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M u
u nnh l loi cy trng ph bin trn ton th gii. Khng th ph nhn gi tr dinh dng v gi tr sinh hc m u nnh v cc sn phm t u nnh lm thc n cung cp cho con ngi cng nh cho cc loi ng vt. Vi hm lng protein cao, giu nguyn t v c, vitamin, t axit bo bo ho v hm lng cholesterol thp, u nnh c th c s dng di dng cc sn phm t u nnh nh u ph, sa u nnh, bt u nnh hay c th b sung vo cc loi thc phm khc lm tng gi tr dinh dng. Tuy nhin, mt nhc im lm hn ch vic s dng u nnh trong cng nghip ch bin thc phm ni chung chnh l nhng mi khng mong mun nh mi u, mi ngi... Cho n nay, Vit Nam cha c mt cng trnh nghin cu no tp trung vo vic gii quyt vn ny. Chnh v vy, ti nghin cu v a ra nhng quy trnh x l nhm hn ch ti a nhng mi khng mong mun trong cc sn phm u nnh. Vic nghin cu quy trnh phn tch v nh gi mi ca u nnh v cc sn phm t u nnh trc v sau khi x l l rt cn thit c th tm ra c quy trnh x l mi hiu qu nht v y cng chnh l nhim v t ra cho n ny.

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Phn I:

Tng quan

I.1. Gii thiu v u nnh: I.1.1. Gii thiu chung v u nnh:

u nnh (tn khoa hc l Glycine max (L.) Merrill, thuc cy h u (Fabaceae), phn h Papilionoidae) bt ngun t ng , c th t Trung v Bc Trung Quc. u nnh c trng nh mt cy lng thc trong hng ngn nm nay Trung Quc, cc nc vng ng v ng Nam v tip tc cho n ngy nay, l mt thnh phn quan trng trong thc n bnh dn truyn thng nhng vng ny. Sau th chin th II, nn nng nghip u nnh mi thc s pht trin M, Brazil, Canadav t vic dng u nnh lm thc phm, trong chn nui, trong cng nghip, y t ngy cng m rng. Theo tnh ton, nu quy ton b lng protein trong cc sn phm t u nnh hng nm trn th gii thnh cc sn phm tiu th ca con ngi th s cung cp cho 1/3 nhu cu protein thc phm trn ton cu. iu ny lm cho u nnh tr thnh mt trong nhng ngun cung cp protein ln nht cho thc n hng ngy [11].

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 n tt nghip I.1.2. Thnh phn ho hc ca ht u nnh :

Thnh phn ca u nnh c th thay i ph thuc vo ging v iu kin trng trt. Bng 1. Thnh phn ha hc ca u nnh Cc phn ca ht Phi V L mm Ton ht % khi lng ton ht 90 8 2 100 % theo khi lng cht kh Protei Lipit Cacbonhydr Cht n 43 9 41 40 23 1 11 20 at 43 86 43 35 tro 5.0 4.3 4.4 4.9

Theo: Cheftel v cc tc gi khc (1985)

I.1.2.1. Protein: Hm lng protein trong ht u nnh chim khong 40%. Phn ln protein u nnh thuc loi globulin, chim 80 90% protein c trong ht. Protein u nnh cha 8 loi axit amin thit yu, l isoleucine, leucine, lysine, methionine, phenylalamine, threonine, trytophane v valine (bng 2). y l nhng loi axit amin m c th con ngi khng th t tng hp c. Mt cch nh gi cht lng ca protein u nnh l dng t s ho hc ca n, thc hin bng cch so snh cc thnh phn axit amin cn thit ca n vi mt protein chun tham kho (v d nh protein tng ca trng). Trong protein u nnh, t l ca chng bng khong 70% so vi cc axit

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amin loi ny trong protein tng ca trng. V vy, t s ho hc ca protein u nnh s xp x 70%. Bng 2. Thnh phn axit amin cn thit ca u nnh Axit amin Isoleucine Leucine Lysine Methionine Meth. + Cyst. Phenylalanine Tyrosine Ph.ala + Tyr Threonine Tryptophan Valine A 4,54 7,78 6,38 1,26 2,59 4,94 3,14 8,08 3,86 1,28 4,80 B 114 110 118 74

133 96 133 97

A; g/16g Nit B: phn trm theo t l mu axit amin tm thi Theo: Thng tin ca FAO (1970) v FAO/WHO (1973)

I.1.2.2. Lipit: Cht bo ca u nnh (du nnh th) bao gm 96% triglycerit, 2% photpho lipit, 1,6% cht khng x phng ha, 0,5% axit bo t do v mt lng nh cc sc t carotenoid. Chng l nhng cht rt cn thit cho s hot ng v pht trin ca con ngi nh tham gia vo qu trnh to mng t bo, vo qu trnh sn sinh hoocmon v m bo s chuyn ho cholesterol v cc vitamin trong c th.

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Thnh phn axit bo trong du u nnh ph thuc vo ging v iu kin trng trt. Bng 3. Thnh phn axit bo trong du nnh Axit bo Tn thng thng Tn khoa hc Lauric Myristic Palmitic Stearic Oleic Linoleic Linolenic Dodecanoic Tetradecanoic Hexandecanoic Octodecanoic 9-Octadecanoic 9,12-Octadecandienoic 0 9,12,15Octadecantrienoic 0 18 : 1 18 : 2 18 : 3
Theo: Whahnon v cc tc gi khc (1988)

K hiu

Phn trm khi lng 12 : 4.5 4.5 14 : 11.6 2.5 16 : 18 : 21.1 52.4 7.1

0 0

I.1.2.3. Cacbonhydrat: u nnh cha khong 30% cacbonhydrat v c th c chia thnh 2 loi: ho tan v khng ho tan. Gluxit ho tan ch yu l 3 loi oligosacharid: sucroza, raffinoza v

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stachyoza. Cc gluxit khng tan trong u nnh l xenlulose, hemixenlulose, cc pectin v tinh bt. I.1.2.4. Khong cht: Hm lng cht khong trong u nnh khong 5%. Khong cht ch yu bao gm kali, canxi v magie. Phn khong cht cn li dng vt l cc nguyn t quan trng i vi dinh dng nh st, km, ng,...

I.1.3. Tnh hnh trng trt v sn xut:

Sn lng u nnh trn th gii khng ngng tng trong nhng nm gn y. Trong M tuy l nc trng u nnh rt mun (nhng nm 30, 40 ca th k 19) nhng li l nc ng u v sn xut u nnh vi khong 50% din tch trng v 75% sn lng ton th gii. Bn nc c sn lng u nnh cao nht trn th gii l M, Braxin, Trung Quc v Achentina, chim khong trn 90% sn lng ton th gii.

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Hnh 1. Biu sn lng v tiu th u nnh trn th gii t nm 1965 n nm 2005
Ngun: USDA report 4/2006

Vit Nam, din tch t trng v sn lng u nnh cng tng theo xu hng ca th gii. Trong lai to c nhiu ging u nnh mi c nng sut cao, thch hp vi cc iu kin a l v thi tit khc nhau, nh m din tch, vng trng cng nh sn lng u nnh khng ngng tng cao, gp phn p ng nhu cu protein cho ngi v gia sc. Bng 4. Din tch v sn lng u nnh ti Vit Nam Nm
Din tch (triu ha) S/lng (triu

1996 110,3 113,8

1998 129,4 146,7

2000 124,1 149.3

2002 158,3 205,6

2004 182,5 242,1

tn) Ngun: Tng cc thng k Vit Nam 2004

I.1.4. Gi tr dinh dng v gi tr y hc ca u nnh:

I.1.4.1. Gi tr dinh dng : Gi tr dinh dng ca u nnh c quyt nh bi cc thnh phn cha trong ht. Protein v lipit l hai thnh phn quan trng, chim khong 60% trng lng ht. Protein u

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nnh c gi tr cao khng ch do c s lng ln m cn v cht lng dinh dng ca n. Protein u nnh cha y cc axit amin cn thit t l cn i hi ho (bng 2). Ngoi ra n cn l ngun b sung lysin c gi tr vo thc phm ng cc. Qua nghin cu lm sng, nhiu tc gi khng nh cng thc protein chit xut t u nnh tng ng vi cng thc sa b v mc tng trng v nng albumin trong huyt thanh, mt khc nu so snh v khi lng gia protein u nnh vi thnh phn dinh dng sa m th u nnh l ngun cung cp protein v nng lng rt di do (bng 5). V vy protein u nnh c ngh thay th cho sa b trong cc cng thc nui tr em. Bng 5. Thnh phn dinh dng ca sa m, sa b v bt u nnh (tnh theo tng lng cht kh) Thnh phn Sa m Sa b Bt u nnh
Theo: Horan (1976)

Protein 8 28 44

Cht bo 27 28 22

Hydratcacb on 64 39 32

Du u nnh cng l loi thc phm qu i vi con ngi, c bit i vi nhng ngi ln tui v huyt p cao v n c h s ng ho cao, cha t axit bo bo ho (bng 3) v hm lng cholesterol thp.

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 n tt nghip
I.1.4.2. Gi tr y hc: Du u nnh cha t axit bo bo ho, trong c axit linolenic (axit omega-3), y l loi axit c kh nng gim thiu s nguy him ca bnh nhi mu c tim v phng nga cc bnh tim mch. Cc sn phm lm t u nnh c th gip ngn nga bnh ung th v trong u nnh c cha 5 nhm cht c c tnh chng li mm ung th, l: protease inhibitors, phytate, phytosterols, saponins, v isoflavones [7]. u nnh cn c bit n nh l mt thc phm chc nng gip ngn nga bnh tiu ng v cha tr cc bnh v thn, ngn chn thoi ha xng v lm cho xng chc kho.

I.2. Mt s sn phm t u nnh:

C th chia cc sn phm t u nnh theo hai nhm chnh sau :

I.2.1. Cc sn phm truyn thng:

V u nnh c ngun gc cc nc ng v Trung Quc nn cc sn phm truyn thng ch yu l nhng nc ny. Sau y l mt vi sn phm truyn thng ng ch . I.2.1.1. Sa u nnh: Sa u nnh l nc chit t nhng ht u nnh nguyn, l dch nh tng trng c, cha cc protein

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v cacbonhydrat ho tan trong nc, v phn ln du c trong ht u. Xay t l phng php sn xut sa u nnh c ph bin rng ri v c truyn thng lu i chu . Quy trnh c tm tt nh sau:

u nnh

Vo i sch
Sa u nnh

Ngm n c un si

Xay t

Lc

Quy trnh sn xut sa u nnh

Sa u nnh sn xut theo phng php ny thng c nhiu mi ngi. Hin nay ngi ta a ra quy trnh sn xut sa u nnh hin i, trong c cc cng on nh tch v, kh mi v x l UHT (tit trng nhit siu cao) gip ci thin c mi v r rt [11]. I.2.1.2. u ph: u ph l sn
Nc Nc chua
u nnh

Ngm, xay Lc, un si Kt ta Cht Hoa u p u ph Quy trnh sn xut u ph

phm rt quen thuc

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i vi nhn dn cc nc Trung Quc, Nht Bn v Vit Nam. Nu da trn mc p bnh u th c th chia u ph thnh 3 loi: loi cng, loi mm v loi u ph la. u ph dng ph bin Vit Nam hin l loi u ph mm. Trung Quc c sn xut c 3 loi nhng loi cng l ch yu, c dng xo vi cc loi rau, tht, trng. u ph la c sn xut quy m ln Nht Bn. Cng ngh sn xut u ph gm 3 khu chnh: xay nghin - lc - gia nhit, kt ta, nh hnh. I.2.1.3. Nc tng: Tng l loi dung dch mn, mu nu en, c mi hng c trng v v m . Thnh phn chnh ca tng l u nnh, bt m, mui v nc. Giai on quan trng nht ca qu trnh ch bin ny l s ln men, ti y cacbonhydrat chuyn ho thnh ru v axit lactic, cn protein b ct mch
Nc tng th u nnh Bt m

Nu Trn, Koji

Rang, ngin
Nm mc

Nc mui

Dch ngm

Vi khun lactic Nm men

Ln men p
Bnh

Thanh trng

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thnh cc peptit v axit amin. Phn ng ho hc gia cc cu t hu c v cc sn phm ln men to ra mu sc v hng v c trng cho nc tng. Qu trnh sn xut nc tng c th c chia thnh 3 giai on chnh: chun b koji, ln men trong nc mui, v tinh ch sn phm. I.2.1.4. Chao: Chao l sn phm ln men t u ph, c gi tr dinh dng cng nh h s tiu ho cao hn u ph v tri qua qu trnh ln men.

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u ph chao Mc bt u ph chao Mc bt

Cy mc Nui mc

Cy mc Nui mc

Dch r u Mui n

Ngm Ln men nng Ln men lnh Bao gi Chao bnh Qu trnh sn xut chao bnh

Nc mui R u

p mui Ln men Chao n c Qu trnh sn xut chao nc

Phn ln protein c chuyn ho thnh cc axit amin. Chao c th s dng ngay hoc cho vo cc thc phm khc nh m n lin, bt canh, bnh lng kh Sn phm chao hin nay gm hai loi chnh l chao bnh v chao nc. Cng ngh sn xut chao gm 3 giai on chnh: sn xut u ph, nui mc, ln men.

I.2.2. Cc sn phm hin i:

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Cc sn phm hin i ch yu c sn xut v ph bin cc nc cng nghip pht trin nh M, Nht Bn, Vit Nam ch yu ph bin cc sn phm u nnh truyn thng nh sa u nnh, u ph, to h, chao II.2.2.1. Cc sn phm u nnh tch bo dng bt v dng tm: Cc sn phm ny u c sn xut t u nnh loi bo v c thnh phn ho hc ging nhau. S khc nhau chnh l kch thc phn t. Sau khi xay th v ry u nnh tch bo, ngi ta thu c sn phm dng tm. thu c bt th th phi xay k hn. u nnh sau khi ra khi h thng chit cht bo cha khong 30% dung mi nn cn c tch dung mi. Qu trnh ny gm 2 giai on. u tin mnh u nnh sau khi chit du c i qua dng hexan nng ly ra phn ln dung mi c trong , sau lng dung mi cn li c tch bng cch cho i qua hi nc. Bng 6. Thnh phn ho hc ca bt v tm u nnh tch bo (Protein, cht bo, cht x v cht tro tnh theo tng lng cht kh) Protein (Nx6,25) u nnh nguyn % 42,6 m % 11,0 Cht Cht x Cht bo % 20,0 % 5,3 tro % 5,0

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ht Bt N tch bo Bt N nguyn bo
Theo: Circle v Smith (1972)

59,0 46,6

7,0 5,0

0,9 22,1

2,6 2,1

6,4 5,2

II.2.2.2.

Protein

nnh

(protein

concentrate): thu c sn phm c hm lng protein cao hn, ngi ta tip tc x l bt hoc tm u nnh tch bo. Bng cch loi b cc ng ho tan trong nc, tro v mt s thnh phn khc, ngi ta thu c protein u nnh c c c hm lng protein khong 60-70%. Quy trnh cng ngh c tin hnh theo s sau:
Bt u nnh tch bo Ra

Thnh phn ho tan (ng, tro, )

Thnh phn khng tan (protein, cacbonhydrat) Protein concentr ate

Quy trnh sn xut protein u nnh c c

C 3 phng php chnh ra bt u nnh tch bo: 1. Ra bng dung dch cn

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2. 3. Ra bng axit long Ra bng nc v nhit Cu t Phn trm khi lng Protein (Nx6,25) 70 Cacbonhydrat khng 20 tan Cht tro Cht bo
Theo: FAO Production Yearbook

Bng 7. Thnh phn ca protein u nnh c c

5-8 1

II.2.2.3. Protein u nnh phn tch (protein isolate): Protein u nnh phn tch l phn chnh cha protein ca u nnh c lm t u nnh bc v bng cch loi b phn ln cc cu t khng protein v phi cha t nht 90% protein tnh theo tng lng cht kh. (theo 90 Soya Bluebook). Nguyn liu ban u l bt u nnh tch bo cha protein c kh nng ho tan cao. Tin hnh chit protein bng dung dch kim long pH t 7,5 n 9,0. Axit ho dch chit n vng ng in pH = 4,5 th protein s kt ta. Thu kt ta protein bng cch lc hoc ly tm, ri ra kt ta bng nc. Sy lun kt ta ta c protein u nnh phn tch ng in. Nu trung ho ri mi sy ta thu c protein u nnh phn tch dng dn xut kim. Phng php sy c dng ph bin l phng php sy phun.

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Bng 8. Thnh phn ca protein u nnh phn tch (tnh theo tng lng cht kh) Cu t Protein Cht bo Cht tro Tng cacbonhydrat
Theo: Kolar v cc tc gi khc (1985)

Phn trm khi lng 90 0,5 4,5 lng 0,3

II.2.2.4. Cc sn phm b sung u nnh: nng cao gi tr dinh dng, ngi ta cn b sung u nnh vo nhiu loi thc phm khc nh c ph, scla, bnh ko, margarin, pho mt, pat, kem,

I.3. Cc hp cht to mi cho u nnh:

Nh trnh by, gi tr dinh dng v tc dng sinh hc ca u nnh c khng nh. Nguyn liu u nnh sn c tim nng ln Vit Nam. Tuy nhin, Vit Nam vic s dng u nnh nh mt thc phm ring l, nht l a vo thnh phn cc loi thc phm khc cn rt hn ch. Nguyn nhn l v u nnh v cc sn phm t u nnh c mi kh chu, m c th l mi ngi v mi u. Vy cc hp cht gy mi cho u nnh tn ti sn trong u hay c sinh ra trong qu trnh ch bin? Trn

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th gii c nhng nghin cu nhm loi b nhng mi khng mong mun ca u nnh. Tuy nhin, Vit Nam cho n nay vn cha c mt nghin cu c th no v vn ny. Da theo nhng nghin cu c h thng v cc hp cht gy mi cho u nnh, ngi ta pht hin ra mi ca u nnh gy ra bi hai nguyn nhn: cc hp cht bay hi c sn trong ht u, v cc sn phm ca qu trnh oxy ho kh lipit bi emzym lypoxygenaza v mt s enzym khc.

I.3.1. Cc hp cht bay hi c sn trong u nnh:

Bng phng php sc k ngi ta tch c nhng nhm cht bay hi sau y: Cc hp cht cacbonyl, cc axit phenollic, cc axit bo v amin bay hi, cc ru v este. Theo Boatright (2002) th hexanal, acetaldehyde v acetone l nhng hp cht cacbonyl bay hi ch yu. Fimaki (1965) cng b rng hexanal trong ht u nnh sng tuy c hm lng rt b, khong 10ppm nhng li c ngng pht hin rt thp v l mt trong nhng thnh phn chnh gy nn mi ngi. Willkens v Lin (1970) cho rng hexanal l loi aldehide c mch trung bnh, c th kt hp vi protein trong u nnh mt cch kh bn vng, nn vic loi b aldehide ny ra khi u nnh l mt vic rt kh khn.

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Ngoi hexanal, nhng hp cht chnh gy ra mi u v mi ngi cho u nnh v cc sn phm u nnh cn l ethyl vinyl xeton, 3-cis hexanal v 2 petyl-furan (Sessa v Rackis, 1977). Theo Evans (1971), ethyl vinyl xeton c ngng pht hin l 0,02ppm, y c th l cht gy ra mi ngi ca u ngm nc. 2 pentyl-furan c pht hin u tin bi Chang v ng s (1966) v l nguyn nhn gy ra mi u cho sa u nnh. Trong s 9 axit phenollic tm thy trong bt u nnh, axit syringgic c nng cao nht, sau l p-coumaric v ferulic. Chng c coi l nguyn nhn gy ra mi kh chu cho bt u nnh. Ch c mt lng nh cc axit bo bay hi c tm thy trong bt u nnh m ch yu l axit iso caproic, ncaproic v n-caprilic. Amoniac methylamin, dimethylamin, piperidin v cadaverin l cc amin bay hi. Vai tr ca chng trong vic to nn mi kh chu cho u nnh cn cha c xc nh r. Cc loi ru nh 3-methyl-1-butanol, n-hexanol, n-heptanol cng gp phn to nn mi u. Cc hp cht to mi c sn trong ht u nm ch yu v. y chnh l mt trong nhng c s xy dng quy trnh x l mi cho u nnh v cc sn phm u nnh.

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 n tt nghip I.3.2. Cc hp cht to mi sinh ra trong qu trnh ch bin:

Trong qu trnh ch bin, lipit trong ht u c th b oxy ho di tc dng ca enzym lipoxygenaza to thnh nhng sn phm khc nhau. Lipoxygenaza hay lipoxydaza l enzym oxy ho kh, xc tc s oxy ho cc axit bo khng no cha 2-3 ni i (hoc nhiu hn) nh axit linoleic, linolenic. im c bit l lipoxygenaza ch oxy ho dng cis-cis cn dng cis-trans v trans-trans th hon ton khng tc dng. C ch tc dng ca lipoxygenaza nh sau: u tin to thnh phc hp gia enzym, oxy v c cht (axit bo). Tip gc kp c to thnh do kt qu ca s chuyn hydro t c cht ti oxy. Mt ni i b chuyn ch to nn h thng ni i lin hp. Phc hp b phn ly gii phng ra enzym v hydro peroxyt c dng cis-trans. T hydro peroxyt s chuyn ho thnh epoxyt, andehit, xeton v nhng vt phm oxy khc, y chnh l mt trong nhng nguyn nhn gy ra mi kh chu cho u nnh v cc sn phm u nnh.
Axit Linoleic (18:2) Axit Linolenic (18:3)

O2

O2

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trans 2 hexenal 2 pentylfuran Hexana l

Hnh 2. S qu trnh oxy ho mt s axit bo thnh cc hp cht gy mi Trong ht u nnh c nhiu lipoxygenaza, pH ti u ca lipoxygenaza trong u nnh l 9,0. c th hn ch s to mi khng mong mun trong qu trnh ch bin, ngi ta a ra nhng bin php nhm c ch hay v hot enzyme ny nh dng nhit, dng Cc hp cht gy mi cho u nnh ch yu tan trong dung mi hu c nh cn v n-Hexan, eter du, diethyl ether... y l c s cho qu trnh chit tch cc hp cht gy mi ra khi ht u v cc sn phm t u nnh c th phn tch thnh phn ca chng.

I.4. Cc phng php phn tch v nh gi cht lng mi:

C hai nhm phng php chnh phn tch v nh gi mi l phng php phn tch ho l v phng php phn tch cm quan. i vi phng php phn tch ho l, ta c th nh tnh, nh lng c thnh phn cc hp cht bay hi. Nh m ta xc nh c bn cht ca nhng s thay i mi trong cc sn phm thc phm sau khi c

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x l. Tuy nhin, phng php ny li khng c c u im nh ca phng php phn tch cm quan, l tm hiu c cm nhn ca con ngi i vi mi ca sn phm. y l mt u im rt ln ca phng php phn tch cm quan v cc sn phm thc phm lm ra cui cng u c con ngi tiu th bng cch n hay ung. Nh phng php phn tch cm quan, ta c th xc nh c cm gic nhn c ca con ngi do cc hp cht bay hi gy ra (con ngi gi l nhng mi g), v liu con ngi c nhn ra s thay i ca cc tnh cht mi hay khng. Nu kt hp c hai phng php phn tch trn phn tch v nh gi mi ca mt sn phm thc phm, ta s ng dng c u im ca c hai phng php.

I.4.1. Phng php phn tch ho l:

c th phn tch thnh phn to mi ca mt sn phm thc phm, trc ht ta phi tch chit cc hp cht mi ra khi cc thnh phn khc, thu chng vo dung mi thch hp, ri sau mi phn tch dch chit. I.4.1.1. Tch chit cc hp cht to mi: Cc hp cht to mi nm trong mt sn phm thc phm c mi lin kt vt l hay ho hc vi cc thnh phn khc trong sn phm nh lipit, protin, gluxit, Tu thuc vo cu trc ca sn phm m cc mi lin kt mnh hay yu, v s c nhng phng php tch chit tng ng. Do vy khi tch chit cc hp cht mi cn ch n cu trc ca sn

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phm . Sau y l mt s phng php tch chit thng s dng. I.4.1.1.1. Phng php c hc : Phng php c hc l phng php thu cc hp cht mi bng cc lc c hc nh p, vt, nghin, Dch thu c c dng huyn ph sau c em i tch nc. Phng php ny thng p dng cho nguyn liu c hm lng cht to mi cao, lin kt t do v phn b gn b mt nguyn liu nh h qu citrus. Tin hnh: ct nguyn liu thnh ming nh t yu cu ri cho qua my p, thu c hn hp dch p gm nc v cc hp cht mi. Sau cc hp cht mi c tch ra khi hn hp nh thit b ly tm. I.4.1.1.2. Phng php chng ct: Chng ct l qu trnh phn tch hn hp lng gm cc thnh phn d bay hi da trn s khc nhau gia p sut ring phn v nhit si ca chng. Trong cng ngh, sau khi cc thnh phn bay hi cun theo hi nc th ngng t hn hp ri tch chng ra khi nc. Phng php ny c p dng khi:
-

Cc cu t to mi bay hi theo hi nc nhng khng hoc t tan trong nc.

- Trong nguyn liu, chng dng t do hoc c gii phng trc khi chng ct. - Chng khng b ph hu nhit cao.

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Tin hnh: Ct hoc nghin nguyn liu n kch thc ph hp ri cho vo bnh chng ct. Tu thuc vo khi lng ring ca hp cht mi ta cn thu m chn loi bnh chng ct tng ng. Cc hp cht phn cc d bay hi b cun theo hi nc trong bnh chng, sau ngng t ti ng ngng v b lu gi . Phn hi lu quay li
ng sinh hi lu v tip tc c chng hn

bnh chng bng ng

ng s h inh n

ct. Qu trnh din ra nh vy cu.

cho n khi t yu

ng hi luu
B c nh u

ng hi luu

B c nh u

3a Hnh 3. S thit b chng ct 3a: Thu cht bay hi c d < 1 hi c d > 1 I.4.1.1.3. Phng php trch ly:

3b 3b: Thu cht bay

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Phng php trch ly l phng php tch mt s cht tan ra khi cht lng hoc cht rn bng mt dung mi da trn tnh ho tan c chn lc ca dung mi . Phng php ny thng p dng cho nguyn liu c hm lng cht to mi dao ng rng, c mi lin kt kh cht vi cc thnh phn khc trong nguyn liu. Tin hnh: ct hoc nghin nguyn liu n kch thc ph hp, cho vo bnh trch ly. Ti y nguyn liu c tip xc vi dung mi v bng vic chn dung mi thch hp, cc hp cht mi s ho tan vo dung mi. Dung mi tip xc vi nguyn liu c th trng thi ng hoc tnh. Sau y l hai phng php trch ly ng vi hai trng thi dung mi thng c s dng. a) Phng php chit
Tr chi t ng h i ng xi phng ng s h inh n

Soxhlet: Nguyn liu sau khi gi, nghin nh n kch thc ph hp c gi vo ng giy ri cho vo tr chit. Dung mi t bnh cu bay hi theo ng hi ri ngng t vo tr chit, ti y dung mi tip xc vi nguyn liu v ho tan cc cht cn tch. Khi dung mi ngng t n mp trn ca ng Hnh 4. Thit b chit Soxhlet Nguyn Th Ngc Hi_ TPN K46
25
B c nh u

 n tt nghip
xiphng th hn hp dung mi v cht cn tch s theo ng xiphng chy xung bnh cu. Ti y dung mi li tip tc bay hi v qu trnh chit c lp li. Nh m nng cht cn tch trong nguyn liu lun cao hn trong dung mi nn lun xy ra qu trnh chuyn cht t nguyn liu vo dung mi. b) Phng php chit siu m: Phng php siu m l mt phng php chit m dung mi tip xc vi nguyn liu dng tnh. Nguyn liu sau khi c nghin n kch thc ph hp, cho vo l cng vi dung mi chit ri t vo b siu m. Cu to ca b siu m rt n gin, l mt b c np, c b phn pht sng siu m bn trong, cc l ng mu c t trn mt khay ngm trong nc (lng nc va ngp ht mu). Sng siu m truyn n mu v vi nng lng ln n ct t hoc lm yu mi lin kt gia cc hp cht mi vi nhng thnh phn khc trong nguyn liu, nh m cc hp cht mi d dng ho tan v dung mi hn. So vi phng php chit Soxhlet, chit siu m c mt nhc im l nguyn liu

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Hnh 5. Phu chit qu l

 n tt nghip
khng c tip xc vi dung mi mi, cn bng sm t c v hiu qu tch s khng cao. Nhc im ny c th khc phc bng cch tng lng dung mi chit. c) Phng php chit lng lng: Chit lng lng l k thut da trn s phn b khc nhau ca cht tan vo hai pha khng trn ln, t tch chit cht phn tch ra khi nn hoc tch cc tp cht ra khi cht phn tch. Nguyn tc ca k thut chit ny l hai pha lng khng trn ln c vo nhau, trong mt dung mi cha cht phn tch, c trong mt dng c chit nh phu chit, bnh chit. Khi lc chit, cht phn tch s c phn b vo hai dung mi, v trng thi cn bng, t l dung mi ca cht phn tch hai pha l mt hng s (h s phn b). i vi cht phn tch, h s phn b cng ln th chit cng trit . Phng php ny p dng cho nguyn liu dng lng. d) Phng php chit pha rn: Chit pha rn s dng pha ng l dung mi di chuyn hn hp cc cht gy mi i qua pha tnh. Cc thnh phn khc nhau ca dch chit s c i lc khc nhau i vi pha ng v pha tnh v xut hin t pha tnh ti nhng thi im khc nhau. Pha tnh v pha ng c la

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chn da vo bn cht ca cc hp cht gy mi t c hiu qu phn tch tt nht. Trong a s cc trng hp, pha tnh thng l silica (SiO2) hay alumina (Al2O3), c trn vi dung mi c s dng lm pha ng. Chit pha rn l phng php lm giu v lm sch mu phn tch t dung dch chit ra bng cch hp ph ln mt ct pha rn, sau cht phn tch c ra gii (gii hp) bng dung mi thch hp. Phng php ny dng tch chit cc mu dng lng, ng thi c th dng lm sch hoc lm giu dch chit. I.4.1.2. Phn tch thnh phn cc cht to mi : Sau khi thu c cc hp cht to mi vo dung mi thch hp, dch chit ny c lm sch nu cn thit ri a i phn tch xc nh thnh phn cc hp cht to mi. C nhiu phng php trong nhm phng php phn tch dng c. V d ta c th dng cc phn ng c trng nh phn ng mu nh tnh v nh lng cc thnh phn, tuy nhin s rt phc tp v tn nhiu cng sc v mi phn ng ch c trng cho mt cht hoc nhm cht nht nh. Hin nay, phng php sc k ang c ng dng rng ri trong phng th nghim phn tch v n c rt nhiu u im nh phn tch n gin (ch cn mt ln chy mu,

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ngi phn tch nh tnh, nh lng cc thnh phn), kt qu chnh xc do nhy ca thit b cao I.4.1.2.1. Nguyn tc chung ca phng php sc k: Tt c cc dng sc k u da trn mt nguyn tc rt n gin. Mu c phn tch (gi l cht tan) c cho tip xc vi 2 pha khc nhau v tnh cht vt l l pha ng v pha tnh. Pha ng y mu qua vng cha pha tnh gi l cht hp ph. Mu phn tch c th cha mt hoc nhiu cu t, khi tip xc vi pha tnh, cc cu t thnh phn ny s t phn b vo 2 pha ng v tnh. Nu cu t no c i lc nhiu vi pha tnh, chng s b gi lu hn trong pha tnh v do vy chng s ra khi h thng sc k mun hn. Ngc li nhng cu t no c i lc t vi pha tnh s b loi ra khi h thng nhanh hn. Cc cu t ny c tch ra l do chng khc nhau v i lc i vi 2 pha ng v tnh. Da theo cc loi pha ng, ta c th phn loi phng php phn tch sc k thnh sc k lng (pha ng l cht lng) v sc k kh (pha ng l cht kh). Trong , phng php sc k kh c ng dng nhiu trong phn tch cc hp cht bay hi. I.4.1.2.2. Phng php sc k kh: Phng php sc k kh c kh nng tch c hon ton nhng cht hu c kch thc ging nhau, n cng c th tch nhng hn hp rt phc tp cha nhiu hp phn. V

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vy m phng php ny c ng dng rng ri. Hn na, thng cc cht to mi l nhng cht d bay hi nn p dng phng php phn tch sc k kh l rt thch hp.

Hnh 6. S thit b sc k kh. Nh c kh mang cha trong bom kh (hoc my pht kh), mu t bung bay hi c dn vo ct tch nm trong bung iu nhit. Qu trnh sc k xy ra ti y. Sau khi ri khi ct tch ti cc thi im khc nhau, cc cu t ln lt i vo detect, ti chng c chuyn thnh tn hiu in. Tn hiu ny c khuch i ri chuyn sang b ghi, tch phn k hoc my tnh. Cc tn hiu c x l ri chuyn sang b phn in v lu kt qu. Vo nhng nm u ca thp k 70, k thut lin hp kn gia phng php sc k kh v ph khi lng c ra i. Cc cu t c tch khi ct sc k s ln lt c a vo ngun ion ca my khi ph. Ti chng c phn mnh v c tch khi nh mt t trng ri i vo b nhn quang chuyn ho thnh tn hiu in. ng vi mi mt pic

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trn sc k s nhn c mt khi ph ring bit v hon chnh.

Hnh 7. S lin hp sc k kh khi ph I.4.1.2.2. Phng php sc k lng: Khi pha ng l cht lng thay cho cht kh th phng php phn tch c coi l php sc k lng (LC). LC bao hm mt phm vi rt rng nhiu khi nim vi nhiu phng php sc k nh sc k lng cao p (HPLC), sc k phng. Trong LC mu c ho tan vo mt dung mi v vic tch s c tin hnh gn hoc ngay nhit phng. Phng php ny c bit c li cho cc hp cht sinh ho c th b phn hu nu phi lm bay hi nh cc lipid trong cc sinh t tan trong m, peptid trong cc mu protein khc nhau

I.4.2. Phng php phn tch cm quan:

So vi cc k thut phn tch trong lnh vc nng sn thc phm, phn tch cm quan c c trng l: con ngi khng ch l k thut vin m cn l thit b phn tch cung cp s liu. S c trng ny gii thch cho vic ra i rt sm ca mn th nm ( t khi loi ngi bt u s

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dng thc n chn) v vic pht trin rt nhanh ca phng php phn tch cm quan trong hai thp k va qua. phn tch v nh gi cht lng mi, v ca thc phm, phng php phn tch cm quan l khng th thay th. Tuy nhin mc nhy cm ca ngi th thay i theo nhiu yu t v rt khc nhau gia nhng ngi th, dn n kt qu c phn tn cao nn vic khai thc chng cn hn ch hn phng php dng thit b. Phn tch cm quan thc phm l k thut s dng cc gic quan ca con ngi tm hiu, m t v nh lng cc tnh cht cm quan vn c ca mt sn phm thc phm nh : mu sc, hnh thi, mi v v cu trc, n khng ph thuc vo ngi th. Trong phn tch cm quan, cc gic quan ca ngi th c s dng nh mt dng c o, gic quan lm nhim v nhn thng tin cm gic, x l v a ra kt qu di dng cc gi tr c lng, so snh v m t. Chnh v vy cc kt qu phn tch cm quan ch c gi tr khi c x l bng cc phng php thng k ton hc. C nhiu php th cm quan, tt c u da trn mt c s chung l khi c kch thch ln c quan cm gic s tip nhn, x l v bn cht v cng ca kch thch . Khi cng kch thch tng th cng cm gic nhn c cng tng theo.

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phn tch, nh gi mi ca u nnh v cc sn phm t u nnh, ta c th p dng mt s php th khi bit cc tnh cht cn so snh sau y: I.4.2.1. Php th so snh cp: a) Mc ch: Php th so snh cp c dng khi mun xc nh xem liu hai sn phm c khc nhau v mt thuc tnh cm quan no khng. b) Nguyn tc: Ngi th c mi tr li liu c s khc nhau gia hai mu A v B i vi mt tnh cht cm quan no khng, nu c th mu no (ngt, chua, thm...) hn. Nu ngi th khng thy mu no hn mu no th h phi chn mt mu bt k trong hai mu gii thiu. I.4.2.2. Php th so hng: a) Mc ch: Php th so hng c s dng khi mun so snh nhiu hn hai sn phm v mt tnh cht cm quan no (v ngt, mi to). Php th ny cng c th s dng vi mc ch xc nh mc a thch ca ngi th i vi mt nhm sn phm cng loi. b) Nguyn tc: Nhim v ca ngi th l sp xp theo mt th t tng dn (hoc gim dn) v mc kch thch nhn c (v

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ngt, v chua) hoc theo mc a thch. Trong trng hp ngi th phi nut mu nh gi ch tiu mi v th khng nn yu cu h sp xp qa 7 mu. Nu ngi th lm vic vi cc kch thch vt l nh so hng ch tiu mu sc th s lng mu c th tng ln. I.4.2.3. Php th cho im: a) Mc ch: Bn cht ca php th l lng ho cng cm nhn ca mt tnh cht cm quan bng im s. V vy, php th ny gip m t mc khc nhau gia cc sn phm. Php th ny i hi mt hi ng phi c xy dng t nhng thnh vin qua hun luyn. b) Nguyn tc: Ngi th s dng mt thang im nh gi cng cm gic. C hai loi thang o thng c s dng l thang c cu trc v thang o khng c cu trc. I.4.2.4. Php th c lng ln: a) Mc ch: Trong thc t sn xut ngi ta thng t vn :
-

Vi cng cm gic xc nh nhn c khi th

nm th mu c nng (hay cng cht kch thch) l bao nhiu?

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-

Nu b sung 1 lng xc nh cht kch thch vo

sn phm th cng cm gic nhn c l bao nhiu? b) Nguyn tc: Cng cm gic nhn c S quan h vi cng kch thch I theo hm: LnS = nlnI + K Trong : S I n : Cng cm gic : Cng kch thch : H s ph thuc php th, bn cht kch thch

K, k: Hng s v thao tc thc hnh Phn ln cc gi tr n u nh hn 1, iu gii thch ti sao cng cm gic nhn c bin i chm hn s thay i ca cng kch thch.

I.5. Mc ch v ni dung nghin cu:

Nhng tng hp trn y cho thy u nnh v cc sn phm t u nnh l ngun thc phm rt c gi tr i vi con ngi, c bit l gi tr dinh dng v tc dng sinh hc. Trn th gii chng loi cc sn phm u nnh rt a dng, t nhng sn phm truyn thng cho n cc sn phm hin i. Ti Vit Nam, mc d din tch v sn lng trng

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u nnh nc ta khng ngng tng ln trong nhng nm gn y, nhng u nnh mi ch c s dng di dng cc sn phm truyn thng nh sa u nnh, u ph, nc tng, chaoNguyn nhn ca hn ch ny l do u nnh c mi kh chu nh mi ngi, mi u. Chnh v vy, ti nghin cu v a ra nhng quy trnh x l nhm hn ch ti a cc mi khng hp dn ny ca u nnh c th m rng phm vi ng dng v a dng ho cc sn phm t u nnh. Xut pht t nhng iu trn, chng ti nhn thy vic phn tch v nh gi mi ca u nnh v cc sn phm t u nnh trc v sau x l l rt cn thit xc nh c quy trnh x l mi hiu qu nht. Mt khc, trn th gii c nhiu nghin cu v vn x l cng nh phn tch, nh gi cht lng mi ca u nnh v cc sn phm t u nnh, in hnh l M v Nht Bn. Tuy nhin, i tng nghin cu ca h l cc ging u nnh trng ph bin nhng vng a l , v h cng ch cng b kt qu nghin cu m cha a ra phng php phn tch v nh gi c th. Hn na ti Vit Nam cho n nay vn cha c mt cng trnh nghin cu no v vn ny. Do vy, trc ht chng ti s nghin cu nhm tm ra quy trnh phn tch v nh gi mi ca u nnh v cc sn phm t u nnh, v i tng m

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chng ti nghin cu l mt ging u nnh trng ti Vit Nam (xem II.1). n ny l mt phn trong ti Nghin cu cc bin php x l mi khng mong mun ca u nnh nhm to ra bt u nnh cao cp ng dng sn xut thc phm chc nng v ung dinh dng. C th tm chia ti ny thnh hai phn chnh l Nghin cu cc phng php x l nhm loi b mi khng mong mun ca u nnh v cc sn phm u nnh v nh gi mi ca u nnh v cc sn phm u nnh trc v sau x l nhm xc nh c phng php x l mi tt nht, trong n m chng ti thc hin l phn 2 ca ti. c th tin theo di n, sau y chng ti xin a ra mc ch v ni dung ca c ti, trong phn ch in nghing l mc ch v ni dung nghin cu ca n ny.

I.5.1. Mc ch:
-

Loi b mi khng mong mun ca u nnh v Xy dng quy trnh phn tch thnh phn

cc sn phm u nnh.
-

to mi ca u nnh v cc sn phm t u nnh trc v sau x l.

-

Xc nh quy trnh x l mi tt nht.

I.5.2. Ni dung:
-

X l u nnh bng cc phng php khc nhau.

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-

Phn tch thnh phn to mi da trn quy

trnh va xy dng v nh gi cm quan cht lng mi ca u nnh v cc sn phm t u nnh.

Kt hp kt qu phn tch ho l v kt

qu nh gi cm quan a ra kt un v quy trnh x l mi tt nht.

Phn II.

Nguyn, vt liu &

phng php nghin cu

II.1. Nguyn, vt liu:
S tng qut quy trnh sn xut bt u nnh ng dng trong cng nghip thc phm (sn xut bt u nnh ung lin, b sung vo ccan phm thc phm khc tng gi tr dinh dng) [11] gm 2 cng on chnh nh sau:

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Ht u nnh

(1)

Sa u nnh

(2)

Bt u nnh

Quy trnh sn xut bt u nnh

Trong : (1): Cng on ch bin thnh sa u nnh. (2): Cng on sy phun thnh bt u nnh. Da trn quy trnh sn xut sa u nnh truyn thng, nhng nghin cu v nguyn nhn gy ra mi khng mong mun cho u nnh v tham kho cc quy trngn xut sa u nnh hin nay ang c p dng, nhm nghin cu thuc phn 1 ca ti a ra cc bin php x l b sung vo quy trnh sn xut sa u nnh truyn thng nhm loi b nhng mi ny, bao gm tch v v x l nhit (hnh 8).
Chn Xay Gia nhit Lc un si
Sa N

Ht u nnh Tch v Ngm

II.1.1. Nguyn liu:

T 7 ging u nnh do Trung tm nghin cu th nghim u Thanh Tr H Ni cung cp vo thng 2 nm 2006, sau khi phn tch thnh phn (bng 9, do phn 1

Sy phun
Bt N

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Hnh 8. S quy trnh sn xut bt u nnh c p dng cc bin php x l [11]

 n tt nghip
ca ti thc hin), ti chn ging u nnh VX 93 nghin cu. VX 93 l ging u nnh c hoa mu trng, thi gian sinh trng 85 - 90 ngy, phn cnh kho, qu khi chn c mu nu. Ht vng sng, khi lng 100 ht 15 - 16 gam, nng sut t t 16 - 20 t/ha. Trong iu kin thm canh t 25 t/ha. y l ging c kh nng chu rt, thch hp cho v Thu ng ng Bng Bc B, thch hp cho v H cc tnh min ni nh: Trng Khnh, Cao Bng.

Bng 9. Thnh phn ho hc ca ging u nnh VX 93 [11] Nh ni, tch v v x l nhit l nhng bin php Thnh phn Protein Lipit X m (%) VX 93 39,43 20,51 5,68 11,85 nhm loi b mi khng mong mun ca u nnh v cc sn phm t u nnh. Vy c th nh gi c hiu qu x l mi ca tng bin php, chng ti tin hnh phn tch v nh gi mi ca nguyn liu ban u l ht u nnh, sn phm trung gian l sa v thnh phm l bt u nnh,c th nh sau: - Ht u nnh nguyn v v tch v - Sa u nnh c v khng gia nhit sau xay

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- Bt u nnh

II.1.2. Thit b, dng c v ho cht:

II.1.2.1. Dng c:
-

B chit Soxhlet B chng ct Phu, cc thuy tinh, l thu tinh Phu chit qu l Giy lc, cc, chn, khay, bnh nha... H thng phn tch sc k kh khi ph GCMS Shimadzu 2010 Thit b chit siu m

- Ci, chy
-

II.1.2.2. Thit b:
-

- My xay sinh t II.1.2.3. Ho cht:

-

n-Hexan Cn 960 Natri sunpht khan Cc loi tinh du

II.2. Phng php nghin cu: II.2.1. Phn tch ho l:

II.2.1.1. Tch chit cc hp cht to mi:

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Nh nu cc phng php phn tch mi, khi tch chit cc hp cht mi cn ch n cu trc ca nguyn liu. Do vy i vi tng loi nguyn liu c cch tch chit ring. II.2.1.1.1. Tch chit cc hp cht mi ca ht u nnh: V y l mu dng rn, kch thc kh ln nn trc khi tch chit, cn tin hnh gi nh ht u nhm tng din tch tip xc vi dung mi. Hn na ta c th nhn thy khi cn dng ht kh, mi ngi v mi u ca ht u nnh khng th hin r. iu ny chng t cc hp cht mi lin kt kh cht ch vi cc thnh phn khc trong ht u. Do vy chng ti s dng cc phng php tch chit nh chng ct, chit siu m v chit Soxhlet tch cc hp cht mi ra khi ht u. Tin hnh nh sau: i vi ht u nnh nguyn v: loi b tp cht (rc, si) v cc ht xu, b hng. i vi ht u nnh tch v: sau khi lm sch ht u nnh nguyn v, sy nhit 45-500C trong khong 30 pht ri mang i c cho v i ht u tch v. Gi ht u (nguyn v hoc tch v) ri tin hnh tch chit theo cc phng php chn. V vic tch chit cc hp cht mi ca u nnh ln u tin c thc hin ti Vit Nam nn cha c ngun ti liu tham kho no c th, do vy phi tin hnh kho st

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kch thc gi ht, khi lng mu tch, loi dung mi v t l dung mi chn ra quy trnh tch hiu qu. Kch thc ht gi: mm. Xay ht u bng my xay sinh t n mn. Khi lng mu tch v t l dung mi (p dng cho phng php chit siu m vi c cc loi dung mi):
-

Gi dp ht u v thnh 4-5 mnh. Gi ht u nh n ht c ng kch khong 2

20 g nguyn liu : 20 ml dung mi 20 g nguyn liu : 40 ml dung mi 10 g nguyn liu : 25 ml dung mi

i vi phng php chit Soxhlet, kho st khi lng nguyn liu trong khong t 3 n 10 g, cn th tch dung mi lun m bo bng khong 3/2 th tch tr chit tnh t y tr n mp trn ca ng xiphng. i vi phng php chng ct, kho st khi lng mu v lng nc s dng nh sau: 150 g nguyn liu : 500 ml nc ct 50 g nguyn liu : 250 ml nc ct 25 g nguyn liu : 250 ml nc ct

Sau khi chn c t l nguyn liu : nc chng ct, chng ti tin hnh kho st chng ct 1 ln v chng ct 2 ln trn t l . Chng ct 2 ln l ly dch ct ln 1 em i chng ct tip.

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II.2.1.1.2. Tch chit cc hp cht mi ca sa u nnh: y l mu dng lng, qua cc cng on ch bin nh xay, x l nhit nn mi lin kt gia cc hp cht mi vi cc thnh phn khc b ct hoc lm suy yu phn no. Do vy, hai phng php tch chit c kho st l chit lng lng v chit pha rn, kho st cc loi dung mi khng tan ln vo sa nh n-hexan, h dung mi n-hexan : diethyl ether (3:1) [13], th tch mu chit, nhit mu chit v t l dung mi. Trc ht kho st phng php tch chit, sau s kho st cc thng s tch chit ca tng phng php, tin hnh nh sau: Nhit mu chit:
-

Ly mu ngay sau khi va ch bin xong sa

u nnh, tc l nhit mu khong 1000C ri chit. mu ngui n nhit phng ri mi ly mu em chit. T l th tch mu chit v dung mi (p dng cho chit lng lng): -

20 ml dch sa : 20 ml dung mi, tin hnh chit 20 ml dch sa : 20 ml dung mi, thu dch chit

mt ln. ln 1. Sau ra li phn cn chit bng 20 ml dung

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 n tt nghip
mi, thu dch chit ln 2. Trn 2 phn dch chit ri a i phn tch. i vi chit pha rn, chng ti kho st lng mu sa cho vo ct, loi v lng dung mi ra gii: -

Lng sa cho vo ct: 1ml, 3ml, 5ml Loi dung mi: n-hexan, h dung mi n-hexan : Lng dung mi ra gii: 50ml (10ml cho fraction 1

diethyl ether (3:1) v 40ml cho fraction 2), 60ml (cho fraction 1 v 50ml cho fraction 2), 70ml (cho fraction 1 v 60ml cho fraction 2) II.2.1.1.3. Tch chit cc hp cht mi ca bt u nnh: y l mu dng rn nhng l bt mn, tri qua nhiu cng on ch bin, c bit l x l nhit nn mi lin kt gia cc hp cht to mi vi nhng thnh phn khc tr nn yu hn. Do , chn phng php chit Soxhlet v chit siu m tch chit cc hp cht mi, s dng loi dung mi c hiu qu tch chit cao i vi cc hp cht to mi trong ht u nnh. II.2.1.2. Phn tch dch chit: Trong cc phng php phn tch dch chit trnh by, do c nhiu u im trong phn tch, c bit l phn tch cc hp cht bay hi v thun li khi s dng thit b nn chn phng php phn tch sc k kh.

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Thit b sc k kh s dng l h thng sc k kh khi ph GC-MS Shimadzu 2010 ti Trung tm gio dc v pht trin sc k EDC thuc trng i hc Bch khoa H Ni. Dch chit thu c bng nhng phng php tch chit trn khng cn phi lm sch hay lm giu na, m em i phn tch ngay. S dng my GC-MS 2010 Shimazu vi cc thng s sau:
-

Kh mang: Hli Th vin ph: NIS T 147, NIS T 107, Wiley 7 Loi ct tch: DB1-MS, DB-5-MS, DB Wax-MS Chng trnh nhit : t 70 n 2500C vi thi

Tin hnh kho st cc thng s sau:

-

- Lu lng dng: 1,0 ml/pht; 1,5 ml/pht

-

gian lu nhit v bc tng nhit khc nhau nh sau: 700C (n pht) k0C/pht 2500C (m pht). Trong : n = (1 10); k = (2 20); m = (4 10)

Hnh 10. H thng sc k kh khi ph GC-MS Shimadzu 2010

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II.2.1.3. X l kt qu: II.2.1.3.1. Phn tch nh tnh: T cc sc k thu c t cc mu, so snh vi ph chun tng ng lu trong cc th vin nu trn xc nh tn cc hp cht. II.2.1.3.2. Phn tch nh lng: Da vo phn trm din tch pic c c nh lng cc hp cht. So snh sc k ca hai mu trc v sau khi x l (c tin hnh theo cng phng php v cng dung mi) rt ra kt lun xem cc thnh phn to mi ca u nnh thay i nh th no sau qu trnh x l.

II.2.2. Phn tch cm quan:

nh gi c hiu qu ca cc cng on x l mi tch v v gia nhit, tin hnh so snh mi ngi v mi u ca cc cp mu trc v sau x l, c th nh sau: - Ht u nnh nguyn v v tch v
-

Sa u nnh c v khng c x gia nhit Bt u nnh sy phun t sa u nnh v sa u nnh

Tiu ch cn nh gi l hiu qu ca qu trnh x l mi, th hin thng qua cng mi u v ngi cn gi li trong sn phm. Do vy php th so snh cp c s dng ch r mi ngi v mi u c gim hay khng sau khi c x l.

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Theo cc ti liu tham kho, ht u nnh v cc sn phm t u nnh c mi u v mi ngi l hai tnh cht th hin mnh nht, chnh v vy m chn nh gi hai ch tiu ny. Vi mc ch nh gi, php th v ch tiu c la chn nh trn, tin hnh la thnh lp hi ng v hun luyn hi ng c th nh gi trn sn phm. II.2.2.1. Phng nh gi cm quan: Cc th nghim u c tin hnh ti phng th nghim nh gi cm quan (PTNGCQ) ca trng i hc Bch khoa H Ni. PTNGCQ m bo c cc yu cu ca mt phng nh gi cm quan nh thong mt, sch, khng c mi l v khng b nh hng bi ting n. Ngi th phi c yn tnh v lm vic c lp. PTNGCQ c thit k 8 ngn lm vic ring bit cho 8 ngi th, mi ngn c h thng n ring m bo yu cu tt nht cho ngi th nh gi mu. Cc bui nh gi c tin hnh trong phng vi nhit ti u l 20 - 250C. II.2.2.2. K hoch th nghim cm quan: Bui 1: La chn hi ng Bui 2: Hun luyn hi ng lm quen vi mi u v mi ngi Bui 3: Hun luyn kh nng nhn bit mi u Bui 4: Hun luyn kh nng nhn bit mi ngi

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Bui 5: Hun luyn kh nng phn bit cng mi u Bui 6: Hun luyn kh nng phn bit cng mi ngi Bui 7: Th nghim nh gi mi ca ht u Bui 8: Th nghim nh gi mi sa u nnh Bui cui: Th nghim so snh cp i mi ca bt u nnh sy phun t sa u nnh (sa ny c ch bin theo quy trnh c p dng bin php x l m cc bi 7, 8 nh gi l hiu qu) vi chnh mu sa II.2.2.3. Cc bc tin hnh: II.2.2.3.1. Mi ngi tham gia hi ng: S ngi c mi l 15 ngi Ngh nghip: sinh vin Vin Cng ngh sinh hc - Cng ngh thc phm, trng i hc Bch Khoa H Ni tui: 23 - 25 tui Gii tnh: 8 nam, 7 n y l nhng ngi tho mn yu cu chung v ngi th. II.2.2.3.2. La chn hi ng: Hi ng cm quan c la chn trong s 15 ngi c mi, thng qua cc php th c bn xc nh mi, v chn ra nhng ngi c kh nng cm gic tt nht tham gia vo hi ng cm quan. Qu trnh la chn gm 4 bi: Bi 1: Phn bit 4 v c bn (theo nguyn l Spencer)

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Yu cu i vi ngi th: tr li ng 100% Bi 2: Phn bit cng v n (theo nguyn l Spencer) Yu cu i vi ngi th: tr li ng 100% Bi 3: Gi tn mi t do [5] Yu cu i vi ngi th: tr li ng 50% Bi 4: Gn tn mi [5] Yu cu i vi ngi th: tr li ng 75% Ni dung la chn c trnh by c th trong ph lc A. Qua 4 bi th, chn c 8 ngi c im s cao nht trong qu trnh la chn trn. II.2.2.3.3. Hun luyn hi ng: Sau khi ka chn c cc thnh vin cho hi ng nh gi, chng ti tin hnh hun luyn hi ng nh gi vi mc ch gip cc thnh vin luyn tp v nng cao kh nng nhn bit cng nh kh nng so snh cng mi u v mi ngi. Qu trnh hun luyn gm 5 bi nh sau: Bi 1: Hun luyn hi ng lm quen vi mi ngi v mi u a) Lm quen mi u: Mc ch: ca bi ny l cho ngi th bit v ghi nh mi u. Do khi a mu mi u cho ngi th th nn trnh by km theo mt s mu mi quen thuc khc h c th nh v phn bit c mi u vi nhng mi khc. Chun b mu mi u:
Cn 100g ht u Ngm nc trong Xay nhuyn Lc ly dch

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Dch mi u

un si

nh lng 600ml dch

Dng cc mu giy lc 1cm x 5cm nhng vo dch va un si ri cho vo l ti mu c np y (trnh by tng t nh cc mu mi tinh du trong bi la chn, nhng s lng mu giy nhng dch cho vo mi l phi nhiu hn mu tinh du), ngui n nhit phng ri ngi. Ngi th nhn c mt khay gm khong 5 l hnh thc ging nhau, ti mu, c np y, ghi r tn mi ca tng l (c th dng li mu mi trong bi la chn), trong c mt l l mu mi u va chun b, mt cc nc lc. H c yu cu ngi tng l ghi nh mi u . Phiu chun b v phiu nh gi (ph lc A) b) Lm quen mi ngi: Mc ch: ca bi ny l cho ngi th bit v ghi nh mi ngi, ng thi cn gip h phn bit c mi ngi v mi u. Sau khi hi ng lm quen vi mi u, tip tc cho hi ng lm quen vi mi ngi. Qu trnh lm quen c thc hin tng t nh bi lm quen mi u. Ch khc ch chun b dch mi ngi. Chun b mu mi ngi:
Cn 50g ht u Ngm nc trong 5-6h nh lng 300ml nc & xay nhuyn

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Dch mi ngi

Gn ly nc

Ngi th nhn c mt khay gm khong 5 l hnh thc ging nhau ti mu, c np y, ghi r tn mi ca tng l (c th dng li mu mi trong bi la chn), trong c mt l l mu mi u, mt l l mu mi ngi v mt cc nc lc. H c yu cu ngi tng l ghi nh mi ngi, ng thi phn bit gia mi u v mi ngi. Phiu chun b mu v phiu nh gi (ph lc A) Cc mu mi chun b t tinh du c th dng li c trong khong 3-4 ngy. Ring mu mi u v mi ngi sau mi bui th, chng ti b i, ra sch l v chun b li trong mi bui th. Bi 2: Hun luyn kh nng nhn bit mi u Mc ch: Kim tra li tr nh ca ngi th v mi u, ng thi gip ngi th cng c li tr nh v mi u.
a)

Xc nh tn mi t do:

Tin hnh: Ngi th nhn c mt khay gm 8 l ti mu c np y v c m ho (trong c 2 l l mu mi u), mt cc nc lc v mt phiu tr li. H c yu cu ngi v xc nh tn mi ca tng l bng 1 hoc 2 t m t c th v chnh xc nht ri vit ln phiu tr li.

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Cc bc chun b mu tip theo tng t nh trn (dng li cc mu mi t tinh du, chun b mu mi u) Yu cu: ngi th phi gi ng c t nht 50% s tn mi, trong bt buc phi gi ng tn 2 mu mi u. b) Gn tn mi: Tin hnh: Ngi th nhn c mt khay gm 8 l mi ti mu c np y v c m ho (trong c 2 l l mu mi u), mt cc nc lc v mt phiu tr li. H c yu cu ngi v xc nh tn mi ca tng l theo danh sch xut ca phiu. Chun b mu: tng t nh bi xc nh tn mi t do (dng li cc mu mi t tinh du, chun b mu mi u) Yu cu: ngi th phi gi ng c t nht 75% s tn mi, trong bt buc phi gi ng tn 2 mu mi u. Phiu chun b mu v phiu nh gi (ph lc A) Bi 3: Hun luyn kh nng nhn bit mi ngi Mc ch: Kim tra li tr nh ca ngi th v mi ngi, ng thi gip ngi th cng c li tr nh v mi ngi. Tin hnh: Cc bi v cc bc tin hnh tng t nh bi hun luyn nhn bit mi u, nhng thay cc l mu mi u bng mu mi ngi. Bi 4: Hun luyn kh nng phn bit cng mi u Mc ch: Gip cc thnh vin phn bit c cng mnh, yu ca mi u bng php th so snh cp i.

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Kh nng phn bit cng ny c nng cao dn nh mc chnh lch v cng mi u ca hai mu gim dn. Tin hnh: Chun b dch mi u nh nu trn, gi l mu A. Trch t mu A (ngay sau khi va un si), pha long mu A n 2 ln bng nc si (VA : Vnc si = 1 : 1), gi l mu B. Trch t mu A (ngay sau khi va un si), pha long mu A n 3 ln bng nc si (VA : Vnc si = 1 : 2), gi l mu C. Chun b 8 l cha mu A, 4 l cha mu B v 4 l cha mu C. Cc l ny ging nhau v hnh thc bn ngoi, l nhng l ti mu, dung tch 100ml, c np y. Mu c ht vo l khi cn nng vi th tch 30ml, m ho, y np v ngui n nhit phng trc khi ngi. Sp xp cc l mu trn thnh cc cp AB v AC nh th t ca phiu chun b mu (ph lc A) Cc cp AC c mc chnh lch v cng mi u ln hn, cn cc cp AB c mc chnh lch v cng mi u nh hn. Mi thnh vin nhn c 4 b mu 1, 2, 3, 4 trnh by theo th t nh trn, h c yu cu ln lt ngi v so snh cng mi u ca cc cp mu ri tr li vo phiu. Sau khi ngh 10 pht, h tip tc nhn c cc cp mu 5, 6, 7, 8 vi yu cu tng t.

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Yu cu: cc thnh vin phi tr li ng t nht 85%, nu khng bi hun luyn s lp li cho thnh vin no cha t. Bi 5: Hun luyn kh nng phn bit cng mi ngi Mc ch: Gip cc thnh vin phn bit c cng mnh, yu ca mi ngi bng php th so snh cp i. Kh nng phn bit cng ny c nng cao nh mc chnh lch v cng mi ngi ca hai mu gim dn. Tin hnh: Vic chun b v trnh by mu tng t nh bi hun luyn 4, ch khc l thay mu mi u bng mu mi ngi. Sau 5 bi hun luyn, hi ng c kh nng nhn bit v phn bit cng mi u v mi ngi. Nh vy hi ng sn sng cho vic so snh cng hai ch tiu mi ny trong u nnh v cc sn phm t u nnh. II.2.2.3.4. nh gi sn phm: a) Th nghim nh gi mi ca ht u nnh: Mc ch: so snh cng mi ngi v mi u ca ht u nnh nguyn v v ht u nnh tch v nh gi hiu qu ca vic tch v. Chun b mu: Ht u nnh nguyn ht c xay kh sao cho lt qua ry 1mm. Cn khong 15g bt u th cho vo l ti mu dung tch 100ml c np y. Thm 20 ml nc m (60-700C) vo

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, quy u cho bt thm nc. y np li v ngui n nhit phng. M ho, ta c mu mi ht u nnh nguyn v (A). Lm tng t vi u nnh tch v. Ch sy ht n khong 10-11% m bng khng kh nng 45-500C, sau xay dp ht u v i v trc v, ri mi ly phn nhn bn chun b tip. M ho, ta c mu mi ht u nnh tch v (B). Chun b 8 cp mu, mi cp gm mt mu A v mt mu B, trong th t ca cc mu A, B c hon i sao cho c 4 cp l AB v 4 cp l BA. Tin hnh: mi php th lp li 8 ln, do vy mi thnh vin s nhn c mt b gm 8 cp mu nu trn, mt phiu tr li v mt cc nc lc. H c yu cu tr li xem trong mi cp mu, mu no c cng mi u mnh hn, mu mo c cng mi ngi mnh hn. b) Th nghim nh gi mi ca sa u nnh: Mc ch: So snh cng mi u v mi ngi ca cc cp sa c v khng c gia nhit nhm nh gi hiu qu loi mi ca vic gia nhit. Chun b mu v tin hnh: Cc mu sa c ch bin theo cc quy trnh nu hnh 8. Ngay sau khi ch bin, ht 30ml dch sa cho vo

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l ti mu dung tch 100ml c np y, m ho, ngui n nhit phng ri ngi. Ngi th cng c yu cu ngi v tr li xem trong mi cp, mu no c cng mi u mnh hn v mu no c cng mi ngi mnh hn. Cch thc m ho mu v lp phiu tr li tng t nh th nghim nh gi mi ht u. c) Th nghim nh gi mi ca bt u nnh: Mc ch: Sau khi nh gi c hiu qu ca hai bin php x l tch v v gia nhit sau xay, chng ti ch bin sa u nnh (A) theo quy trnh c p dng bin php x l c nh gi l hiu qu, ri tin hnh so snh cng mi u v mi ngi ca bt u nnh sy phun t mu sa A vi chnh mu sa A nh gi hiu qu loi mi ca vic sy phun. Chun b mu: Ch bin sa u nnh theo quy trnh c p dng bin php x l c nh gi l hiu qu (theo kt qu th nghim nh gi mi sa u nnh), ri cng chun b mu sa ngi tng t nh trn. c th nh gi c mi ca bt u, chng ti pha bt u thnh dch theo cng thc 30g bt : 200ml nc si. Sau cng ht ly 30ml dch pha bt cho vo cc l ging nh chun b mu sa.

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Ngi th nhn c 8 cp mu v cng c yu cu ngi v tr li xem trong mi cp, mu no c cng mi u mnh hn v mu no c cng mi ngi mnh hn. Cch thc m ho mu v lp phiu tr li tng t nh th nghim nh gi mi ht u. II.2.2.3.5. X l s liu: Dng chun kim nh thng k:
2

Cng thc tnh : Trong : Q : Cc gi tr quan st c T : Gi tr l thuyt vi gi thuyt l A = B

2 Nu gi tr tnh c ln hn hoc bng gi tr tc (khi
2

(Q - T) 2 T

bnh phng tiu chun) mt mc ngha no th

2 hai mu c coi l khc nhau mc ngha ( tc ).
2

2 Gi tr tc c tra bng 2, ph lc A, [5].

Phn III: Kt qu & tho lun

III.1. Quy trnh phn tch thnh phn to mi ca u nnh v cc sn phm t nnh: Nguyn Th Ngc Hi_ TPN K46
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 n tt nghip III.1.1 Quy trnh tch chit cc hp cht to mi:

Vi cc thng s kho st v loi dung mi, lng mu, t l dung mi nu phn phng php, chng ti tin hnh phn tch trn my sc k kh khi ph (GC-MS) v thu c cc sc thnh phn to mi trong ht u tng ng. Do cc dch chit c chy trn cng mt ct vi cng chng trnh nhit nn c th nh gi hiu qu tch chit ca cc thng s kho st da vo s lng pic pht hin c, mc phn gii ca chng v cc thnh phn nh tnh c. nh gi cc sc thu c theo nhng tiu ch nu trn, chng ti a ra c cc quy trnh tch chit cc hp cht mi tt nht i vi tng loi nguyn liu. III.1.1.1. Quy trnh tch chit cc hp cht mi ca ht u: Trong s cc sc phn tch thnh phn mi ht u nhn c, chng ti nhn thy:
-

Vi kch thc ht c gi nh n kch thc 2 mm, sc xut hin nhiu pic v phn gii cao. C th gii thch nh sau: kch thc ht nh lm tng din tch tip xc gia nguyn liu vi dung mi nn qu trnh chuyn khi cc hp cht bay hi t ht u vo dung mi xy ra kh trit , ng thi kch thc ny cng

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khng qu nh nn trnh c hin tng hn hp nguyn liu-dung mi bin thnh dch cho.
-

Sc phn tch dch chit c x l bng phng

php chit Soxhlet vi dung mi cn (hnh 1a, ph lc B) c phn gii tt nht, s lng pic c pht hin nhiu (16 pic).
-

Bn cnh , sc phn tch dch chit x l

bng phng php chit siu m vi dung mi cn (hnh 1b, ph lc B) cng c s lng pic kh nhiu (11 pic), v mc d phn gii cha cao nhng li pht hin c mt s pic c thi gian lu khng trng so vi thi gian lu ca cc pic trn sc hnh 1 ph lc B.
-

Sc phn tch dch chng ct (lm sch bng

phng php chit pha rn) (hnh 1c, ph lc B) c s lng pic cng nh phn gii thp nht. Kt lun: T chng ti c th kt lun chit Soxhlet bng dung mi cn l phng php tch chit cc hp cht mi ra khi ht u nnh tt nht. Ngoi ra c th s dng phng php chit siu m phn tch nhm b sung kt qu phn tch cho nhau.

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Sau y chng ti a ra quy trnh tch chit cc hp cht mi t ht u nnh bng phng php chit Soxhlet v
Ht u (8g, gi nh 2mm) Ht u (10g, gi nh 2mm)

Cn 960 Chit Soxhlet

Dch chit

Cn 960
Chit siu m (5h, 500C, 10g ht: 25ml cn) Dch chit

Quy trnh tch chit (chit Soxhlet, dm cn 960)

Quy trnh tch chit (chit siu m, dm cn 960)

chit siu m vi dung mi cn: Hnh 11. Cc quy trnh tch chit hp cht mi ca ht u nnh III.1.1.2. Quy trnh tch chit cc hp cht mi ca sa u:

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Nh trnh by phn phng php nghin cu, chng ti kho st hiu qu tch chit i vi cc yu t phng php chit, nhit mu chit, loi dung mi v t l dung mi chit (khi chit, chng ti lc hn hp trong phu chit 1 pht, sau yn 5 pht hn hp tch lp [13], p dng cho tt c th nghim tch chit hp cht mi ca sa). Da vo cc tiu ch nh gi sc nu trn, chng ti nhn thy:
-

Sc phn tch dch chit sa bng phng

php chit lng lng (hnh 4a, ph lc B) pht hin c nhiu pic hn (19 pic) so vi sc phn tch dch chit sa bng phng php chit pha rn (hnh 4b) (14), ng thi phn gii ca cc pic ca sc hnh 4a cng tt hn so vi sc hnh 4b. Do chng ti khng kho st trn phng php chit pha rn na m ch kho st cc thng s i vi phng php chit lng lng.
-

Nu chit cc thnh phn to mi ca sa ngay sau

khi un si th cho sc c nhiu pic hn (14 pic) so vi sa ngui ri mi chit (9 pic) (hnh 2a v 2b, ph lc B). iu ny c th gii thch nh sau: ti nhit cao, qu trnh chuyn khi xy ra vi tc ln hn. Nu a nhit chit ln cao hn th tc bay hi ca cc hp cht mi li tng nhanh hn so vi tc chuyn khi cc hp cht mi t sa vo dung mi. Nu ngui ri

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mi chit th trong thi gian ngui , cc hp cht mi cng bay hi i phn no, lm gim lng hp cht mi chit c.
-

Vi h dung mi n-hexan: diethyl ether [13], ngay khi

chit chng ti nhn thy hn hp chit kh phn lp, gn nh ton b hn hp chuyn thnh dng dch st ng nht rt kh tch. ng thi khi dng dung mi nhexan, hn hp li phn lp r rng. Do vy, chng ti chn n-hexan lm dung mi chit.
-

Vi dung mi chit l n-hexan, kho st vi cc t l

chit khc nhau l 20ml sa : 20ml dung mi chit 1 ln, v 20ml sa : 40 ml dung mi chit 2 ln trong 20ml dung mi cho ln chit 1 v 20ml dung mi cn li cho ln chit 2, chng ti nhn thy vi t l chit 20ml sa : 20ml n-hexan chit 1 ln, sc thu c l tt nht (pht hin c nhiu pic, cc pic phn gii tt, khng chng ln nhau) (hnh 15a v 15b). Kt lun: T nhng nhn xt trn y, chng ti a ra c quy trnh tch chit cc hp cht mi ca sa u nnh bng phng php chit lng lng, s dng dung mi nhexan nh sau:

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Sa u nnh (ngay sau ch bin)

nhexan Chit tnh (20ml sa:20ml dm, lc 1 pht, yn 5 pht) Dch chit Hnh 12. Quy trnh tch chit hp cht mi ca sa u nnh bng phng php chit lng lng s dng dung mi nhexan

III.1.1.3. Quy trnh tch chit cc hp cht mi ca bt u: Da theo kt qu kho st loi dung mi v phng php chit cc hp cht mi ca ht u nnh, chng ti tin hnh kho st hai loi dung mi l cn v n-hexan i vi c 2 phng php tch chit l chit Soxhlet v chit siu m. Cc sc thu c cho thy phng php chit Soxhlet tch chit tt hn, v kh nng tch l nh nhau i vi c hai loi dung mi. C th nh sau: Sc thnh phn to mi ca bt u nnh c tch chit bng phng php chit Soxhlet s dng dung mi nhexan (hnh 3a, ph lc B) pht hin c nhiu pic (19 pic), cc pic phn gii tt, khng chng ln nhau.

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Sc thnh phn to mi ca bt u nnh c tch chit bng phng php chit siu m, cng s dng dung mi cn (hnh 3b, ph lc B) pht hin c t pic hn (13 pic) v phn gii ca cc pic rt km. Kt lun: T y, chng ti a ra quy trnh tch chit cc hp cht mi ca bt u nnh bng phng php chit Soxhlet vi hai loi dung mi l cn v n-hexan nh sau:
Bt u nnh

Chit Soxhlet (5g, cn hoc nhexan) Dch chit

Dung mi cn hoc n-hexan

Hnh 13. Quy trnh tch chit hp cht mi ca bt u nnh

III.1.2. Xy dng phng php phn tch trn my sc k kh khi ph: Cc thng s kho st chng ti trnh by phn phng php. Chng ti dng 1 mu dch chit l dch chit thnh phn to mi ca ht u nnh nguyn v kho st cc thng s. nh gi cc sc thu c theo nhng tiu ch nu (s lng pic, phn gii, thnh phn nh tnh c), chng ti chn thng s chy my sc k kh khi ph (GC-MS) nh sau:

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-

Loi ct: Chng ti chy kho st trn 3 ct l DB1-

MS, DB5-MS v DBWax-MS, y l cc ct c phn cc tng dn v thng c dng phn tch thc phm v cc hp cht to mi. Kt qu cho thy ct DB5-MS l ph hp nht.
-

Lu lng dng kh mang: i vi kh Hli, khi lu lng

dng nm trong khong t 2 n 20 ml/pht th hiu sut ct tch s nm trong khong cho php, trong ti khong 10 ml/pht th hiu sut ct tch l cao nht. Chng ti kho st cc lu lng 2; 4; 6; 10 ml/pht v nhn thy hiu sut tch ti cc lu lng ny xp x nhau. Do vy chng ti chn lu lng 2 ml/pht tit kim kh mang.
-

Chng trnh nhit : kho st chng trnh nhit

700C/(n pht) - tng k0C/pht - 2500C/(m pht), trong n = (1 10) pht, k = (2 20)0C, m = (4 10) pht. Da vo mc xut hin v phn tch cc pic, chng ti a ra c chng trnh nhit nh sau: 700C/(1 pht) - tng 100C/pht - 2500C/(4 pht).

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0C 250

100 70 1 18 24 phut

Chng trnh nhit phn tch thnh phn mi u nnh v cc sn phm u nnh trn my sc k kh khi ph (GC-MS) III.1.3. Kt lun: Tng hp cc quy trnh tch chit v phn tch trn my sc k kh khi ph cc hp cht to mi ca u nnh v cc sn phm u nnh, chng ti c quy trnh phn tch sau y:

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Ht u (gi nh 2mm) Cn 960 Chit Soxhlet (8g, 10h) Chit siu m (5h, 500C, 10g ht: 25ml cn)

Sa u
nnh (ngay sau ch bin) nhexan Chit tnh (20ml sa:20ml dm, lc 1 pht, yn 5 pht)

Bt u nnh Dm cn; nhexan Chit Soxhlet (5g, cn hoc n-hexan)

Dch chit

Phn tch trn my GC-MS

X l kt qu

Quy trnh phn tch thnh phn to mi cho u nnh v cc sn phm u nnh

III.2. Kt qu phn tch sc k & nh gi cm quan: III.2.1. Kt qu phn tch, nh gi cht lng mi ht u:
III.2.1.1. Kt qu phn tch sc k: Nhn xt hai sc (S) thnh phn to mi trong ht u nnh nguyn v (hnh 14a) v ht u nnh tch v (hnh 14b):

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-

C 17 pic trn S thnh phn to mi ca ht u

nnh nguyn v, cn trn S thnh phn to mi ca ht u nnh tch v ch c 10 pic.

-

Trong s c 6 pic trn hai S c thi gian lu trng

nhau, ti cc thi gian 9,1; 10,9; 13,6; 15,3; 15,4; 15,7 (pht) nhng din tch ca a s cc pic trn S ht u tch v l nh hn so vi cc pic trn S ht u tch v.
-

Bng phng php nh tnh v nh lng nu

phn phng php, chng ti nh tnh v nh lng c 7 nh pic sau: Bng 10. Mt s hp cht xc nh c trn S ht u (nguyn v v tch v) St t 1 2 Tg lu (pht ) 7,1 9,1 isododecane octanoic acit, 8hydroxy-, metyl 3 4 5 6 10,9 12,6 13,6 13,9 este isotetradecane pentadecane metyl hexadecanoate isobutyl o-phtalate C14H30 C15H32 C17H34O
2

Tn

CTPT*

% din tch pic Nguyn Tch v -** 1,85 v 1,98 2,12

C12H26 C9H18O3

1,99 2,19 4,69 5,23

0,53 0 8,58 0

C16H22O
4

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7
* **

15,3

metyl cis-9, cis-12-

C19H34O
2

3,96

1,27

octadecadienoic : Cng thc phn t : Khng pht hin

Hnh 14a. Sc thnh phn to mi ht u nnh nguyn v (chit Soxhlet dm cn)

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Hnh 14b. Sc thnh phn to mi ht u nnh tch v (chit Soxhlet dm cn) Trong s cc pic nh tnh v nh lng c, a s chng u c nng gim hoc mt i sau khi ht u c tch v, isododecane, pentadecane, isobutyl o-phtalate u mt i, cn octanoic acit, 8-hydroxy-, metyl este, isotetradecane, metyl cis-9, cis-12-octadecadienoic c nng gim i ng k. Ch ring c metyl hexadecanoate th nng trong ht u tch v li tng, iu ny c th do sai s khi thao tc th nghim. Quan st cc hp cht trn, chng ti nhn thy chng u l cc hydrocacbon mch h, thuc nhm cc hp cht gy mi ngi cho thc vt. III.3.1.2. Kt qu phn tch cm quan: Sau khi tng hp cc phiu nh gi ca 8 thnh vin, mi thnh vin lp li php th 8 ln, chng ti c bng s

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cu tr li so snh cng mi ngi v mi u ca ht u nguyn v v tch v nh sau: Bng 11. Kt qu so snh cng mi u, mi ngi ca ht u nnh Mu Cng mi ngi Mnh Yu hn hn 54 10
2

Cng mi u Mnh Yu hn 35 29 hn 29 35

Ht N nguyn v Ht N tch v Kt qu tnh Q1 = 54;

2

10 54

cho mi ngi: Q2 = 10; +

(10 32 ) 2 32

T = 32; +
2 tc

(54 32 ) 2 32
2 n

(10 32 ) 2 32

(54 32 ) 2 32

= 60,5

Gi tr Gi tr
2 tc

= 60,5 ln hn gi tr

= 6,64 tra theo bng

vi bc t do l 1 mc ngha l = 1%

Vy hai mu ht u ny khc nhau v cng mi ngi mc ngha 1% v ht u nguyn v c mi ngi mnh hn ht u tch v. Kt qu tnh
(35 32 ) 2 32
2

cho mi u: Q2 = 29; +
(29 32 ) 2 32

Q1 = 35;
2 tc 2

T = 32; +
(35 32 ) 2 32

(29 32 ) 2 32

= 1,125 <

= 6,64 vi = 1%.

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Vy hai mu ny khng khc nhau v cng mi u mc ngha = 1%. III.3.1.3. Kt lun: Kt hp c kt qu phn tch ho l v kt qu phn tch cm quan, c th nhn xt nh sau:
-

Thnh phn cc hp cht to mi trong ht u sau Kt qu phn tch cm quan cng cho thy mi ngi So snh kt qu ca hai phng php phn tch, nh gi cm quan. l cc nhm cht

khi loi b v gim c v lng v cht.

-

ca ht u tch v c gim i.
-

chng ti thy c s tng ng gia phn tch ho l v hydrocacbon trong ht u tch v gim hoc mt i tng ng vi cng mi ngi ca ht u tch v gim i. Chng t cc hydrocacbon ny l mt phn nguyn nhn gy ra mi ngi cho u nnh v cc sn phm t u nnh, v y l nhng hp cht c sn trong ht u, nm ch yu v. Nh vy vic tch v l mt cng on hiu qu gip loi b mi khng mong mun ca u nnh. iu ny c ngha l bc tip theo chng ti s nh gi mi ca hai mu sa lm t ht u tch v, trong c mt mu sa c gia nhit sau khi xay v mt mu sa khng gia nhit sau xay.

III.2.2. Kt qu phn tch, nh gi cht lng mi sa u: Nguyn Th Ngc Hi_ TPN K46
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III.2.2.1. Kt qu phn tch sc k: T sc k ca 2 mu sa tch v, khng gia nhit sau khi xay (A) (hnh 15a) v mu sa tch v, gia nhit sau khi xay (B) (hnh 15b) ta c th nhn xt nh sau:
-

S lng pic trn sc k ca mu sa A l 19 i vi 10 pic c thi gian lu trng nhau ti cc thi

nhiu hn so vi mu sa B (c 14 pic)
-

gian 5,0; 5,4; 5,9; 12,3; 13,5; 13,6; 13,9; 15,3; 15,4 v 16,7 pht th din tch pic ca mu sa B gim i rt nhiu so vi mu sa A.
-

Chng ti nh tnh v nh lng c 4 nh

pic nh sau: Bng 12. Mt s hp cht xc nh c trn sc sa u nnh (c v khng gia nhit sau khi xay) St t 1 2 3 4 Tg lu (pht ) 5,0 13,6 13,9 15,3 l-limonene n-methyl--phenyl ethyl amine di-n-butylphtalate methyl 8,11octadecadienoate C10H16 C19H23N O C16H22O
4

Tn

CTPT

% din tch pic khng gia nhit 15,56 6,45 4,49 4,43 C gia nhit 7,77 3,30 4,19 3,35

C19H34O
2

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Hnh15a. Sc cc thnh phn mi ca sa u nnh

tch v, khng gia nhit sau xay (chit lng lng dug mi nhexan) Hnh15b. Sc cc thnh phn mi ca sa u nnh tch v, c gia nhit sau xay (chit lng lng dung mi nhexan) T bng trn chng ti nhn thy, nng cc hp cht trong sa u nnh c gia nhit sau khi xay gim hn (th hin phn trm din tch). Trong , l-limonene gim nhiu nht. ng ch , trong s cc hp cht nh tnh c ngoi cc hydrocacbon, cn c mt hp cht cha nhm amin, l n-methyl--phenyl ethyl amine. y l mt cht thuc nhm amin bay hi, c th gp phn gy ra mi u cho sa u nnh.

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III.3.2.2. Kt qu phn tch cm quan: Bng 13. Kt qu nh gi mi ca sa u nnh Mu Sa N A Sa N B Cng mi ngi Mnh hn Yu hn 38 26 26 38
2

Cng mi u Mnh hn Yu hn 59 5 5 59

Kt qu tnh Q1 = 38;
2 tc 2

cho mi ngi: Q2 = 26;

(26 32 ) 2 32

T = 32; +
(38 32 ) 2 32

(38 32 ) 2 32

(26 32 ) 2 32

= 4,5 <

= 6,64 vi = 1%.

Vy hai mu ny khng khc nhau v cng mi ngi mc ngha = 1%. Kt qu tnh

(59 32 ) 2 32
2

cho mi u: Q2 = 5;
(5 32 ) 2 32

Q1 = 59; >
2 tc 2

T = 32; +
(59 32 ) 2 32

(5 32 ) 2 32

= 91,125

= 6,64 vi = 1%.

Vy hai mu ny khc nhau v cng mi u mc ngha = 1% v mu sa u nnh A c cng mi u mnh hn so vi mu sa u nnh B. III.3.2.3. Kt lun: Kt hp c kt qu phn tch ho l v kt qu phn tch cm quan, c th nhn xt nh sau:

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- Thnh phn cc hp cht to mi trong sa u nnh c gia nhit sau khi xay gim c v s lng v cht lng.
-

Vic phn tch cm quan cng ch ra s thay

i ny, l cng mi u ca sa u nnh c gia nhit sau khi xay gim so vi sa u nnh khng gia nhit.
-

Hai kt qu phn tch ho l v phn tch cm

quan trn i tng v sa u nnh cng cho thy s tng ng, th hin: khi nng cc hp cht pht hin c gim i th cng mi u cng gim i. T y c th rt ra cc hp cht nh tnh c, nht l n-methyl--phenyl ethyl amine c th l nguyn nhn gy ra mi u cho sa u nnh. Ngoi ra, chng ti cho rng cc hp cht ny a s c hnh thnh trong qu trnh ch bin v chng khng h xut hin trong sc phn tch thnh phn mi ht u.
-

Kt hp c kt qu phn phn tch v nh gi mi

ht u vi kt qu phn ny, chng ti nhn thy: mi ngi ch yu do cc hp cht c sn trong ht u m c th l c trong v u gy ra; cn mi u ch yu do cc hp cht c sinh ra trong qu trnh ch bin, c th c sinh ra cng on xay t sau khi ngm.

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Nh vy vic gia nhit sau khi xay l mt cng on hiu qu gip loi b mi u ca sa u nnh. iu ny c ngha l bc tip theo s nh gi mi ca bt u nnh sy phun t sa u nnh lm t ht u tch v v c gia nhit sau khi xay.

III.3.3. Kt qu phn tch v nh gi mi bt u:

III.3.3.1. Kt qu phn tch sc k: Hnh 16a v 16b l sc k ca mu bt u nnh lm t sa u nnh lm t ht u tch v v c gia nhit sau khi xay (sa N B). Quan st trn hai sc ny, c th nhn xt nh sau:
-

a s pic c thi gian lu trng nhau xut hin trn c

hai sc , l 7 pic vi thi gian lu 13,9; 15,0; 15,1; 15,9; 17,5; 21,0 v 23,7 pht
-

Din tch cc pic hnh 16a ln hn so vi hnh

16b - Ngoi ra c hai sc cn c nhng pic c thi gian lu khng trng nhau nh cc pic c thi gian lu 8,2; 11,2; 13,7; 18,0 trn sc 16a v 5,8; 6,3; 6,6; 7,7 v 9,9 trn sc 16b. T ta cho thy c hai loi dung mi cn v n-hexan u c kh nng tch chit cc hp cht to mi c trong bt

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u nnh. Do vy chng ti dng c hai phng php tch chit b sung kt qu phn tch cho nhau. Chng ti nh tnh v nh lng c cc hp cht to mi sau:

Hnh 16a. Sc thnh phn to mi ca bt u nnh (chit Soxhlet dung mi cn)

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Hnh 16b. Sc thnh phn to mi ca bt u nnh (chit Soxhlet dung mi n-hexan)

Bng 14. Mt s hp cht xc nh c trn sc bt u (tch chit bng hai dung mi cn v n-hexan) St t Tg lu (pht ) 1 2 3 4 5 6,3 6,6 7,7 8,2 9,9 disulfide,di-2-propenyl C6H10S2 butane,2,2-dimetyl C6H14 2,3-epithiopropyl allyl C6H10OS ether undecane C11H24 trisulfide,di-2-propenyl C6H10S3 Cn 5,38 1,8 Tn CTPT* % din tch pic nhexan 2,5 2,01 1,61 4,84

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6 7 11,2 13,7 tetradecane 1,2benzendicarboxylic 8 18,0 acid,diethyl ester 1,2benzendicarboxylic
*

C14H30 C12H14O4

8,66 3,4

1,09

C16H22O4

7,73

12,6

acid,dibuthyl ester : Cng thc phn t III.3.3.2. Kt qu phn tch cm quan: Bng 16. Kt qu nh gi mi bt u nnh v sa u nnh Mu Cng mi ngi Mnh Yu hn Cng mi u Mnh Yu hn hn 58 6 T = 32; +
(40 32 ) 2 32

hn Sa N 4 40 24 Dch bt N 24 40 2 Kt qu tnh cho mi ngi: Q1 = 40;

2 tc 2 n

6 58

Q2 = 24; +
(24 32 ) 2 32

(40 32 ) 2 32

(24 32 ) 2 32

= 8 >

= 6,64 vi = 1%.

Vy hai mu ny khc nhau v cng mi ngi mc ngha = 1% v dch pha bt c cng mi ngi yu hn. Kt qu tnh
2

cho mi u: Q2 = 6; T = 32;

Q1 = 58;

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2 tc 2

(58 32 ) 2 32

(6 32 ) 2 32

(6 32 ) 2 32

(58 32 ) 2 32

= 84,5 >

= 6,64 vi = 1%.

Vy hai mu ny khc nhau v cng mi u mc ngha = 1% v dch pha bt c cng mi u yu hn. III.3.3.3. Kt lun: Bng vic sy phun sa u nnh, mt ln na cc hp cht gy mi khng mong mun cho u nnh v cc sn phm u nnh b loi b. iu ny th hin trn kt qu phn tch cm quan, l cng mi ngi v mi u ca bt u nnh gim i so vi chnh mu sa lm nn bt u nnh. Vy, vic kt hp c 3 bin php x l l tch v, gia nhit v sy phun s gip loi b c mi ngi v mi u ca u nnh v cc sn phm u nnh mt cch c hiu qu. T y chng ti c th a ra quy trnh x l hiu qu nhm loi b mi ngi v mi u ca u nnh v cc sn phm t u nnh nh sau (hnh 17).
Hnh 17. Quy trnh sn xut bt u x l hiu qu mi u v mi ngi

Ht u nnh Tch v Ngm Chn Xay Gia nhit Lc un si

Sa N

Sy phun
Bt N

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Phn IV:

Kt lun

1. Qua qu trnh nghin cu, chng ti a ra c quy

trnh tch chit cc hp cht mi tng ng cho tng loi nguyn liu, l phng php chit Soxhlet s dng dung mi cn p dng cho ht u nnh, phng php chit lng lng s dng dung mi n-hexan p dng cho sa u nnh v phng php chit Soxhlet s dng hai loi dung mi cn 960 v n-hexan p dng cho bt u nnh. ng thi chng ti cng xc nh c thng s cho my sc k kh khi ph (GC-MS) khi phn tch cc hp cht mi ca u nnh v cc sn phm t u nnh.
2. Kt qu phn tch ho l v kt qu phn tch cm

quan cho thy: Ht u tch v c cng mi ngi gim so vi ht u nguyn v; Sa u nnh c gia nhit sau khi xay gim v cng mi u so vi sa u nnh khng c gia nhit; v Bt u nnh sau khi sy phun gim c v cng mi u v mi ngi.
3. Trn c s phn tch v nh gi cht lng mi ca

u nnh v cc sn phm u nnh trn y, chng

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ti xc nh c quy trnh x l mi tt nht cho u nnh v cc sn phm t u nnh.

Mt s kin ngh:
-

Trn sc chng ti khng pht hin thy

Hexanal, y l mt hydrocacbon m theo cc ti liu tham kho c coi l nguyn nhn chnh gy ra mi ngi cho ht u. C th v hm lng hexanal c trong ht u ging VX 93 qu t, hay do quy trnh tch chit cha thu c hon ton hexanal nn khng pht hin c khi phn tch nn nhng nghin cu sau cn ch n vn ny.
-

i vi sa u nnh, khi chit lng lng chng ti

khng th s dng dung mi tan ln vo sa nh cn tch chit cc hp cht mi, m ch c th dng nhng dung mi khng tan ln nh n-hexan, diethyl ether. iu ny c th lm cho qu trnh tch chit khng trit v n-hexan v diethyl ether l nhng dung mi phn cc thp, m cc hp cht to mi a s l phn cc. C th p dng phng php sc k lng cao p phn tch

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v phng php ny cho php bm trc tip mu dng lng vo my phn tch, tuy nhin trong khun kh thi gian c hn khi lm n ny, chng ti cha thc hin c. Hi vng nhng nghin cu tip theo s c th trin khai tng ny.
-

Trong phn tch cm quan, chng ti dng php

th so snh cp so snh mi ca u nnh v cc sn phm t u nnh trc v sau x l xc nh c phng php x l mi tt nht. C th s dng php th cho im nh gi cht lng mi, php th ny c u im l ch r c mc thay i cng mi, t nh gi c mc hiu qu ca tng phng php x l. Tuy nhin, c th s dng php th ny phi mt rt nhiu thi gian v cng sc hun luyn hi ng nh gi. Do iu kin v thi gian, con ngi v phng th nghim nn trong khong thi gian hn hp, chng ti chn php th so snh cp.

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