1

T VN
Chit xut dc liu c vai tr cc k quan trng trong vic quyt nh n
cht lng, an ton v hiu qu ca cc ch phm ng dc. c bit, hin
nay xu hng pht trin thuc c ngun gc t nhin ang ngy cng ph bin
v hiu qu tt m li an ton nn vn chit xut cng c quan tm.
Vit Nam thuc vng kh hu nhit i gi ma vi thm thc vt phong ph
vi tr lng ln, c nhiu tim nng v cy thuc. Bn cnh nc ta cng
c nn y hc c truyn lu i, cng c rt nhiu kinh nghim v s dung dc
liu lm thuc. l nhng li th ngnh cng nghip Dc v dc liu
php trin. Tuy nhin, c khng t kh khn trong vic s dng thuc c ngun
gc t nhin nh: mt s c tc dng nhanh, mnh dng trong cp cu nh
nhn sm, ph t, qu nhc, c c dc chng cng c c tnh cao. C ch
tc dng ca nhiu thuc vn cha c lm sng t nhiu khi s dng theo
kinh nghim Yu cu t ra ca ngnh Dc l lm sao tm ra c cht hoc
nhm cht gy tc dng sinh hc trong dc liu v chit xut, phn lp c
hot cht .V vy, chit xut dc liu l mt k thut quan trng pht trin
thuc theo hng ny. Trong , vic p dng cc cng ngh-k thut mi
trong chit xut c ngha then cht. Gn y trong cng ngh chit xut dc
liu c nhiu k thut mi ra i nh: chit xut h tr bi vi sng , sng
siu m, chit xut lng siu ti hn, chiu di p lc cao th hin tnh u
vit, tin li v hiu qu so vi phng php chit xut truyn thng.
Xut pht t thc tin trn, em la chn chuyn K thut chit xut
dc liu nhm tm hiu k hn v ni dng ny, gp phn nng cao kin thc
bn thn, ng thi c cch nhn khi qut v vn nghin cu. Bi bo co
gm cc ni dung chnh sau:
1. Tng quan v chit xut dc liu.
2. Mt s phng php chit xut thng thng.
3. Mt s phng php chit xut hin i.
4. V d minh ha nghin cu v chit xut dc liu.

I.TNG QUAN V CHIT XUT DC LIU

Chit xut l phng php s dng dung mi ly cc cht tan ra khi cc
m thc vt. Sn phm thu c ca qu trnh chit xut l mt dung dch ca
cc cht ha tan trong dung mi. Dung dch ny c gi l dch chit. C ba
qu trnh quan trng ng thi xy ra trong chit xut l:
- S ha tan ca cht tan vo dung mi.
- S khuych tn ca cht tan trong dung mi.
- S dch chuyn ca cc phn t cht tan qua vch t bo thc vt.
Cc yu t nh hng ln ba qu trnh ny (bn cht ca cht tan, dung mi,
nhit , p sut, cu to ca vch t bo, kch thc tiu phn bt dc liu...)
s quyt nh cht lng v hiu qu ca qu trnh chit xut.
Mt quy trnh chit xut dc liu in hnh gm cc bc sau:
1. Chun b dc liu:
Lm kh, chia nh dc liu hoc lm ng nht cc b phn ti (hoa,l,)
hay ngm ton b cc phn ca cy trong mt dung mi.
2. La chn dung mi chit:
Dung mi chit ph thuc vo bn cht ca cht cn chit, cc thnh phn cn
chit cng nh tp cht trong dc liu v phng php chit xut. C 3 nhm
dung mi chnh sau (Da vo phn cc):
- Dung mi phn cc cao: nc, ethanol, methanol
- Dung mi phn cc trung bnh: ethyl acetat, dichloromethane
- Dung mi km phn cc hoc khng phn cc: n-hexan, ether du ha,...
3. La chn phng php chit xut:
C rt nhiu phng php, k thut chit xut khc nhau, c la chon ty
thuc vo tnh cht ca cht cn chit, dung mi chit, c im ca dc liu,
v iu kin c s vt cht sn c.
- Ngm
- Ngm kit
- Chit siu ti hn
- Hm, sc
- Thng hoa
- Soxhlet
1.1.

Nguyn liu chit xut

Nguyn liu chit xut l nhng b phn ca dc liu hay ang c

nghin cu cha cc cht cn chit xut. C th l hoa qu, l, cnh hay ton
b cy thuc. Nguyn liu trc khi chit cn phi kim tra v mt thc vt xem
c ng loi, ng loi cn chit hay khng. Cn ch trng n ni thu hi, thi
v thu hi v thi gian thu hi m bo hm lng hot cht cn chit cao
nht. C th chit dc liu ti hoc dc liu sau khi c lm kh, ch
cc dc liu m hot cht d b bin i khi lamg kh hoc ngay c khi ti
(cn x l dit enzyme), kch thc dc liu em chit cng nh hng rt
nhiu n hiu qu qu trnh chit xut.
1.2. Dung mi chit xut
Cn la chn dung mi sao cho c kh nng ha tan ti a cc cht c tc dng
iu tr v ti thiu cc tp cht trong dc liu. Mt s yu cu v dung mi
chit xut khi la chn nh:
- D thm vo dc liu ( nht thp, sc cng b mt nh)
- Ha tan chon lc (ha tan nhiu hot cht, t tp cht)
- Tr v mt ha hc (khng lm bin i hot cht, khng gy kh khn
cho qu trnh bo qun, khng b bin i nhit cao)
- Phi d dng bay hi khi cn c c dch chit
Mt s dung mi thng s dng nh: nc, ethanol, cc dung mi hu
c khc nh ether, chloroform
Ty theo tng loi dc liu v hot cht cn chit m la chn dung mi
thch hp. V nguyn tc, chit c cc cht phn cc (alkaloid,
glycoside, polyphenol,) th phi s dng cc dung mi phn cc, chit
cc cht km phn cc (cht bo, tinh du, carotenoid, steroid) th phi s
dng cc dung mi km phn cc. Trn thc t, ethanol cc nng khc
nhau l dung mi c s dng nhiu nht do ha tan c nhiu nhm hot
cht, khng c, r tin, d kim.
V d v la chn dung mi trong chit xut c th hin bng1.
Bng 1. Dung mi khc nhau dng trong chit xut cc nhm hot cht t
dc liu (Houghton v Raman, 1998)

phn cc
Thp

Trung bnh

Dung mi
Phn nhm ha hc chit
Chloroform
Terpenoid, Flavonoid, Alcaloid, Aglycogen
Cyclohexan
Sp, cht bo
Hexan
Sp, cht bo
Dicloromethan Terpenoid, Flavonoid, Alcaloid, Aglycogen
Diethylether
Alcaloid, Aglycogen
Ethylacetat
Alcaloid, Aglycogen, Glycosid
Aceton
Flavonoid, Alcaloid, Aglycogen
Ethanol
Tannin, Polyphenol, Flavonoid, Terpenoid,
Methanol

Sterol, Alcaloid, Polyacetylen

Saponin, Tanin, Phenon, Flavon, ng,
Aminoacid, Anthrocyanidin, Terpenoid,

Nc

Xanthoxyllin, Totarol, Lactone, Polyphenol

ng, Aminoacid, Saponin, Tanin, Lectin,
Terpenoid, Athrocyanin, Tinh bt,
Polypeptid

Cao
1.3. Qu trnh chit xut
Qu trnh chit xut din ra ln lt nh sau:
- Dung mi thm nhp vo bn trong dc liu.
- Dung mi ha tan cc cht bn trong dc liu.
- Khuch tn cc cht tan trong dung mi (khuch tn phn t v khuch
tn i lu) bao gm:
+ Khuch tn cc cht tan t trong dc liu qua mng t bo ra mt ngoi
dc liu (khuch tn qua l xp v khuch tn phn t)
+ Khuch tn cc cht t mt ngoi dc liu ra lp dung mi xa hn
(khuch tn phn t).
+ Khuch tn cc cht theo dng chuyn ng ca dung mi (khuch tn i
lu).
1.4. Cc yu t nh hng n qu trnh chit xut
1.4.1.Cc yu t lin quan n dc liu.

- Mng t bo l mng thm tch, cho dung mi thm vo bn trong t bo

v cho cc cht tan phn t nh i qua, gi li cc cht c phn t ln.
Nhng dc liu mng t bo c cu trc mng manh, dung mi d thm
(hoa,l) th qu trnh chit xut xy ra d dng. Nhng dc liu mng
t bo rn chc, s nc (nha, sp, bn) th kh chit xut hn.
- Cht nguyn sinh trong t bo c tnh bn thm, ch cho dung mi i vo
bn trong t bo v vy khi dc liu cn ti th rt kh chit cc cht
tan trong t bo m phi lm kh hay s dng cn cao ph hy
mng nguyn sinh cht, to iu kin, cho cht tan i qua mng t bo.
- Mt s tp cht trong dc liu lm nh hng n qu trnh chit xut
nh pectin, gm, cht nhy, tinh bt, cht bo, sp, nhacn phi c cc
bin php khc phc khi khng mong mun chit xut cc cht ny.
- Ty tng loi dc liu m cn phi c chia nh n mn thch hp
tng din tip xc vi dung mi, tng qu trnh khuch tn nhng ng
thi li khng qu nh trnh mng t bo b v nhiu lm tp cht ln
nhiu vo dch chit.
1.4.2.Cc yu t thuc v dung mi
- T l dc liu v dung mi: nu dng qu nhiu dung mi dch chit b
long, ln nhiu tp cht, nu dng qu t dung mi s khng chit kit
c hot cht.
- pH dung mi
- nht v sc cng b mt
- phn cc ca dung mi
1.4.3.Cc yu t thuc v k thut
- Nhit chit xut
- Thi gian chit xut
- iu kin khuy trn
1.5. Phng php chit xut
Phng php chit xut l yu t quan trng gp phn vo s thnh cng ca
qu trnh nghin cu thuc ngun gc t nhin, bao gm cc phng php
truyn thng v phng php hin i.
Phng php chit xut truyn thng s dng dung mi kt hp vi gia nhit
v khuy trn. Cc phng php chit xut hay s dng nh:
a. Phng php chit lnh:

- Phng php ngm

b. Phng php chit nng:
- Phng php sc.
- Phng php hm.
- Phng php chit Soxhlet.

- Phng php ngm kit

- Phng php hm.
- Phng php ct ko hi nc.

Ngoi ra, hin nay c nhiu k thut chit xut hin i mi c p dng vi
nhiu u vit trong chit xut cc hp cht t nhin nh:
-

Chit xut vi s h tr ca sng siu m (Chit siu m- UAE).

Chit xut vi s h tr ca vi sng (Chit vi sng- MAE).
Chit xut lng siu ti hn (Chit siu ti hn SPE).
Chit xut bng dung mi di p lc cao (Chit dung mi nhanh- ASE).

II. MT S PHNG PHP CHIT XUT THNG THNG.

2.1. Phng php ngm.
L phng php cho dc liu chia nh ti mn thch hp, tip xc
vi dung mi trong thi gian nht nh, sau gn, p, lng, lc thu ly dch
chit. Ty theo nhit chit xut, chia thnh
- Ngm lnh: ngm nhit phng, c th khuy trn, thng p dng
vi nhng dc liu cha hot cht d b phn hy bi nhit.
- Hm: cho dung mi vo dc liu chia nh trong mt thi gian xc
nh, c th khuy trn, thng dng cho hp cht d tan trong thi gian
ngn nhit cao.
- Hm: ngm dc liu chia nh vi dung mi trong mt bnh kn, gi
nhit thp hn nhit si ca dung mi nhng cao hn nhit
phng trong mt thi gian nht nh, thnh thong c khuy trn, thng
p dng vi nhng dc cht t tan nhit thng, d phn hy nhit
cao.

- Sc: un si u v nh nhng dc liu vi dung mi trong mt thi gian

nht nh.
Ty theo s ln ngm chia thnh:
- Ngm mt ln vi ton b dung mi.
- Ngm nhiu ln vi tng phn on dung mi.
u im: n gin, d thc hin.
Nhc im:
- Nng sut thp, thao tc th cng.
- Mt nhiu thi gian, c th t vi gi n vi tun.
- Chit mt ln th cha kit, nhiu ln th tn dung mi.
2.2. Phng php ngm kit.
L phng php cho dung mi chy rt chm qua khi dc liu ng trong
mt dng c ngm kit theo quy nh (Hnh 1), trong sut qu trnh khng
khuy trn. Dc liu lun c tip xc vi dung mi mi, lun to ra s
chnh lch nng hot cht cao nn c th chit kit c hot cht.
u im: Dc liu c chit kit, dch chit u m c.
Nhc im: Nng sut thp, lao ng th cng.

Hnh 1. Bnh ngm kit hnh nn ct.

2.3. Phng php chit hi lu v ct ko hi nc.
Dc liu c ngm cng dung mi trong mt bnh cu y trn c ni
vi h thng ngng t. un nng bnh cu cha dc liu v dung mi n nhit
si, dung mi bc hi s ngng t v quay tr li bnh chit.

Ct ko hi nc l phng php ct mt hn hp hai cht lng bay hi c

khng trn ln vo nhau (nc v tinh du). Khi p sut hi bo ha bng p
sut kh quyn, hn hp bt u si v hi nc ko theo hi cht lng cn li
(tinh du). Hi nc c th a vo t bn ngoi t cc ni cung cp hi hoc t
to trong ni ct.
2.4. Phng php chit Soxhlet.
Dc liu c cho vo mt ng giy lc ri t vo ngn chit. Dung mi
mi c cho vo bnh cu v un hi lu. Dung mi bc hi ln c ngng t
xung ngn chit v khi trn s chy qua ng xi phng xung bnh cu bn di,
mang theo cc cht ha tan t dc liu. bnh ct, cht tan c gi li, dung
mi bc hi ln c ngng t xung bnh chit v i qua lp dc liu ha
tan cc cht tan cn li. C nh vy cho n khi dc liu c chit kit.

u im:
- Qa trnh chit xut lin tc.
-Tn t dung mi hn cc phng php trn.
-Dch chit khng cn phi lc

Nhc im:
- Dch chit lun nhit si ca dung mi nn cc
cht khng bn vi nhit d b ph hy.
- Khng thc hin c s khuy trn.

Hnh 2. Bnh chit soxhlet

III. MT S PHNG PHP CHIT XUT HIN I

3.1 Phng php chit siu m (UAE)
Nguyn l ca phng php :Siu m l mt dng sng in t cao tn (>20
KHz) tai ngi khng nghe c (20 KHz > 1-16 KHz). Tn s: 10.000 KHz (>
10 MHz) dng trong y hc, 20 - 100 KHz s dng trong k thut nh v, 20 - 40
KHz s dng ty sch (nha, kim hon).

Siu m lm dung mi (ti cc hc b mt tip xc) b si bt, y tp

cht ra khi b mt mu. Bn cht sng siu m khc vi sng in t.
Nguyn tc, c ch hot ng ca siu m
Khi xuyn qua c th, ch mt lng rt nh ca sng siu m b cc m
hp th v chuyn thnh nhit nng. S ta nhit ny khng ko di, khng lm
tng b mt nhit ti ch.

10

Phn ln nng lng ca sng siu m chuyn thnh c nng (lm rung).
S rung ko di s lm v cc bt kh ti ch, gy tn thng ti ch, i khi
nghim trng.
Di tc dng ca siu m: dung mi ti cc hc nh/dc liu b si bt,
y cht cn chit ra khi dc liu, cht tan vo trong dung mi (chit xut).
u im:
- Thit b tng i n gin, bo qun v vn hnh n gin, thit b
khng qu t tin.
- Chit c nhiu nhm hot cht, dung mi chit kh a dng
-Lng mu: c th ln n hng trm gam.
- Gim c nhit v p sut, u im ny c u tin p dng
chit cho cc hot cht khng bn vi nhit.
- Tng c khi lng dch chit v rt ngn c thi gian chit, v
nh vy cng ko theo tit kim nng lng u vo. Nh trong nghin
cu ny, lng phenolic ton phn thu c bng UAE trong thi gian 15
pht cao hn ng k so vi chit bng SE trong thi gian 60 pht.

Nhc im:
- Dung mi kh c lm mi trong sut qu trnh chit xut, v vy hiu
lc ca n l mt hm s ph thuc vo h s phn ly.

11

- Thi gian lc v ra dch chit ko di, v vy s tn nhiu dung mi,

lm mt mt lng dch chit hoc dch chit c th b nhim bn.
- S thoi ha b mt ca u d theo thi gian s nh hng n hiu
sut chit.
Phm vi ng dng ca siu m
Phm vi ng dng ca siu m l kh rng.
- Chit xut nhiu nhm hp cht t nhiu dc liu khc nhau t
bit l nhng cht dng cho thc phm.
- Lm sch b mt

- Ph hy cu trc t bo
- Phn tn

3.2 Phng php chit siu ti hn SFE

Mi cht mt iu kin nht nh tn ti trong mt trng thi no
trong 3 trng thi rn, lng v kh. Nhng khi nhit v p sut ca mt cht
c nng ln trn gi tr ti hn ca n, cht s ri vo mt vng trng thi

12

c bit gi l trng thi siu ti hn. Ti im siu ti hn, hp cht lc

c goi l lng siu ti hn, p sut v nhit c cc gi tr c gi l p
sut ti hn (Pc) v nhit ti hn (Tc), hai gi tr ny c trng cho tng cht.
Cht trng thi siu ti hn mang nhng c tnh u vit ca c cht kh v
cht lng: linh ng nh cht kh, kh nng chuyn khi ln hn cht kh, c
kh nng ha tan cc cht nh cht lng nhng nht, sc cng b mt thp
hn cht lng

Gin 3 pha c th hin hnh3.

Phng php chit lng siu ti hn l phng php chit xut s dng dng
dung mi c bit l dung mi trng thi siu ti hn. Dung mi thng dng
nht l CO2 , khng phn cc, im siu ti hn l 31C/74 atm nn d t, d
duy tr, an ton, khng c hi.

13

Hnh4. H thng siu ti hn

Nguyn tc hot ng:

- Np dc liu vo bnh chit, ng np li.
- M dng CO2 lng i qua b phn lm lnh ri qua bm nn. Sau qua
b tng nhit. Khi t nhit v p sut, CO2 tr thnh dng siu ti hn.
- Dng ny vo bnh chit. Hot cht theo dng CO2 qua b phn lm lnh.
Ti y CO2 ha lng v c a vo bnh tch.
- iu chnh nhit v p sut thch hp, CO2 bin thnh dang kh, sn
phm s lng xung, thu c ring.
- CO2 dng kh c a qua b phn nn lnh, ha lng v tr li bnh
cha. Qu trnh chit li tip tc.
u im:
- Hiu sut chit cao.
- im siu ti hn ca CO2 d t.
- chn lc cao vi loi hp cht cn thit.
- Tc (CO2)=31C chit c cht d phn hy nhit cao.

14

- Dung mi an ton, c th ti s dng nn chi ph r hn.

Nhc im:
-Thit b chuyn dng, t tin.
- Khng thch hp vi mu chit dng lng.
- Dung mi CO2 siu ti hn khng phn cc, ch chit c cc cht km
phn cc; c th phi hp vi mt lng dung mi phn cc (MeOH) thay
i tnh phn cc ca dung mi, chit cc cht phn cc hn.
3.3 Phng php chit vi sng (MAE)
C s: Vi sng l sng cc ngn hay sng siu tn, c tn s t 0.3GHz
n 300 GHz, tng ng vi di sng trong khong 100cm n 1 cm,
thng s dng bc x in t tn s 2450MHz.
Vi sng c tc dng lm tng nhit ca vt cht mt cch c bit, khng
ph thuc vo s dn nhit ca bnh cha hay vt cht. Nhit sinh ra theo 2 c
ch dn truyn ion v quay lng cc. 2 c ch ny lm sinh nhit trong lng
khi vt cht lm cho vic ra nhit nhanh v hiu qu hn rt nhiu. Nhng hp
cht cng phn cc th cng mau nng di s chiu x ca vi sng, c bit l
nc.
Trong chit xut, khi chiu x vi sng vo mi trng chit cha dc liu v
dung mi phn cc, cc phn t dung mi v cc cht phn cc s dao ng v
nng ln nhanh chng lm tng kh nng ha tan cc cht vo dung mi. Hn
na, vi sng cng lm ph hy cu trc vch t bo thc vt, to iu kin cho
cht tan gii phng vo mi trng d dng, lm cho vic chit xut nhanh hn
nhng cng lm cho dch chit ln nhiu tp cht hn.
u im ca vi sng:
- Khng c qun tnh nhit.
- Hiu sut chit cao hn so vi mt s phng php chit thng
thng.
- Sn phm trch ly cht lng tt.

15

- Thit b d s dng, an ton v bo v mi trng (Nng lng

sch,d ch to v d kim sot.
- Thi gian chit nhanh.
- C tc dng c bit vi cc phn t phn cc.
Nhc im:
- Khng p dng cho cc phn t khng phn cc.
- Kh p dng cho quy m cng nghip v u t cho thit b to vi sng
l khng nh c cng sut.
- Nhit si ca cc dung mi t c rt nhanh, c th gy n
ng dng ca vi sng:
Vi nhng tnh nng vt tri m n c c, vi sng c ng dng
rng ri v tin cy. c bit, trong cc phn ng cn cp nhit, cc phn
ng gia cc pha d th. Vi sng cn c tc dng tng cng khuy trn,
tng tip xc pha lm cho hiu sut phn ng c ln hn.
Vi sng c ng dng trong cc lnh vc:
- H tr chit xut.
- Tng hp hu c (kch hot phn ng):
Gim thi gian phn ng.
Gim phn ng ph.
Tng hiu sut.
Tng chn lc.
- Phn tch (ha v c, o m, thy gii,...)
H tr cng vic phng th nghim:
Sy kh cc vt dng thy tinh.
Tng hot sc k bn mng.
Hot ha ti to cht hp thu sc k, cht ht m, ry
phn t, cht mang rn.
- Trong thng tin lin lc: Bluetooth v cc chun IEEE 802.11g,
802.11b v 802.11a.
- Trong cng nghip:
Sy kh thc phm.
St trng thc phm.
Sy g.
3.4. Phng php chit di p sut cao -ASE

16

L phng php s dng cc dung mi thng thng nh cc phng php

chit xut c in khc nhng hot ng p sut v nhit tng i cao
(150 bar/ 100-180C). Nhit v p sut cao lm tng kh nng ha tan v
khuych tn dung mi cho vic chit xut hiu qu hn. Trong chit xut,
ngi ta c xu hng tng nhit tng hiu sut chit nhng khi n nhit
si, dung mi ha hi khng cn kh nng ha tan cc cht na. Da trn
nguyn tc, nhit si ca cht lng tng khi p sut tng, ngi ta tng p
sut nhit dung mi c a ln cao nhng cha n vng ti hn.

u dim:
- S dng c nhiu loai dung mi, chit c nhiu cht phn cc v
khng phn cc hn phng php chit siu ti hn.
- Dung mi chit gim, thi gian chit gim, hiu sut tng.
Nhc im:
- Thit b chuyn dng, t tin.
3.5 So snh cc phng php chit xut
So snh mt s phng php chit xut c m t tm tt bng 2.
T bng di cho thy, cc phng php chit xut hin i th hin nhiu u
im hn cc phng php chit truyn thng ( soxhlet) nh: thi gian chit
ngn hn, lng dung mi tiu th t hn, chn lc v hiu qu chit cao hn,
mt s phng php an ton v thn thin vi mi trng hn ( chit siu ti
hn). Tuy nhin, cc phng php chit xut hin i cng tn ti mt s nhc
im nh vn u t cao, mt s phng php kh trin khai khi nng cp ln
quy m cng nghip.

17

Bng 2 : So snh cc k thut chit truyn thng v hin i

Tn

M t

Thi

Dc

Dung

Chi

u im

Nhc im

Soxhlet

Dc liu t

gian
3-48h

liu
1-30g

mi
150-500

ph
Thp

-thao tc n

- Thi gian

gin.

lu.

lc v cho dung

-Dch chit

- Lng dung

mi qua

khng cn lc

mi nhiu

D s dng

- Lng dung

trong ng giy

UAE

SFE

ml

Dc liu c

10-60

ngm trong dung

pht

1-30g

50-200

Thp

ml

mi nhiu

mi v nhng

- Phi lc

siu m

dch chit

Dc liu c

10-60

trong bnh

pht

1-5g

2-5ml

Cao

-chit nhanh.

Nhiu thng

(rn)

-t dung mi.

s cn theo

chu p sut cao

30-60ml

-Khng cn

di

v lin tc a

(lng)

lc.

dng mi siu ti
MAE

- chn lc

hn i qua.
Dc liu c

3-30

Va

cao.
-D thao tc.

Dung mi

ngm trong dung

pht

phi

-Chit nhanh

chit phi hp

mi v chiu x

-Tiu th

th vi sng

vi sng

lng dung

1-10g

10-40ml

18
mi va phi
ASE

Dc liu c

10-20

lm nng v cho

pht

1-30g

15-60ml

Cao

-Chit nhanh
-t dung mi.

dung mi i qua

-Khng cn

di p sut cao

lc dch chit

IV. V D MINH HA V CHIT XUT DC LIU

4.1. Nghin cu chit xut polyphenol trong cy ch Camellia sinenssis
O.Ktze ( Thea chinensis Seem.)
4.1.1. Nguyn liu v phng php.
a. Nguyn liu
Ch xanh (bp v 2,3 l non) vng Bo Lc, Lm ng, c cung cp
bi cng ty ch Minh Rong. Ch c bt hot enzyme bng cch hp khong
90 giy trc khi s dng.
Thuc th Folin Ciocalteu (Merk), cc ha cht cn li s dng trong cc
th nghim u t tinh khit ca cht dng phn tch.
b. Phng php chit c s h tr ca vi sng.
S dng l vi sng gia dng (Whirpool Oven, cng sut 450, 600, 800W)
c thit k li cho ph hp[7].

19

A household microwave oven (National, Japan, full power 700 W) was modified
in our laboratory with the addition of a magnetic stirrer [14], water condenser,
temperature measurement and time controlling
100g ch ( c bt hot enzyme) c ct v xay nh, sau trn vi
dung mi vi t l thch hp. Sau chiu vi sng (mt pht chiu x, hai pht
ngng) gi cho nhit khng tng ln hn 800C. Sau ngui nhit
phng, lc, v bo qun t lnh (40C) s dng nh lng Polyphenol ton
phn.
Cc phng php chit xut khc nh: un hi lu, chit siu m, chit
nhit phng u thc hin nh vy nhng thay i thi gian chit c th nh
sau: un hi lu (60 pht), chit siu m (60 pht), chit nhit phng (24
gi).
4.1.2. Kt qu.
a. nh hng ca nng ethanol trong quy trnh chit xut polyphenol.
Ethanol c s dng trong th nghim ny v tnh cht khng c v l
dung mi khng t tin. Kt qu trnh by trong Biu 1 cho thy kh nng
chit polyphenol t ch xanh chu nh hng ca nng ethanol trong nc.

20

Khi nng ethanol tng t 0 n 60% hm lng cc polyphenol chit ra c

tng ng k. S tng nng ethanol sau nh hng khng ng k n s
tng ln ca hm lng polyphenol. Kt qu ny mu thun vi nghin cu ca
[8] , hiu sut chit gim xung khi nng ethanol tng ln.
Bn cnh s tng ln v nng ca cc polyphenol, mu sc ca dch
chit cng thay i t vng chanh sang xanh m khi c s tng ln v phn
trm th tch ca ethanol cao hn 60%, iu ny cho thy nhiu hp cht khng
mong mun v cc chlorophyl cng c chit xut. V th, y chnh l l do
chn ethanol 60% lm dung mi chit xut trong tin trnh th nghim.

Biu 1: nh hng ca nng ethanol trong qu trnh chit xut cc

polyphenol. T l nguyn liu/ dung mi: 1:6 (g/ml), th tch dung mi 150ml,
thi gian chiu x 6 pht, ngun cng sut 800W.
b. nh hng ca t l nguyn liu/dung mi.
Kt qu thc nghim cho thy rng t l nguyn liu/dung mi khng nh
hng ng k n hiu xut chit. Kt qu c trnh by trong Biu 2 ch
ra rng hiu sut chit tng khi tng t l nguyn liu/ dung mi t 1:4 n 1:6,
sau , hiu sut chit gim nh, t 19.36 n 18.10 khi t l tng ln t 1: 6 n
1:8, sau li tng ln t 18.10 ln 21.05 khi t l nguyn liu/ dung mi tng

21

n 1:10. S dng mt t l nguyn liu /dung mi cao c th t n hiu sut

cao nh mong mun (v d nh t l 1 : 10). Tuy nhin, dch chit polyphenol
thu c c mt lng ln dung mi l khng cn thit. Do , t l nguyn
liu/dung mi c la chn l 1 :6. Trong mt cng trnh nghin cu trc
[8], t l nguyn liu /dung mi c la chn s dng l 1 :20 cao hn nhiu so
vi gi tr thc nghim trong thit k th nghim ny. iu ny c th gii thch
c l do trong thit k th nghim[8] nguyn liu s dng l ch c sy
kh ( m 5-75), cn trong thit k th nghim ny s dng nguyn liu ti
( m 75-80%).

Biu 2: nh hng ca t l nguyn liu/dung mi trong qu trnh

chit xut polyphenol. Dung mi ethanol 60%, th tch s dng 150 ml, thi
gian chiu x: 6 pht, cng sut ngun 800W.
c. nh hng ca thi gian chiu x n qu trnh chit xut polyphenol.
Nng polyphenol trong dch chit tng ln nhanh chng trong khong 6
pht u tin ca qu trnh chit xut (c th hin trong Biu 3). Sau thi
gian ny, vic chiu x tip tc ch lm tng nh lng polyphenol chit c.
V vy, vic tip tc chiu x trong quy trnh chit xut sau khong thi gian 6
pht l khng cn thit. Cng trong mt nghin cu trc y [8], thi gian
chiu x ti u c la chn chit xut polyphenol vi mc cao nht lng

22

hot cht thu c ch din ra trong 4 pht. Kt qu thc nghim cng ch ra

rng s dng l c cng sut cng cao th cng d t c hiu sut chit ti u
trong khong thi gian ngn. Chiu x trong khong thi gian 4 pht vi my
c cng sut 800W cho hiu sut tng ng vi chiu x 6 pht my c cng
sut 450W.

Biu 3: nh hng ca thi gian chiu x ln qu trnh chit xut

polyphenol. Dung mi ethanol 60%, th tch dung mi 150ml, t l nguyn
liu/dung mi 1:6 g/mg.
d. Hiu sut chit vi thi gian lin tc.

Biu 4: Hiu sut chit trong thi gian lin tc. Dung mi: ethanol
60%, th tch dung mi 150ml, t l nguyn liu/dung mi 1:6 g/ml, thi gian
chiu x 6 pht, cng sut my 800W.

23

Quy trnh chit xut c lp li vi ln n khi dch chit khng i mu

khi thm vi git dung dch thuc th FeCl3 3%/HCl 0.1N (mu xanh ru s
xut hin nu c s hin din ca polyphenol trong dch chit) chit kit ton
b polyphenol trong nguyn liu. Biu 4 cho thy hiu xut chit trong ln
chit u tin khong 82,4% cao hn nhiu so vi nhng ln chit tip theo sau
(8.91%, 6.55%, 2.06% trong ln th 2, ln 3, ln 4). Kt qu ny chng minh
c rng phng php chit xut c s h tr vi ba cho hiu sut chit cao
trong khong thi gian ngn.
e. Hm lng Catechin trong dch chit polyphenol t ch.
Dch chit polyphenol bng phng php chit c h tr vi ba c s
dng kho st thnh phn hot cht v nh lng bng phng php HPLC
vi cc iu kin sc k c khai bo trn.
Trong qu trnh sc k, nhng peak tng ng vi Catechin (C),
Epicatechin (EC), Epicatechin gallate (ECG), Epigallocatechin gallate (EGCG)
v cafein c phn tch v nh tnh v nh lng. Trong sc k HPLC,
bn cnh 5 peak c ni trn, mt vi peak ph cng c pht hin, iu
cho thy qu trnh chit xut cng chit ra mt vi thnh phn catechin
khc t trong cy ch.

Sc k HPLC ca dch chit xut t cy ch ghi nhn bc sng 230nm.

24

Hot cht

EC

ECG

EGCG Cafein

Thi gian lu tR

6.331

7.070

8.651

7.264

6.634

0.39

0.93

1.92

12.85

3.3

Hm lng chit c (%
tnh theo nguyn liu kh)

4.1.3. So snh kt qu chit xut bng phng php chit c h tr vi

sng vi cc phng php chit xut khc.
Nhng quy trnh chit xut khc nh : phng php chit bng cch un
hi lu, chit xut c h tr siu m, v ngm kit nhit phng c tin
hnh so snh kt qu vi phng php chit xut c h tr vi sng. Kt qu
c trnh by trong Bng 1 cho thy rng hiu sut chit 82.46% (tnh theo
khi lng nguyn liu kh) vi thi gian chiu x l 6 pht v chit xut 18 ln
s cho hiu sut cao hn cc phng php chit xut khc : phng php chit
xut c h tr siu m (tin hnh trong vng 60 pht ) chit xut un hi lu
tron 60 pht v chit xut ngm kit nhit phng trong vng 24 gi vi
hiu sut chit ln lt l 63.3%, 65.64% v 49.39%.
Phng php

Thi

Lng Catechin

Hiu sut

gian

(% so vi khi lng dc

(%)

Chit c h tr

6 pht

liu kh)
16.90

82.46

vi sng
un hi lu
Chit c h tr

60 pht
60 pht

13.45
12.97

65.64
63.30

siu m
Ngm kit

24 gi

10.12

49.39

nhit phng
Bng 1 : So snh kt qu chit xut gia chit c h tr vi sng v cc
phng php chit xut khc.
Dung mi ethanol 60%, th tch dung mi 150ml, t l nguyn liu/dung
mi 1 :6 g/ml. Chit c h tr vi sng : thi gian chiu x 6 pht, cng sut my
800W ; un hi lu tin hnh nhit si ca dung mi s dng my khuy

25

t ; chit xut ngm kit nhit phng : 30C ; chit c h tr siu m

30C.

26

4.1.4. Kt qu v nhn nh
a. Kt lun
Phng php chit xut c h tr vi sng c s dng chit
Polyphenol t ch. Cc thng s lin quan nh nng ethanol (%), t l
nguyn liu/dung mi (g/ml), thi gian chit, cng sut l c kho st
xc nh u kin thch hp cho qu trnh chit. Kt qu cho thy :
Nng cn: 60% (thay i ty theo cht cn chit).
T l nguyn liu/dung mi: 1 : 6 (thay i ty theo cht cn chit).
Thi gian chiu x: 6 pht (hm lng cht cn chit tng ng k trong
mt khong thi gian u, sau khng tng nhiu khi ta tng thi gian
chiu x).
Cng sut my : 800W (cng sut cng ln hiu sut chit cng cao).
Phng php chit c h tr vi sng cho hiu sut chit cao (82,46% sau
6 pht chiu x), cao hn phng php un hi lu (65,64% sau 60 pht),
phng php chit c h tr siu m (63,30% sau 60 pht) hay phng php
chit ngm kit nhit phng (49,39% sau 24 gi).
Lng catechin c trong lng polyphenol ton phn c chit xut
bng phng php chit xut c h tr vi sng l C : CE : ECG : EGCG 0,39 :
0,93 : 1,92 : 12,85 tnh theo phn trm khi lng nguyn liu kh
b. Nhn nh
Phng php chit xut c h tr vi sng c kho st chit sut
polyphenol ton phn t cy ch cho thy c nhiu trin vng ng dng rng
ri trong quy m cng nghip bi hiu sut chit cao, thi gian nhanh, tit kim
dung mi v nng lng.
4.2. Nghin cu chit xut Capsaicin t cy t Capsicum frutescens (L.)
Bail
Chun b: Tri t c mua t mt ch a phng trong huyn Mae Chan,
Chiangrai, Thi Lan dng t kh. Dc liu c ng gi trong ti nha
c ty vi nit, v c lu tr 40C trc khi s dng.

27

4.2.1. Ngm:
25g bt dc liu c ngm kit vi 200ml ethanol 95% (v/v) trong bnh cha
dung tch 250ml. Tin hnh 450C, 250 vng/pht trong 15h.
4.2.2. Chit bng Soxhlet:
Chun b dng c chit:
Soxhlet dung tch 250ml: lm 1 ti bng giy lc hnh tr
c kch thc va vi thn Soxhlet, u trn ca ti khng
thp hn u trn ca ng dn dung mi (ng nh) v
khng cao hn l thng ca ng dn hi dung mi (ng
ln).
Cn khong 25g dc liu. Nghin nh dc liu trong ci
s, c th thm ct trung tnh tng xp, nu dc liu c m cao th nn
nghin vi Na2SO4 khan.
Cho dc liu nghin nh vo ti giy, dng bng gn tm dung mi lau sch
ci v cho ln pha trn lp dc liu. Ph ln trn mt dc liu mt lp giy
lc ngn khng cho dc liu ni ln trn v ri xung bnh chit (c th thm
vi vin bi thy tinh dn ln trn ming giy lc).
Lp dng c trn bp cch thy, dng phu rt ethanol 95% qua sinh hn
(khong 200ml).
Kim tra ln cui ton b dng c, nc sinh hn. Khng ch nhit 78.1 0C.
un trn bp cch thy trong 5h.
Thu dch chit.
4.2.3. Chit vi s h tr ca sng siu m:
Th nghim c tin hnh 3 ln trong bn siu m vi tn s 35kHz vi mc
nng lng l 600W (Bandelin Sonorex Super RK 1050), nhit 450C trong 3h
Bnh cha hnh ch nht (50cm: 60cm: 20cm) phi hp vi bn iu nhit
(Polyscience 9610, USA). Erlen 250ml gm 25g dc liu vi kch c 3mm
cng vi dung mi (ethanol 95%) c nhng chm vo bn siu m c kim
sot theo mc nc khong 10mm t di y bn. Kch c dc liu ng vai

28

tr quan trng trong qu trnh chit xut, nu kch c nh hn 3mm s gy bt

tin khi lc.
4.2.4. Kt qu v bn lun:
Hiu qu v thi gian chit bng siu m
Hiu qu ca siu m v thi gian chit xut trong vic thu hi capsaicinoid
c th hin trong hnh 1.

Tc

chit

capsaicinoid rt cao trong thi gian 5 pht u tin. Sau , vic thu hi
capsaicinoid tng dn vi thi gian chit xut.
Phng

Thi

Nhit

php
Ngm
Soxhlet
UAE
UAE
UAE

gian
15h
5h
3h
3h
3h

(0C)
45.0
78.1
45.0
45.0
45.0

Dc
mi
1:8
1:8
1:8
1:6
1:5

liu:dung T l phc hi
(%)
79.4
92.0
87.4
84.5
84.3

Thu hi Capsaicinoid bng UAE trong 3 gi c so vi ngm (15 h) v

Soxhlet (5 h).
Phn trm thu hi capsaicinoid tng ng bi ngm, Soxhlet v UAE l
79,4, 92,0 v 87,4. UAE thu hi hn 10% so vi capsaicinoid so vi phng

29

php ngm nhng thp hn khong 5% so vi dng Soxlet. Chit xut bng
Soxhlet cho vic thu hi t l capsaicinoid cao nht, v qu trnh chit xut c
thc hin ti nhit si (78.10C) v dung mi lun c lm mi.
UAE ci thin ng k sn lng chit xut. UAE c th y nhanh s
trng n v hydrat ha, gy ra m rng l rng bn trong cc t bo thc vt.
S ph v cc t bo thc vt bng siu m sau khi bt cc hc b ph v c
th tng t l dung mi thm nhp vo t bo thc vt . T l phng thch
capsaicinoid rt cao khi bt u do hiu ng ca gradient nng capsaicinoid
gia dung mi v dc liu v d dng chit xut t phn bn ngoi ca cc
ht giai on sm. Sau t l phng thch capsaicinoid gim ng k do
gradient nng thp hn v v capsaicinoid cn li c nm phn bn
trong.
Hiu qu ca nhit chit xut
Hiu qu ca nhit trong phng thch capsaicinoid khi chit xut bng
95% ethanol v acetone lm dung mi c hin th tng ng trong Fig 2 v 3.

Khi

dng

ethanol

95% lm dung

mi, s

gia tng nhit

t 300C n 450C tng cng vic thu hi capsaicinoid.

Nhit nh hng nhiu c tnh vt l nh nht, khuch tn, ha
tan, p sut hi v sc cng b mt.

30

Tc dng chnh ca siu m l lm dung mi trong hc ca dc liu b

si bt v ph v bt, gip y cc cht cn chit ra khi dc liu . Thc t l
hiu qu cao hn ti 45 0 C thay v 60 0C. Ti 450C, s lng bt cc hc tng
v b ph v, mnh thc hin chit xut. Ti 600 C, mc d s lng bt
cao hn, nhng s ph v chng l km hiu qu hn. Do , hiu ng ti hc
450C th hin cng mnh hn 60 0C.
Dng aceton lm dung mi, khng nng cao ng k hiu qu khi nhit
c nng ln t 30 0C n 45 0C (hnh 3).

Ethanol
thng c coi l km hiu qu hn so vi dung mi aceton do nht cao v
khuch tn thp hn. Tuy nhin, i vi siu m, aceton l t hiu qu hn so
vi ethanol. Trong nghin cu ny, nng sut UAE tng i tng (v/v) ethanol
75-95% c s dng nh mt dung mi so vi aceton. p sut hi ca aceton
l cao hn nhiu so vi ethanol v nc, do nhiu bt nhng hiu qu ph v
khng cao, c bit l nhit chit xut. Do , vic tng cng hiu qu
chit xut bi hiu ng ti khoang s dng aceton thp hn so vi ethanol hoc
nc.

31

Hiu lc ca t l ca nc trong ethanol

T l nc trong dung mi ethanol c mt nh hng quan trng trong
chit xut capsaicinoid (hnh 4).

i vi tt
c nhit kim sot, thy rng ethanol 50% khng hiu qu cho chit xut
capsaicinoid. iu ny do s khc bit trong phn cc gia cc dung mi v cc
hp cht chit xut. S phn cc ca capsaicinoid l thp hn nhiu so vi nc,
iu khng tt khi ha tan trong ethanol 50% (v/v). Ti 45 0C, s thu hi
capsaicinoid yu hn nhiu khi s dng ethanol 95% so vi ethanol 70%. Trong
khi , ti 600 C, ethanol 75% l dung mi c hiu qu hn. Nc c mt vai tr
quan trng trong chit xut. S trng n ca dc liu trong nc nng cao
hiu qu chit xut. Cng ca siu m ti khoang trong hn hp ethanol vi
s hin din ca nc cng c tng ln nh l mt kt qu ca vic tng sc
cng b mt v gim nht. Tuy nhin, t quan im kinh t, chi ph s
dng ethanol 75% (v / v) nh mt dung mi t vic loi b cc dung mi
bc cui cng cn nng lng tiu th cao. Do , ethanol 95% (v/v) dng lm
dung mi cho vic nghin cu tng lai.
Hiu lc ca t l dung mi - vt liu
Hiu qu chit xut ti cc t l khc nhau ca t bt (g) vi 95% ethanol
(ml) c hin th trong bng 1.
Phng

Thi

Nhit

php

gian

(0C)

Dc
mi

liu:dung T l phc hi
(%)

32

Ngm
Soxhlet
UAE
UAE
UAE

15h
5h
3h
3h
3h

45
78.1
45
45
45

1:8
1:8
1:8
1:6
1:5

79.4
92
87.4
84.5
84.3

T l c tm thy c nng ln t 01:05 n 01:08, vic thu hi tng

nh ln. Tuy nhin, kt qu c th khc nhau nu mt s lng ln cc dung mi
c dng. Mt khc, bng cch s dng mt s lng ln cc dung mi khng
c coi l hiu qu do chi ph vn hnh cao ca cc dung mi v nng lng
tiu th. Do , t l trng lng kh ca bt t vi ethanol 95% (v / v) c
chn 01:05.
4.2.5. Kt lun:
Phng php ngm thi gian tin hnh lu nht (15h) v t l phc hi
capsaicinoid thp nht (79.4%) do qu trnh ngm t nhin cho qu trnh
thm thu, ha tan v khuch tan din ra hon ton m khng c s tr gip ca
cc thit b khc. Mt khc qu trnh ngm c th t hiu sut cao hn nhng
phi dng lng dung mi ln hn gp nhiu ln do d ta khng dng.
Qu trnh chit bng Soxhlet thu c t l capsaicinoid cao nht (92%) v
qu trnh chit xut c tin hnh 78.10C v dung mi lun c lm mi
trong qu trnh chit xut.
Qu trnh chit vi s h tr ca sng siu m c t l capsaicinoid cao hn
phng php ngm 12.6%, nhng thp hn dng Soxlet khong 5%. Tuy nhin
ta khng th thc hin qu trnh chit xut vi s h tr ca sng siu m
nhit cao do theo thc nghim hiu qu ca qu trnh chit sut cao hn khi
tin hnh 450C, nhit ny s lng bt kh to thnh tng ln v nng
lng thu c t s nh v cc bt kh ny cho qu trnh chit xut xy ra
nhanh chng. Mt khc nhit cao hn (khong 60 0C), s lng bt kh
c to thnh tng ln tuy nhin qu trnh lm v cc bt kh ny li gim.

33

Thi gian chit xut vi s h tr ca siu m ngn hn do trong qu trnh

ny nng lng ca sng siu m chuyn thnh c nng (lm rung) lm v cc
bt kh ti ch, y cc cht cn chit ra khi dc liu nhanh hn.

KT LUN
T ni dung ca bi bo co, rt ra mt s kt lun sau:
Mi phng php chit xut c u, nhc im, phm vi ng dng nht
nh. Ty thuc tnh cht, c im ca dc liu v iu kin c s vt

34

cht sn c la chn cc phng php chit xut ph hp em li hiu

qu cao nht.
Chit xut l mt giai on quan trng trong nghin cu thuc c ngun
gc t nhin v vy vic la chn phng php chit xut c ngha ln
n cht lng sn phm
Cc phng php chit xut truyn thng vn c p dng rng ri, th
hin hiu qu chit xut nht nh, d thc hin, ph hp vi c s c vn
u t t nc ta.
Khi la chn c phng php chit xut cn kho st cc thng s,
yu t nh hng n chit xut ( kch thc dc liu, nhit , thi
gian, p sut,) tm ra mt quy trnh chit xut ti u cho hot cht
trong dc liu

Cc phng php chit xut hin i ang rt c ch trng pht trin

v c nhiu u im nh thi gian chit xut ngn hn, lng dung mi
tiu th t hn, chn lc v hiu qu chit cao, mt s thn thin vi
mi trng ( chit siu ti hn). Tuy nhin, cc phng php ny li cn
vn u t ban u cao, mt s phng php kh trin khai khi nng cp
quy m cng nhip (vi sng).

MC LC
T VN .
I.TNG QUAN V CHIT XUT DC LIU
1.1. Nguyn liu chit xut.

35

1.2. Dung mi chit xut.

1.3 Qu trnh chit xut.
1.4 Cc yu t nh hng n qu trnh..
1.5 Phng php chit xut...
II. MT S PHNG PHP CHIT XUT THNG THNG
2.1.Phng php ngm
2.2. Phng php ngm kit
2.3. Phng php chit hi lu v ct ko hi nc
2.4. Phng php chit soxhlet..
III.MT S PHNG PHP CHIT XUT HIN I.
3.1Phng php chit siu m (UAE)
3.2. Phng php chit siu ti hn- SFE..
3.3. Phng php chit bng vi sng (MAE).
3.4. Phng php chit p sut cao ASE..
3.5. So snh cc phng php chit xut
IV. MT S NGHIN CU V CHIT XUT DC LIU.
4.1. Nghin cu chit xut polyphenol trong cy ch Camellia sinenssis O.Ktze
( Thea chinensis Seem.)........................
4.2. Nghin cu chit xut Capsaicin t cy t Capsicum frutescens (L.)
Bail.........................................................................................
KT LUN..

B GIO DC V O TO

B QUC PHNG
HC VIN QUN Y

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CHUYN
NGHIN CU THUC NGUN GC T NHIN

K THUT CHIT XUT DC LIU

H v tn: Trn Th Oanh

Lp:

Dc 5

H Ni - 2015

TI LIU THAM KHO

1.B Y T, Dc liu, tp 1, tp 2, Nh xut bn Y hc.

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2. B Y T (2009), K thut bo ch v sinh dc hc cc dng thuc, tp 1,

Nh xut bn Y hc.
3. B Y T (2007), K thut sn xut dc phm, tp 2, Nh xut bn Y hc.
4. http://www.duoclieu.org/
5. http://luanvan.co/luan-van/ung-dung-vi-song-trong-chiet-xuat-polyphenol-tucay-che-189/
5. Supercritial fluid extraction bamgbose Timothy Oluwaseyi First Year M.Sc.