i

LI CM N

Trc ht, ti xin by t lng bit n n Ban Gim hiu, Ban Ch

nhim khoa, v quy thy c ging vin khoa Cng ngh Thc phm trng
i hc Nha Trang to mi iu kin v c s vt cht, cung cp cho ti
nhng kin thc v k nng cn thit trong sut thi gian hc tp ti trng,
gip ti hon thnh chng trnh o to C nhn ngnh Cng ngh Thc
phm.
Ti xin chn thnh cm n cn b ph trch Phng th nghim b mn
Ha to iu kin tt nht v dng c v trang thit b trong qu trnh thc
hin n tt nghip.
Ti cng xin chn thnh cm n TS. Trn Th Hong Quyn quan
tm hng dn v gip nhit tnh v mt kin thc v ti liu nghin cu
trong sut thi gian thc hin ti tt nghip.
c bit, ti xin gi li bit n chn thnh v su sc nht n
TS. Hong Th Hu An dnh rt nhiu thi gian v tm huyt gip ti
trong vic nng cao kin thc, rn luyn k nng thc nghim v to mi iu
kin v trang thit b, vt t cho ti tin hnh nghin cu.
Xin cm n gia nh v bn b lun ng vin, gip v ng gp
cho ti nhiu kin qu bu trong sut qu trnh hc tp v lm n tt
nghip ny.
Nha Trang,

thng 7 nm 2012

Sinh vin
V TH THIT

ii

MC LC
LI CM N ............................................................................................ i
DANH MC CC CH VIT TT ................................................. iv
DANH MC CC BNG ..................................................................... v
M U................................................................................................... 28
PHN 1. TNG QUAN ............................................................................ 31
1.1.

Tng quan v cht mu thc phm ..................................................... 31

1.1.1. Tm quan trng ca cht mu trong ch bin thc phm ...............................31

1.1.2. Phn loi cht mu thc phm .........................................................................31
1.1.3. Nguyn tc s dng cht mu thc phm ........................................................32

1.2.

Tng quan v hoa cc vn th ............................................................ 33

1.2.1. Tn gi ..............................................................................................................33
1.2.2. c im hnh thi v sinh trng ...................................................................33
1.2.3. Phn loi cc vn th .......................................................................................34
1.2.4. Tnh hnh sn xut v tiu th cc vn th Vit Nam v trn th gii ........35

1.3.

Gii thiu v cht mu lutein, lutein ester .......................................... 37

1.3.1. Khi nim v cht mu lutein ..........................................................................37

1.3.3. So snh kh nng hp th ca lutein v lutein ester ........................................44

1.4.

Tng quan v cc phng php chit xut cht mu t nhin .......... 45

1.4.1. Khi nim chit xut ........................................................................................45

1.4.2. C ch qu trnh chit xut ..............................................................................45
1.4.3. Nguyn tc chit xut .......................................................................................46
1.4.4. Cc phng php chit xut .............................................................................46
1.4.5. Cc yu t nh hng ti qu trnh chit xut .................................................47
1.4.6. Vi k thut chit hin i dng chit xut cht mu t nhin ..................47
1.4.7. Tnh hnh nghin cu chit xut lutein ester trn th gii ...............................49

PHN2. I TNG V PHNG PHP NGHIN CU ................... 51

2.1. i tng nghin cu ............................................................................ 51
2.1.1. Vt liu nghin cu ..........................................................................................51
2.1.2. Dng c v thit b ...........................................................................................51

iii

2.2. Phng php nghin cu ....................................................................... 52

2.2.1. Xc nh mt s thnh phn ha hc ca cnh hoa cc vn th .....................52
2.2.2. Quy trnh d kin sn xut lutein ester t hoa cc vn th. ............................53
2.2.3. Xc nh iu kin thch hp chit lutein ester t hoa cc vn th ................56
2.2.4. Th nghim quy trnh chit nh gi hiu sut chit ..................................59
2.2.5. Tinh ch v nh gi cht lng sn phm......................................................59
2.2.6. Nghin cu phng php bo qun dch chit lutein ester .............................59
2.2.7. X l s liu .....................................................................................................61

PHN 3. KT QU V THO LUN ....................................................... 62

3.1. Mt s thnh phn ha hc ca hoa cc vn th ................................... 62
3.2. Xy dng quy trnh chit lutein ester t hoa cc vn th ....................... 63
3.2.1. Chn dung mi chit ........................................................................................63
3.2.2. Chn t l dung mi:nguyn liu .....................................................................64
3.2.3. Chn thi gian chit v s ln chit .................................................................65

3.3. Hon thin quy trnh chit sn xut cht mu lutein ester ..................... 67
3.3.1. Hon thin quy trnh chit sn xut cht mu lutein ester .67
3.3.2. Kt qu th nghim quy trnh - nh gi cht lng sn phm .....................71

3.4. Kt qu kho st bn mu ca lutein ester ........................................ 73

KT LUN .............................................................................................. 76
KIN XUT ................................................................................ 77
TI LIU THAM KHO ................................................................... 78
PH LC .................................................................................................. A

iv

DANH MC CC CH VIT TT

VIT TT

TING ANH

TING VIT

CVT

Marigold

Cc vn th

EtOH

Etanol

Ru etanol

Hour

Gi

H2 O

Water

Nc

TB

Average

Trung bnh

TL

Weight

Trng lng

UV-Vis

Ultraviolet-Visible

T ngoi-kh kin

v/v

Volume/volume

Th tch / th tch

v/w

Volume/weight

Th tch/khi lng

w/v

Weight/volume

Khi lng/th tch

DANH MC CC BNG

Bng 1.1. Hm lng -caroten, lutein, lycopen trong mt s thc vt .... 41

Bng 2.1. Danh mc ha cht s dng trong ti ................................... 51
Bng 3.1. Mt vi thnh ha phn ha hc c bn ca hoa cc .............. 62
Bng 3.2. Kt qu th nghim quy trnh sn xut lutein ester ................... 71
Bng 3.3. Kt qu phn tch hm lng carotenoid trong sn phm .....72

vi

DANH MC CC HNH

Hnh 1.1. Cu to phn t ca lutein [38] .................................................. 37

Hnh 1.2. Cu to phn t ca lutein ester [30].......................................... 42
Hnh 2.1. S quy trnh d kin sn xut lutein ester ............................. 55
Hnh 2.2. B tr th nghim xc nh iu kin chit lutein ester ............... 58
Hnh 2.3. B tr th nghim nh gi bn mu ca dch chit ............... 61
Hnh 3.1. nh hng ca dung mi n hiu sut chit lutein ester ......... 63
Hnh 3.2. nh hng ca t l dung mi:nguyn liu n hiu sut chit 65
Hnh 3.3. S ph thuc ca hiu sut chit vo thi gian ngm chit ....... 66
Hnh 3.4. S ph thuc ca hiu sut chit vo s ln ngm chit ............ 67
Hnh 3.5. S quy trnh sn xut cht mu lutein ester ......................................70

Hnh 3.6. Sn phm lutein ester tinh ch .................................................... 72

Hnh 3.7. bn mu ca lutein ester khng b sung 0,1% BHT(w/v) ..... 73
Hnh 3.8. bn mu ca lutein ester khi b sung 0,1 % BHT (w/v) ........ 74
Hnh 3.9. Mu dch chit khng b sung BHT khi bo qun ..................... 75
Hnh 3.10. Mu dch chit b sung BHT sau khi bo qun ....................... 75

28

M U
Hin nay, nhu cu con ngi ngy cng i hi p ng cao hn trong
vn sc khe, n ung v chm sc sc p.
Chnh v vy, lutein - mt sc t carotenoid mu vng-vng cam c
ngun gc t nhin hin ang c quan tm ng dng trong lnh vc cng
nghip cht mu thc phm nhm thay th cho mt s cht mu tng hp
mu vng c nguy c gy bnh ung th cho con ngi (nh Tartrazine,
Sudan, Yellow Suset,). Ngoi kh nng to mu, lutein cn c tc dng
chng tia cc tm, do gip bo v trnh tn thng t bo da, vim da, ung
th da, chng lo ha nn lutein cng c quan tm ng dng trong cng
nghip m phm. Lutein cng c trong thnh phn ca cc loi dc phm b
sung nhm h tr iu tr v phng chng suy thoi vng mc do tui gi, h
tr ngn nga bnh thoi ha im vng, c thy tinh th, gip ci thin th
lc i vi nhng ngi lm vic nhiu vi my tnh hay tip xc nhiu vi
nh sng cng cao (th hn), [6]
Kt qu kho st cho thy cnh hoa cc vn th chu Phi (Tagetes
erecta L.) l ngun nguyn liu lutein t nhin l tng cho vic thu nhn
lutein ester do cha hm lng carotenoid kh cao (khong 1,6% trng lng
kh) v gn nh tinh khit (trn 95% carotenoid l lutein ester, cn li l dng
ng phn zeaxanthin [7]. y l loi thc vt rt thch hp vi cc vng c
kh hu nhit i v hin ang c trng nhiu Peru, n , Mexico,
Trung Quc, Thi Lan... cung cp nguyn liu cho cng nghip tch chit,
tinh ch lutein [39]. Cy cc vn th (CVT) cng sinh trng rt tt v c th
c trng quanh nm nhiu a phng ca nc ta.
V vy, vic nghin cu chit xut cht mu lutein t hoa CVT nhm
ng dng trong cng nghip thc phm, m phm v dc phm l mt yu
cu cn thit hin nay.

29

Hin nay trong nc c mt s nghin cu tch chit v tinh ch

lutein t hoa cc vn th [30] nhng cc quy trnh ny a s ch ch trng ti
vic to ra sn xut lutein t do ng dng trong cng nghip dc phm. Vi
mc ch ng dng lm cht mu thc phm hay ph gia to mu cho thc n
chn nui, thc t ch cn s dng lutein dng ester v kh nng to mu ca 2
dng ny cng gn tng ng. Mt trong nhng vn cn quan tm i
vi cht mu t nhin l bn mu bi v a s cht mu t nhin rt d b
nht mu trong qu trnh s dng v bo qun.
Chnh v vy, n tt nghip "Chit lutein ester t bt hoa cc vn
th (Tagetes erecta L.) - nh gi bn mu ca sn phm" c chng
ti thc hin.
Trong n ny, k thut ngm chit c p dng tch lutein ester
t bt hoa cc vn th kh, nhng khc vi cc tc gi trc, chng ti
nghin cu kh nng s dng h dung mi hexane-ethanol thay th cho
hexane nhm gim gi thnh sn phm v quy trnh chit thn thin hn vi
mi trng.
Mc tiu nghin cu ca ti: Xy dng quy trnh tch chit lutein
ester t cnh hoa CVT (Tagetes erecta L.) bng phng php ngm chit s
dng h dung mi hexane-ethanol.
Ni dung nghin cu ca ti
- Xc nh iu kin thch hp chit lutein ester t bt hoa CVT bng
phng php ngm chit s dng h dung mi hexan-etanol.
- Th nghim quy trnh chit - nh gi hiu sut chit
- Tinh ch sn phm. nh gi bn mu ca sn phm trong qu trnh
bo qun.

30

Nhng kt qu nghin cu ca ti ny c th xem l c s ban u

sn xut cht mu lutein ester ng dng trong cng nghip thc phm, m
phm.
Do kin thc v kinh nghim nghin cu cn hn ch cng nh kh
khn v iu kin thc nghim nn mc d rt c gng ti ny khng trnh
khi nhng thiu st.
Rt mong c s ch bo ca qu thy c, cng nh s gp t cc
bn sinh vin ti ny c th c hon thin hn.

31

PHN 1. TNG QUAN

1.1.

Tng quan v cht mu thc phm

1.1.1. Tm quan trng ca cht mu trong ch bin thc phm

Mu sc l mt trong nhng ch tiu cm quan c s dng nh
gi cht lng ca sn phm, lm tng gi tr cm quan ca thc phm (ko,
nc gii kht....). T mu sc ta c th c lng phm cht ca thc phm
Theo mt nghin cu ca nh thc phm hc Johnson vi ti Bo v thc
phm (1983) cho thy mu sc nh hng n ngt ca sn phm,
ngt tng 212% do vic s dng ng mu sc. [35]
Do vy, vic s dng b sung cht mu trong thc phm c ngha rt
quan trng [37]:
- Gip phc hi li cht mu t nhin ban u ca sn phm, khi cht
mu t nhin ny b mt i trong qu trnh ch bin hay trong qu trnh
bo qun;
- Xc nh r hay nhn mnh cho ngi tiu dng ch n mi t
nhin rt nhiu thc phm;
- Gip ngi tiu dng xc nh r c nhng thc phm c xc
nh theo thi quen tiu dng;
- Gia tng mu sc c hiu ca thc phm c cng mu km;
- Lm ng nht mu sc ca thc phm;
- To thc phm c mu sc hp dn hn.
1.1.2. Phn loi cht mu thc phm
Cht mu v c
Cht mu v c c sn xut rt nhiu, tuy nhin trong thc phm ch
cho php s dng CuSO4 gi mu cho hoa qu. Phn ln cc cht muv c
c tnh c nn cn phi thn trng khi dng trong thc phm.

32

Cht mu tng hp
Cc phm mu c to ra bng cc phn ng tng hp ho hc nh:
Tatrazin, Azorubin, Amaran, xanh lc sng BS,...
Tt c cc cht mu tng hp u c i vi con ngi nn khi s dng
phi tun theo s ch dn trong ti liu k thut.
Cht mu c du n t nhin
Cc cht c tng hp gn ging nh cc cht mu t nhin
Cht mu t nhin
Cc cht mu c chit xut ra hoc c ch bin t cc nguyn liu hu
c (thc vt, ng vt) sn c trong t nhin nh: sc t clorofin, sc t antoxian,
sc t carotenoid, riboflavin, axit cacminic, caramel, polyphenol,.
Tt c cc cht mu t nhin u khng c, to ra mu rt p cho thc
phm, ng thi chng li c mi, v gn ging nh ngun t nhin ca chng.
Mt s cht mu t nhin cn c tc dng tt i vi sc khe (nh c kh nng
chng oxy ha, tng cng sc khng ca c th,). Do vy, ngi tiu dng
ngy cng c xu hng a chung nhng loi thc phm dng cht mu t nhin.
[24]
1.1.3. Nguyn tc s dng cht mu thc phm
V nguyn tc, b sung cht mu nhm phc hi mu vn c ca mt
s thc phm b tn tht trong qu trnh ch bin. Khng c dng cht mu
che y khuyt im ca thc phm hoc ngi tiu dng nhm ln v
s c mt khng thc ca mt vi thnh phn cht lng trong thc phm.
Trc khi quyt nh s dng cht mu thc phm b sung vo mt sn
phm thc phm no chng ta cn quan tm cc yu t sau:
- Trng thi ca cht mu: Ngi tiu dng vn a dng cht mu
dng lng hn l dng bt;

33

- S chuyn mu mi nh th no: Thng phi trn nhiu mu vi nhau

to ra mu mong mun ph hp vi thc phm ca mnh;
- Thnh phn cc cht cha trong thc phm cn phi trn cht mu: S
c mt ca protein, nc, tanin hay cht bo s anh hng ti trong hay
vn c ca cht mu phi trn vo trong thc phm ;
- pH: Hin tng to ta c trong thc phm do pH cht mu gy ra;
- iu kin ch bin: c bit l thi gian v nhit nh hng ti cht
lng mu phi trn;
- iu kin bao gi: quyt nh n mc xm nhp ca khng kh v
nh sng vo thc phm, c th lm bin i cht mu;
- iu kin bo qun v thi gian s dng: Cht mu s dng ch bn
di nhng iu kin bo qun thch hp v trong mt thi hn nht nh [32].
1.2.

Tng quan v hoa cc vn th

1.2.1. Tn gi
Tn ting Vit: Cc vn th.
Tn ting Anh: Marigold.
Tn khoa hc: Tagetes spp.
H: Cc (Asteraceae)
1.2.2. c im hnh thi v sinh trng
Chi CVT (Tagetes) l mt chi ca khong 60 loi cy thn tho mt
nm v lu nm trong h cc. Chng c ngun gc ti khu vc ko di t ty
nam Hoa K qua Mxic v v pha nam ti khp Nam M.
Cc loi khc nhau c kch thc cao t 0,052,2 m. Chng c cc l
lng chim mu xanh lc vi hoa t trng, vng kim, da cam, vng ti gn nh
, ng knh khong 0,1 ti 46 cm. Tn l ca CVT c mi thm nh x
v hng, mc d cc ging, th sau ny c to ra l khng c mi.

34

Cy hoa CVT sinh trng nhanh, tt trong iu kin nh sng y .

Cy chu nng tt. Cy d trng bng cch tch bi hay gieo ht. CVT rt d
trng, trng bng cy con t ht hoc nhn ging bng chi, ngn cy trng
bng chi mm thng nh, khng cao v mau ra hoa.
Cy CVT khng kn t trng, thch hp nht l tt tht nh, nhng
nu l t ct pha, t st pha, t c ln si , cy hoa ny vn sng
c. iu i hi l t phi cao ro, m v khng ng thu. t nhim
phn v nhim mn cng khng trng c CVT. [27]
1.2.3. Phn loi cc vn th
Trn th gii, CVT chia ra lm ba loi nguyn v loi lai sau y: [14]
Loi cc vn th chu Phi
Tn khoa hc l Tagetes erecta L., ting Anh gi l African Marigold.
y thng l ging vn th cy cao nht v hoa cng to nht.
Loi nh Nguyt (Moonlight) hoa kp to, n trn xoe khng ci, cao
chng 40 cm v mc dy kht nhau. Loi Gold-n Vanilla cao khong 5070
cm, cho nhiu hoa mu sc khc nhau ln ln t cam n vng, vng kim,
vng chanh, vng b. Loi Tui Vng (Golden Age), cy cao hn 75 cm, loi
Doublon cao n 1,50 m cho hoa rt to, c ng knh 12,5 cm [29].
Loi cc vn th Php
Tn khoa hc l Tagetes patula, ting Anh gi l French Marigold.
Loi ny thng hp dn hn loi chu Phi, hoa cng nh hn. Hoa c
mu kiu, dn chng u M thng trng ging hoa n, mt lp cnh
hoa di, c ci.
Ging Oai V (Majestic) cao 30 cm, hoa vng n, cnh sc nu
hay sc mu g , ci vng. Loi Janie cao chng 20 cm hoa c ba mu
c a thch l vng, la v vng kim, nhng cng cn c mu g ,
mu cam m, mu qut tiu son, hay ln ln nhiu mu Loi ln Naughty

35

Marietta, ch cao 25 cm, hoa n, cnh bn trong im vt nu Loi Mt Cp

(Tiger eyes), cao 3035 cm l mt vn th l v l cnh n huyt vin
ba ngoi hoa, cn bn trong n nh l cc vng cam. Loi N Hong (Queen
series) hoa n ta hoa tr mi, hi ng, cy ln 2530 cm,...Cng nh mi
loi CVT Php khc, nhng vng c kh hu nng nh nc ta, cy c th
cao hn 60 cm nh ging K Hoa Sc (Striped Marvel) thn cao n 75
cm, hoa mu sc [25].
Loi hoa cc vn th nh
Tn khoa hc l Tagetes tenuifolia, hay Tagetes signata. Hoa n cnh,
c ci v nh 12 cm. Loi hay trng u M l Stafire Mix, c c im l
l thm mi chanh bi, nht l khi tri nng nc .
Loi lai c tn l American Marigold
Loi lai Antigua Yellow l loi CVT vng ti, hoa kp to c ng
knh 78cm, nc ta c trng lng hoa G Vp. Cy mc kht v cao
3050 cm. Loi lai Inca Hybrid hoa kp c ng knh 1013 cm, cy cao
5070 cm chu nhit n 3940oC. Ging tam nhim lai triploid, thuc
nhm F1 l ging phi hp cy ln ca vn th Php v hoa kp to ca vn
th Phi Chu, chu c lnh v nng.
1.2.4. Tnh hnh sn xut v tiu th cc vn th Vit Nam v trn th
gii
Trong nc
Hin nay, c hai loi CVT c trng ph bin nc ta l CVT Php,
hay CVT ln (Tagetes patula L.) v CVT chu Phi (cng c tn thng gi
l CVT cao; tn khoa hc l Tagetes erecta L.)
CVT Php thng gi l CVT c cung c hoa mu vng sm, ging
hoa Nht cnh hoa va c mu vng thm li va c mu vng nht (Tagetes

36

patul L.). CVT chu Phi, hoa vng ngh v vng hong yn (Tagetes erecta
L.).
Bn cnh vic chi hoa, trang tr v cng ving, cy CVT cn c s
dng nh mt dc phm cha bnh. L v hoa c dng trong iu tr cc
bnh v ng tiu ha, kch thch tun hon mu. Hoa CVT c cha
nhiu vitamin C, protein v flavonoid [41].
Ngoi nc
CVT rt ph bin, c trng hu ht cc nc trn th gii vi
nhiu tn gi khc nhau. Mi tn gi gn lin vi vng t sinh ra n nh
CVT chu Phi (African Marigold), CVT Php (French Marigold),... v nhiu
ging hoa lai khc.
Nghin cu ti Yamagata Prefecrural Institute of Public Health, Nht
(Phytotherapy Research S 16-2002) ghi nhn dch chit bng metanol t hoa
Tagetes patula c hot tnh c ch c cc phn ng sng-vim cp tnh v
kinh nin ni chut nht v chut nh b sng ph bng -carrageenin [11].
Nghin cu ti i hc Hamdard, Karachi, Pakistan (Archives of
Pharmacy Research S 27 2004) v tc dng ca r CVT Php trn huyt p
ghi nhn: Dch chit bng metanol t r cho cc cht citric, malic acid gy
h huyt p v pyridine hydrochloride gy tng huyt p cng mt hp cht
mi 2-hydroxy,5-hydroxymethylfuran [3].
Carotenoid trch t hoa CVT c s dng b sung vo thc n nui
c hi, gip to mu tht mu cam ca c [5].
Trong Cng nghip thc phm, sc t chit t hoa CVT cng c
dng trong k ngh nui tm, nghin cu ti i Hc Universidad Autonoma
Metropolitana-Iztapalapa, Mexico (1996) so snh tc dng to mu ca
dch chit hoa CVT v astaxanthin tng hp pha trn trong thc phm nui
tm trng (ging Panaeus vannamei). Sau 14 ngy cho n, tm nui bng

37

dch chit vn th c mu vng ca tht p hn astaxanthin gp nhiu ln, kt

qu cng cho thy cc carotene trong hoa CVT nh lutein v zeaxanthin
c chuyn bin thnh astaxanthin trong c th tm. [24].
Mt s loi hoa CVT (Tagetes) c dng trong cc cng thc nu n
ti chu M. Trong dn gian Trung Quc cng c mt s bi thuc s dng
hoa CVT tr bnh (au rng, au mt, ho g, kh mt, mi mt). Ti n ,
l hoa CVT dng p tr mn nht, nc p t l tr au sng tai. Hoa tr bnh
mt, lot bao t, thanh lc mu cho c th [40].
1.3.

Gii thiu v cht mu lutein, lutein ester

1.3.1. Khi nim v cht mu lutein

1.3.1.1. Cu to phn t
-

Lutein c coi l tin thn ca vitamin A, l dn xut 3,3-diol

ca beta, epsilon-caroten, cng thc phn t l C40H56O2.

-

Lutein l mt dng oxy ha ca carotenoid, cn gi l xanthophyll, cu

trc mch carbon C40 gm 8 n v isoprene cha 2 vng 6 cnh mi u

phn t v mch chnh polyene gm nhiu ni i lin hp, to ra nhiu ng
phn khc nhau ng thi vi 2 nhm hot ng hydroxyl gn 2 u to
tnh nng oxy ha cho phn t. Lutein trong t nhin thng tn ti cu
hnh all-trans [26]
16 17

19
6

21

HO

20
11

9
8

18'

10

15

13
12

14

14'
15'

12'
13'
20'

18

10'
11'

8'
9'
19'

Hnh 1.1. Cu to phn t ca lutein [38]

1.3.1.2. Tnh cht l-ha

Tnh cht vt l
- Lutein tn ti dng bt chy lng c mu vng cam.

5'

4'

6'

3'
2'

7'
17'

16'

OH

38

- Nhit nng chy cao: 190oC.

- Tnh tan: l cht mu khng phn cc nn khng tan trong nc, tan
trong cc dung mi khng phn cc.
- hp th nh sng cao nht bc sng 445 nm.
- nh hng n bn mu trong iu kin nhit , nh sng, khng
kh, acid. D b oxy ha mt mu.
Tnh cht ha hc
- Nhm hydroxyl hai u phn t lutein hot ng mnh, d dng tham
gia vo cc phn ng oxy ha nn lutein c kh nng oxy ha mnh.
- H thng polyen cng l nguyn nhn lm cho cc lutein dng t
do rt d b oxy ha v ng phn ha bi cc tc nhn nh sng, nhit ,
acid. Do , cn bo qun lutein dng t do trong kh tr, chn khng, bo
qun nhit thp, nn bao kn v trnh nh sng mt tri. [26]
1.3.1.3. Hot tnh sinh hc. ng dng
Tc dng to mu
Mu carotenoid l do nhm mang mu (chromophore) l h thng lin
hp lin kt i C=C. Trong phn t carotenoid c th c n 15 lin kt i,
nhng thng gp t 711 lin kt i. Nhm mang mu c t 7 lin kt i
tr ln c kh nng hp th nh sng vng thy c v lm cho carotenoid
c mu vng, cam, [31]. Chnh v vy, lutein l cht c kh nng to mu
vng cam nh chc nng sinh hc ca n. H thng lin kt i ny gi l
polyene ca carotenoid. Chng rt d b phn hy di tc ng ca nh sng,
nhit , oxy v acid (Li et al, 2007).[16]
Cht mu t nhin ang c khuynh hng thay th dn cht mu nhn
to bi nhng kh nng vt tri v c tnh sinh ha, m bo an ton tnh
trng c hi ca cht mu tng hp trong thc phm hin nay, ng thi m

39

ra nhiu hng nghin cu mi v cht chng oxy ha cho ngnh cng

nghip dc, m phm v kh nng to mu trong chn nui. Tnh hnh khai
thc v s dng cht mu t nhin ngy cng c quan tm nghin cu.
Trong bi bo co v nh gi ha cht v k thut (CTA) ca t chc
lng thc v thc phm (FAO) nm 2004 (D tho ln u tin do Richard
Cantrill chun b) a ra phng php tch chit lutein trong du nha ca
cnh hoa CVT bng dung mi hu c v s dng n nh mt cht ph gia b
sung cht mu vng t nhin vo trong thc phm. [7]
Nh c tnh sinh hc c kh nng to mu v tan trong hu ht cc
cht bo, trong nghin cu lutein c s dng b sung vo thc n chn nui
to mu vng cho da, chn v lm tng lp lng trng g cng nghip. [4],
[5], [8], [24]
Theo PGS-TS L Vn Knh, Vin ph Vin Khoa hc K thut Nng
nghip min Nam, cho bit nhng cht to mu c php s dng sn
xut thc n chn nui hin nay c chit xut t hoa CVT nn kh c kh
nng nh hng n sc khe ngi s dng.
Kh nng chng oxy ha
Kiu c trng ca cc lin kt n v i xen k trong khung phn t,
ng thi s c mt ca nhm hydroxyl hai u phn t gip lutein hp th
nng lng tha t cc phn t khc, gii thch cho tnh oxy ha mnh ca
cc carotenoid ni chung v lutein ni ring trong chc nng sinh hc
(Oliver et al., 1998)[16]. L cht d b oxy ha, cc lutein c kh nng
chng oxy ha mnh, ng vai tr quan trng i vi sc khe con ngi.
Trn th trng Chu u, lutein c bit n nh mt dc phm c
s dng rng ri trong y dc v m phm nh vo cc c tnh sinh hc sn
c ca n. Lutein v zeaxanthin c bit n nh mt cht chng oxy ha, c

40

ch s peroxy lipid, l tc nhn gy bnh v vng mc v bnh tim mch, c

ch s dy ln ca thnh ng mch gy t qu, v mt s bnh ung th,
Ngoi ra, trong lnh vc cng ngh m phm, lutein c bit n vi
vai tr bo v da, ngn chn s tn thng t bo v chng ung th da di
tc hi ca tia UV bng cch dp tt oxy n phn t hoc ha gii cc tc
nhn quang hc. [1], [12], [13], [15], [28]
Theo quyt nh ca th tng chnh ph s 61/2007/Q-TTG ngy 07
thng 05 nm 2007 v vic ph duyt Chng trnh nghin cu khoa hc
cng ngh trng im quc gia pht trin cng nghip ho dc n nm
2020, trong ni dung 5, cp vn nghin cu tch chit lutein,
zeaxanthin t hoa CVT lm nguyn liu bo ch thuc Vicuva.
Theo nghin cu ca trng i hc Khoa hc T nhin, i hc Quc
gia H Ni (2006) hm lng lutein c tm thy trong nhiu loi rau qu
lm thc n hng ngy v c nghin cu ng dng lm thuc cha
bnh [30].
Vin Y hc Qun s Vit Nam a ra mt s cc nghin cu mi y
cho thy, trong l inh Lng c hm lng lutein nhiu nht trong s cc loi
rau xanh.
Trong nc, vic khai thc lutein ch mi dng li mc nghin
cu v hot tnh dc hc v kh nng to mu trong chn nui m cha thc
s quan tm n kh nng to mu ca n trong thc phm.

41

Bng 1. Hm lng -caroten, lutein, lycopen trong mt s thc vt [30]

% cc cht carotenoid
Mu

lutein lycopen
caroten

% cc cht carotenoid
Mu

lutein lycopen
caroten

Xoi

50,41

L u

57,06

11,86

t vng

28,19

9,27

L dip c

29,58

25,94

Hoa hng
vng

22,66

18,91

7,54

L b ng

35,87

23,60

6,50

Hoa t
mui

3,44

9,17

L ngi
cu

33,03

12,57

C chua

20,57

18,83

22,48

L
tng

11,44

Hoa ip

17,05

1,85

3,36

L rau m

12,47

22,19

Rau dn
cm

50,69

24,66

3,07

L rau du

52,92

12,50

Rau dn
ta

42,29

17,72

L din

42,57

21,61

28,86

19,02

L tru

35,60

22,96

8,58

16,36

L rau ngt

50,05

13,98

L mp
ng

24,64

6,82

18,38

50,76

Cn ty
Rau ta
t
Rau b

37,99

21,55

L hnh

20,82

33,33

L inh
lng

Rau ay

19,54

28,99

L rau sam

55,31

12,78

L m

49,40

12,10

Tht mi
mt

12,73

14,60

L u

57,06

11,86

Tht b

52,70

15,21

42

1.3.2. Khi nim v cht mu lutein ester

Lutein ester l cht mu vng cam trong cht du nha lutein
(oleoresin) ca cnh hoa CVT, tn ti trong t nhin di dng ester ca
lutein vi cc acid bo nh acid palmitic (C16H32O2), acid myristic
(C14H28O2), acid stearic (C18H36O2). Trong dch chit lutein ester bng dung
mi, chim hn 80% l carotenoid tng s, trong , lutein chim 7078%,
zeaxanthin (29%) v mt s carotenoid khc, ngoai ra, cn tm thy cht sp
(14%) v acid bo (1%) trong oleoresin cha qua ch bin [7].
1.3.2.1. Cu to phn t ca lutein ester
Lutein ester l mt dn xut ca lutein, c mt hoc nhiu chui axit
bo lin kt vi lutein thng qua cc nhm hydroxyl hai u ca lutein.
Trong t nhin, lutein tn ti di dng ester, do , cn phi qua bc x
phng ha lutein ester to lutein.
S phn b cc ester bo trong hoa CVT ch yu l lutein dipalmitate.
Ngoi ra, cn c lutein dimyristate, lutein palmitatestearate, lutein myristate
palmitate, lutein distearate v mt lng nh lutein myristatestearate.[30]

Hnh 1.2. Cu to phn t ca lutein ester [30]

1.3.2.2. Tnh cht l-ha ca lutein ester
Tnh cht vt l
- Lutein ester cho sn phm dng rn mu vng cam.

43

- Khng tan trong nc, tan trong cht bo, cc dung mi clo ha nh:
chloroform, dichloromethane v cc dung mi khng phn cc khc.
- hp th nh sng cao nht bc sng 445 nm.
- Gim bn mu trong iu kin nhit , nh sng, oxy khng kh.
Tnh cht ha hc
Lutein ester bn hn, t b oxy ha hn v nhm hydroxyl ca lutein b
bt hot ha;
Tham gia phn ng x phng ha bng KOH to lutein;
Tham gia phn ng thy phn di tc dng ca enzyme trong c th
ngi to lutein c hot tnh chng oxy ha cao [2].
1.3.2.3. Hot tnh sinh hc. ng dng ca lutein ester
Kh nng chng oxy ha
Lutein ester l mt ngun cung cp lutein phong ph v cn thit v c
th khng t to lutein m tiu th lutein qua cc ngun thc phm cha
lutein ester. S chuyn i lutein ester t nhin thnh lutein trong c th lm
tng nng lutein ti im vng vng mc, huyt thanh trong c th ngi,
nh hot tnh chng oxy ha cao, lutein gip lm chm thoi ha im vng
ngi cao tui, gim kh nng c thy tinh th, tng nhn p, chng cc
bnh thoi ha nh ung th v, ung th rut kt, ung th phi, ung th d
dy,, ci thin sc khe tim mch, chng x va ng mch v t qu.
Tc dng to mu
C th x phng ha lutein ester bng KOH v tinh ch to sn phm
lutein dng bt b sung cht mu, thnh phn dinh dng vo cc thc phm
nh bnh quy gin t go, cookies v bnh kem xp, thc phm ln men nh
bnh m v sa chua, cc loi ung dinh dng giu protein, thc phm
dinh dng, thc phm chc nng; to ch phm dc phm dng vin nn,

44

vin nang gelatin cng, vin nang gelatin mm hoc trong nhiu trng hp
pha trn vi cc thnh phn khc.
Vi kh nng bn mu hn lutein nh cu to phn t, lutein ester to
ph gia dng lng c th b sung to mu cho cc thc phm nng nh
bnh m v cc loi bnh nng,
Lutein ester tn ti trong cnh hoa CVT, l ngun nguyn liu d tm
v cng l dc liu quen thuc vi tt c mi ngi, c Hoa K cng
nhn an ton v sc khe.
Bo v da, chng tia UV
Lutein c kh nng hp th mt cch chn lc nng lng nh sng
xanh nh tia UV gy tn thng da nh bng da do chy nng, kch thch sc
t nu lm bin mu da, nng hn na l vim da v ung th da.[6]
1.3.3. So snh kh nng hp th ca lutein v lutein ester
Lutein v lutein ester khc nhau v cu to phn t v khi lng, dn
n tnh cht l ha cng khc nhau, v th, kh nng hp th ca lutein v
lutein ester cng khng ging nhau. hp th ph thuc vo khong thi
gian b sung, cu to phn t, thnh phn cht bo trong khu phn n v
chc nng sinh l ca tui.
- Theo nghin cu cho thy s hp th lutein ph thuc phn ln vo
khong thi gian b sung lutein. Ty thuc vo liu lng a ra, t c
nng lutein n nh trong huyt tng c th mt 6 8 tun b sung lutein
v mt 6 thng tng mt sc t quang ti im vng ca mt. Do , sau
6 tun c th xc nh c nng lutein trong huyt thanh so snh kh
nng hp th ca lutein v lutein ester.
- Trong t nhin lutein tn ti dng mono hoc diesters lutein, lm
gim hot tnh dc hc ca lutein ester, do , trong dc phm b sung
lutein ester: lutein theo t l 1:1 hoc 2:1 bng cc ch phm lutein ester dng

45

vin nang cng v lutein dng vin nang mm. T l b sung tnh theo gi
thit thy phn 100% lutein vi s c mt ca cc enzyme thy phn trong
ng rut, tuy nhin, thc t ch thy phn 95% hm lng lutein ester.
- S c mt ca cht bo gip tng cng kh nng hp th carotenoid
trong ng rut [J.W. Erdman,1988]. Do yu cu hm lng cht bo cao
(khng vt qu quy nh) trong khu phn n y mnh qu trnh thy phn
lutein ester v kh nng hp th lutein trong c th. Theo bo co ca
Roodenburg et al.(2000) khi tng hm lng cht bo ln 12 ln th kh nng
hp th lutein t lutein ester tng ln gp i [Roodenburg AJ, Leenen R, van
het Hof KH, Weststrate JA and Tijburg LB., 2000]
- tui ng vai tr quan trng vic hp th lutein v qu trnh lo ha
nh hng trc tip n kh nng hot ng ca men tiu ha. Trong mt nghin
cu ca tin s Johnson E. (2004), a ra mi lin quan gia tui tc v hm
lng lutein trong huyt thanh nhm ngi c s dng lutein ester. Pht hin
ny cho thy kh nng thy phn lutein ester gim khi tui tc cng cao. y l
iu kin xem xt b sung liu lng lutein cao hn cho ngi cao tui [2].
1.4.

Tng quan v cc phng php chit xut cht mu t nhin

1.4.1. Khi nim chit xut

Chit xut l phng php dng mt dung mi (n hay hn hp)
tch ly mt cht hay mt nhm cc cht t hn hp cn nghin cu. Chit
xut nhm mc ch iu ch hay phn tch.
1.4.2. C ch qu trnh chit xut
Qu trnh chit xut bng dung mi c bit n nh l mt qu trnh
bao gm 4 c ch vt l ca cc qu trnh sau y:
- S tng tc ca dung mi trong qu trnh trao i cht trn b mt vt
liu chit xut;

46

- Qu trnh truyn dung mi bn trong sn phm c thc hin th

lng bi nhng qu trnh khc nhau nh: s mao dn, khuch tn phn t v
gradient ca nng dung mi l ng lc cho qu trnh ny;
- Qu trnh truyn cht tan vo dung mi xy ra bn trong sn phm,
n c thc hin bng qu trnh khuch tn bn trong vt liu. Gradient ca
nng cht tan l ng lc ca qu trnh ny;
- Qu trnh vn chuyn cht tan t b mt vt liu ra ngoi mi trng
dung mi, qu trnh ny c thc hin bng qu trnh khuch tn i lu. [34]
1.4.3. Nguyn tc chit xut
Phng php chit xut l bao gm vic chn dung mi, dng c chit v
cch chit. Mt phng php chit xut thch hp ch c th c hoch nh
mt khi bit r thnh phn ca cc cht cn li trch trong cy ra. Mi loi hp
cht c ha tan khc nhau trong tng loi dung mi. V vy, khng th c
mt phng php chit xut chung p dng cho tt c hp cht thin nhin.
Phng php c in chit xut mt hp cht thin nhin l dng mt
dy dung mi bt u t khng phn cc n phn cc mnh li trch, phn
on cc hp cht ra khi hp cht thin nhin. Da vo tnh phn cc ca
dung mi v ca cc nhm hp cht ta c th d on s c mt ca cc cht
trong mi phn on li trch. [36]
1.4.4. Cc phng php chit xut
Chit gin on:
Theo phng php ny ta ngm nguyn liu vo dung mi. Sau mt
thi gian nht nh, khi gia dung mi v nguyn liu t nng cht cn
thit mc cn bng, tin hnh dung mi c ra, thay dung mi mi
vo. C nh th cho n khi chit ht cht cn chit. Phng php ny c u
im l n gin, d thc hin, khng tn my mc, thit b cng nh chi ph

47

nng lng. Nhc im l tn cng, tn thi gian cng nh tn dung mi

chit nn khng kinh t, khng ph hp vi quy m sn xut ln.
Chit bn lin tc:
Nguyn l ca phng php ny l dng nhiu thit b chit gin on
b tr thnh mt h thng lin hp tun hon, nhm mc ch gim thi gian
chit, t tn cng hn, tit kim c nhiu dung mi hn. i vi phng
php ny, qu trnh chit thc hin theo nguyn tc dung mi i t ni c
nng cht chit cao n nng cht chit thp.
Chit lin tc:
Nguyn l l ngm dung mi trong dng chuyn ng cng chiu hay
ngc chiu ca dung mi. u im ca phng php ny l cho hiu qu
kinh t cao, thch hp cho sn xut cng sut ln, p dng cho quy m cng
nghip. Tuy nhin, nhc im l thit b kh phc tp, chi ph u t ln.[36]
1.4.5. Cc yu t nh hng ti qu trnh chit xut
- Chnh lch nng cht cn chit trong nguyn liu v dung mi;
- Hnh thi, tnh cht v cu to ca t chc nguyn liu;
- nh hng ca thi gian v nhit chit;
- Dung mi chit [34]
1.4.6. Vi k thut chit hin i dng chit xut cht mu t nhin
Chit nh siu m (Ultrasound-assisted extraction)
Nguyn liu c trn vi dung mi thch hp ri chit bng siu m.
Nhiu nghin cu cho thy rng siu m c kh nng ph v mng t bo ca
nguyn liu, t gip dung mi xm nhp vo bn trong t bo d dng hn.
Ngoi ra, siu m cn c tc dng khuy trn mnh dung mi, gia tng s tip
xc ca dung mi vi cht cn chit v ci thin ng k hiu sut chit. [9], [22]
Chit siu ti hn (SFE: Supercritical Fluid Extraction)

48

y l phng php chit c quan tm nhiu nht hin nay trong

lnh vc chit cc hp cht c hot tnh sinh hc t nguyn liu t nhin,
nhm ng dng trong cng nghip dc phm v thc phm. Phng php
ny cho php t ng ha qu trnh chit v hn ch vic s dng cc dung
mi hu c c hi. Dung mi chit l mt cht lng trng thi siu ti
hn. trng thi ny, cht lng c nhng tnh cht c bit nh c tnh chu
nn cao, khuch tn nhanh, nht v sc cng b mt thp, Do , n c
kh nng khuch tn mnh vo nn nguyn liu tt hn nhiu so vi cc
dung mi thng thng, v th lm tng hiu sut chit ln nhiu ln. Trong
phng php ny, thng dng CO2 trng thi siu ti hn lm dung mi
chit (i khi trn vi vi % dung mi phn cc no nh etanol, metanol,
2-propanol lm tng kh nng ha tan carotenoit ca CO 2), do , n cho
php chit nhanh, chn lc, khng lm oxy ha carotenoit v an ton trong
vn hnh.[19]
Chit dung mi tng tc (ASE: Accelerated Solvent Extraction) hay
chit di p sut cao (PFE: Pressurized Fluid Extraction)
y cng l mt phng php chit mi, cho php chit rt nhanh, t
ng ha, hiu qu v tit kim dung mi. Nguyn tc ca n tng t nh
phng php chit Soxhlet c in, ngoi tr vic qu trnh chit c thc
hin nhit v p sut cao (nhng vn di im ti hn ca dung mi s
dng). Trong phng php ASE, nguyn liu cn chit c xay nh, lm
kh (thng l ng kh), ri nhi vo mt ng chit (extraction cell). ng
chit ny c t trong l duy tr nhit thch hp (c th iu chnh t
40200oC). Dung mi c bm vo ng chit v gi p sut 10 20 MPa
trong vi pht (static time), sau dch chit c y vo mt bnh hng
bng mt th tch dung mi mi (flush volume). Qu trnh c lp li vi

49

ln. Cui cng, ton b dch chit c y ra bng mt dng kh tr

N2.[10]
1.4.7. Tnh hnh nghin cu chit xut lutein ester trn th gii
Vic ng dng lutein vo cc ngnh cng nghip m phm, dc phm
pht trin mnh m. Kt qu kho st cho thy, CVT chu Phi l ngun
nguyn liu lutein l tng cho vic thu nhn lutein: cnh hoa c hm lng
lutein ester kh ln (1,01,6% carotenoid tng s tnh theo trng lng kh),
trong , khong 90% lng carotenoid ny l lutein v 5% l zeaxanthin.
Trong dch chit lutein ester bng dung mi [FAO(2004)], chim hn
80% l carotenoid tng s, trong lutein chim 7078%, zeaxanthin (29%)
v mt s carotenoid khc, ngoi ra, cn tm thy cht sp (14%) v acid bo
(1%) trong oleoresin cha qua ch bin [7].
Trong nghin cu Philip, T. (1977) nghin cu chit xut lutein ester
t bt hoa CVT sy kh bng ete du m nhit phng vi t l 1:3 (w/v).
Dch chit c c ui dung mi di p sut thp 50oC thu lutein
oleoresin, sau , em ha tan trong isopropanol nng 75oC ri lc b tp cht
khng tan. Dch lc c lm lnh xung 15o C hay thp hn. Lutein ester tch
ra c lc v lm kh di p sut thp 30o C thu lutein ester tinh th.[23]
Levy, L. W. (2001) ci tin quy trnh ca philip T. (1977)
thu sn phm oleoresin c t l trans-ester cao bng cch bng cch ngm
bt hoa CVT kh trong hexan (8 l/kg) nhit phng. Tch ly dch chit,
c ui dung mi thu lutein ester th. Sau , thm isopropanol vo lutein
ester th nhit phng ha tan cis-lutein ester v cc tp cht khng
thuc nhm xanthophyll. Sn phm sau c c chn khng loi b
isopropanol. Kt qu thu c sn phm cha 69% lutein ester trong t l
ng phn trans:cis l 90:10.[20]

50

Bt hoa CVT cng c s dung lm nguyn liu trong nghin cu

ca Kumar S.T.K.(2004). Tc gi dng 2-propanone hoc 2-butanone thay
cho isopropanol trong cng on loi b tp cht v ng phn cis-lutein ester c
trong oleoresin. Nh vy, d lng dung mi c th c ui nhit phng.
Kt qu tng t l ng phn trans:cis trong sn phm ln t nht 18:1. [18]
Ngoi ra, cng xut hin nhiu hng nghin cu hn ch s thy phn
lutein ester trong cng on chit, gim thi gian chit v tng hiu sut chit
bng cch dng cc k thut hin i nh chit vi s h tr ca sng siu
m, chit bng CO2 siu ti hn hay chit vi p sut cao.
Theo nghin cu ca Peter Amala Sujith, T.V. Hymavathi and P.
Yasoda Devi (2010) s dng phng php chit bng CO2 siu ti hn vi p
sut l 27,5 MPa, nhit l 60oC dch chit thu c t bt hoa CVT chim
t 23 40% hm lng lutein ester. [17]
Theo nhm nghin cu ca Trng K thut Du kh v Ha cht, i
hc Khoa hc v Cng ngh Lan Chu, Trung Quc (2010), thc hin chit
lutein ester t hoa CVT (Tagetes erecta) vi dung mi h tr sng siu m
thu c hm lng lutein ester 17,17 mg/kg. [21]
Mc d cc k tht chit hin i dc p dng kh ph bin trong
quy m phng th nghim cc nc trn th gii, tuy nhin, vic p dng
rng ri trong thc t sn xut cn hn ch, do chi ph u t thit b cao,
khng kinh t. Trong thc t, lutein ester ch yu vn c chit bng
phng php ngm chit ca hoa CVT kh trong hexan. Chnh v vy, trong
n tt nghip ny thc hin chit lutein ester t bt hoa CVT (Tagetes
erecta L.) bng phng php ngm chit gin on.
.

51

PHN 2.
I TNG V PHNG PHP NGHIN CU
2.1.

i tng nghin cu

2.1.1. Vt liu nghin cu

Nguyn liu chnh:
Trong nghin cu ny, nguyn liu dng chit lutein l hoa CVT
(Tagetes erecta L.) c sy kh 70oC v nghin thnh bt mn
Ha cht:
Bng 2.1. Danh mc ha cht s dng trong ti
Ha cht

tinh khit

Etanol

99,7%

Hexan

97%

Na2SO4 khan

99%

Isopropanol

99,7%

BHT
Nc ct 1 ln

Xut x

Trung Quc

99%

Merck (c)

Chng ct ti phng th
nghim

2.1.2. Dng c v thit b

Dng c:
- Bnh lng gn;
- ng nghim;
- Bnh tam gic 250 ml;

52

- Bnh nh mc 10ml, 25ml; 50ml, 100 ml

- Cc thy tinh 500ml, 50ml, 100ml;
- Phu lc 8;
- ng nghim 18;
- Pipet 1ml, 5ml;
- Phu lc thy tinh;
- Giy lc, giy nhm, bng ht nc;
- ng ong;
- Cc t thy tinh.
Thit b:
- Quang ph k Genesys 20 (THERMO, M );
- Quang ph k UV-VIS Carry 50 (VARIAN, M);
- T lnh;
- Cn phn tch in t chnh xc

10-4 g (SATORIUS, M);

- T sy mu UNB 400 (Memmert, c);

- My ct nc 2 ln WSC/4D (Hamilton, Anh);
- H thng ct quay chn khng gm:
My c quay chn khng RV10 (IKA, c);
Bm ht chn khng N 026.3 AN 18 (KNF, c);
B lm lnh tun hon VS 1902 WF (VISION, Hn quc).
2.2.

Phng php nghin cu

2.2.1. Xc nh mt s thnh phn ha hc ca cnh hoa cc vn th

a) Xc nh hm lng cht kh: Sy nhit 105110oC n khi
lng khng i (Ph lc 1)
b) Xc nh hm lng lutein tng s: Chit hon ton lutein ester bng
acetone, phn b dch chit trong eter du m, nh mc v o hp th ca
dung dch 445 nm (Ph lc 2).

53

2.2.2. Quy trnh d kin sn xut lutein ester t hoa cc vn th.

Trn c s kt qu ca mt s nghin cu lin quan, chng ti xut
quy trnh d kin sn xut cht mu lutein t cnh hoa CVT nh hnh 2.1
Gii thch quy trnh
- Nguyn liu
Bt hoa kh thu nhn t cnh hoa CVT ti bng cch sy kh 70oC
ri nghin thnh bt. Bt hoa c bo qun 4oC v bao b kn, ht chn
khng
- Ngm chit
Ngm chit nhm mc ch chit xut cht du nha oleoresin cha
cht lutein ester cn chit ra khi bt hoa CVT bng dung mi hu c.
Tin hnh ngm bt hoa CVT trong iu kin kn kh theo phng php
ngm chit vi cc iu kin thch hp (t l dung mi ethanol:hexane (v/v);
t l dung mi/nguyn liu, thi gian ngm; s ln chit) ngm nhit
phng, trnh nh sng t nhin. Dung mi chit c b sung 0,1% BHT
(w/v) ngn s oxy ha v phn hy lutein ester. Thu dch chit, lm khan
bng Na2SO4 .
- Lm khan
Thu dch chit qua phu lc bng bng y t c Na2SO4 dng rn
ht lm khan nc trong dch chit, ng thi gip loi phn cnh hoa cn st
li trong dch chit to iu kin thun li cho qu trnh c ui dung mi tip
theo.
- C ui dung mi
C ui dung mi nhm mc ch thu hi cht du nha lutein oleoresin
dng c c ng thi thu hi dung mi, em chng ct v ti s dng.

54

Tin hnh lm kh dung mi iu kin p sut thp 400600 mbar,

nhit khng qu 40oC, trnh nh sng t nhin hn ch kh nng oxy
ha v phn hy lutein ester.
- Tinh ch
Tinh ch lutein ester thu sn phm dng tinh th.
Sau khi c ui ht dung mi thu c lutein ester th. Tinh ch lutein
ester bng cch ha tan trong isopropanol 75oC nhm mc ch ha tan
lutein ester. Tip theo, lc dung dch qua phu lc ht chn khng lc b
nhng tp cht khng tan. Dch lc sau c lm lnh xung 15oC, qua
m trong t mt to iu kin kt tinh cc lutein ester. Sau , lc qua
phu lc ht chn khng mt ln na thu sn phm lutein ester tinh th.
- Sy chn khng
Thc hin sy chn khng sn phm kt tinh 50oC thu lutein ester
dng tinh th kh. ui dung mi cn st li bng dng kh N2 (30 pht)
Sau khi thu hi hn hp dung mi hexan-ethanol, cn tin hnh chng
ct hn hp ny tch ring tng dung mi c thu hi v tip tc dng
cho cc m sau.
Dung dch cha isopropanol sau khi lc ly tinh th cng c c chn
khng v chng ct thu hi isopropanol tinh khit v tip tc s dng cho
ln sau.
Tt c cc thao tc trn nn thc hin trong bng ti.
Tt c cc dung mi v isopropanol thu hi s c lc qua Na2SO4
dng rn loi nc trong dung mi sau thu hi.

55

Bt hoa cc vn th (G gam)

Ngm chit

Dung mi

- Dung mi?
- Dung mi:Nguyn liu?
- Thi gian ngm?.
- S ln chit?

C ui dung mi

Lc
Chng ct

Thi
isopropanol

Du nha lutein

Tinh ch

Isopropanol

Lutein ester
tinh th

Sy chn khng

Bt mu lutein
ester

Hnh 2.1. S quy trnh d kin sn xut lutein ester t hoa cc vn th

56

2.2.3. Xc nh iu kin thch hp chit lutein ester t hoa cc vn th

Cn chnh xc khong G g nguyn liu cho vo bnh cha.
t c hiu sut chit cao, cn chn c iu kin tt nht
chit lutein bng cch tin hnh cc l th nghim sau:
a) Chn dung mi chit (L TN1):
Vic la chn dung mi chit trc ht cn da vo kh nng ha
tan cc sc t, tc da trn s tng thch gia tnh phn cc ca dung mi v
ca cht cn chit. Lutein trong hoa CVT tn ti dng ester, c tnh phn
cc thp. Do vy, cc dung mi c tnh phn cc thp s ha tan tt sc t
ny. Trong mt s nghin cu trn th gii v tch chit cht mu lutein ester.
Cc dung mi thng c s dng l ete du m (Philip, T., 1977), hexan
(Levy L. W., 2001), hay acetone, ethanol hoc hn hp dung mi hexaneacetone (Serena Lim Sue Lynn, 2003) [19], [ 22], [25], trong thng dng
nht l dng hexane, Tuy nhin, hexane l dung mi kh t tin nn tit
kim chi ph dung mi, chng ti th nghim thay th hexane bng hn hp
hexane-ethanol. Do vy, cn kho st h dung mi hexane-ethanol thch hp
t hiu sut chit cao nht. Th nghim tin hnh nh sau:
C nh t l dung mi trn nguyn liu l 10:1(v/w). Tin hnh chit 1
ln bng phng php ngm chit (24 h, nhit phng, trong ti).
Dung mi chit s dng l dung dch ethanol-hexane, trong t l
hexane trong hn hp dung mi (X1) thay i ln lt t 0 100% (v/v):
X1 = 0; 20; 40; 60; 80; 100 % (v/v).
Dch chit sau c chit sang dung mi hexan, pha long n th
tch thch hp bng hexan ri o quang 445 nm xc nh hiu sut chit
lutein (%H) theo cng thc:

57

%H =

lng lutein chit c

100%
lng lutein tng s

T , chn t l hexan thch hp.

Lu :
Lutein km bn nhit v nh sng nn tt c cc th nghim ngm chit
u tin hnh chit nhit phng, trong bng ti.
b) Chn t l dung mi/ nguyn liu (L TN2):
Dng dung mi chit thch hp chn. Tin hnh chit 1 ln bng
phng php ngm chit (24 h, nhit phng, trong ti), trong t l
dung mi/nguyn liu (X2) thay i nh sau:
X2 = 8:1; 10:1; 12:1; 14:1; 20:1 (v/w)
Gn ly dch chit, tin hnh o quang xc nh hiu sut chit. T
, chn t l dung mi:nguyn liu thch hp.
c) Chn thi gian chit (L TN3)
Ngm nguyn liu trong dung mi chn vi t l dung mi:nguyn
liu thch hp , trong thi gian ngm chit (X3) thay i nh sau:
X3 = 1; 2; 3; 4; 5 (ngy)
Sau nhng thi gian trn, ly mu tng ng ra, gn ly dch chit v
xc nh hiu sut chit. T , xc nh thi gian chit thch hp nht.
d) Chn s ln chit (L TN4)
Tin hnh ngm chit vi cc thng s c xc nh (dung mi, t
l dung mi:nguyn liu, thi gian ngm chit), trong thay i s ln chit
i vi tng mu (X4) nh sau chn s ln chit thch hp:
X4 = 1; 2; 3; 4 (ln)
S b tr th nghim xc nh iu kin thch hp chit lutein t
cnh hoa CVT bng phng php ngm chit trnh by hnh 2.2.

58

L TN 1

Ngm chit (X2 = 10/1 v/w; X3 = 24 h; X4 =1 ln)

X1 = % Hexane (v/v)

20

40

60

80

100

Tnh hiu sut thu hi lutein

% Hexane thch hp (X1 opt)
L TN 2

Ngm chit (X1 opt; X3 = 24 h, X4 = 1 ln)

X2 = Dmi/N.Liu (v/w)

8/1

10/1

12/1

14/1

20/1

Tnh hiu sut thu hi lutein

T l dung mi/nguyn liu thch hp (X2 opt)

L TN3

Ngm chit (X1 opt ; X2 opt ; X4 = 1)

3
X3 = t/gian (ngy)

Tnh hiu sut thu hi lutein

Thi gian chit thch hp (X3 opt )

L TN4

Ngm chit (X1 opt; X2 opt ; X3 opt )

X4 = s ln chit

Tnh hiu sut thu hi lutein

S ln chit (X4 opt)

Hnh 2.2. B tr th nghim xc nh iu kin chit lutein ester t hoa

cc Vn th

59

2.2.4. Th nghim quy trnh chit nh gi hiu sut chit

Sau khi a ra c cc thng s, iu kin chit thch hp cho quy
trnh chit lutein ester t hoa CVT, tin hnh th nghim chit xut lutein
ester vi quy m phng th nghim. nh lng lutein tng s chit c
xc nh hiu sut chit ca quy trnh.
2.2.5. Tinh ch v nh gi cht lng sn phm
Dch chit lutein ester sau c c ui dung mi thu sn phm
lutein ester th (du nha lutein) v thu hi dung mi.
Sn phm th cn cha nhiu tp cht, do to ra sn phm c
kh nng ng dng lm cht mu thc phm hay ph gia thc n chn nui,
cn tin hnh tinh ch. thc hin iu ny, chng ti p dng quy trnh
tinh ch lutein ester ca Phillip (1997) [23]. Theo quy trnh ny, ha tan sn
phm th trong mt lng ti thiu isopropanol c un nng n 75 o C sao
cho khng tan c na. Lc dung dch nng qua phu lc ht chn khng
loi b tp cht khng tan. Dch lc c ngui 15 oC qua m kt
tinh lutein ester. Lc ht chn khng thu tinh th lutein ester. Lp li qu
trnh tinh ch mt ln na. Tinh th lutein ester thu c em sy chn khng
50o C ri ui bng dng kh N2 loi b d lng dung mi cn st li.
nh gi cht lng sn phm lutein ester theo quy nh ca B Y t v
cht mu thc phm carotenoid (Ph lc 23. QCVN 4-10 : 2010/BYT) thng
qua xc nh cc ch tiu: cm quan; tan; quang ph UV-Vis; phn ng
mu; % carotenoid tng s (Ph lc3).
2.2.6. Nghin cu phng php bo qun dch chit lutein ester
Sn phm lutein ester c th b phn hy bi tc ng ca cc yu t bn
ngoi (nhit , nh sng, khng kh), lm gim cng v c tnh mu sc
ca sn phm. V vy, cn nghin cu bn mu ca lutein ester trong cc

60

iu kin bo qun khc nhau nhm a ra phng php bo qun thch hp

cng nh iu kin ng dng cho sn phm.
Ha tan khong 0,01 g sn phm lutein trong 25 ml dung mi hexan.
- Ht 10 ml dung dch thu c cho vo bnh nh mc 250 ml ri pha
long v nh mc bng hexan (k hiu dung dch: KBHT)
- Cng lm tng t nh vy nhng pha long v nh mc bng hexan
c cha 0,1% BHT (k hiu dung dch: BHT).
o quang xc nh nng ban u ca lutein ester trong cc dung
dch trn.
Cho cc dung dch trn ln lt vo cc l nh ng mu c np vn
ri bo qun cc iu kin khc nhau nh sau (xem hnh 2.3):
a) Dung dch khng c BHT:
- nhit phng, di nh sng t nhin (k hiu mu: KBHT-TS)
- nhit phng, trong bng ti (k hiu mu: KBHT-TT)
- Bo qun lnh 4o C, trong ti (k hiu mu: KBHT-4)
b) Dung dch c BHT:
Cng bo qun trong cc iu kin nh trn v c k hiu tng ng
l: BHT-TS; BHT-TT v BHT-4
C sau 1 tun, ly cc mu ra (mi loi mu ly 2 l) em o hp
th ca dung dch bc sng 445 nm (dng hexan lm dung dch so snh)
v xc nh % lutein cn li theo cng thc:
% lutein cn li =

lng lutein cn li
100%
lng lutein ban u

61

Dung dch lutein ester

Bo qun lnh

Bo qun nhit phng

Ti

(40 C), trong ti

Sng

Thi gian (tun):

o quang - Quan st
nh gi bn mu

Hnh 2.3. B tr th nghim nh gi bn mu ca dch chit

lutein ester

Tin hnh nh gi bn mu ca dch chit lutein ester trong cc

iu kin bo qun ni trn. T , xc nh phng php thch hp cho vic
bo qun v iu kin s dng cht mu lutein este thu c.
2.2.7. X l s liu
X l s liu v v th hai chiu bng phn mm MS Excel 2003.

62

PHN 3.
KT QU V THO LUN
3.1. Mt s thnh phn ha hc ca hoa cc vn th
Kt qu xc nh hm lng cht kh (bng PL4.9) v hm lng
lutein tng s (bng PL4.8) ca nguyn liu bt hoa cc vn th c lit k
trong bng 3.1 sau y:
Bng 3.1. Mt vi thnh ha phn ha hc c bn ca hoa cc vn th
Thnh phn

Hm lng

Cht kh (%)
Lutein tng s, mg/kg TL ti

88,6
7.308

Lutein tng s, mg/kg TL kh

8.247

T kt qu bng 3.1, ta thy:

- Nguyn liu CVT chng ti s dng thu c t cnh hoa CVT ti
qua cng on sy kh ( 70oC n va kh gin) v c bo qun hn 1
nm 4oC. Tuy vy, kt qu phn tch cho thy hm lng lutein ester trong
nguyn liu vn cn kh cao, c th dng lm ngun nguyn liu tt chit
xut lutein. Nh vy, vic sy kh hoa cc vn th v bo qun dng bt
kh vn l gii php thun tin, gip ch ng c ngun nguyn liu trong
sn xut.
- Hm lng cht kh ca hoa cc vn th khong 88,6%, ng vi
m nguyn liu l 11,4%. Vi m ny cc vi sinh vt (vi khun, nm mc)
vn c th pht trin c nu bo qun nhit phng trong mi trng
m. Do , bo qun trong thi gian di, sau khi sy xong nn ng
nguyn liu trong bao b ht chn khng hay c cht chng mc.

63

3.2. Xy dng quy trnh chit lutein ester t hoa cc vn th

3.2.1. Chn dung mi chit
Kt qu thu c khi tin hnh chit lutein ester t hoa CVT bng
phng php ngm chit (Bng PL4.1 v hnh 3.1) cho thy:
- Khi tng nng hexan trong dung mi ln t 0 60% th hiu sut
chit lutein ester tng ln. iu ny c gii thch bi s gim dn phn
cc ca dung mi khi tng t l hexan, do tng hiu qu ha tan v chit
xut cht mu. Nhng sau , tip tc tng nng hexan (80 100%) th
hiu sut chit tng khng ng k, v mc nng hexan 60% th kh nng
chit xut lutein ester gn nh hon ton. Do , mc d nng hexane c
tng ln nhng lng lutein cn li trong nguyn liu rt t dn n hiu sut
chit lutein ester v sau khng cao.
- Vi t l ethanol:hexane 40:60 th hiu sut chit lutein ester ln nht.
Do , dung dch ethanol:hexane vi t l 40:60 (v/v) c chn lm dung
mi chit.

90.00

nh hng ca dung mi n hiu sut chit lutein

ester
74.88

Hiu sut chit(%)

80.00
70.00

78.00

80.00

20/80

0/100

57.86

60.00
50.00

37.50

40.00
30.00
20.00

13.01

10.00
0.00
100/0

80/20

60/40

40/60

Ethanol / Hexane (v/v)

Hnh 3.1. nh hng ca dung mi n hiu sut chit lutein ester

64

3.2.2. Chn t l dung mi:nguyn liu

Biu (hnh 3.2) v bng PL 4.2 trnh by kt qu kho st nh hng
ca t l dung mi:nguyn liu n hiu sut chit lutein ester t nguyn liu
bt hoa CVT sy kh, cho thy t l dung mi:nguyn liu tng nhng hiu
sut chit lutein ester tng khng ng k.
- Khi t l dung mi:nguyn liu ny nh hn 12:1 (v/w), hiu sut chit
tng ln khi tng t l dung mi:nguyn liu. Tuy nhin, nu tip tc tng t
l dung mi:nguyn liu th hiu sut chit gn nh khng thay i. iu ny
c th gii thch nh sau: Vi t l dung mi:nguyn liu nh hn 12:1 (v/w),
lng dung mi cha ngp ht nguyn liu, do , hiu sut chit kh thp
(ch khong gn 40%). Khi t l ny t n ngng gi tr trn, ton b khi
nguyn liu ngp ht trong dung mi nn hiu sut chit tng ng k.
- Vi t l dung mi:nguyn liu t 12:1 (v/w) tr ln, ton b khi
nguyn liu c tip xc hon ton vi dung mi, hm lng lutein ester gn
nh tch hon ton ra khi nguyn nn d t l tng nhng hiu sut gn nh
l khng thay i.
Mc d t l 12:1 cho hiu sut chit tng i cao trong thi gian
ngn nhng hiu sut tng khng ng k so vi chi ph dung mi (Bng
PL4.10), iu ny nh hng trc tip n mc ch kinh t. Do , t l
dung mi:nguyn liu thch hp nht c chn l 10:1 (v/w)

65

Chn t l dung mi/ nguyn liu

109.53

60.00

58.80

76.87

67.09

66.54

66.93

66.27

80.00

59.05

100.00
56.10

Hiu sut chit lutein (%)

120.00

Hiu sut chit lutein

49.84

Chi ph dung mi

40.00
20.00
0.00
8/1

10/1

12/1

14/1

20/1

T l dung mi/nguyn liu

Hnh 3.2. nh hng ca t l dung mi:nguyn liu n hiu sut chit

lutein ester
3.2.3. Chn thi gian chit v s ln chit
3.2.3.1. Xc nh thi gian ngm chit
Qua th biu din s thay i hiu sut chit lutein ester vo thi
gian ngm chit (hnh 3.3 v bng PL4.4) ta thy:
Khi tng thi gian chit t 13 ngy th hiu sut chit tng mnh (t
55,79 70,86%). Nhng vi thi gian chit di hn, hiu sut chit li gim
S gim hiu sut chit trong trng hp ny c th gii thch bi s tc ng
ca yu t nhit , v s oxy ha lm mt mu v gim hm lng lutein.
Vi thi gian ngm chit l 3 ngy, hm lng lutein thu c l cao nht. Do
, thi gian ngm chit c chn l 3 ngy.

66

Chn thi gian ngm chit

Hiu sut chit Lutein (%)

80.00

70.86

70.00
60.00

64.82

67.35

67.25

55.79

50.00
40.00
30.00
20.00
10.00
0.00
1

Thi gian ngm chit (ngy)

Hnh 3.3. S ph thuc ca hiu sut chit vo thi gian ngm chit
3.2.3.2. Xc nh s ln ngm chit
Kt qu kho st s thay i hiu sut chit theo s ln ngm chit
(hnh 3.4 v bng PL4.3) cho thy:
Khi tng s ln chit t 1 n 3 ln th hiu sut chit tng ln
(66,27 76,47%), nhng sau 4 ln chit hiu sut tng rt t.
Nguyn nhn: Sau 3 ln chit hm lng lutein cn li trong nguyn
liu rt t, do , gradient nng gia b mt phn cch pha rn (nguyn
liu) vi dung mi rt nh, dn n tc chit khng ng k. Nh vy,
gim chi ph sn xut ta chn s ln chit l 3 ln.

67

Hiu sut chit lutein (%)

Chn s ln ngm chit

80.00

76.47

75.00
70.00

78.28

78.83

72.05
66.27

65.00
60.00
55.00
1

S ln ngm chit (ln)

Hnh 3.4. S ph thuc ca hiu sut chit vo s ln ngm chit

Tng hp ton b qu trnh nghin cu cho thy, iu kin thch hp
chit lutein ester t hoa cc nh sau:
- T l thnh phn dung mi chit ethanol:hexane = 40:60 (v/v);
- T l dung mi:nguyn liu 10:1 (v/w);
- Thi gian ngm chit: 3 ngy/ln chit
- S ln chit: 3 ln
- iu kin ngm chit: nhit phng, trnh nh sng t nhin.
3.3. Hon thin quy trnh chit sn xut cht mu lutein ester t hoa cc
vn th - Sn xut th nghim v nh gi cht lng sn phm
3.3.1. Hon thin quy trnh chit sn xut cht mu lutein ester t hoa
cc vn th

68

T nhng kho st trn , c th xut quy trnh sn xut cht mu

lutein ester t hoa CVT nh sau (hnh 3.5):
Thuyt minh quy trnh
- Nguyn liu
S dng nguyn liu l bt hoa cc vn kh (thu c bng cch st
cnh hoa ti 700C cho n khi kh gin, ri nghin thnh bt mn).
Nguyn liu c bo qun trong bao b kn, ht chn khng, trong mt.
- Ngm chit
Cn chnh xc lng nguyn liu (G kg) cho vo bn ngm chit c
np y kn hn ch s bay hi ca dung mi. Thm vo mt th tch
hn hp ethanol: hexane 40/60 v/v (c 4 lt ethanol trn vi 6 lt hexane) vi
t l dung mi: nguyn liu l 10/1 v/w (c 1 kg nguyn liu thm vo 10
lt dung mi). y kn h thng, yn nhit phng.
C sau 3 ngy chit rt dch chit mt ln, thm dung mi mi vo. Lp
li nh vy 3 ln. Tt c dch chit c gom li vo mt bn cha.
- C ui dung mi
Dch chit sau c ht vo bnh cha dung mi ca h thng c
chn khng. Tin hnh c dch chit di p sut thp sao cho nhit si
khng qu 40oC, trnh nh sng t nhin.
Sau khi kt thc qu trnh c dc chn khng, thu c du nha lutein
(lutein ester th) trong bnh c quay, Dung mi tch ra c em chng ct
thu ethanol v hexane ti s dng cho cng on ngm chit cc m
nguyn liu tip theo.
-

Tinh ch
Du nha lutein c c c tinh ch bng cch ha tan trong mt th

tch ti thiu isopropanol c un nng 75oC ha tan lutein ester.

Lc ly dung dch qua phu lc ht chn khng loi b nhng tp cht

69

khng tan. Dch lc sau c lm lnh n 15oC, qua m trong t mt

kt tinh lutein ester. Hn hp li c lc qua phu lc ht chn khng
mt ln na thu tinh th lutein ester nm trn phu lc. Dch lc c c
chn khng thu hi isopropanol ti s dng. Lp li qu trnh tinh ch va
ri mt ln na vi tinh th lutein ester.
- Sy chn khng
Sn phm lutein ester tinh th thu c sau khi tinh ch c em sy
chn khng 50oC trong 3 h ri ui dung mi cn st li bng dng kh N 2.
Sn phm thu c l bt mu lutein ester
Tt c cc thao tc trn nn thc hin trong bng ti.
Tt c cc dung mi v isopropanol thu hi s c lc qua Na2SO4
dng rn loi nc trong dung mi sau thu hi.

70

Bt hoa cc vn th

Ngm chit
Dung mi
C ui dung mi
Lc
Chng ct

Thi
isopropanol

- Dung mi etanolhexan 40:60 (v/v) + BHT

0,1% (w/v)
- Dung mi/ nguyn liu
10:1 (v/w)
- Thi gian ngm: 3
ngy/ln chit.
- S ln chit: 3 ln

Du nha lutein

Tinh ch

Isopropanol

Lutein ester tinh

ch

Sy chn
khng

Bt mu
lutein ester

Hnh 3.5. S quy trnh sn xut cht mu lutein ester

t hoa cc vn th

71

3.3.2. Kt qu th nghim quy trnh - nh gi cht lng sn phm

Tin hnh th nghim quy trnh vi 200 g bt hoa CVT kh (tin hnh 3
ln), thu c kt qu nh sau:
Bng 3.2. Kt qu th nghim quy trnh sn xut lutein ester
Mu

Lng

Hiu sut thu nhn

Lng du nha

Lng sn phm

mu (g)

lutein ester (%)

lutein ester (g)

lutein ester (g)

200

70,49

13,19

4,46

200

66,62

12,15

4,10

200

69,34

13,12

4,39

T.Bnh

200

68,82

12,82

4,32

Kt qu sn xut thc nghim bng 3.2 cho thy:

S dng h dung mi ethanol:hexane cho hiu sut chit lutein ester
trung bnh gn 70% vi lng sn phm lutein ester tinh ch l 4,32 g/ 200g
bt hoa kh, ng vi 21,6 g sn phm/kg bt hoa kh. Hiu sut ny hi thp
hn kt qu thu c khi tin hnh chit vi cc lng mu nh trong qu
trnh kho st (khong 76%) do s hao ht cht mu trong qu trnh lc, chit
(bm dnh trn cc dng c cha) tuy nhin lng tinh th lutein ester thu
c li cho kt qu cao hn so vi hm lng lutein ester thu c 21g/kg
bt hoa kh trong nghin cu ca Philip T. (1977).
Kt qu phn tch % Carotenoid trong sn phm lutein tinh th (bng
PL4.7) cho thy tinh khit ca sn phm thu c khng cao: 39,76%
carotenoid tng s, thp hn so vi nghin cu 51% ca Philip T. (1977). Nu
xem ton b lng carotenoid ny l lutein v gi thit lutein ester trong mu
tn ti di dng lutein dipalmitat th c th tnh c hm lng lutein ester
c trong sn phm cht mu nhn c l 72,34%. Mt khc, thc nghim

72

cho thy sn phm thu c khng hn dng bt kh m hi do, st v c

mu cam hi sm. iu ny chng t sn phm cn mt s tp cht kh loi
b c trong qu trnh tinh ch.
S khc nhau v tinh khit gia sn phm chng ti thu c v ca
Philip T. (1997) c l xut pht t nguyn nhn l do Philip dng hoa cc
vn th qua xi-l ph v thnh t bo sc t cha cellulose, pectin.
ri sy kh trc khi chit, nguyn liu chng ti s dng khng qua bc x
l nh trn nn cn cha cc tp cht pectin c th b chit vo dung mi. Tp
cht pectin ny b nha ha trong qu trnh c c sn phm, bm dnh vo
cc tinh th lutein ester trong qu trnh kt tinh nn kh tch, lm gim
tinh khit ca sn phm.
Bng 3.3. Kt qu phn tch hm lng carotenoid trong sn phm lutein ester

Ch tiu

Kt qu

% Carotenoid tng s

39,76

% Lutein dipalmitat

72,34

Hnh 3.6. Sn phm lutein ester tinh ch

73

3.4. Kt qu kho st bn mu ca lutein ester

Hnh 3.7 v hnh 3.8 trnh by kt qu kho st bn mu ca dch
chit lutein ester trong cc iu kin bo qun khc nhau.
Kt qu o cng mu ca dch chit lutein ester bng PL4.5 v
bng PL4.6 cho thy:
- Trong iu kin khng cng nh c b sung BHT, mc suy gim
hm lng lutein cn li trong cc mu bo qun u theo quy lut l: mu
bo qun 40C bn mu hn bo qun nhit phng trong ti; mu bo
qun nhit phng trong ti bn mu hn mu bo qun di nh sng t
nhin.
- Mu c b sung BHT bn hn mu khng cha BHT. Chng hn, i
vi mu c BHT bo qun 4oC (BHT-4) sau 7 tun th hm lng
carotenoid gim i cn khong 85% nhng mu khng cha BHT gim i
cn khong 79%.
BN MU CA LUTEIN ESTER TRNG
HP KHNG B SUNG BHT

% lutein cn li

120.00

100.00

KBHT TS

93.85
89.62

90
.77
87.15

80.00

88
.54
80.77

87.54

85
.69
80.15

85.38

83
.08
78.46

KBHT TT

83.85

81
.54
77.69

79.31

78
.62

78.77

76
.38

72.31

71.62

KBHT -4

60.00
0

Thi gian(tun)

Hnh 3.7. bn mu ca lutein ester khng b sung 0,1% BHT(w/v)

74

BN MU CA LUTEIN ESTER TRNG

HP B SUNG BHT

% lutein cn li

120.00

100.00

94.92

92.38

BHT TS
90.77

92.00
87.69

90.46

80.00

84.62

89.23

87.69

83.08

87.69

84.62

86.62

83.85

85.77

83.69

BHT TT

80.54
81.54

78.92

78.77

BHT - 4
76.08

60.00
0

Thi gian( tun)

Hnh 3.8. bn mu ca lutein ester khi b sung 0,1 % BHT (w/v)
Tuy nhin, v cm quan s khc bit v mu sc ca cc mu trn trong
cng iu kin khng thy s khc bit ng k. cho kt qu chnh lch nhau
khng ng k. Tht vy, qua quan st bng mt cho thy: Cc mu bo qun
iu kin thng trong ti, iu kin thng c nh sng v 4oC cho mu
sc khng khc bit nhiu sau 7 tun bo qun (hnh 3.7).
Nh vy, so vi cc carotenoid khc, lutein ester t nhy cm hn nhiu
vi oxy khng kh v nh sng hn cc carotenoid khc. Cc thc phm c
to mu bng lutein ester c th bo qun nhit phng hay 4 oC m
khng cn b sung BHT. Tuy nhin, nu bo qun lu di th nn gi trong ti
nhit thp nht c th.

75

i vi nguyn liu bt mu lutein ester th m bo % carotenoid

nh cng b, cn bo qun trong ti, nhit thp.

Mu KBHT-TS

Mu KBHT-TT

Mu KBHT-4

Hnh 3.9. Mu dch chit khng b sung BHT khi bo qun

cc iu kin khc nhau

Mu BHT-TS

Mu BHT-TT

Mu BHT-4

Hnh 3.10. Mu dch chit b sung BHT sau khi bo qun

cc iu kin khc nhau

76

KT LUN

T nhng kt qu nghin cu trn y, c th i n nhng kt lun sau:

1. iu kin thch hp chit cht mu lutein t bt hoa cc vn th
kh bng phng php ngm chit vi h dung mi ethanol:hexane nh sau:
- T l ethanol:hexane l 40:60 (v/v);
- T l dung mi: nguyn liu l 10/1 (v/w)
- Thi gian ngm chit: 3 ngy/ln chit
- S ln chit: 3 ln
(Tin hnh ngm chit nhit phng, trong ti, trong h kn)
Hiu sut chit lutein ester ca quy trnh khong 69%.
2. Sn phm lutein ester thu c sau khi tinh ch c hm lng
carotneoid tng s khong 40%, ng vi hm lng lutein dipalmitat c
chng 72%.
3. Sn phm cht mu lutein kh bn trong qu trnh bo qun. Tuy
nhin, cht mu b phn hy nhiu hn di nh hng nh sng, nhit v
khi khng c mt cht chng oxy ha.

77

KIN XUT
1. Cn nghin cu ci tin phng php ngm chit rt ngn thi gian
v nng cao hiu sut thu hi lutein ester (thc hin ngm chit c khuy
trn).
2. Cn x l nguyn liu bng phng php enzyme ph v thnh t
bo sc t trc khi chit, nhm d dng loi b cc tp cht trong qu trnh
tinh ch, nng cao tinh khit ca sn phm cht mu.
3. Cn nh gi y cc ch tiu cn t i vi sn phm kt lun
v kh nng ng dng lm cht to mu trong cng nghip thc phm.

78

TI LIU THAM KHO

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T.K.(2004),

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PH LC
Ph lc 1
Phng php xc nh hm lng cht kh ca cnh hoa.
a) Nguyn tc :
Dng nhit cao lm bay hi nc ht trong mu phn tch.
Da vo hiu khi lng ca mu trc v sau khi sy tnh hm
lng nc v hm lng cht kh c trong mu phn tch.
b) Tin hnh:
- Chun b cc sy : em cc i ra ro, sau sy 105110o C n
khi lng khng i. ngui trong bnh ht m ri em cn v xc nh
khi lng cc sy.
- Chun b mu: Nguyn liu bt hoa CVT em cn xc nh khi lng
G (g) cho mi mu (khong 1 g/mu) cho vo 3 cc sy ( c xc nh
khi lng trn). Cn cc c cha mu xc nh khi lng G1. a cc
c cha mu sy 6070o C trong vng 2 h, sau nng nhit ln
105110o C v sy trong vng 2 h (thnh thong trn u). Ly mu ra,
ngui trong bnh ht m v em cn. Lp li qu trnh sy vi ln n khi
chnh lch khi lng gia 2 ln cn lin tip khng qu 0,2 mg. Ghi khi
lng cui cng (G2).
Hm lng nc trong nguyn liu tinh theo cng thc:
%H2O =

G1

G2
G

.100%

Trong :
G: khi lng mu
G1: khi lng mu trc khi sy; G2: khi lng mu sau khi sy

Ph lc 2
Phng php xc nh hm lng lutein tng s
a) Nguyn tc:
Chit lutein ester t mu phn tch bng dung mi thch hp. o hp
th ca dch chit lutein ester trong dung mi chit 445 nm v xc nh hm
lng lutein bng nh lut Lambert-Beer.
b) Tin hnh
Cn chnh xc G (g) mu phn tch (khong 0,1g) cho vo 3 ng
nghim, thm 5ml axeton c cha 0.1% cht chng oxy ha BHT, vn nt
kn. Ngm mu nhit phng, trnh nh sng, thi gian khong 24h, lc
ly dch. Lp li qu trnh trn vi phn b cho n khi dch chit nhn
c khng c mu. Gp cc dch li em i chit v nh mc bng axeton
thnh VmL (Nu cn, pha long dch o quang vi h s pha long D); o
hp th ca dch 445nm, cuvet 1cm trn quang k UV VIS dng axeton
lm dch so snh hiu chnh my. Hm lng lutein tng s (Lutein TS) trong
mu phn tch (tnh theo trng lng kh) c tnh theo cng thc:
Lutein TS (mg/kg) =

Trong :

A V D 10000
2500 d G

A hp th ca dung dch o quang

V Th tch axeton dch chit sau khi nh mc (ml)
D H s pha long
2500 : H s hp th trung bnh ca dung dch lutein 1% (w/v) o vi
cuvet 1cm 445nm
d Chiu dy cuvet (d = 1cm)
G Khi lng kh tuyt i ca mu (g)

Ph lc 3. Phng php xc nh % carotenoid trong sn phm lutein ester

(da theo Ph lc 23-QCVN 4-10 : 2010/BYT)

Cn G gam mu (khong 27 33 mg) mu cht mu cho vo mt bnh
nh mc 100 ml, ha tan bng hn hp hexane:etanol:aceton:toluee
10:6:7:7 v/v/v/v ri nh mc n vch, lc u trong 10 min.
Ly 1 ml dung dch cho vo bnh nh mc 100 ml, pha long bng etanol
n vch mc 100 ml, lc u dd trong 20 sec. o quang 446 nm, cuvet 1
cm (dng ethanol lm dung dch so snh). Ghi gi tr hp th A.
Lm 3 ln, ly kt qu trung bnh.
Tnh kt qu: %Carotenoid

A.10000
G( g ).2550

Trong :
2550: H s hp th ca dung dch cha 1% carotenoid trong etanol
10000: H s pha long.

Ph lc 4. S liu phn tch

Bng PL4.1. nh hng ca bn cht dung mi n hiu sut chit
lutein ester.
T l Ethanol/ Hexane

Hiu sut chit(%)

100/0

13,01

80/20

37,50

60/40

57,86

40/60

74,88

20/80

78,00

0/100

80,00

Bng PL4.2. nh hng ca t l dung mi/nguyn liu n hiu

sut chit lutein este
T l dung mi/ nguyn liu

Hiu sut chit TB (%)

8/1

56,10

10/1

59,05

12/1

66,27

14/1

66,93

20/1

67,09

Bng PL4.3. S ph thuc ca hiu sut chit lutein ester vo s ln

ngm chit

S ln ngm chit(ln)

Hiu sut chit TB(%)

66,27

72,05

76,47

78,28

78,83

Bng PL4.4. nh hng ca thi gian ngm chit n hiu sut

chit lutein ester

Thi gian ngm chit (ngy)

Hiu sut chit TB(%)

55,79

64,82

70,86

67,35

67,25

Bng PL4.5. Kt qu kho st bn mu ca lutein ester trng

hp b sung 0.1% BHT
IU KIN

Tun

Nhit
phng, nh

sng t nhin
(TS)

Nhit

phng, trong
ti (TT)

hp
th (A)

Lutein (g/l)

%lutein cn
li

0,65

2,95

100,00

0,65

2,95

100,00

0.57

2,59

87,69

0,61

2,75

93,23

0,56

2,54

86,15

0,54

2,45

83,08

0,54

2,45

83,08

0,54

2,45

83,08

0,53

2,41

81,54

0,53

2,41

81,54

0,50

2,26

76,46

0,53

2,40

81,38

0,51

2,29

77,69

0,52

2,36

79,85

0,47

2,13

72,00

0,52

2,37

80,15

0,65

2,95

100,00

0,65

2,95

100,00

0,58

2,62

88,62

0,62

2,82

95,38

%lutein
TB cn
li
100,00

90,46

84,62

83,08

81,54

78,92

78,77

76,08

100,00

92,00

Lnh 4oC
3

0,50

2,26

76,46

0,53

2,40

81,38

0,57

2,59

87,69

0,57

2,59

87,69

0,55

2,50

84,62

0,55

2,50

84,62

0,55

2,50

84,62

0,54

2,45

83,08

0,58

2,62

88,62

0,62

2,82

95,38

0,52

2,38

80,46

0,52

2,38

80,62

0,65

2,95

100,00

0,65

2,95

100,00

0,64

2,90

93,10

0,60

2,70

91,54

0,57

2,59

87,69

0,61

2,75

93,23

0,51

2,29

77,69

0,52

2,36

79,85

0,53

2,41

81,54

0,53

2,41

81,54

0,57

2,59

87,69

0,57

2,59

87,69

87,69

87,69

84,62

83,85

83,69

80,54

100,00

94,92

92,38

90,77

89,23

87,69

0,56

2,54

86,15

0,54

2,45

83,08

0,54

2,45

83,08

0,54

2,45

83,08

86,62

85,77

Bng PL4.6. Kt qu kho st bn mu ca lutein ester trng

hp khng b sung 0.1% BHT
IU
KIN

Tun

Nhit

phng, nh
sng t

nhin (TS)
4

%lutein

%lutein

cn li

TB cn li

hp
th (A)

Lutein (g/l)

0,65

2,95

100,00

0,65

2,95

100,00

0,55

2,48

84,00

0,59

2,67

90,31

0,53

2,41

81,54

0,52

2,36

80,00

0,52

2,34

79,38

0,53

2,39

80,92

0,50

2,27

76,92

0,52

2,36

80,00

0,51

2,32

78,46

0,50

2,27

76,92

0,46

2,09

70,62

0,48

2,19

74,00

100,00

87,15

80,77

80,15

78,46

77,69

72,31

Nhit
phng,

trong ti
(TT)

Lnh 4oC

0,45

2,05

69,54

0,48

2,18

73,69

0,65

2,95

100,00

0,65

2,95

100,00

0,62

2,82

95,38

0,56

2,54

86,15

0,56

2,56

86,77

0,59

2,67

90,30

0,61

2,77

93,69

0,51

2,29

77,69

0,54

2,45

83,08

0,54

2,45

83,08

0,54

2,45

83,08

0,52

2,36

80,00

0,52

2,34

79,38

0,51

2,30

77,85

0,50

2,28

77,08

0,49

2,24

75,69

0,65

2,95

100,00

0,65

2,95

100,00

0,63

2,86

99,20

0,59

2,68

90,77

0,57

2,59

87,69

0,60

2,70

91,54

71,62

100,00

90,77

88,54

85,69

83,08

81,54

78,62

76,38

100,00

93,85

89,62

0,55

2,48

84,00

0,59

2,69

91,08

0,55

2,50

84,62

0,56

2,54

86,15

0,55

2,50

84,62

0,54

2,45

83,08

0,54

2,44

82,77

0,49

2,24

75,85

0,52

2,37

80,15

0,50

2,29

77,38

87,54

85,38

83,85

79,31

78,77

Bng PL4.7. Xc nh % carotenoid tng s trong sn phm kt tinh

STT

Lng mu (g)

Vm (ml)

% CAROTENOID

0,0312

100

100

0,328

41,23

0,0303

100

100

0,317

39,85

0,0298

100

100

0,304

38,21

Trung bnh

39,76

Bng PL4.8. Xc nh hm lng lutein tng s trong nguyn liu

STT

Mu (g)

1
2
3

0,10
0,10
0,10

Dm (ml)

25
50
25
50
25
50
TRUNG BNH

Lutein tng
s
(mg/kg ti)

Lutein tng s
(mg/kg kh)

7270
7100
7554
7308

8205
8013
8525
8247

0,15
0,14
0,16

Bng PL4.9. Xc nh hm lng cht kh trong nguyn liu

STT Chn (g) Hoa kh (g)

Chn + hoa
(sau sy)

%TL
Kh

33,17

1,00

34,06

89,05

32,88

1,00

33,77

88,88

34,98

1,00

35,86

87,90

%TL Kh TB

88,61

Bng PL4.10. Tnh chi ph dung mi cho nguyn liu

T l
dung mi/ nguyn liu

Lutein (mg/ kg kh)

Chi ph dung mi
(trm ng/ mg lutein)

8/1

4815

49,84

10/1

5102

58,80

12/1

5410

66,54

14/1

5464

76,87

20/1

5478

109,53