luan van hoan chinh

Kha lun tt nghip

V Thu Quyn BQCBBK52

TRNG I HC NNG NGHIP H NI KHOA CNG NGH THC PHM

KHO LUN THC TP TT NGHIP TI:BC U NGHIN CU QUY TRNH TRCH LY GAMMA ORYZANOL T CM GO CH BIN THC PHM CHC NNGSinh vin thc hin: Lp: Gio vin hng dn: V THU QUYN BQCBBK52 1. ThS. NGUYN C TIN Vin C in nng nghip v Cng ngh sau thu hoch 2. TS. NGUYN TH BCH THY B mn Cng ngh sau thu hoch

H Ni 2011

Trng i hc Nng NghipHN

Khoa Cng ngh thc phm 1

Kha lun tt nghip

V Thu Quyn BQCBBK52

LI CAM OAN

Ti xin cam oan rng, s liu v kt qu nghin cu trong kha lun ny l trung thc. Ti xin cam oan rng, mi s gip cho vic thc hin kha lun ny c cm n v cc thng tin c trch dn trong kha lun ny c ghi r ngun gc. H Ni, thng 5 nm 2011

V Thu Quyn


Khoa Cng ngh thc phm

i

Kha lun tt nghip LI CM N

V Thu Quyn BQCBBK52

Kha lun tt nghip ny c thc hin ti Vin C in Nng nghip v Cng ngh sau thu hoch di s hng dn ca ThS. Nguyn c Tin Vin C in nng nghip v Cng ngh sau thu hoch v TS. Nguyn Th Bch Thy B mn Cng ngh sau thu hoch, Trng i hc Nng nghip H Ni. Trc ht ti xin gi li cm n chn thnh ti TS. Nguyn Th Bch Thy cng ton th cc thy c gio khoa Cng ngh thc phm tn tnh gip ti trong sut qu trnh hoc tp v lm kha lun. Ti xin by t lng bit n su sc i vi s hng dn tn tnh, chu o ca Th.S. Nguyn c Tin cng tt c cn b Phng Tn dng ph ph phm Vin C in Nng nghip v Cng ngh sau thu hoch to mi iu kin gip ti hon thnh kha lun ny. Cui cng, ti xin gi li cm n su sc ti gia nh v bn b quan tm, ng vin, gip ti trong sut thi gian qua. Ti xin chn thnh cm n! H Ni, thng 5 nm 2011 Sinh vin

V Thu Quyn


Khoa Cng ngh thc phm ii

Kha lun tt nghip

V Thu Quyn BQCBBK52

MC LCMC LC.......................................................................................................iii DANH MC BNG......................................................................................viii DANH MC HNH.........................................................................................ix DANH MC CH VIT TT V K HIU.................................................x Vit tt HSTL Phn I M U...........................................................................................................1 1.1. T VN .........................................................................................1 1.2. MC CH YU CU.........................................................................2 1.2.1. Mc ch...................................................................2 1.2.2. Yu cu....................................................................2 Phn II TNG QUAN TI LIU.................................................................................3 2.1. GII THIU V CM GO...................................................................3 2.1.1. Thnh phn ha hc ca cm go...........................3 2.1.2. Cng dng ca cm go..........................................7 Theo phn tch khoa hc, trong cm go cha rt nhiu cc vitamin nh B1, B6, PP v axt folic... Cm go c dng b sung vitamin B, c bit l B1 v b sung axt folic cho khu phn n ca ph n c thai, gip cho s pht trin h thn kinh ca thai nhi. Lng cht bo trong cm go rt cao (15-22%), thng dng chit xut du cm; cht m trn 12%, cht st trn 14%. Do cm c th cung cp nhiu cht dinh dng, tt cho sc khe con ngi [14]....................................................................7 Vitamin E tocopherol v tocotrienol, gamma oryzanol v phytosterol gip bo v chng li ung th, c cha cc cht t nhin c th chng li cc gc t do l nguyn nhn ca ung th, c hiu qu nh duy tr h thn kinh v kh nng no. Vitamin E trong cm go c th bo v v chng li bnh alzheimer, iu chnh h thng ni tit t cho ngi gi.....................................................................8 c l................................................................................................x Hiu sut trch ly................................................x


Khoa Cng ngh thc phm iii

Kha lun tt nghip

V Thu Quyn BQCBBK52

Nhiu nghin cu trn th gii chng minh s dng cm go c li cho sc khe: chng lo ha, n nh huyt p, tng sc khng, cn bng ng huyt, iu chnh h thng ni tit t cho ngi gi, hn ch c s pht trin ca t bo ung th Qureshi v cng s ti Vin nghin cu Y hc ti Madison Wisconsin cho thy bng cch tiu th 20 gam cm go tinh ch / ngi mi ngy, trong 60 ngy, c hai loi bnh i tho ng Type I v Type II, bnh nhn tiu ng c th gim mc ng huyt ca h xung 30% v 20%................................................8 2.2. GII THIU CHUNG V GAMMA ORYZANOL.................................9 2.2.1. Khi nim Gamma oryzanol...................................................................9 2.2.2. Tc dng ca Gamma oryzanol.............................11 2.3. TNH HNH SN XUT, TIU TH V NGHIN CU GAMMA ORYZANOL T CM GO TRN TH GII...........................................13 2.4. TNH HNH SN XUT, TIU TH V NGHIN CU GAMMA ORYZANOL T CM GO TRONG NUC.............................................15 2.7. CC PHNG PHP TRCH LY HOT CHT [1].............................17 2.8. CC YU T NH HNG N QU TRNH TRCH LY CM GO .....................................................................................................................19 2.8.1. 2.8.2. 2.8.3. 2.8.4. 2.8.5. Phn III S lng dung mi.................................................19 Loi dung mi........................................................19 Mt ca cm go...............................................19 m....................................................................19 Nhit .................................................................20

I TNG - NI DUNG V PHNG PHP NGHIN CU............................................................21 3.1. I TNG, VT LIU, A IM & THI GIAN NGHIN CU..21 3.1.1. i tng nghin cu............................................21 3.1.2. Vt liu nghin cu................................................21 3.1.3. a im v thi gian nghin cu..........................22 3.2. NI DUNG NGHIN CU...................................................................22


Khoa Cng ngh thc phm iv

Kha lun tt nghip

V Thu Quyn BQCBBK52

3.2.1. Nghin cu quy trnh trch ly Gamma oryzanol t cm go...........................................................................22 3.3. PHNG PHP NGHIN CU...........................................................23 3.3.1. B tr th nghim..................................................................................23 3.3.1.1. X l cm go nguyn liu trc khi trch ly.....23 3.3.1.2.Xc nh loi dung mi nh hng n hiu sut trch ly..............................................................................23 Qu trnh trch ly nh hng rt nhiu yu t cng ngh: loi dung mi trch ly, nng dung mi, nhit , t l dung mi, thi gian trch ly Mt trong s nhng yu t nh hng l loi dung mi no c nh hng ti hiu sut trch ly gamma oryzanol. Nhng dung mi dng chit xut du cn phi c mt im si tng i thp v khng d chy, khng c trong c hai cht lng v hi nc. Tuy nhin, cn chn loi dung mi an ton cho sc khe ngi s dng, r tin, khng c nguy c chy n. 23 3.3.1.3. Xc nh nng trch ly nh hng n hiu sut trch ly......................................................................24 3.3.1.4.Xc nh nhit trch ly nh hng n hiu sut trch ly..............................................................................25 3.3.1.6. Xc nh thi gian trch ly n hiu sut trch ly .........................................................................................26 .........................................................................................................................27 3.3.2. Cc ch tiu nghin cu.........................................28 3.3.2.1. Xc nh hm m ca cm go nguyn liu theo phng php sy kh n khi lng khng i.............28 Do cm go trng thi xp, t bo ph v, c h enzym lipase ni ti, cng s xm nhp ca mi trng, nn phn ng thy phn v oxi ha ca du trong cm pht trin nhanh. S thy phn du to ra cc acid bo t do cao lm cho cm b i hng, chua kht khng thun li cho qu trnh ch bin sau ny. Chnh v vy, tng tr cm go cho ch bin cng nh chit xut gamma oryzanol cn c phng php bo qun cm go cho trch ly gamma oryzanol bng phng php sy kh n khi lng khng i...............................................................28 * Cch tin hnh..............................................................28 Cn 100 g cm nguyn liu trong hp nhm bit trc trng lng. t hp nhm c cha mu vo t sy


Khoa Cng ngh thc phm v

Kha lun tt nghip

V Thu Quyn BQCBBK52

nhit 105oC. Sau 3 gi em i lm ngui trong bnh ht m v cn. Tip theo li t hp cng mu vo t sy v sy tip khong t 30 60 pht ri li em l ngui trong bnh ht m. Sau em cn li, nu sai s gia hai ln cn khng qu 0.01 g th qu trnh sy xem nh kt thc..................................................................................28 * Cch tnh.......................................................................28 Hm m ca nguyn liu c tnh theo cng thc sau: 28 W = 100%......................................................................28 Trong :..........................................................................28 a: Khi lng hp nhm cng nguyn liu trc khi sy (g) .........................................................................................28 b: Khi lng hp nhm cha nguyn liu sau khi sy (g) .........................................................................................28 c: Khi lng hp nhm kh khng cha nguyn liu (g)28 3.3.2.2. Xc nh hm lng gamma oryzanol................28 3.3.3. Phng php x l s liu......................................30 Phn IV KT QU V THO LUN.........................................................................31 4.1. X L CM GO NGUYN LIU TRC KHI TRCH LY .........................................................................................32 4.1.1. Nghin cu xc nh phng php x l cm nguyn liu cho trch ly gamma ozyzanol t cm go....32 Dung mi ho tan cc cht cn trch ly v gi li cc tp cht, dung mi c nh hng ln ti cht lng ca dch chit v thnh phm, v vy vic la chn dung mi thch hp l ht sc quan trng. Da trn tnh cht vt l ( nht, sc cng b mt, phn cc) v c tnh ha tan chn lc ca dung mi i vi hot cht la chn dung mi. Nhng dung mi dng chit xut du cn phi c mt im si tng i thp v khng d chy, khng c trong c hai cht lng v hi nc. Tuy nhin, cn chn loi dung mi an ton cho sc khe ngi s dng, r tin, khng c nguy c chy n. Ngoi ra qua ti liu cho bit gamma oryzanol l hp cht khng phn cc nn chng ti tin hnh th nghim kho st vi cc loi dung mi khng phn cc: n hexan, isopropyl alcohol, cn ethanol 96o...............................................................34 Bng 4.2. nh hng ca loi dung mi n hiu sut trch ly..............................................................................34


Khoa Cng ngh thc phm vi

Kha lun tt nghip

V Thu Quyn BQCBBK52

.........................................................................................................................43 - Dch trch ly thu c em i lc qua vi mn v qua giy lc bn to, loi b ht b cm go, tp cht v cn xt li trong dch. Sau a vo h thng c chn khng, thu hi dung mi quy m 70 lt/m. Thu c dch gamma oryzanol c ti khi lng khng i chnh l du th gamma oryzanol v thu hi cn ethanol.................................................................................................45 Phn V.............................................................................................................46 KT LUN V NGH.............................................................................46 5.1. KT LUN...............................................................................................46 Sau thi gian thc hin ni dung ca ti chng ti rt ra kt lun sau:.....46 1. X l cm go nguyn liu thch hp cho qu trnh trch ly dit men lipase 100 150oC/ 10 pht, sau sy 60o/ 2 gi t hm m 5.3% ri em trch ly. ....................................................................................................................46 TI LIU THAM KHO..............................................................................48 14. Hegsted, M., and M.M. Windhauser, Reducing Human Heart Disease Risk with Rice Bran, Louisiana Agriculture (1993)..............................................................................49 15. Nicolosi, R.J., L.M. Ausman, and D.M.Hegsted, Rice Bran Oil Lowers Serum Total and Low Density Lipoprotein Cholesterol and Apo B Levels in Nonhuman Primates, Atherosclerosis (1991).....................................................49 16. Rukmini, C., Chemical, Nutritional and Toxicological Studies of Rice Bran Oil, Food Chem (1988)....................49 17. Sharma, R.D., and C. Rukmini, Rice Bran Oil and Hypocholesterolemic in Rats, Lipids 21:715-717 (1986). 50 Du th trch ly t cm go.............................................................................51 ..................................................................................................................51 Qu trnh kt tinh Gamma oryzanol...............................................................51 nh hng ca thi gian sy ti cht lng cm sau sy................................55


Khoa Cng ngh thc phm vii

Kha lun tt nghip

V Thu Quyn BQCBBK52

DANH MC BNG Bng 2.1. Thnh phn x v gi tr dinh dng ca cc nguyn liu............................................................4 Bng 2.3. Thnh phn ca du cm go th...............5 Bng 4.1. nh hng thi gian sy n cht lng cm go sau sy........................................................33 Bng 4.3. nh hng nng dung mi cn ethanol n.............................................................................36 hiu sut trch ly........................................................36 Bng 4.4. Nhit trch ly nh hng n hiu sut trch ly........................................................................38 Bng 4.5. nh hng ca t l dung mi cn ethanol 96o n hiu sut trch ly................................................39 Bng 4.6. Thi gian trch ly nh hng ti hiu sut trch ly........................................................................41


Khoa Cng ngh thc phmviii

Kha lun tt nghip

V Thu Quyn BQCBBK52 DANH MC HNH

Hnh 2.1. S lp cm go trong ht la Hnh 2.2. Cm go............................................................4 Hnh 2.3. Cu trc phn t gamma oryzanol...........9 Hnh 2.4. Cu trc phn t ca 24-methylene cycloartenyl ferlulate..............................................10 Hnh 2.5. Cu trc phn t ca ferlulate cycloartenyl ...............................................................................10 Hnh 2.6. Cu trc phn t ca ferlulate campesteryl............................................................10 Hnh 2.7. Gamma oryzanol.....................................15 th 4.1. nh hng thi gian sy n cht lng cm go sau sy.....................................................52 th 4.2. nh hng ca loi dung mi n hiu sut trch ly.............................................................52 ..............................................................................53 th 4.4. nh hng ca nhit n hiu sut trch ly....................................................................53 th 4.5. nh hng ca t l dung mi cn ethanol 96o n.....................................................54 hiu sut trch ly.....................................................54 th 4.6. nh hng ca thi gian trch ly n hiu sut trch ly.............................................................54


Khoa Cng ngh thc phm ix

Kha lun tt nghip

V Thu Quyn BQCBBK52

DANH MC CH VIT TT V K HIU Vit tt HSTL LHCT LHCNL c l Hiu sut trch ly Lng hot cht tinh Lng hot cht c trong nguyn liu


Khoa Cng ngh thc phm x

Kha lun tt nghip Phn I

V Thu Quyn BQCBBK52

M U1.1. T VN Vit Nam l mt nc c nn nng nghip lu i gn lin vi nn vn minh la nc nn t lu cy la tr thnh cy lng thc ch yu c ngha quan trng trong i sng v nn kinh t x hi nc ta. Trong nhng nm gn y, nc ta tr thnh mt trong ba nh xut khu go hng u th gii. Qu trnh ch go to ra mt lng ln ph ph phm l cm go. Cm go c gi thnh thp thng c ngi dn s dng lm thc n chn nui, cha khai thc c ht tim nng v gi tr vn c ca n. Nu lu s b i hng, gy lng ph ln. Cm go c cha nhiu cht dinh dng qu gi nh protein, lipit, glucocide, vitamin nhng khng nhiu ngi bit rng cm go cha nhiu cht chng oxi ha c bit quan trng l gamma oryzanol c li cho sc khe cha bnh v c s dng lm m phm c gi tr kinh t cao. Gamma oryzanol c trong cm go, thc ny mm, cm i mch, ng ny mm nhng ch trong cm go mi c hm lng gamma oryzanol nhiu hn c, cn cm i mch, ng ny mm c lng gamma oryzanol rt thp. Hin nay ch phm gamma oryzanol c s dng cho sn xut cc sn phm thc phm, m phm, dc phm cha gamma oryzanol u phi nhp khu. Vit Nam hu nh cha c cng trnh nghin cu no v tch chit gamma oryzanol t cm go, nguyn liu ny phn ln vn cn nhp khu t Trung Quc v cc nc khc. Do vy vic nghin cu s dng cm go sn xut gamma oryzanol phc v cho cng ngh thc phm, ch bin thc phm chc nng l ht sc quan trng.



Kha lun tt nghip

V Thu Quyn BQCBBK52

Nhn thy c s cn thit ca vn nu trn, c s ng ca Vin C in nng nghip Cng ngh sau thu hoch v b mn Cng ngh sau thu hoch Khoa Cng ngh thc phm Trng i hc Nng nghip H Ni, chng ti tin hnh nghin cu ti: Bc u nghin cu quy trnh trch ly gamma oryzanol t cm go ch bin thc phm chc nng. 1.2. MC CH YU CU 1.2.1. Mc ch Xy dng quy trnh trch ly v thu nhn gamma oryzanol t cm go ch bin thc phm chc nng. 1.2.2. Yu cu - Nghin cu ch x l cm go nguyn liu trc khi trch ly. - Xc nh loi dung mi, nng dung mi n hiu sut trch ly gamma oryzanol t cm go. - Nghin cu nh hng ca nhit , thi gian trch ly n hiu sut trch ly gamma oryzanol. - Xy dng quy trnh trch ly gamma oryzanol t cm go.



Kha lun tt nghip Phn II

V Thu Quyn BQCBBK52

TNG QUAN TI LIU2.1. GII THIU V CM GO 2.1.1. Thnh phn ha hc ca cm go Cm go l ph phm chnh thu c t la sau khi xay xt v thng chim khong 10% trng lng la. Cm go c hnh thnh t lp v ni nh, mm phi ca ht, cng nh mt phn t tm. Cm go c mu sng v mi thm c trng. T l protein trong cm go mn c th t 12 14 %. Lng protein th cm go cao hn so bp ht. Lng cht bo trong cm go rt cao (15 22 %), thng dng chit xut du cm, cht m trn 12%, cht st trn 14 %. Trong cm go ngoi nhng thnh phn c gi tr dinh dng nh protein, lipit, glucocide, vitamin trong cm go cn cha hn hp cht c bit quan trng l gamma oryzanol. Gamma oryzanol c trong cm go, thc ny mm, cm i mch, ng ny mm ch trong cm go mi c hm lng gamma oryzanol nhiu hn c (1.0 4.0 % trng lng), cn cm i mch, ng ny mm c hm lng gamma oryzanol rt thp [7,15, 19].



Kha lun tt nghip Hnh 2.1. S lp cm go trong ht la

V Thu Quyn BQCBBK52 Hnh 2.2. Cm go

Bng 2.1. Thnh phn x v gi tr dinh dng ca cc nguyn liu Cm go Cm go Cm la nguyn Trch La m Bt m m du du 13 3.100 8 19 15 5 4 9 (96) 15 2.250 11 27 21 7 6 11 (97) 12 3.350 2,5 10,5 9,5 2,5 1 5,5 (77) 16 2.520 11 44 38,2 11 5,8 21 (99) 16 2.965 9 27 23,5 8 3,5 15 (97)

Ch tiu Protein (%) DE (Kcal/kg) X th (%) X tng s (%) NSP tng s (%) Cellulose (%) Lignin (%) Arabinoxylan (%) (% khng ho tan)

Bp 8 3.525 2,2 9,5 9 2,0 0,5 3,7 (94)

(ngun: Gene v cng s, 2002)



Kha lun tt nghip

V Thu Quyn BQCBBK52

Bng 2.2. Thnh phn ha hc ca cm go Thnh phn Protein th (% N.6,25) Cht bo th (%) Si th (%) Carbohydrate (%) Tro th (%) Cm 12,0 15,6 15,0 19,7 7,0 11,4 31,1 52,3 6,6 9,9 Cht khong Canxi (mg/g) Magie (mg/g) Photpho (mg/g) Phytin photpho (mg/g) Silica (mg/g) Km (mg/g) 0,3 1,2 5,0 13,0 11,0 25,0 9,0 11,0 6,0 11,0 43,0 25 Vitamin Vitamin B1 Vitamin B2 Niaxin 12,0 24,0 1,8 4,3 267 499 3,0 19,0 1,7 2,4 224 389 0,5 0,7 6,0 7,0 10,0 22,0 12,0 17,0 2,0 3,0 17,0 90,0 Cm trng 11,8 13,0 10,1 12,4 2,3 3,2 51,1 55,0 5,2 7,3

Bng 2.3. Thnh phn ca du cm go th Thnh phn Lipid c th ha x phng Trung ha lipit Phn trm (%) 90 96 88 89



Kha lun tt nghip

V Thu Quyn BQCBBK52

Cht bo Diglycerides Cht bo Monglycerides Acid bo t do Cc loi sp Glycolipids Phospholipid Cht khng x phng ha lipid Phytosterol Este sterol Ri Triterphene Hydrocacbon Tocopherols (ngun: Cheruvanky v cng s, 2003)

34 67 24 34 67 45 4,2 43 10 28 18 1

Theo bng 2.2, cm go c lng dinh dng rt cao vi lng cht bo cha bo ha cao, vitamin nhm E, nhm B, phylate, km, canxi, kali u rt cao. Ngoi ra trong cm cn c cht bo Omega 3 kh cao. Thnh phn ca cm go c nhiu vitamin v cht bo, cn i v c nhiu x d tiu rt tt cho con ngi. Tuy nhin, cm thng c dng lm thc n cho cc loi gia sc v thy sn ch khng dng cho ngi. Nguyn nhn l do cm c mt s enzym ni ti hot ng rt mnh m lm oxi ha cc cht bo v cm go bi i ha c mi kh chu. Thm vo , do cng ngh xay xt go cha cao li t c u t theo hng thu cm sch nn cm thng ln rt nhiu tp cht (v tru, sn ). V nhng l do trn m hu ht lng cm go thu c thng c nng dn s dng nui cc loi gia sc, gia



Kha lun tt nghip

V Thu Quyn BQCBBK52

cm. Hng nm trn th gii c khong 40 45 triu tn cm c sn xut v 90% l chu . Cm go cng nh cc nguyn liu c ngun gc thc vt khc thng cha hm lng pht pho kh cao dng phytate. Mc khc, gc pht pht t phytate thng to lin kt vi cc cht nh axt amin v cht khong lm gim s tiu ha cc dng cht ny khi b sung vo khu phn. Thng thng c khong 2/3 hm lng pht pho c trong nhng loi nguyn liu th c s dng lm thc n gia sc. Cm go c lng pht pho kh cao nhng trn 50% l dng phytate. ng vt c d dy n kh tiu ha cht ny do khng sn xut lng enzyme phytase ni sinh cn thit. Mc d nhiu bo co khoa hc cho thy khu phn x cao mang nhiu tin li nh lm gim s lot d dy v pht trin vi sinh vt, gip tha mn cn i (i vi ln ni trong thi k mang thai), sn xut nhiu acid bo bay hi c bit l acid acetic c tng hp thnh m sa v cung cp sa nng lng cao hn, x cn l cht n gii quyt khi lng vt cht kh trong khu phn, kch thch tiu ha thc n v bi thi cht c hi ra ngoi c th, kch thch s pht trin ca ng tiu ha. 2.1.2. Cng dng ca cm go Theo phn tch khoa hc, trong cm go cha rt nhiu cc vitamin nh B1, B6, PP v axt folic... Cm go c dng b sung vitamin B, c bit l B1 v b sung axt folic cho khu phn n ca ph n c thai, gip cho s pht trin h thn kinh ca thai nhi. Lng cht bo trong cm go rt cao (15-22%), thng dng chit xut du cm; cht m trn 12%, cht st trn 14%. Do cm c th cung cp nhiu cht dinh dng, tt cho sc khe con ngi [14].



Kha lun tt nghip

V Thu Quyn BQCBBK52

Vitamin E tocopherol v tocotrienol, gamma oryzanol v phytosterol gip bo v chng li ung th, c cha cc cht t nhin c th chng li cc gc t do l nguyn nhn ca ung th, c hiu qu nh duy tr h thn kinh v kh nng no. Vitamin E trong cm go c th bo v v chng li bnh alzheimer, iu chnh h thng ni tit t cho ngi gi. Nhiu nghin cu trn th gii chng minh s dng cm go c li cho sc khe: chng lo ha, n nh huyt p, tng sc khng, cn bng ng huyt, iu chnh h thng ni tit t cho ngi gi, hn ch c s pht trin ca t bo ung th Qureshi v cng s ti Vin nghin cu Y hc ti Madison Wisconsin cho thy bng cch tiu th 20 gam cm go tinh ch / ngi mi ngy, trong 60 ngy, c hai loi bnh i tho ng Type I v Type II, bnh nhn tiu ng c th gim mc ng huyt ca h xung 30% v 20%. t rt lu cm go c coi l mt b quyt sc p ca ngi ph n ng ni chung, ngi Nht Bn, Hn Quc v c bit l ngi Vit Nam ni ring. iu lm nn iu k diu ca cm go l gamma oryzanol, mt cht chng oxi ho hng u, gip ngn nga qu trnh lo ho trong c th. gamma oryzanol l thnh phn c bit ca cm go, c c t chit xut du th ca cm go (v la ca ht go). Ngi ta bit n gamma oryzanol u tin t nhng nm 1950 v vo u nhng nm 1960, ngi Nht bit s dng gamma oryzanol nh mt loi thuc cha bnh.Cng dng ni bt nht ca gamma oryzanol l cht chng oxi ho mnh, gip lm chm ng k qu trnh lo ho ca c th. Mc d vitamin E ni ting l mt cht chng oxi ho hiu qu nhng cc nghin cu gn y chng minh gamma oryzanol cn c tc dng gp 4 ln vitamin E trong vic ngn chn cc m b oxi ho. Ngoi ra, gamma oryzanol cn bo v c th khi tia cc tm, do ngi ta cn s dng gamma oryzanol trong cc loi m phm nh



Kha lun tt nghip

V Thu Quyn BQCBBK52

kem chng nng, gip bo v ln da khi cc h tn do oxi ho gy ra di tc ng ca mi trng bn ngoi [12]. 2.2. GII THIU CHUNG V GAMMA ORYZANOL 2.2.1. Khi nim Gamma oryzanol Gamma oryzanol c cng thc phn t l: C40H55O4, trng lng phn t 602,89 g/mol, b nng chy nhit 135 137 oC, gamma oryzanol bao gm 2 phn, phn mch di l sterol v phn cn li l axit ferulic [4].

Hnh 2.3. Cu trc phn t gamma oryzanol Trng thi vt l: gamma oryzanol dng bt, khng mi, ha tan trong nc, t tan trong ete diethyl v heptane. Tan nhiu trong isopropyl v ha tan trong clorofom. gamma oryzanol c mu trng hoc vng.



Kha lun tt nghip

V Thu Quyn BQCBBK52

Hnh 2.4. Cu trc phn t ca 24-methylene cycloartenyl ferlulate Hnh 2.5. Cu trc phn t ca ferlulate cycloartenyl Hnh 2.6. Cu trc phn t ca ferlulate campesteryl Ba trong s cc phn phn on, cycloartenyl ferulate, 24methylenecycloartenyl ferulate v campesteryl ferulate, l thnh phn chnh ca gamma oryzanol [4, 8]. Gamma oryzanol l cht chng oxi ha mnh m ch tm thy trong cm go. Gamma oryzanol khng phi l mt cht chng oxy ha n nhng l hn hp ca 20 thnh phn c tnh cht chng oxi ha. Trong cm go ngoi nhng thnh phn c gi tr dinh dng nh protein, lipit, glucocide, vitamin , trong cm go cn cha hn hp cht c


Khoa Cng ngh thc phm10

Kha lun tt nghip

V Thu Quyn BQCBBK52

bit quan trng l gamma oryzanol. Gamma oryzanol c trong cm go, thc ny mm, cm i mch, ng ny mm ch trong cm go mi c hm lng gamma oryzanol nhiu hn c (1,0 4,0 % trng lng), cn cm i mch, ng ny mm c lng gamma oryzanol rt thp [8]. 2.2.2. Tc dng ca Gamma oryzanol Khoa hc hin i khm ph gamma oryzanol - vi cht nm lp mng bao quanh ht go - c tc dng gim cholesterol tha trong c th. y c xem l mt pht hin quan trng gp phn ngn nga nguy c bnh tim mch. T nhng nm 1950, nhiu nghin cu khoa hc ch ra rng b n ca ht go lt nm chnh lp mng go bao quanh n, vn cha mt lng ln vitamin, khong cht, cht bo c li v c vi cht gamma oryzanol - c tc dng lm chm qu trnh lo ha ca t bo, xy dng c bp, chng trm cm... Bc s ni ting ngi Nht Sakurazawa Nyoichi cn ra hn phng php thc dng - cha bnh bng go lt, sau ny c T chc Y t th gii (WHO) cng nhn l mt phng php phng v cha bnh hiu qu. n nhng nm 1980, gii khoa hc khm ph gamma oryzanol trong lp mng go thm ch cn c kh nng gim cholesterol tha trong mu, bo v c th khi bnh tim mch. Pht hin mi ny tht s c ngha vi cng ng bi tha cholesterol ang l thc trng ng bo ng chung trong cuc sng hin i. Theo iu tra mi nht ca Vin Dinh dng quc gia va cng b ti hi tho "Tnh trng cholesterol cao ngi Vit Nam v gii php d phng" gn y, bnh qun t l ngi Vit Nam c lng cholesterol cao l 29,1%, trong t l ny ngi dn thnh th ln ti 44,3% [10].



Kha lun tt nghip

V Thu Quyn BQCBBK52

2.2.2.1. Tc dng gim cholesterol tha ca gamma oryzanol Tha cholesterol xp vo nguy c hng u gy ra bnh tim mch, v tng c nghin cu cho thy gim cholesterol tha trong c th s gim c 30% t l t vong do bnh ng mch vnh. iu tr tha cholesterol trong mu s gip hn ch s hnh thnh v tin trin ca cc mng va x, t gim cc tai bin v t vong do bnh l tim mch. Nhiu nghin cu lm sng trn th gii khng nh gamma oryzanol c kh nng ngn chn rut hp thu cholesterol t thc n v cholesterol do gan tit ra, sau o thi chng khi c th. Cc chuyn gia tim mch trn th gii cng khuyn ngi tha cholesterol nn dng thc phm cha gamma oryzanol nh mt cch t nhin a cholesterol v mc "chun", ngn nga nguy c bnh tim mch. y l mt bin php n gin v an ton, d thc hin. Cc nghin cu lm sng cho thy c hiu qu gim cholesterol tha th mt ngi nn dng t nht 50 mg gamma oryzanol/ngy [11]. 2.2.2.2. Tc dng ngn nga qu trnh lo ha ca gamma oryzanol Li ch ca gamma oryzanol, b quyt ngn nga qu trnh lo ha trong du go gamma oryzanol, thnh phn c bit ca du go, c c t chit xut du th ca cm go (v la ca ht go). Ngi ta bit n gamma oryzanol u tin t nhng nm 1950 v vo u nhng nm 1960, ngi Nht bit s dng gamma oryzanol nh mt loi thuc cha bnh. Ban u, gamma oryzanol c dng cha tr cho cc bnh nhn c biu hin lo lng. n cui nhng nm 1980, tr liu gamma oryzanol c cng nhn cho php dng trong vic cha tr cc nhng trng hp b cholesterol trong mu cao...[13].



Kha lun tt nghip

V Thu Quyn BQCBBK52

2.2.2.3. Tc dng chng oxi ha ca gamma oryzanol Gn y, nhiu nghin cu cho thy cng dng ni bt nht ca gamma oryzanol l cht chng xy ha mnh, gip lm chm ng k qu trnh lo ha ca c th. Mc d Vitamin E ni ting l mt cht chng xy ha hiu qu nhng cc nghin cu gn y chng minh gamma oryzanol cn c tc dng gp 4 ln Vitamin E trong vic ngn chn cc m b xy ha. Ngoi ra, gamma oryzanol cn bo v c th khi tia cc tm, do ngi ta cn s dng gamma oryzanol trong cc loi m phm nh kem chng nng, gip bo v ln da khi cc h tn do xy ha gy ra di tc ng ca mi trng bn ngoi [15, 17, 18]. Ngoi nhng cng dng tt cho sc khe, gamma oryzanol c trong du go cn c tc dng d tiu ha v gip ngon ming khi trn vi salad; to hng v nh nhng c bit thch hp cho mn rn, nng m khng lm thay i mi v ca thc phm sau khi nu. Du go c sc chu nhit v im bc khi rt cao (khong 246oC), do vy rt ph hp rn, chin, xo m khng lm thc n tr nn kh cng, du b bc khi, hay si bt. Loi du go c th tm thy trn th trng l Neptune 1:1:1 [3]. 2.3. TNH HNH SN XUT, TIU TH V NGHIN CU GAMMA ORYZANOL T CM GO TRN TH GII Hin nay trn th gii nh: Nht Bn, M, Trung Quc, n , Thai Lan, cng ngh sn xut gamma oryzanol a tao ra ch phm gamma oryzanol thng mai, lam nguyn liu cho ch bin thc phm, dc phm va my phm. c nhiu cng trnh nghin cu v gamma oryzanol cng nh v hot cht sinh hc ca n, v kh nng s dng gamma oryzanol trong vic bo v, phng chng mt s bnh tt cho con ngi. Gamma oryzanol c tc dng chng oxy ho, chng vim lot, c bit l vim lot d dy, lm gim lng m trong mu, c hot tnh ging nh proestrogen, gamma oryzanol



Kha lun tt nghip

V Thu Quyn BQCBBK52

cn c kh nng lm tng vic lu thng mu, gim cholesterol trong mu, giam m trong c th, l hot cht c tc dng chng acid ha, ngn chn c s xm nhp ca tia cc tm, cn tr hot ng bi tit sc t melanin trong biu b, co tc dng phng chng nm daHin nay oryzanol t cm go c ng dng lm nguyn liu trong dc phm, my phm, sn xut sn phm thc phm b sung chng lao hoa cho phu n, lam ep, lam min mang da, lam giam cac triu chng bc hoa va cng thng, s thay i bt thng cua phu n thi ky man kinh va thi ky sau o (vin gamma oryzanol thc phm b sung, sa dng da, kem chm sc da) [7, 14, 18]. Sn phm gamma oryzanol ch t cm go dng thc n ca hng Sunfarmnhi Australia v mt s hng khc trn th gii c bn rng ri ti th trng Ty u. Cc nc khng trng c la nc hin ang nhp khu cm go sn xut cc ch phm c gi tr cao. Nh vy r rng cm go tr thnh mt mt hng c gi tr. Cng trnh khoa hc ca Ishibaski Kyohide (Nht Bn) nghin cu dng gamma oryzanol sn xut m phm bi da c cha 3% gamma oryzanol chng vim da v chng lo ho da cng l bng chng na ni ln ng dng rng ri ca gamma oryzanol cm go i vi sn xut v i sng [16]. Hin nay a c hn 40 bng sng ch v chit xut gamma oryzanol cam gao a a ra t cac nc san xut lua gao nh Trung Quc, M, Nht bn, n , Thi Lan. Nhng hiu ng sinh l hc c ch ra lin quan n gamma oryzanol lam gim hm lng colesteron, s kt t tiu cu, s tng hp sinh hc cholesteron v s hp th cholesteron. Gamma oryzanol cng c s dng tng s bi tit axit trong mt v trong iu chnh s khng cn bng dy thn kinh v nhng s mt cn bng cua thi k mn kinh.



Kha lun tt nghip

V Thu Quyn BQCBBK52

Trn th trng hin nay gamma oryzanol c bn di dng vin nang dng du hoc bt xut x ch yu t Nht Bn c tn thng mi gama oryzanol, gama-O 32[7].

Hnh 2.7. Gamma oryzanol 2.4. TNH HNH SN XUT, TIU TH V NGHIN CU GAMMA ORYZANOL T CM GO TRONG NUC Hin nay ch phm gamma oryzanol c s dung cho san xut my phm, dc phm va cac san phm thc phm cha gamma oryzanol u phai nhp khu. S thay i v li sng, ch n ung khin ngi dn phi i mt nhiu hn vi cc bnh mn tnh nh tim mch, ung th, tiu ng Ch dinh dng nhiu cht bo, gim cht x cng vi s nhim mi trng ngy cng tng ln l nguyn nhn lm cho c th ca chng ta d gp phi cc nguy c bnh tt. Mt khc, do nn kinh t pht trin, ngi dn, c bit l ngi dn thnh ph H Ni c mc sng cao nn h khng ch quan tm n cc san phm lam ep, thc phm tng cng sc kho, hoc c tc dng i vi chc nng no vi c th. Chnh v vy, vi nhng sn phm c qung co l cha bch bnh, ko di tui thanh xun, nh nc ung Noni,... c ngi tiu dng rt hng ng mc d cc loi thc phm ny c



Kha lun tt nghip

V Thu Quyn BQCBBK52

gi thnh rt cao m tc dng tht ca sn phm cha hn ng vi qung co v thc phm chc nng khng c tc dng cha bnh nh thuc hoc cc loi dc phm [12]. Ch phm gamma oryzanol hin s dung lam nguyn liu cho san sut my phm, dc phm va thc phm trong nc la phai nhp khu. Trong nc cha co c s nao san xut ch phm gamma oryzanol. Trong khi o ngun cam gao nguyn liu co th s dung cho san xut ch phm gamma oryzanol di dao. Cam gao trong nc chu yu s dung lam thc n chn nui va xut khu. Cha khai thac c gia tri tim nng to ln cua no. Cha co iu tra anh gia ham lng gamma oryzanol trong cam gao nguyn liu cac loai khac nhau trong nc. Tuy khng nhiu bng Trung quc, Nht ban, My nhng chng ta cng c mt s cng ngh trich ly hoat cht sinh hoc t nng san tao ra cac ch phm cho san xut thc phm, to ra c sn phm thng mi. Cc sn phm ny c hiu qu mt s chc nng nh: chc nng gim bo, chng mt mi, sng mt, li tiu, mt gan, gim hm lng ng trong mu. Mt s cc loi nguyn liu c tc dng chng oxy ho, c sn trong nc nh: cy tt l, mp ng, c rt, cy nhu, ti, gc hoc cc loi nguyn liu c tc dng tng cng sinh lc nh g c, cu gai, nhm bin, hi sm c khai thc sn xut thc phm chc nng, ch phm san xut my phm. Cng ngh trch ly cc thnh phn chc nng t nguyn liu to ra cc ch phm c hot cht sinh hc cao nh ch phm isoflavon, glycosid hoc cng ngh s dng enzim thu phn protein thnh cc peptit chc nng, hoc cng ngh sn xut cc ng chc nng nh FOS, xyliton theo con ng vi sinh vt c tin hnh trong phng th nghim [10, 12, 13]. Vit Nam cha co cng ngh san xut gamma oryzanol t cam gao, cam gao mi dng lai cho ch bin thc n chn nui, san xut du cam



Kha lun tt nghip

V Thu Quyn BQCBBK52

gao cha khai thac c gia tri tim nng vn co cua no. Nm 2008 Vin C in nng nghip va cng ngh sau thu hoach, Nguyn c Tin va cng s trong phong thi nghim, trich ly cam gao co c dich ch phm oryzanol quy m phong thi nghim. 2.7. CC PHNG PHP TRCH LY HOT CHT [1]. Trch ly l qu trnh rt cht ha tan trong cht lng hay cht rn bng mt cht lng khc. Trch ly cht ha tan trong cht lng c gi l trch ly lng, cn trch ly ha tan trong cht rn gi l trch ly rn [1]. L thuyt v qu trnh trch ly c nghin cu v pht trin t rt sm khi ngnh cng nghip ha cht bt u pht trin. Ngy nay, cng ngh trch ly c s dng rng ri trong cng nghip, c bit trong cng ngh chit xut cc hp cht thin nhin nh tinh du, nha du. Bn cht ca qu trnh trch ly hot cht trong dc liu bng dung mi l qu trnh vn chuyn vt cht (s chuyn khi) trong h pha rn lng, trong dung mi l pha lng, dc liu l pha rn. Qu trnh chuyn khi rt phc tp v c s c mt ca mng cht nguyn sinh. Ngoi ra hin tng ha tan, khuch tn cn l hin tng thm thu, thm tch. Khi cho dc liu thc vt kh chia nh tip xc vi dung mi, qu trnh chit xut din ra theo c ch sau: - Dung mi thm qua cc mao qun vo cc t bo dc liu, thi gian thm dung mi ph thuc vo ng knh, chiu di mao qun, p lc khng kh, bn cht dung mi. - Thm t dung mi ca cc cht trong t bo dc liu. Qu trnh ny ph thuc vo tnh cht ha l ca cc cht, bn cht dung mi. - Ha tan cc cht trong t bo vo dung mi. Qu trnh ha tan ph thuc vo bn cht ha hc ca cc cht tan v dung mi, cc cht c nhiu nhm phn cc (- OH, - COOH) d tan trong



Kha lun tt nghip

V Thu Quyn BQCBBK52

dung mi phn cc (nc, ethanol, isopropanol thp ). Cc cht c nhiu nhm thn du (cht bo, CH3 C2H5 v ng ng) d tan trong dung mi khng phn cc. Khuch tn cc cht tan trong t bo dc liu vo dung mi gm hai loi: - Khuch tn cht tan trong t bo dc liu qua mng t bo gi l khuch tn phn t hay khuch tn ni. Lng cht khuch tn ph thuc vo yu t: s chnh lch nng , din tch b mt, thi gian, nhit , nht dung mi, b dy ca lp khuych tn. - Khuch tn cht tan t dc liu vo dung mi gi l khuch tn i lu hay khuch tn ngoi. Khuch tn i lu nhanh hn khuch tn phn t 2 3 ln v khng c s tham gia ca mng t bo. Nh vy, chit rt cc hot cht t dc liu mt cch hon ton v nhanh nht th ngoi vic la chn dung mi phi to cc iu kin ti u nht cho qu trnh khuch tn. Mt h trch ly c th l: - Trch ly lng rn - Trch ly lng lng Qu trnh trch ly Gamma oryzanol t cm go, cm go c ngm chit nhit , t l dung mi v thi gian xc nh, chuyn Gamma oryzanol t cm go vo dung mi bng cch khuch tn phn t v khuch tn i lu, trong thi gian trch ly cm go c lin tc ngm trong dung mi, hn hp trch ly c thu hi nhiu ln, ui dung mi thu hi du cm go th Gamma oryzanol.



Kha lun tt nghip

V Thu Quyn BQCBBK52

2.8. CC YU T NH HNG N QU TRNH TRCH LY CM GO 2.8.1. S lng dung mi Cc dung mi c s dng chit xut du cn li c thi ra bng cch s dng khai thc c hc v qu trnh chit dung mi em li li ch lng du cao hn ng k so vi chit xut c kh. Nhng trong vic s dng lng ln dung mi n i hi thi gian di cho dung mi bay hi, cn li chit xut du. Nh vy cm go c chit xut n nh t l dung mi ti cm thch hp 1,77 : 1,00 [9]. 2.8.2. Loi dung mi Nhng dung mi chit xut du cn phi c mt im si tng i thp v khng d chy, khng c trong c hai cht lng v hi nc. Tuy nhin, hexane c s dng nh mt dung mi chit xut du t thng mi cm go mc d hexane gy chy, tim nng, sc khe v cc mi nguy him mi trng, nhng n c im si thp v khng tn km [6, 5]. Cn ethanol xut lm dung mi trch ly, do an ton, r tin hn v n cng c s dng trch ly cm go [9]. 2.8.3. Mt ca cm go Ngoi dung mi ra th mt cm go cng l nhn t nh hng n hiu sut trch ly. Rt kh trch ly du t cm go vi mt cao, nu mnh cm go c mt thp, cc vy d dng b ph v. V vy, c c khai thc du ti u, n l cn thit duy tr dy thch hp v gim nhng ht cm go di [5]. 2.8.4. m m vy cm go cn c m khng qu 10% v cc dung mi kh xm nhp vo hai mt khi vy cm go c m cao hn. Bn cnh hiu



Kha lun tt nghip

V Thu Quyn BQCBBK52

qu ca vic khai thc du ph thuc vo ngun nc hoc hi m trong dung mi. 2.8.5. Nhit Nhit trong qu trnh khai thc du l khong 60oC rt ngn thi gian khai thc. C gii hn trn cho vic khai thc nhit , dung mi an ton trng thi lng. Phm vi si ca dung mi hexan thng mi thng l 64 69oC [9]. 2.8.6. Thi gian Cc chit xut t du cm go c th s dng thi gian t 1 2 gi [6].



Kha lun tt nghip Phn III

V Thu Quyn BQCBBK52

I TNG - NI DUNG V PHNG PHP NGHIN CU3.1. I TNG, VT LIU, A IM & THI GIAN NGHIN CU 3.1.1. i tng nghin cu - Cm t cc loi go ly t cc c s xay st an Phng H Ni. Cm go c bo qun 20oC cho n khi s dng lm mu th nghim. 3.1.2. Vt liu nghin cu 3.1.2.1. Ha cht - Cn thc phm (C2H5OH) 96o - NaOH 10% - Clorofom (CHCl3) 99% - Ete - Acid clohydric (HCl) 36 37 % - Acid ascobic 10-2M - HCl 2%; 6N - Phenolphatalein 1% - Acid ecetic (CH3COOH) 10% - FeSO4 10-3 - Ha cht dng nh tnh sterol: anhydrit acetic, CHCl 3, H2SO4, tricloacetic, dung dch brom trong CHCl3. 3.1.2.2. Thit b v dng c



Kha lun tt nghip - B trch ly Soxhlet - Ni p sut

V Thu Quyn BQCBBK52

- H thng trch ly, c chn khng, thu hi dung mi quy m 70 lt/m - My c c - Cn k thut, cn phn tch nh tnh - T sy t ng khng ch nhit - Bp in, bp cch thy - Vi mn, phu lc, cc ong, ng ong v bnh tam gic cc loi - a thy tinh, buret, pipet cc loi, giy lc, nhit k cc loi - Bnh nh mc 100, 250, 500 v 1000 ml 3.1.3. a im v thi gian nghin cu - a im: Vin C in nng nghip v Cng ngh sau thu hoch a ch: S 126 ph Trung Knh Trung Ha Cu Giy H Ni. - Thi gian nghin cu: 15/1/2011 n 15/5/2011. 3.2. NI DUNG NGHIN CU 3.2.1. Nghin cu quy trnh trch ly Gamma oryzanol t cm go - Nghin cu ch x l cm go nguyn liu trc khi trch ly. - Xc nh loi dung mi, nng dung mi n hiu sut trch ly Gamma oryzanol t cm go. - Nghin cu nh hng ca nhit , thi gian trch ly n hiu sut trch ly Gamma oryzanol. - Xy dng quy trnh trch ly Gamma oryzanol t cm go.



Kha lun tt nghip 3.3. PHNG PHP NGHIN CU 3.3.1. B tr th nghim

V Thu Quyn BQCBBK52

3.3.1.1. X l cm go nguyn liu trc khi trch ly Cm go ly t cc c s xay st go an Phng H Ni. Cm go c ng gi vo ti v bo qun nhit 20oC. Do cm go trng thi xp, t bo ph v, c h enzym lipase ni ti, cng s xm nhp ca mi trng, nn phn ng thy phn v oxi ha ca du trong cm pht trin nhanh. Cm go rt d b i hng, chua kht Nn ta cn x l cm go nguyn liu trc khi a vo qu trnh trch ly gamma oryzanol qu trnh trch ly xy ra thun li hn m khng gy ra cc nh hng xu n hiu sut thu hi gamma oryzanol. - Tin hnh cn 100 g cm go, cho vo cc thy tinh em sy trong t sy t ng khng ch nhit , 100oC 105oC trong thi gian 10 pht. - Trong qu trnh sy tin hnh o trn cm nhit tip xc cm u hn. - Sau sy tip, h nhit ca t sy xung 60oC khong thi gian 3 gi, o trn u tay cho ti khi hm m ca cm t khong 5 %. Ta thu c cm go nguyn liu thc hin cc th nghim tip theo. 3.3.1.2. Xc nh loi dung mi nh hng n hiu sut trch ly Qu trnh trch ly nh hng rt nhiu yu t cng ngh: loi dung mi trch ly, nng dung mi, nhit , t l dung mi, thi gian trch ly Mt trong s nhng yu t nh hng l loi dung mi no c nh hng ti hiu sut trch ly gamma oryzanol. Nhng dung mi dng chit xut du cn phi c mt im si tng i thp v khng d chy, khng c trong c hai cht lng v hi nc. Tuy nhin, cn chn loi dung mi an ton cho sc khe ngi s dng, r tin, khng c nguy c chy n.



Kha lun tt nghip

V Thu Quyn BQCBBK52

- Sau khi x l cm go nguyn liu c m thch hp l 5%, tin hnh trch ly 3 mu song song vi cc loi dung mi khc nhau: n hexan, isopropyl alcohol, cn ethanol 96o. - Tin hnh cn 100 g cm go c x l, cn tip 300 g dung mi mi loi. nhit 65oC trong thi gian 3 gi, trch ly 3 mu song song bng b trch ly Soxhlet. - Dch trch ly c x l ui dung mi n trong lng khng i. Lng hot cht sinh hc th em tinh ch thu c hot cht tinh. Da vo hiu sut trch ly xc nh loi dung mi no cho hiu sut trch ly cao nht. Kt qu u c x l thng k theo chng trnh IRRISTAT 4.0. 3.3.1.3. Xc nh nng trch ly nh hng n hiu sut trch ly Sau khi lm th nghim xc nh loi dung mi nh hng n hiu sut trch ly la chn c loi dung mi thch hp. Tin hnh th nghim dung mi chn trn cc nng dung mi l 65o, 75o, 96o cng iu kin cng ngh xc nh nng dung mi ph hp cho vic tch chit hot cht. - Tin hnh cn 100 g cm go c x l, cn tip 300 g dung mi chn mc 3.3.1.2. - nhit 65oC trong thi gian 3 gi, trch ly 3 mu song song bng b trch ly Soxhlet. - Dch trch ly c x l ui dung mi n trong lng khng i. Lng hot cht sinh hc th em tinh ch thu c hot cht tinh. Da vo hiu sut trch ly xc nh nng dung mi l bao nhiu th cho hiu sut trch ly cao nht. Kt qu u c x l thng k theo chng trnh IRRISTAT 4.0.



Kha lun tt nghip

V Thu Quyn BQCBBK52

3.3.1.4. Xc nh nhit trch ly nh hng n hiu sut trch ly Ngoi cc yu t nh loi dung mi trch ly, nng dung mi th nhit cng l yu t quan trng c nh hng ti hiu sut trch ly gamma oryzanol. Sau khi lm th nghim xc nh nng dung mi nh hng n hiu sut trch ly la chn c nng dung mi thch hp. xc nh nhit trch ly nh hng ti hiu sut trch ly ta tin hnh th nghim cc mc nhit l 45oC, 55oC, 65oC, 70oC, 75oC. - Tin hnh cn 100 g cm go c x l, cn tip 300 g dung mi vi nng chn th nghim mc 3.3.1.2 v mc 3.3.1.3. - nhit 65oC trong thi gian 3 gi, trch ly 5 mu song song bng b trch ly Soxhlet. - Dch trch ly c x l ui dung mi n trong lng khng i. Lng hot cht sinh hc th em tinh ch thu c hot cht tinh. Da vo hiu sut trch ly xc nh nhit trch ly l bao nhiu th cho hiu sut trch ly cao nht. Kt qu u c x l thng k theo chng trnh IRRISTAT 4.0. 3.3.1.5. Xc nh t l dung mi nh hng n hiu sut trch ly T l dung mi cng l mt trong nhng yu t quyt nh ti hiu sut trch ly cao nht. Cht lng ca dch chit ph thuc vo t l nguyn liu v dung mi. Nu dng t dung mi c th khng chit ht c hot cht, nhng nu dng nhiu dung mi qu th lng hot cht trong dch chit khng tng m li tng lng tp cht v gy lng ph dung mi. Chnh v vy ta cn xc nh t l dung mi v nguyn liu hiu sut trch ly gamma oryzanol l cao nht.



Kha lun tt nghip

V Thu Quyn BQCBBK52

- Th nghim tin hnh vi cc t l dung mi / cm go nguyn liu khc nhau l: 1:1, 2:1, 3:1, 4:1, 5:1 xc nh t l dung mi / nguyn liu ph hp nht cho vic tch chit hot cht. - Tin hnh cn 100 g cm go c x l. Cn ln lt cc mu 100 g, 200 g, 300 g, 400 g, 500 g dung mi vi nng chn mc 3.3.1.2 v mc 3.3.1.3. - nhit 65oC trong thi gian 3 gi, trch ly 5 mu song song bng b trch ly Soxhlet. - Dch trch ly c x l ui dung mi n trong lng khng i. Lng hot cht sinh hc th em tinh ch thu c hot cht tinh. Da vo hiu sut trch ly xc nh nhit trch ly l bao nhiu th cho hiu sut trch ly cao nht. Kt qu u c x l thng k theo chng trnh IRRISTAT 4.0. 3.3.1.6. Xc nh thi gian trch ly n hiu sut trch ly Sau khi xc nh c cc yu t hiu sut trch ly cao nht nh thi gian sy nguyn liu, loi dung mi, nng dung mi, nhit dung mi. Tin hnh th nghim vi cc ngng thi gian cho mi ln trch ly l: 2.5, 3, 3.5, 4, 4.5 gi xc nh thi gian trch ly ph hp cho hiu sut trch ly cao nht. - Tin hnh cn 100 g cm go c x l, cn tip 300 g dung mi v nng dung mi chon cc mc trn. - nhit 65oC trong thi gian 4 gi, trch ly 4 mu song song bng b trch ly Soxhlet. - Dch trch ly c x l ui dung mi n trong lng khng i. Lng hot cht sinh hc th em tinh ch thu c hot cht tinh. Da vo hiu sut trch ly xc nh thi gian trch ly l bao nhiu th cho hiu sut



Kha lun tt nghip

V Thu Quyn BQCBBK52

trch ly cao nht. Kt qu u c x l thng k theo chng trnh IRRISTAT 4.0. 3.3.1.7. Xy dng quy trnh trch ly gamma oryzanol

Cm go nguyn liu X l cm go: Sy 100 105oC/10 pht, sy tip 60oC/ khong 4 gi, hm m khong 5%

Trch ly Gamma oryzanol Ethanol 96o/ cm go = 3/1 Trch ly 65 70oC/4 gi B cm go trch ly

C dch trch ly Thu hi ethanol

Du cm go th, gamma oryzanol



Kha lun tt nghip 3.3.2. Cc ch tiu nghin cu

V Thu Quyn BQCBBK52

3.3.2.1. Xc nh hm m ca cm go nguyn liu theo phng php sy kh n khi lng khng i Do cm go trng thi xp, t bo ph v, c h enzym lipase ni ti, cng s xm nhp ca mi trng, nn phn ng thy phn v oxi ha ca du trong cm pht trin nhanh. S thy phn du to ra cc acid bo t do cao lm cho cm b i hng, chua kht khng thun li cho qu trnh ch bin sau ny. Chnh v vy, tng tr cm go cho ch bin cng nh chit xut gamma oryzanol cn c phng php bo qun cm go cho trch ly gamma oryzanol bng phng php sy kh n khi lng khng i. * Cch tin hnh Cn 100 g cm nguyn liu trong hp nhm bit trc trng lng. t hp nhm c cha mu vo t sy nhit 105oC. Sau 3 gi em i lm ngui trong bnh ht m v cn. Tip theo li t hp cng mu vo t sy v sy tip khong t 30 60 pht ri li em l ngui trong bnh ht m. Sau em cn li, nu sai s gia hai ln cn khng qu 0.01 g th qu trnh sy xem nh kt thc. * Cch tnh Hm m ca nguyn liu c tnh theo cng thc sau: W = 100% Trong : a: Khi lng hp nhm cng nguyn liu trc khi sy (g) b: Khi lng hp nhm cha nguyn liu sau khi sy (g) c: Khi lng hp nhm kh khng cha nguyn liu (g) 3.3.2.2. Xc nh hm lng gamma oryzanol



Kha lun tt nghip

V Thu Quyn BQCBBK52

Nguyn liu c sy ti hm m khong 5%. Cn 100 g cm go nguyn liu ri em trch ly bng dung mi cn ethanol 96o vi t l ethanol 96o / cm go = 3 / 1, trch ly nhit 65 70oC trong thi gian l 4 gi. Dch trch ly thu c em lc qua vi mn v giy lc kh to loi b ht cn v tp cht thu c du cm go th. Du cm go th c x phng ha vi 1.7 % NaOH 90oC trong thi gian l 1 gi. Trung ha kim d bng Na 2CO3. Dch x phng ha em sy i lu v to si x phng kh 115 oC trong 2 gi. Si x phng kh trch ly vi ethyl acetate trong khong 4 gi. Thu hi dung mi cn v lc tch sp thu c dch lc. Sau c ti khi lng khng i ta xc nh c hm lng gamma oryzanol thu c. 3.3.2.3. Xc nh hiu sut trch ly gamma oryzanol nh gi hiu qu tch chit hot cht gamma oryzanol ngi ta dng phng php xc nh hiu sut trch ly gamma oryzanol. Hiu sut trch ly (HSTL): l hiu sut thu gamma oryzanol theo hm lng gamma oryzanol c trong nguyn liu. HSTL c tnh theo cng thc sau:

HSTL (%) = 100% Trong : HSTL: Hiu sut trch ly (%) LHCT: Lng hot cht tinh (g) LHCNL: Lng hot cht c trong nguyn liu (g)



Kha lun tt nghip Lng hot cht c trong nguyn liu l 0.32 g. 3.3.3. Phng php x l s liu

V Thu Quyn BQCBBK52

S liu c x l thng k theo chng trnh IRISTAT 4.0.

S tin hnh cc bc th nghim ca ti



Kha lun tt nghip

V Thu Quyn BQCBBK52

Nguyn liu

X l cm go nguyn liu

Chn loi dung mi thch hp

Chn nng dung mi thch hp Lng dung mi Xc nh ch trch ly thch hp Tinh ch gamma oryzanol Nhit trch ly Thi gian trch ly

gamma oryzanol

Phn IV

KT QU V THO LUN



Kha lun tt nghip

V Thu Quyn BQCBBK52

4.1. X L CM GO NGUYN LIU TRC KHI TRCH LY 4.1.1. Nghin cu xc nh phng php x l cm nguyn liu cho trch ly gamma ozyzanol t cm go Cm go trng thi xp, t bo ph v, c h enzym lipase ni ti, cng s xm nhp ca mi trng, nn phn ng thy phn v oxi ha ca du trong cm pht trin nhanh. S thy phn du to ra cc acid bo t do cao lm cho cm b i hng, chua kht khng thun li cho qu trnh ch bin sau ny. Chnh v vy, tng tr cm go cho ch bin cng nh chit xut gamma oryzanol cn c phng php bo qun cm go cho trch ly gamma oryzanol. Tc gi Phm c Thi chng trnh 20A (Gim tn tht v nng cao hiu qu s dng lng thc) a ra c quy trnh cng ngh dit men lipasa bng phng php sy cm go nhit 100 105oC/10 pht cho hiu qu dit men lipase cao trong cm go. ti k tha quy trnh ny dit men lipase trong x l cm go nguyn liu cho trch ly Gamma oryzanol t cm go. 4.4.2. Nghin cu xc nh hm m cm go nguyn liu cho trch ly gamma oryzanol Tnh cht ca nguyn liu c nh hng ln n hiu sut trch ly gamma oryzanol. Nu hm m nguyn liu gim th tc trch ly tng ln, v lng nc trong nguyn liu tc dng vi protein v cc cht ho nc khc ngn cn s chuyn dch ca dung mi thm su vo trong nguyn liu, lm chm qu trnh khuch tn. Chng ti tin hnh xc nh hm m ca cm go nguyn liu sau sy cm go nguyn liu t ti hm m nht nh ri tin hnh trch ly bng dung mi cn ethanol 96o trong thi gian 3 gi, thu c dch trch ly, em xc nh hm lng gamma oryzanol bng phng php o mt quang bc sng 280 400 nm. Kt qu phn tch c trnh by bng 4.1.



Kha lun tt nghip

V Thu Quyn BQCBBK52

Bng 4.1. nh hng thi gian sy n cht lng cm go sau sy Thi gian sy (gi) Ch tiu nghin cu 0 (Cm go mi xay st) 13.8 0.16 1 7.1 0.21 2 5.3 0.243 4 2.6 0.245

Hm m cm go (%) Hm lng gamma oryzanol (g/100 g cm) Hiu sut trch ly gamma oryzanol (%)

50.00c

65.60b

75.93a

76.56a

(Ghi ch: Cc s liu theo ct c ch s m khc nhau l c gi tr khc nhau c ngha mc ngha = 0.05) Cm go nguyn liu sau khi sy, trch ly theo t l cm go/ dung mi ethanol 96o theo t l 1: 3 nhit 65oC cc mc thi gian 0 gi (cm go mi xay st), 1 gi, 2 gi, 4 gi c th hin qua bng 4.1 nh sau: Chng ti nhn thy khi thi gian sy tng th hm m cm go gim dn t 13.8 % > 7.1 % > 5.3 % > 2.6 %, hm lng gamma oryzanol tng vi lng nh t 0.16 g < 0.21 g < 0.243 g < 0.245 g. Lc cha sy cm go c hiu sut trch ly thp nht l 50%. Sau khi sy cm c 1 gi thi hiu sut trch ly tng ln r rt l 65.6%. Hiu sut trch ly t gi tr ti u sau khi sy nguyn liu 2 gi vi hiu sut cao t 75.93%. Khi tng thi gian sy ln 4 gi th hiu sut trch ly c tng nhng tng khng ng k l 76.56% khng ph hp vi iu kin kinh t. Qua th nghim ny chng ti nhn thy rng, nguyn liu cng kh th cng to iu kin thun li cho qu trnh trch ly, song rt kh khn sy nguyn liu n m qu thp v chi ph nng lng cho nng lng sy



Kha lun tt nghip

V Thu Quyn BQCBBK52

nguyn liu cao. Mc d, thi gian sy l 4 gi cho hiu sut trch ly cao hn nhng cao hn khng ng k so vi hiu sut trch ly thi gian sy l 2 gi. Nu ta sy cm go nguyn liu trong 4 gi s rt mt nhiu thi gian v gy lng ph ln. Qua tin hnh kho st, chng ti nhn thy x l cm go nguyn liu thch hp cho qu trnh trch ly gamma orynazol l cm go nguyn liu sau khi dit men lipase 100oC 105oC trong thi gian 10 pht, tip tc sy 60oC trong thi gian 2 gi t hm m khong 5.3% ri em trch ly. 4.2. XC NH LOI DUNG MI NH HNG N HIU SUT TRCH LY Dung mi ho tan cc cht cn trch ly v gi li cc tp cht, dung mi c nh hng ln ti cht lng ca dch chit v thnh phm, v vy vic la chn dung mi thch hp l ht sc quan trng. Da trn tnh cht vt l ( nht, sc cng b mt, phn cc) v c tnh ha tan chn lc ca dung mi i vi hot cht la chn dung mi. Nhng dung mi dng chit xut du cn phi c mt im si tng i thp v khng d chy, khng c trong c hai cht lng v hi nc. Tuy nhin, cn chn loi dung mi an ton cho sc khe ngi s dng, r tin, khng c nguy c chy n. Ngoi ra qua ti liu cho bit gamma oryzanol l hp cht khng phn cc nn chng ti tin hnh th nghim kho st vi cc loi dung mi khng phn cc: n hexan, isopropyl alcohol, cn ethanol 96o. Bng 4.2. nh hng ca loi dung mi n hiu sut trch ly Hm lng gamma Loi dung mi oryzanol (g/100g cm go) n Hexan 0.254 79.37a Hiu sut trch ly (%)



Kha lun tt nghip

V Thu Quyn BQCBBK52

Isopropyl alcohol Cn ethanol 96o

0.261 0.243

81.56a 75.93b

(Ghi ch: Cc s liu theo ct c ch s m khc nhau l c gi tr khc nhau c ngha mc ngha = 0.05) T bng 4.2 chng ti nhn thy, hiu sut trch ly ca Isopropyl alcohol cao nht l 81.56%, sau ti hiu sut trch ly ca n Hexan l 79.37%, v cui cng l hiu sut trch ly ca cn ethanol 96 o l thp nht 75.93%. Hiu sut trch ly ca Isopropyl alcohol > n Hexan > Cn ethanol 96o. Mc d trch ly bng dung mi Isopropyl alcohol cho hiu sut trch ly l cao nht song vic chn dung mi cho vic trch ly cc sn phm dng cho thc phm v dc phm th ngoi cc yu t tnh ha tan chn lc cao, nht thp, sc cng b mt nh ngi ta cn lu n tnh c hi ca dung mi, vn kinh t, r tin. Cn ethanol l dung mi c th ha tan tt hu ht cc cht trong nguyn liu. Ngoi ra, cn cng l nguyn liu d kim, s dng rt tin li, c th tch cn ra khi dung dch chit mt cch d dng bng cch un nng nh sau mt thi gian s thu c dch chit khng cn. Vic trch ly bng cn s c d dng do cn c kh nng lm mt nc nguyn sinh cht t bo, lm ng t protein v pectin. Do thnh t bo mt tnh thm thu v cht ha tan khuch tn t do. Cn ethanol p ng tng i y cc yu t ny, chnh v vy, chng ti chn cn ethanol 96oC l dung mi thch hp cho qu trnh trch ly hot cht v s dng dung mi ny cho cc nghin cu tip theo. Qua kt qu x l thng k cho thy gi tr hiu sut trch ly cc dung mi khc nhau l khc nhau mc ngha = 0.05. 4.3. NGHIN CU XC NH NNG DUNG MI CN ETHANOL NH HNG N HIU SUT TRCH LY



Kha lun tt nghip

V Thu Quyn BQCBBK52

Do cn ethanol t c hi ca dung mi, vn kinh t, r tin v d kim nn ta chn cn l dung mi trch ly. Cn ethanol s dng tch chit hot cht phi t cc tiu chun quy nh an ton thc phm. Kh nng trch ly gamma oryzanol t cm go thay i theo nng , do cn phi xc nh nng ca ethanol t hiu sut trch ly hot cht cao nht. Khi nng cc cht trch ly c tng ln dn th s chnh lch nng gim dn v t ti cn bng, qu trnh khuch tn xy ra rt chm. Do chng ti tin hnh th nghim kho st cc ngng nng dung mi (vi dung mi l cn chn trn) nh sau: 65o, 75o, 96o, cm go nguyn liu dit men v c hm m 5% c trch ly vi t l cm go nguyn liu / cn ethanol theo t l 1: 5; trch ly trong 3 gi. Kt qu th nghim nghin cu xc nh nng dung mi cn ethanol nh hng n hiu sut trch ly c th hin qua bng 4.3.

Bng 4.3. nh hng nng dung mi cn ethanol n hiu sut trch ly Hm lng gamma Nng dung mi () oryzanol (g/100g cm go) 65 0.11 34.37c Hiu sut trch ly (%)



Kha lun tt nghip

V Thu Quyn BQCBBK52

75 96

0.14 0.243

43.75b 75.93a

(Ghi ch: Cc s liu theo ct c ch s m khc nhau l c gi tr khc nhau c ngha mc ngha = 0.05) T bng 4.3 chng ti thy rng khi nng dung mi cng cao th hiu sut trch ly cng ln. Nng dung mi 65 o, 75o, 96o ln lt cho hiu sut trch ly l 34.37%, 43.75%, 75.93%. Nng ethanol l 96o cho hiu sut rt cao so vi 2 loi nng cn li, thch hp s dng trch ly gamma oryzanol. Theo chng ti gii thch iu ny c th l do nguyn liu ha tan rt it trong nc v ha tan nhiu trong cn m c. Trong cn ethanol 65% th cha nhiu nc hn cn ethanol 75% v cn 96%. M cycloartenyl ferulate, 24-methylenecycloartenyl ferulate v campesteryl ferulate, l thnh phn chnh ca gamma oryzanol, l cht phn cc yu nn ha tan nhiu trong cn 96o. Qua kt qu x l thng k nhn thy cc gi tr hiu sut ba nng cn khc nhau mc ngha = 0.05 v gi tr hiu sut nng cn 96 o l cao nht. Do vy, m bo hiu sut trch ly l cao nht chng ti chn cn c nng 96o s dng cho nghin cu tip theo. 4.4. NGHIN CU NH HNG CA NHIT N HIU SUT TRCH LY GAMMA ORYZANOL T CM GO BNG CN ETHANOL Nhit l mt trong nhng yu t c nh hng rt ln n qu trnh trch ly. Khi nhit trch ly cng cao s lm cho xp v linh ng ca nguyn liu tng ln (do nguyn liu trng n) v hot cht ha tan d dng Trng i hc Nng NghipHN Khoa Cng ngh thc phm37

Kha lun tt nghip

V Thu Quyn BQCBBK52

trong dung mi. Tuy nhin, nhit l mt yu t c gii hn v khi nhit qu cao c th xy ra cc phn ng khc khng cn thit nh tng tan ca mt s tp cht, kh khn cho qu trnh lc v s thc y cc bin i ha hc ca cc thnh phn c trong nguyn liu dn n cht lng dch chit b thay i theo chiu hng xu i. Bng 4.4. Nhit trch ly nh hng n hiu sut trch ly Hm lng gamma Nhit (oC) oryzanol (g/100g cm go) 45 55 65 70 75 0.285 0.317 0.320 0.322 0.325 75.00c 83.42b 84.21ab 84.73ab 85.52a Hiu sut trch ly (%)

(Ghi ch: Cc s liu theo ct c ch s m khc nhau l c gi tr khc nhau c ngha mc ngha = 0.05) T kt qu bng 4.4 chng ti thy gi tr hiu sut tng dn v t l thun vi nhit trch ly (75% 45oC, 83.42% 55oC, 84.21% 65oC, 84.73% 70oC, 85.52% 75oC). Vic thu c kt qu ny c gii thch nh sau: Gia hai cht lng bt k lun c chuyn ng nhit (chuyn ng Brown) v chuyn ng tng h. Khi nhit thp 45 oC chuyn ng nhit gim lm cho linh ng ca cc cu t gamma oryzanol trong nguyn liu thp v vy s lng gamma oryzanol khuch tn vo dung mi t nn hiu sut trch ly nh (75%). Khi nhit tng ln 55oC th hiu sut trch ly cng tng ln l 83.42%. Khi nhit cao 65oC v 70oC th chuyn ng nhit tng Trng i hc Nng NghipHN Khoa Cng ngh thc phm38

Kha lun tt nghip

V Thu Quyn BQCBBK52

lm cho cc cu t gamma oryzanol chuyn ng nhanh hn nn d dng c chit tch ra khi nguyn liu. Hn na, nhit cng cao s thay i cu trc m t bo cng ln, lm cho vch t bo b ph hy nn dch bo d dng thot ra ngoi, hiu sut tng r rt (84.21% 65 oC, 84.73% 70oC). Cn nhit 75oC th hiu sut trch ly c tng nhng khng ng k t 85.52%. Hiu ng ny cho thy khi tng nhit th lm gim nht v tng khuch tn, dn n hiu sut trch ly tng. Qua kt qu x l thng k cho thy hiu sut trch ly cc nhit 45oC, 55oC, 75oC l khc nhau mc ngha = 0.05 v hiu sut trch ly nhit 65oC v 70oC l khng c s khc nhau mc ngha = 0.05. Cn c vo kt qu v iu kin th nghim cho thy 65 70oC l ph hp trch ly gamma oryzanol t cm go bng cn ethanol 96o. 4.5. NGHIN CU NH HNG CA T L DUNG MI CN ETHANOL 96o N HIU SUT TRCH LY GAMMA ORYZANOL Cht lng ca dch chit ph thuc vo t l nguyn liu v dung mi v nu dng t dung mi c th khng chit ht c hot cht nhng nu dng nhiu dung mi lng hot cht trong dch chit khng tng m li tng lng tp cht. V vy cn nghin cu t l nguyn liu v dung mi. Qua cc ti liu tham kho chng ti tin hnh kho st vi cc t l dung mi / nguyn liu c nu trong bng sau. Bng 4.5. nh hng ca t l dung mi cn ethanol 96o n hiu sut trch ly Hm lng gamma T l dung mi oryzanol (g/100g cm go) 1:1 0.265 27.63d Hiu sut trch ly (%)



Kha lun tt nghip

V Thu Quyn BQCBBK52

2:1 3:1 4:1 5:1

0.328 0.330 0.336 0.340

69.98c 86.54b 87.42b 89.47a

(Ghi ch: Cc s liu theo ct c ch s m khc nhau l c gi tr khc nhau c ngha mc ngha = 0.05) Kho st vi cc t l dung mi / nguyn liu, kt qu th 2 cho thy vi cng thi gian, nhit v lng cm go trch ly, khi tng lng dung mi trch ly th gamma oryzanol ton phn trong dch trch ly cng tng. T kt qu bng 4.5 chng ti nhn thy t l dung mi cn ethanol 96oC / cm go nguyn liu = 1 : 1 cho hiu sut trch ly thp nht l 27.63% do lng dung mi nh khi s chnh lch nng cht tan trong dung mi so vi lng cu t gamma oryzanol thp nn s khuch tn cc cu t gamma oryzanol v dung mi chm, do vy hiu sut trch ly l nh. T l cn ethanol 96o / cm go = 2 : 1, hiu sut trch ly tng ng k l 69.98%. t l cn ethanol 96o / cm go = 3 : 1 v 4 : 1 cho hiu sut trch ly gamma oryzanol gn nh nhau (tng ng 86,54 % v 87,42 % ), qu trnh trch ly c thc hin d dng. Tng t l dung mi ln 5 : 1 v 6 : 1 so vi cm go nguyn liu cho hiu sut trch ly tng ln khng ng k t 87.42% v 89.47% do khi dung mi qu nhiu gy cn tr cho qu trinh trch ly nn lng gamma oryzanol chit tch ra khng nhiu. Qua kt qu x l thng k cho thy gi tr hiu sut trch ly t l dung mi cn ethanol 96oC / cm go nguyn liu l 1 : 1, 2 : 1, 5 : 1 l khc nhau mc ngha = 0.05. Nhng hiu sut trch ly t l dung mi cn ethanol 96oC / cm go nguyn liu l 3 : 1, 4 : 1 l khng c s khc nhau.



Kha lun tt nghip

V Thu Quyn BQCBBK52

Cn c vo kt qu th nghim m bo hiu sut trich ly cng nh hiu qu kinh t, t l cn ethanol 96o / cm go = 3 / 1 l ph hp cho trch ly gamma oryzanol. 4.6. NGHIN CU NH HNG CA THI GIAN N HIU SUT TRCH LY GAMMA ORYZANOL T CM GO BNG CN ETHANOL Gamma oryzanol trong cm go nguyn liu khi khuch tn vo dung mi vi thi gian nht nh s t n cn bng. iu d nhn thy thi gian trch ly cng di th lng gamma oryzanol thu c cng ln. Tuy nhin nu ko di thi gian trch ly, t l hot cht Gamma oryzanol trong dch chit khng tng m s tng t l tp cht khuch tn vo dch chit v ng thi dn ti chi ph cho qu trnh trch ly cao. V vy, thi gian chit xut nh hng rt ln ti cht lng dch chit v hiu sut trch ly, cn tin hnh th nghim xc nh c thi gian thch hp cho trch ly gamma oryzanol t hiu qu cao. Bng 4.6. Thi gian trch ly nh hng ti hiu sut trch ly Hm lng gamma Thi gian (gi) oryzanol(g/100g cm go) 2.5 3 3.5 4 4.5 1.35 0.32 0.33 0.332 0.334 62.34d 69.98c 79.65b 86.84a 86.89a Hiu sut trch ly(%)



Kha lun tt nghip

V Thu Quyn BQCBBK52

(Ghi ch: Cc s liu theo ct c ch s m khc nhau l c gi tr khc nhau c ngha mc ngha = 0.05) Kt qu t bng 4.6 cho thy hiu sut trch ly tng dn theo thi gian trch ly th hin: Khi trch ly vi thi gian 2.5 gi, hm lng gamma oryzanol trong cm go l 1.35 g v hiu sut trch ly nh nht l 62.34 %. Nhng khi tip tc tng thi gian trch ly ln 3 gi th cc thng s tng ln r rt (0.32 g; 69.98% ), thi gian trch ly l 3.5 gi c cc thng s (0.33 g; 79.65% ). Hiu sut trch ly t ti u thi gian l 4 gi vi hm lung gamma oryzanol 0.332 g, hiu sut trch ly l 86.84%. Khi tip tc ko di thi gian n 4.5 gi th hiu sut trch ly hu nh khng tng ng k, iu ny cn c th hin r t kt qu thng k. Kt qu c th gii thch do cu trc thnh t bo ca cm go kh vng chc nn cn ngm trong thi gian di. Thi gian trch ly cng di, th cng trch ly kit hot cht gamma oryzanol c trong nguyn liu, song thi gian trch ly tng n mt lc no th gamma oryzanol khuch tn vo dung mi s tng khng ng k. V th tip tc tng thi gian ln 4.5 gi th hiu sut tng ln khng nhiu. Qua th nghim ny chng ti nhn thy tin hnh trch ly thi gian 4 gi l thch hp nht v mang li iu kin kinh t cao. 4.7. QUY TRNH CNG NGH TRCH LY GAMMA ORYANOL T CM GO BNG CN ETHANOL 96o T kt qu nghin cu mt s yu t ca cng ngh trch ly gamma oryzanol t cm go bng cn ethanol nh nh hng ca thi gian sy, loi dung mi, nng dung mi, nhit trch ly, t l dung mi, thi gian trch ly. Ta xy dng c quy trnh cng ngh trch ly gamma oryzanol t cm



Kha lun tt nghip

V Thu Quyn BQCBBK52

go theo s quy trnh cng ngh trch ly gamma oryzanol t cm go bng cn ethanol 96o di y. Vi t l dung mi cn ethanol 96o / cm go nguyn liu = 3 / 1 65-70oC trong thi gian l 4 gi, cho hiu sut trch ly gamma oryzanol t khong 86,84%.

Cm go nguyn liu X l cm go: Sy 100 105oC/10 pht, sy tip 60oC/ khong 4 gi, hm m khong 5%

Trch ly gamma oryzanol Ethanol 96o/ cm go = 3/1 Trch ly 65 70oC/4 gi B cm go trch ly

C dch trch ly Thu hi ethanol

Du cm go th, gamma oryzanol

Quy trnh cng ngh trch ly gamma oryzanol t cm go bng cn ethanol 96o



80 70 Kha lun tt nghip 60 50 Hiu sut 40 trch ly (%) 30 20 10 0 0 1 2 4

V Thu Quyn BQCBBK52

* Thuyt minh quy trnh cng ngh trch ly gamma oryzanol t cm go bng cn ethanol 96o 1. X l cm go - Mc ch: Do cm go trng thi xp, t bo ph v, c h enzym lipase ni ti, cng s xm nhp ca mi trng, nn phn ng thy phn v oxi ha ca du trong cm pht trin nhanh. Cm go rt d b i hng, chua kht Nn ta cn x l cm go nguyn liu trc khi a vo qu trnh trch ly gamma oryzanol. - Thit b: t sy t ng khng ch nhit . - Cm go nguyn liu c a vo t sy t ng khng ch nhit nhit 100oC 105oC trong thi gian l 10 pht, sau mang em sy tip nhit 60oC trong khong 2 gi, sy ti khi cm go nguyn liu c hm m t khong 5.3% ri em i trch ly gamma oryzanol. 2. Trch ly gamma oryzanol - Mc ch: qu trnh trch ly to ra du cm go th cha gamma oryzanol, c th tip tc cho qu trinh tinh ch gamma oryzanol tip theo. - Thit b: s dng b trch ly Soxhlet. - Trch ly gamma oryzanol bng dung mi l cn ethanol 96 o vi t l dung mi v cm go l 3:1. S dng b trch ly Soxhlet. Trch ly nhit 65 70oC trong thi gian l 4 gi. Ta thu c dch trch ly gamma oryzanol. 3. C dch trch ly

Thi gian sy (gi)



Kha lun tt nghip

V Thu Quyn BQCBBK52

- Mc ch: ui ht nc v dung mi cn ethanol 96 o c trong dch trch ly tip tc qu trnh tinh ch tip theo. - Thit b: h thng c chn khng, thu hi dung mi quy m 70 lt/m. - Dch trch ly thu c em i lc qua vi mn v qua giy lc bn to, loi b ht b cm go, tp cht v cn xt li trong dch. Sau a vo h thng c chn khng, thu hi dung mi quy m 70 lt/m. Thu c dch gamma oryzanol c ti khi lng khng i chnh l du th gamma oryzanol v thu hi cn ethanol.



Kha lun tt nghip

V Thu Quyn BQCBBK52

Phn V

KT LUN V NGH5.1. KT LUN Sau thi gian thc hin ni dung ca ti chng ti rt ra kt lun sau: 1. X l cm go nguyn liu thch hp cho qu trnh trch ly dit men lipase 100 150oC/ 10 pht, sau sy 60o/ 2 gi t hm m 5.3% ri em trch ly. 2. Xc nh c mt s yu t cho cng ngh trch ly gamma oryzanol t cm go bng ethanol 96o l: - T l dung mi ethanol 96o / cm go = 3 / 1. - Trch ly 65 - 700C. - Thi gian trch ly 4 gi l ph hp. - Hiu sut trch ly gamma oryzanol t khong 86,84%. 3. a ra c quy trnh cng ngh trch ly gamma oryzanol t cm go bng ehanol 96o. 5.2. NGH Trong khun kh ca kha lun tt nghip ti c mt s kt qu thnh cng bc u. Sau khi tt nghip chng ti hy vng:



Kha lun tt nghip

V Thu Quyn BQCBBK52

- ti s c hon thin thm c cng ngh sn xut gamma oryzanol t ph ph phm cm go. - Nghin cu tip quy trinh tinh ch gamma oryzanol thu c gamma oryzanol kt tinh t tin hnh ch bin to vin thc phm chc nng t gamma oryzanol thu c quy m phng thi nghim. - Sn xut th thc phm chc nng quy m cng nghip.



Kha lun tt nghip

V Thu Quyn BQCBBK52

TI LIU THAM KHO

Ti liu Ting Vit 1. B mn ha sinh trng H Dc HN Ha sinh hc NXB Y hc. 2. ThS. hu Phng - c san khoa hc k thut thc n chn nui - s 4/2004. Ti liu nc ngoi 3. Cicero, A. F. G., and Gaddi, A. (2001) Rice bran oil and -oryzanol in the treatment of hyperlipoproteinaemias and other conditions. Phytotherapy Research, 15(4) : 277-289. 4. Cheruvanky, R. (2003) Phytochemical products: rice bran. In Phytochemical functional foods (Johnson, I. T., and Williamson, G., eds), pp. 347-376. CRC Press; Woodhead Publishing, Boca Raton, Fla; Cambridge. 5. Johnson, L.A., and E.W. Lusas, Comparison of Alternative Solvents for Oils Extraction, J. Am. Oil Chem. Soc. 60:229-242 (1983). 6. Health and Safety Guide No.59: n-Hexane Health and Safety Guide, World Health Organization, Geneva, 1991. 7. Narumon N., Panvipa K., Chitkavee P., Aeumporn S., Rassmidara H., Weena J., (2004). Preparation and evaluation of crude extract containing oryzonol from rice bran oil.



Kha lun tt nghip

V Thu Quyn BQCBBK52

8. Seetharamaiah, G.S., and J.V. Prabhakar, Oryzanol Content of Indian Rice Bran Oil and Its Extraction from Soapstock, J. Food Sci. Technol. 23:270273 (1986). 9. Talwalkar, R.T., N.K. Grag, and C.R. Krishna Murti, Rice Bran a Source Material for Pharmaceuticals, J. Food Sci. and Technol. 2:117-119 (1965). 10. Kahlon, T.S., R.M. Saunders, R.N. Sayre, F.I. Chow, M.M. Chiu, and A.A. Betschart, Cholesterol Lowering Effects of Rice Bran and Rice Bran oil Fractions in Hypercholesterolemic, Cereal Chem. 69:485 489 (1992). 11. Kim. J. S., Godber, J. S., King, J. M., and Prinyawiwatkul, W. (2001) Inhibition of cholesterol autoxidation by the nonsaponnifiable fracion in rice bran in an aqueous model system. JAOCS, Journal of the American Oil Chemists Society. 12. Kaimal, T.N.B., Oryzanol from Rice Bran Oil, J. Oil Technol. Assoc. India (1999). 13. Lloyd, B.J., Siebenmorgen, T.J and Beers, K.W.2000. Effect of commercial processing on antioxidants in rice bran. Cereal Chemistry (2000). 14. Hegsted, M., and M.M. Windhauser, Reducing Human Heart Disease Risk with Rice Bran, Louisiana Agriculture (1993). 15. Nicolosi, R.J., L.M. Ausman, and D.M.Hegsted, Rice Bran Oil Lowers Serum Total and Low Density Lipoprotein Cholesterol and Apo B Levels in Nonhuman Primates, Atherosclerosis (1991). 16. Rukmini, C., Chemical, Nutritional and Toxicological Studies of Rice Bran Oil, Food Chem (1988).



Kha lun tt nghip 17. Sharma, R.D., and C.

V Thu Quyn BQCBBK52 Rukmini, Rice Bran Oil and

Hypocholesterolemic in Rats, Lipids 21:715-717 (1986). 18. Seetharamaiah, G. S., and Chandrasekhara, N. (1989) Studies on hypocholesterolemic activity of rice bran oil. Atherosclerosis, 78(2-3): 219223. 19. Shin, T., J.S. Godber, D.E. Martin, and J.H. Wells, Hydrolytic Stability and Changes in E Vitamers and Oryzanol of Extruded Rice Bran During Storage, J. Food Sci. 62:704708 (1997). 20. Yukihisa Tanaka, Ikuko Sakaki, Takeshi Ohkubo. Extraction of phospholipids from salmon roe with supercritical carbon dioxide and an entrainer. Journal of Oleo Science. 53, 417-424, 2004 (CA 141:394384). Ti liu tham kho t internet



Kha lun tt nghip

V Thu Quyn BQCBBK52

PH LC

H thng thit b trch ly, c v sy chn khng c s dng sn xut Gamma oryzanol t cm go Cm go sau khi trch ly gamma oryzanol

Du th trch ly t cm go

Qu trnh tch sp

Qu trnh kt tinh Gamma oryzanol

80 70 60 50 Hiu sut 40 trch ly (%) 30 20 10 0

Trng i hc Nng NghipHN 0

1

Khoa 2

Cng ngh thc phm 51 4

Thi gian sy (gi)

Kha lun tt nghip

V Thu Quyn BQCBBK52

th 4.1. nh hng thi gian sy n cht lng cm go sau sy

82 81 80 79 Hiu sut 78 trch ly (%) 77 76 75 74 73

n Hexan

Isopropyl alcohol

Cn ethanol 96

Loi dung mi

th 4.2. nh hng ca loi dung mi n hiu sut trch ly



Kha lun tt nghip

V Thu Quyn BQCBBK52

80 70 60 50 Hiu sut 40 trch ly (%) 30 20 10 0 65 75 Nng dung mi 96

th 4.3. nh hng nng dung mi cn ethanol n hiu sut trch y

86 84 82 80 Hiu sut 78 trch ly (%) 76 74 72 70 68 45 55 65 Nhit ( C) 70 75

th 4.4. nh hng ca nhit n hiu sut trch ly



Kha lun tt nghip

V Thu Quyn BQCBBK52

90 80 70 60 Hiu sut 50 trch ly (%) 40 30 20 10 0 1:1 2:1 3:1 4:1 5:1

T l dung mi

th 4.5. nh hng ca t l dung mi cn ethanol 96o n hiu sut trch ly

90 80 70 60 Hiu sut 50 trch ly (%) 40 30 20 10 0 2.5 3 3.5 Thi gian (gi) 4 4.5

th 4.6. nh hng ca thi gian trch ly n hiu sut trch ly



Kha lun tt nghip

V Thu Quyn BQCBBK52

nh hng ca thi gian sy ti cht lng cm sau syBALANCED ANOVA FOR VARIATE KQ FILE HS 14/ 5/** 10: 4 ---------------------------------------------------------------- PAGE 1 Anh huong cua thoi gian say toi chat luong cam sau say VARIATE V003 KQ LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ===================================================== ======================== 1 CT$ 3 936.103 312.034 66.16 0.002 2 * RESIDUAL 4 18.8649 4.71623 ----------------------------------------------------------------------------* TOTAL (CORRECTED) 7 954.968 136.424 ----------------------------------------------------------------------------TABLE OF MEANS FOR FACTORIAL EFFECTS FILE HS 14/ 5/** 10: 4 ---------------------------------------------------------------- PAGE 2 Anh huong cua thoi gian say toi chat luong cam sau say MEANS FOR EFFECT CT$ ------------------------------------------------------------------------------CT$ 0 1 2 4 NOS 2 2 2 2 KQ 50.0000 64.1000 75.9300 76.5600

SE(N= 2) 1.53561 5%LSD 4DF 6.01928 ------------------------------------------------------------------------------ANALYSIS OF VARIANCE SUMMARY TABLE FILE HS 14/ 5/** 10: 4 ---------------------------------------------------------------- PAGE 3 Anh huong cua thoi gian say toi chat luong cam sau say F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL. SECTION - 1 VARIATE GRAND MEAN STANDARD DEVIATION C OF V |CT$ (N= 8) -------------------- SD/MEAN | | NO. BASED ON BASED ON % | | OBS. TOTAL SS RESID SS | | 8 66.648 11.680 2.1717 3.3 0.0018 |

KQ



Kha lun tt nghip

V Thu Quyn BQCBBK52

nh hng ca loi dung mi ti hiu sut trch lyBALANCED ANOVA FOR VARIATE KQ FILE QUYEN 15/ 5/** 9:25 ---------------------------------------------------------------- PAGE 1 Anh huong cua loai dung moi toi hieu suat trich ly VARIATE V003 KQ LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ===================================================== ======================== 1 CT$ 2 32.2177 16.1089 16.12 0.024 2 * RESIDUAL 3 2.99861 .999538 ----------------------------------------------------------------------------* TOTAL (CORRECTED) 5 35.2164 7.04327 ----------------------------------------------------------------------------TABLE OF MEANS FOR FACTORIAL EFFECTS FILE QUYEN 15/ 5/** 9:25 ---------------------------------------------------------------- PAGE 2 Anh huong cua loai dung moi toi hieu suat trich ly MEANS FOR EFFECT CT$ ------------------------------------------------------------------------------CT$ NOS n - hexan 2 Isopropyl al 2 Con ethanol 2 KQ 79.3700 81.5600 75.9300

SE(N= 2) 0.706943 5%LSD 3DF 3.16807 ------------------------------------------------------------------------------ANALYSIS OF VARIANCE SUMMARY TABLE FILE QUYEN 15/ 5/** 9:25 ---------------------------------------------------------------- PAGE 3 Anh huong cua loai dung moi toi hieu suat trich ly F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL. SECTION - 1 VARIATE GRAND MEAN STANDARD DEVIATION C OF V |CT$ (N= 6) -------------------- SD/MEAN | | NO. BASED ON BASED ON % | | OBS. TOTAL SS RESID SS | | 6 78.953 2.6539 0.99977 1.3 0.0244 |

KQ



Kha lun tt nghip

V Thu Quyn BQCBBK52

nh hng ca nng dung mi ti hiu sut trch lyBALANCED ANOVA FOR VARIATE KQ FILE NDDM 15/ 5/** 9:36 ---------------------------------------------------------------- PAGE 1 Anh huong cua nong do dung moi toi hieu suat trich ly VARIATE V003 KQ LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ===================================================== ======================== 1 CT$ 2 1900.51 950.257 ****** 0.000 2 * RESIDUAL 3 2.26140 .753800 ----------------------------------------------------------------------------* TOTAL (CORRECTED) 5 1902.77 380.555 ----------------------------------------------------------------------------TABLE OF MEANS FOR FACTORIAL EFFECTS FILE NDDM 15/ 5/** 9:36 ---------------------------------------------------------------- PAGE 2 Anh huong cua nong do dung moi toi hieu suat trich ly MEANS FOR EFFECT CT$ ------------------------------------------------------------------------------65 75 96 CT$ NOS 2 2 2 KQ 34.3700 43.7500 75.9300

SE(N= 2) 0.613922 5%LSD 3DF 2.75121 ------------------------------------------------------------------------------ANALYSIS OF VARIANCE SUMMARY TABLE FILE NDDM 15/ 5/** 9:36 ---------------------------------------------------------------- PAGE 3 Anh huong cua nong do dung moi toi hieu suat trich ly F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL. SECTION - 1 VARIATE GRAND MEAN STANDARD DEVIATION C OF V |CT$ (N= 6) -------------------- SD/MEAN | | NO. BASED ON BASED ON % | | OBS. TOTAL SS RESID SS | | 6 51.350 19.508 0.86822 1.7 0.0000 |

KQ



Kha lun tt nghip

V Thu Quyn BQCBBK52

nh hng ca nhit ti hiu sut trch lyBALANCED ANOVA FOR VARIATE KQ FILE ND 15/ 5/** 10: 2 ---------------------------------------------------------------- PAGE 1 Anh huong cua nhiet do toi hieu suat trich ly VARIATE V003 KQ LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ===================================================== ======================== 1 CT$ 4 148.170 37.0425 83.40 0.000 2 * RESIDUAL 5 2.22081 .444163 ----------------------------------------------------------------------------* TOTAL (CORRECTED) 9 150.391 16.7101 ----------------------------------------------------------------------------TABLE OF MEANS FOR FACTORIAL EFFECTS FILE ND 15/ 5/** 10: 2 ---------------------------------------------------------------- PAGE 2 Anh huong cua nhiet do toi hieu suat trich ly MEANS FOR EFFECT CT$ ------------------------------------------------------------------------------45 55 65 70 75 CT$ NOS 2 2 2 2 2 KQ 75.0000 83.4200 84.2100 84.7300 85.5200

SE(N= 2) 0.471255 5%LSD 5DF 1.71185 ------------------------------------------------------------------------------ANALYSIS OF VARIANCE SUMMARY TABLE FILE ND 15/ 5/** 10: 2 ---------------------------------------------------------------- PAGE 3 Anh huong cua nhiet do toi hieu suat trich ly



Kha lun tt nghip

V Thu Quyn BQCBBK52

F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL. SECTION - 1 VARIATE GRAND MEAN STANDARD DEVIATION C OF V |CT$ (N= 10) -------------------- SD/MEAN | | NO. BASED ON BASED ON % | | OBS. TOTAL SS RESID SS | | 10 82.576 4.0878 0.66646 0.8 0.0003 |

KQ

nh hng ca t l dung mi ti hiu sut trch lyBALANCED ANOVA FOR VARIATE KQ FILE TLDM 15/ 5/** 9:47 ---------------------------------------------------------------- PAGE 1 Anh huong cua ty le dung moi toi hieu suat trich ly VARIATE V003 KQ LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ===================================================== ======================== 1 CT$ 4 5469.96 1367.49 ****** 0.000 2 * RESIDUAL 5 1.84003 .368007 ----------------------------------------------------------------------------* TOTAL (CORRECTED) 9 5471.80 607.978 ----------------------------------------------------------------------------TABLE OF MEANS FOR FACTORIAL EFFECTS FILE TLDM 15/ 5/** 9:47 ---------------------------------------------------------------- PAGE 2 Anh huong cua ty le dung moi toi hieu suat trich ly MEANS FOR EFFECT CT$ ------------------------------------------------------------------------------1:1 2:1 3:1 4:1 5:1 CT$ NOS 2 2 2 2 2 KQ 27.5400 69.9800 86.5400 87.4200 89.4700

SE(N= 2) 0.428956 5%LSD 5DF 1.55820 ------------------------------------------------------------------------------ANALYSIS OF VARIANCE SUMMARY TABLE FILE TLDM 15/ 5/** 9:47



Kha lun tt nghip

V Thu Quyn BQCBBK52

---------------------------------------------------------------- PAGE 3 Anh huong cua ty le dung moi toi hieu suat trich ly F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL. SECTION - 1 VARIATE GRAND MEAN STANDARD DEVIATION C OF V |CT$ (N= 10) -------------------- SD/MEAN | | NO. BASED ON BASED ON % | | OBS. TOTAL SS RESID SS | | 10 72.190 24.657 0.60664 0.8 0.0000 |

KQ

nh hng ca thi gian ti hiu sut trch lyBALANCED ANOVA FOR VARIATE KQ FILE TG 15/ 5/** 10:13 ---------------------------------------------------------------- PAGE 1 Anh huong cua thoi gian toi hieu suat trich ly VARIATE V003 KQ LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ===================================================== ======================== 1 CT$ 4 931.516 232.879 331.13 0.000 2 * RESIDUAL 5 3.51641 .703283 ----------------------------------------------------------------------------* TOTAL (CORRECTED) 9 935.033 103.893 ----------------------------------------------------------------------------TABLE OF MEANS FOR FACTORIAL EFFECTS FILE TG 15/ 5/** 10:13 ---------------------------------------------------------------- PAGE 2 Anh huong cua thoi gian toi hieu suat trich ly MEANS FOR EFFECT CT$ ------------------------------------------------------------------------------CT$ 2.5 3 3.5 4 4.5 NOS 2 2 2 2 2 KQ 62.3400 69.9800 79.6500 86.8400 86.8900



Kha lun tt nghip

V Thu Quyn BQCBBK52

SE(N= 2) 0.592994 5%LSD 5DF 2.15407 ------------------------------------------------------------------------------ANALYSIS OF VARIANCE SUMMARY TABLE FILE TG 15/ 5/** 10:13 ---------------------------------------------------------------- PAGE 3 Anh huong cua thoi gian toi hieu suat trich ly F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL. SECTION - 1 VARIATE GRAND MEAN STANDARD DEVIATION C OF V |CT$ (N= 10) -------------------- SD/MEAN | | NO. BASED ON BASED ON % | | OBS. TOTAL SS RESID SS | | 10 77.140 10.193 0.83862 1.1 0.0001 |

KQ



luan van hoan chinh

Documents