyanin. Dich chiet anthocyanin dflde hda tan 10 lan trong cac dung dich dem KCI pH 1 va CHjCOONa pH 4,5. Do dp ha'p thu ciia cac dung dich nay tai 520 va 700 nm. Ham Ifldng anthocyanin dflde tinh theo cdng thfle sau:
, ,• ,- TAC (mg/g) = A*MW*f*V/m/s/I Trong dd:
A = (A520 — A7oo)pHl " (A520 ~ A7oo)pH4.5
f: he sd' pha loang
1: ehieu day cuvette (cm) V: the tich dich chie't (1) m: khd'i Ifldng vd vai (g)
MW = 449,2 g/mol va s = 26.900 1/mol/em la kh6'i Ifldng phan tii va dp ha'p thu phan tii ciia cyanidin-3-glueoside (anthocyanin phd bie'n trong tfl nhien).
Ham Ifldng anthocyanin dflde bieu dien bang mg dfldng Ifldng eyanidin-3-glucoside trong 1 g cha't khd vd vai (mg CGE/g CK- mg Cyanidin-3-Glucoside Equivalent/g cha't khd).
2.4.3. Khd ndng khdng oxi hoa
Kha nang khang oxi hda dflde xae dinh bang phfldng phap 1,1 - diphenyl - 2 - pieryl hydrazyl radical (DPPH) (Tabart va cs., 2009). 100 pi m i u dflde phan flng vdi 2900 |il DPPH 0,2 mM pha trong methanol d 25°C trong 20 phut. Do dp ha'p thu eua hdn hdp tai 517 nm. M l u control dflde tie'n hanh tfldng tfl nhflng thay 100 ^1 dich chie't bang nflde ca't.
% kim h a m = (Acontrol - Amiu)/
AcontroPlOO.
Trong dd: Acontrol va AmIu la dp ha'p thu tai 517 nm cua mSu control va cua dich chie't.
Trolox - mpt d i n xua't cua vitamine E dflde dflng lam chat chuan (y = 0,0806x - 0,2453; R' = 0,9997). Kha nang khang 0x1 hda dflde bieu diln bang moi dUdng Ifldng Trolox tren 1 gam chat khd vd vai (moi TE/g CK - moi Trolox Equivalent / g cha't khd).
Mo hinh hoa qua trinh chiet polyphenol tiy vo vai M6 hinh hda bang phan mem Nemrowd.
Cac thi nghiem dflde lap lai 3 lan. Ke't qua dflde xii ly bang Excel 2003 va SAS 9.0.
3. KET QUA vA THAO LUAN
3.1. Mot so chl t i e u ciia vo vai nguyen lieu va vo vai dong kho
Bdt vd vai dflde chie't bang triple extraction (chie't lan Ifldt bang acetone, hdn hdp methanol : nflde : acid acetic, hdn hpp ethanol : nflde : acid acetic). Dich chie't dflde cat dudi dung m6i, len the tieh 10 ml va dflde dung de xac dinh ham Ifldng polyphenol t6ng sd'va kha nang khang oxi hda.
.Mpt s6' ehi tieu cu the eiia vd vai tfldi va vd vai nguyen lieu dflde xae dinh:
- Ty le vd qua: 13,77% khdi Ifldng qua tfldi.
- Ham Ifldng chat kh6 tong s6' eua vd tfldi: 27,00%.
- Ham Ifldng chat kh6 t6ng s6' eiia mau ddng khd: 93,28%.
- Mau ddng kh6 ed mau hdng tia va mfli dae trflng ciia vd vai.
- Ham Ifldng polyphenol tdng s6' ciia vd vai: 96,25 mg' GAE/g CK hay 25,99 mg GAE/g vd vai tfldi.
- Kha nang khang eiia oxi hoa eiia vd vai 894,26 moi TE/g CK
- Ham Ifldng anthocyanin cua vd vai:
0,61 mg eyanidin-3-glucosid/g CK hay 16,47 mg cyanidin-3-glueosid /lOOg vd vai tfldi.
Ham Ifldng polyphenol cung nhfl anthocyanin xae dinh dflpc eua vd vai thu hai tai Trfldng Dai hpe Ndng nghiep lan Ifldt la 96,25 mg GAE/g CK va 0,61 mg CGE/g CK.
Ke't qua nay gan vdi cae ke't qua dflde cdng bd' trflde day. Ruenroengkhn va cs. (2008) xac dinh ham Ifldng polyphenol thu dflde tfl vd vai la 80-120 mg GAE/g CK tfly thupc vao dieu kien tach chie't. Theo Hu va es. (2010), tdng Ifldng acid phenohc, flavanoid, proanthocyanin ciia vd vai la 41 - 106 mg/g CK. Ham Ifldng anthocyanin ciia vd vai dflde Duan va cs.
(2007) xac dinh la 18,6 mg/100 g chat tfldi.
So vdi cac phe pham khac eua nganh cong nghiep thflc pham, vd vai cd ham Ifldng
polyphenol cao hdn ba tao (phe pham cua cdng nghiep san xua't nflde ep tao, 7,16 mg GAE/g) (Sudha va cs., 2007), eao hdn bdt ba nho (phe pham eua cdng nghiep san xua't rfldu vang, 2,78 - 5,33 mg/g CK) (Chun Yi va es., 2009) tha'p hdn so vdi vd qua Iflu (phe pham cua che bie'n Iflu - 140 mg GAE/g CK) (Shabtay va es., 2008). So vdi cac loai la cay rflng Amazon hien dang dflpc quan tam nhfl ngudn polyphenol ddi dao ed ham Ifldng polyphenol bien ddi trong khoang 44,2 - 63,0 mg GAE/g CK (Souza va cs., 2008), vd vai cd ham Ifldng polyphenol cao hdn. Cae ke't qua nay eho tha'y, vd vai la ngudn polyphenol thflc vat ddi dao. Viec tan dung vd vai - mdt loai phe pham cua cdng nghiep thfle pham gdp phan tang gia tri kinh te eua cay vai, tao them san pham ed ngudn gd'c tfl nhien flng dung dflpc trong cdng nghiep thfle pham va cdng nghiep dflde nhfl chat chd'ng oxi hda.
3.2. Khao sat stf bo anh hifding cua mot so' yeu to' cong nghe de'n hieu qua chie't polyphenol txi vo vai
3.2.1. Anh hudng cua dung moi
Hieu qua chie't polyphenol tfl nguyen lieu thfle vat phu thudc vao loai dung mdi sii dung dac biet dp phan cue cua dung mdi.
Viee sii dung duy nha't dung mdi ed dp phan cue eao nhu nflde hay eae dung mdi hflu ed kem phan cfle nhfl hexan hay ehloroforme khdng cho hieu qua thu polyphenol cao vi thanh phan polyphenol thflc vat ra't da dang (phan cfle va khdng phan cfle). Khdng cd loai dung mdi hay he dung mdi chuan nao dflng chung de tach polyphenol ciia thfle vat.
Methanol va hdn hdp methanol thfldng dflde sii dung trong eae thi nghiem chie't polyphenol (Silva va cs., 2007; Souza va cs., 2008). Cae dung mdi khae nhfl ethanol, acetone, ethylacetate eflng dflde sii dung de chie't polyphenol (Chirinos va cs., 2007).
Tien hanh chie't polyphenol tfl vd vai bang methanol, ethanol, hdn hdp methanol : nflde, hdn hdp ethanol : nflde (Bang 1).
Lai Thi Ngoc Ha, Nguyin Thi Thu Huong, Phan Thi HSng
Bang 1. Anh hxidng cxia loai d u n g moi d e n h i e u s u a t c h i e t p o l y p h e n o l tU vo vai
Loai dung moi Polyphenol tong so (mgGAE/g CK) Hieu suat thu hoi polyphenol (%) Methanol ..,,,
Ethanol
Methanol/nu'O'c, 70/30, v/v Ethanol/nu-o-c, 70/30, v/v
41,93' 19,76' 28,94 ' 38,55 '
43,56 20,53 30,07 40,05 Cdc so vai cdc chir khdc nhau thi khdc nhau a miicy nghia 0,05.
': Hieu suat thu hdi = Polyphenol tdng sd thu duac (mgGAE/g CK)/Polyphenol tdng sd ciia vd vdi (mgGAE/g CK)*100 Bang 2. Anh hifdfng
Nhiet do (°G) 40 50
cua n h i e t do de'n h i e u s u a t c h i e t p o l y p h e n o l tU vo vai Polyphenol tong s6 (mg GAE/g CK)
44,74°
60,23^
44,62"
Hieu suat thu hoi polyphenol (%) 46,48
62,58 46,36 Cdc: so vai cdc chU khdc nhau thi khdc nhau a mirc y nghia 0,05.
': Hieu sudt thu hdi = Polyphenol tdng sd thu duac (mgGAE/g CK)/Polyphenol long sd ciia vo vai (mgGAE/g CK)*IOO Ket qua xii ly bang phan mem SAS9.0
cho ta tha'y cae loai dung mdi khae nhau thi eho kha nang trieh ly polyphenol khac nhau b mflc y nghia P = 0,05. Trong dd, methanol la loai dung mdi eho kha nang chie't ldn nha't, cho ham Ifldng polyphenol tdng so' la 41,93 mgGAE/gCK, hieu sua't thu hdi 43,56%, ethanol cho kha nang chiet tha'p nha't. Viec them nflde vao ethanol lam tang hieu qua chie't polyphenol tfl vd vai. Nflde them vao ethanol eho phep tang hieu qua chie't cae hdp cha't phenol b dang glycoside vd'n de hda tan trong nflde. Xet ve mat hieu sua't thu hdi, viee dflng hdn hdp ethanol:
nflde cho ke't qua nhd hdn so vdi viec dung methanol nhflng xet ve mat an toan, ethanol an toan hdn eho ngfldi sii dung. Vdi muc dich san xua't dich chie't polyphenol tfl vd vai flng dung trong cdng nghiep thue pham va cdng nghiep dflde, hdn hdp ethanol: nflde dflde chpn de chie't polyphenol tfl vd vai va dflde dflng trong thi nghiem md hinh hda.
3.2.2. Anh hUdng cua nhiet do
Nhiet dp la yeu to' quan trpng anh hfldng khdng nhflng tdi hieu sua't trich ly ma cdn anh hfldng tdi chi phi va cha't Ifldng ciia dich chie't polyphenol. Nhiet dp lam giam dp nhdt trong dung dich, tang td'e dp tham tha'u dung mdi vao te bao va tang hieu sua't trieh ly. Tuy nhien, tie'n hanh chie't d nhiet dp qua eao thi vfla ton ehi phi dn nhiet vfla tang nguy ed giam chat Ifldng dieh chie't do phan flng nau hda.
Tie'n h a n h chie't polyphenol tfl vd vai d cac nhiet dp khac nhau, ke't qua dUdc gidi thieu b bang 2.
Ke't qua xii ly thd'ng ke bang phan mem SAS9.0 cho tha'y b mfle y nghia P = 0,05, nhiet dp anh hfldng de'n kha nang tach chie't polyphenol tfl vd vai. Nhiet dp cho ham Ifldng polyphenol ldn nha't d 50°C la 60,23 mgGAE/gCK vdi hieu sua't thu hdi la 62,58%. Tfl 40"C khi nhiet dp tang tfl 40 de'n 50°C, kha nang chie't polyphenol tang do nhiet dp eao thuc day sfl xam nhap ciia dung mdi vao nguyen lieu va chie't r u t polyphenol.
Tuy nhien khi nhiet dp tang len, phan flng oxi hda polyphenol bdi khdng khi tang len do dd Ifldng polyphenol thu dflde giam. Mau ciia dieh chie't d 60°C cd mau sSm nha't cho tha'y d nhiet dp nay phan flng oxi hda polyphenol vd vai xay ra manh nha't.
Ket qua anh hudng eua nhiet dp de'n hieu sua't chie't polyphenol vd vai thu dflde phfl hdp vdi cdng bdciia Todaro va cs. (2009):
nhiet dp anh hfldng ra't rd ret de'n hieu qua chie't anthocyanin tfl vd qua ea tim; khi nhiet dp tang tfl 0 de'n 40°C, hieu qua chie't tang nhflng khi nhiet dp tang tren 40°C, Ifldng anthocyanin thu dflde giam. Ruenroengkhn va cs. (2008) khi nghien cflu anh hfldng ciia nhiet dp de'n qua trinh chie't polyphenol vd vai cho tha'y khi nhiet dp tang tfl 40°C den 60°C, hieu qua chie't polyphenol tang nhflng sau dd hieu qua chie't polyphenol khdng tang khi tang nhiet dp tfl 60°C de'n 70°C.
B a n g 3. A n h Thai g an (phut)
M6 hinh hoa
hifd?ng c i i a t h d i Polyphenol
qua trinh chiet polyphenol tu" vd vai
gian de'n h i e u suat tong so (mgGAE/g CK)
chie't polyphe nieu suat th
noi txi vo u hoi polyphe
v a i
;nol (%) 60
90 120 150 180
59,01' 55,47' 58,08' 58,27' 61,68'
61,31' 57,63' 60,34' 60,54' 64,09' Cdc: sd vai cdc chir khdc nhau thi khdc nhau a mirc y nghia 0,05.
': Hieu sudt thu hoi = Polyphenol tdng sd thu duac (mgGAE/g CKJ/Polyphenol tdngsd cua vo vai (mgGAE/g CK)*IOO 3.2.3. Anh hudng cua thdi gian —
Thdi gian chie't c u n g la mdt t r o n g nhflng yeu to' q u a n trpng, a n h hUdng de'n h i e u sua't, cha't lfldng dieh chie't polyphenol c u n g nhfl tinh kinh te cua q u a t r i n h . Ne'u thdi gian qua ngan, khdng dii de d u n g moi x a m n h a p vao trong te bao, hda t a n polyphenol va chie't r u t ra ngoai it thi Ifldng polyphenol t h u dflde se tha'p, ngflde lai thdi gian chie't qua dai lam giam hieu sua't sii d u n g t h i e t bi va polyphenol cd the bi oxi hda. Tie'n h a n h chie't polyphenol tfl vd vai trong cac k h o a n g thdi gian k h a c nhau, ke't qua cho d b a n g 3. So' lieu dflde xii ly ke't qua b a n g p h a n m e m SAS9.0 eho tha'y thdi gian chie't t h a y ddi tfl 60 de'n 120 p h u t khdng CO a n h hfldng tdi qua t r i n h tach chiet polyphenol tfl vd vai d mflc ,y nghla - 0,05.
Dieu dd cho tha'y dd'i vdi vd vai, thdi gian 60 phut dli d^ tach chie't polyphenol trong cac dieu kien da dUde xae dinh.
Xet sd bp a n h hfldng ciia mdt sd' yeu td' den hieu sua't t h u hdi polyphenol tfl vd vai t a thay, thdi gian 60 p h u t dii de chiet polyphenol tfl vd vai. N h i e t dp t r o n g k h o a n g 40°C - 60"C anh hfldng de'n h i e u sua't chiet. T h e m vao do, vd vai cdn chfla n h i e u a n t h o c y a n i n , cac hdp chat nay ben d p H acid, n h i e u ke't q u a chi r a r a n g pH ed a n h hfldng de'n h i e u q u a chie't polyphenol tfl vd vai ( R u e n r o e n g k h n va cs., 2008; Zhong va cs., 2007). Xua't p h a t tfl eae ket qua n g h i e n cflu sd bd va t a i lieu t h a m khao ve eae yeu to' a n h hfldng d e n h i e u sua't chie't polyphenol tfl vd vai, n g h i e n eflu n a y da tie'n h a n h md h i n h h d a q u a t r i n h chie't polyphenol tfl vd vai vdi cae yeu td': n d n g dp ethanol, n h i e t dp chie't va p H ciia dich chie't.
3 . 3 . M o h i n h h o a q u a t r i n h chie't p o l y p h e n o l txi v o v a i
Tien h a n h t h i n g h i e m t h e o 3 yeu td' a n h hfldng:
Xi: N d n g dp e t h a n o l (% v/v) Xg: N h i e t dp xii ly ("C)
X3: p H d u n g dieh chie't ; H a m muc tieu la Y: H a m lfldng polyphenol t h u dflde tfl 1 g chat khd vd vai (g GAE/g DW).
D a n g md h i n h :
Y = bo + biXi + baXj + baXa + h.^X.X^ +
^"- «':• b,3X1X3 -H b23X2X3
Cac mflc t h i n g h i e m gidi t h i e u 6 b a n g 4.
Vdi cac mflc t h i n g h i e m nhfl t r e n , b a n g ma t r a n thfle n g h i e m (Bang 5) dflpc xay dflng. Mdi t h i n g h i e m dflde lap lai 3 lan, ket qua eho d b a n g 6.
T r o n g dd: cac bie'n X^i la cae bie'n e h u a n . Cdng thfle c h u y e n cac bie'n thflc t h a n h bie'n c h u a n nhfl s a u : X®, = 2 * (X; - mfle gd'c) / (mfle t r e n - mfle dfldi).
M a t r a n ke't q u a dflde dfla vao p h a n m e m N e m r o w d de xae d i n h md h i n h . Ke't qua dflde ehi r a d b a n g cac he sd' eua md hinh (Bang 7).
Vay md h i n h mieu t a a n h hfldng ciia eae yeu td' n d n g dp e t h a n o l , n h i e t dp va pH den h i e u sua't chie't polyphenol vd vai nhfl sau:
Y= 62745,17 + 3946,87XS, + 6643,00XS2 - 9949,96X83 - 1392,63XSiXS2 H- 2718,14XSiXS3 + 9880,41X82X83 T r o n g dd:
Cac bie'n XSi la cae bien c h u a n .
R^ - 0,934 cho tha'y md h i n h p h a n a n h 93,4% thflc te.
Lai Thi Ngoc Ha, Nguyin Thi Thu Huong, Phan Thi Hang
Bang 4. Cac mvfc thi nghiem
MLFC thi nghiem X, - N6ng dp ethanol (%v/v) X2 - Nhiet dp (°C) X3 - pH
MifC g6c 50 50 4 Khoang bien doi 10 -'.::- 10 2 MLPCtren 60 '"::'•'' 60 6 Mi>c du'O'i 40 " 40 2
Bang 5. Ma t r a n thxic nghiem
thi nghiem 1 2 3 4 5 6 7 8
x^
-1 +1 -1 +1 -1 +1 -1 +1
Biln chl
x^
-1 -1 +1 +1 -1 -1 +1 +1
uan
X ^ -1 -1 -1 -1 +1 +1 +1 +1
X,- N6ng do ethanol (%v/v)
40 60 40 60 40
60 ,,1 40
60
Biln th
X2- LfC
Nhiet do (°C)
40 40 60 60 40 40 60 60
X j - p H 2 2 2 2 6 6 6 6
Bang 6. Ma t r a n ket qua
Thi nghiem 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
N6ng do ethanol (% v/v)
40 40 40 60 60 60 40 40 40 60 60 60 40 40 40 60 60 60 40 40 40 60 60 60
Nhiet do (°C)
40 40 40 40 40 40 60 60 . 60 60 60 60 40'"t-^:
40 40 40 40 40 60 60 60 60 60 60
pH 2 2 ' 2 2 , .,!:;2. :,;
2 2 2 2 2 2 2 6 6 6 6 6 6 6 6 6 6 6 6
Polyphenol tpng s6 (pg GAE/g CK)
77941,56 74894,70 78151,81 72679,79 76905,21 75022,19 63576,69 66131,90 68101,80 69069,67 79047,51 70818,77 25319,42 24709,25 23559,31 45261,42 47844,26 50937,12 70599,25 66635,34 65958,65 73611,97 68190,26 70916,31
Mo hinh hoa qua trinh chiet polyphenol tU" vo vai
Bang 7. Cac he so' ciia mo hinh
0 6 lech chuan cua ham R^
Bac tlf do
muc tieu 2825,037
0,934 16
He s6 bo b, b2 b3 b,2 b,3 b23
Gia tri 62745,174
3946,867 6643,004 -9949,960 -1392,628 2718,143 9880,412
F 1,00 1,00 1,00 1,00 1,00 1,00 1,00
€)6 lech chuan 576,658 576,658 576,658 576,658 576,658 576,658 576,658
t 108,81
6,84 11,52 -17,25 -2,41
4,71 17,13
MLPC y nghTa %
<,0,01 * "
<0,01 " *
< 0,01 * "
< 0,01 * "
2 , 6 9 * 0,0269 ***
< 0,01 ***
*; Ynghia 0,05: ***:y nghTa 0,001
Bang 8. So s a n h gia t r i thiic t e va gia tri tinh t o a n txi mo hinh
Thi nghiem
1 2 3 4
Nong do ethanol (% v/v) 52 54 56 58
B i i n thu'C
Nhiet do (°C)
53 56 59 62
pH
3,5 3 2,5
2
X ^
0,2 0,4 0,6 0,8
Bien chuan
x^
0,3 0,6 0,9 1,2
X=3
-0,25 -0,5 -0,9 -1
Polyphenol tong so (pg GAE/g CK)
Tinh toan
67054,44 69442,72 69910,00 68456,30
Thi/c te
71235,82 75948,58 79750,35 67727.99
Sai khac*
94,13 91,43 87,66 101,08
.• Sai khdc = Gid tri tinh lodn/Gid trj thuc te*IOO Bang 7 eho tha'y cac yeu to' nong do ethanol, nhiet do va pH deu anh hfldng den ham muc tieu d mfle y nghia = 0,001 trong dd pH la yeu to' anh hfldng manh nha't roi tdi nhiet do va nong do ethanol. Tuy nhien, chieu hfldng anh hfldng cua eae yeu td' nay de'n ham mue tieu thi khae nhau: trong gidi han nghien cflu, nong do ethanol va nhiet do anh hfldng dflOng de'n ham mue tieu cd nghia la khi tang nong do ethanol hoae tang nhiet do chiet, IflOng polyphenol thu dflOc tang; pH anh hfldng am de'n ham muc tieu, khi pH giam, lfldng polyphenol thu dflOe tang. Khi nhiet do tang, do nhdt eua dung dich giam, kha nang tham tha'u ciia dung mdi vao te bao va chie't polyphenol ra khdi te bao tang.
pH tha'p mdt mat giup lam ben cac hdp chat anthocyanin, mat khac kim ham sfl boat
ddng eiia enzyme polyphenol oxidase (Ruenroengkhn va es., 2008). Nong do ethanol tang lam tang hieu sua't thu polyphenol cd le dflde quye't dinh bdi ty le cac hdp chat phenol fla nflde va ky nflde cua vd vai.
Sfl tfldng tae gifla hai yeu to' trong eae yeu td' nghien cflu deu anh hfldng cd y nghia de'n ham muc tieu, dac biet sfl tfldng tae gifla pH va nong do ethanol, sfl tfldng tac gifla pH va nhiet do deu anh hfldng de'n ham muc tieu vdi mfle y nghia 0,001. Tfldng tae gifla nhiet do va pH anh hfldng manh nha't de'n ham muc tieu.
3.4. Kiem tra mo hinh
Tie'n hanh thi nghiem kiem tra tinh dung dSn ciia mo hinh d cac dieu kien trong khoang nghien cflu ciia cac yeu to' (Bang 8).
Lai Thj Nggc Ha, Nguyin Thi Thu Huong, Phan Thi HSng Ke't qua eho tha'y gia tri tinh toan bie'n
doi 87% de'n 101% so vdi gia tri thu dflde tren thflc te. Dieu nay phfl hdp vdi gia tri R^
thu dflde eiia md hinh.
4. KET LUAN
Vd vai ed ham Ifldng polyphenol cao (9,625% chat khd). Day la nguon nguyen lieu ddi dao cho san xua't dieh chie't polyphenol flng dung trong dflde pham va cdng nghiep thflc pham.
Cae yeu to' nhiet do, nong do ethanol va pH anh hfldng de'n hieu qua tach chie't polyphenol tfl vd vai. Md hinh md ta qua trinh tach chie't theo eae yeu td'tren nhfl sau:
Y = 62745,17 + 3946,87XSi -t- 6643,00X82 - 9949,96X83 - 1392,63X81X82 + 2718,14X81X83 + 9880,41X82X83.
Md hinh nay ed the sii dung dflde de tie'n hanh td'i flu hda dieu kien tach chie't polyphenol tfl vd vai.
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