Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

MC LC
M U 3
NI DUNG ....4
Chng 1. C S L THUYT V SC K 4
1. Lch s sc k . 4
2. nh ngha sc k 4
3. Cc nguyn tc c bn ca k thut sc k 5
4. Phn loi sc k... 6
4.1. Phn loi theo bn cht ca hai pha s dng 6
4.2. Phn loi theo bn cht tng tc.. 7
4.3. Phn loi theo cu hnh . 8
Chng 2: SC K CT H ....... 10
1. DNG C - HA CHT .. 10
1.1. Ct...10
1.2. Cc loi pha tnh dng nhi ct . 10
1.2.1. Silica gel ... 10
1.2.2. Alumina .... 14
1.2.3. Kieselguhr Celite ... 14
1.2.4. Gel .14
1.3. Dung mi ... 15
1.3.1. Cc dung mi thng dng cho sc k ct .. 15
1.3.2. Cch chn dung mi thch hp.. 15
1.4. Mu sc k . 15
2. K THUT TIN HNH . 16
2.1. Chun b ct .. 16
2.2. Np mu cn tch ln ct sc k 17
2.2.1. Np mu dng dung dch .. 17
2.2.2. Np mu dng bt kh .. 18
2.3. Gii ly cht ra khi ct .. 19
2.3.1. Cc phng php gii ly ... 19
2.3.2. Dung mi gii ly v k thut tng dn tnh phn cc cho dung mi gii
ly 20
1

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

2.3.3. Vn tc gii ly .. 21
2.3.4. Theo di qu trnh gii ly ct 22
2.3.5. Ghi nhn kt qu sc k 23
2.4. Xc nh cu trc ha hc ca mt hp cht 23
3. U, NHC IM V NG DNG CA SC K CT H 24
3.1. u im . 24
3.2. Nhc im ... 24
3.3. ng dng ... 24
Chng 3: MT VI K THUT SC K CT KHC .. 25
1. Sc k ct kh 25
2. Sc k ct nhanh . 27
3. Sc k nhanh ct kh .. 29
KT LUN 32
TI LIU THAM KHO ..... 33

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

M U
Thc vt l kho tng v cng phong ph cc hp cht thin nhin, hng trm
nghn cc hp cht thin nhin c tm ra v c nghin cu phc v cho
nhiu lnh vc ca cuc sng. Thin nhin khng ch l ngun nguyn liu cung cp
cc hot cht qu him to ra cc bit dc m cn cung cp cc cht dn ng
tng hp ra cc loi thuc mi. T nhng tin cht c phn lp t thin nhin, cc
nh khoa hc chuyn ha chng thnh nhng hp cht c kh nng tr bnh rt cao.
V vy vic tch chit, c lp hp cht thin nhin l cng vic tht s cn thit.
C nhiu phng php tch chit, c lp cc hp cht thin nhin t cy c. Mt
trong nhng phng php thng hay dng nht l phng php sc k. Phng php
sc k ra i cung cp cho ha hc mt cng c tch chit hiu qu v nhanh
chng, m ra mt giai on pht trin rc r ca ngnh ha hc, c bit l ha hc
cc hp cht thin nhin. Mt c im ca phng php sc k l tnh a dng, cho
php ta ng dng n mi hon cnh, mi iu kin. Ngy nay, phng php sc k
c s dng tch tt c mi hp cht d c mu hay khng mu, d trng lng
phn t nh hay ln.
Do cc phn t sinh hc rt thin hnh vn trng vi trng lng phn t ln
nh khc nhau, tnh phn cc nhiu t khc nhau nn khng th no c mt k thut
sc k chung cho cc loi hp cht khc nhau. Trong bi bo co ny, ti xin gii thiu
mt phng php sc k thng dng trong phng th nghim l Phng php Sc
k ct. i vi ngi nghin cu v hoa hoc hp cht thin nhin thi vic tim hiu s
b v sc ky ct cung nh vic ng dung ky thut sc ky ct la mt vic v cung cn
thit, co th s dung hiu qua vao trong cng tac nghin cu cua minh.

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

NI DUNG
Chng 1: C S L THUYT V SC K
1. Lich s sc ky
T ng sc ky trong ting Anh la chromatography co xut x t ch chroma
trong ting La Tinh co nghia la cht mau. Nm 1903 nh thc vt hc ngi Nga
Mikhail Semyonovich Tsvett dng ct nhm oxit (c ti liu ni ct canxi
cacbonat) lm pha tnh v pha ng l ete du ho tch thnh cng chlorophyl t l
cy. ng gii thch hin tng bng i lc hp ph khc nhau ca cc sc t v t
tn phng php ny l phng php sc k (chromatography), sc k ngha l ghi
mu v tch c nhng cht c mu.
Ky thut sc ky pht trin nhanh chng trong sut th k 20. Cc nh nghin cu
nhn thy nguyn tc nn tng ca sc ky Tsvet c th c p dng theo nhiu cch
khc nhau, t xut hin nhiu loi sc ky khc nhau. ng thi, k thut thc hin
sc ky cng tin b lin tc, cho php phn tch cc phn t tng t nhau. S pht
trin phong ph ca sc k lm cho danh t sc k khng cn ng theo ngha
ban u ca n. Tuy nhin tt c cc phng php sc k u c nhng nt chung
nht: qu trnh tch da trn s chuyn dch ca hn hp cht phn tch qua lp cht
bt ng (pha tnh) l cht rn hoc cht lng mang trn cht rn hoc giy v s
chuyn dch c thc hin bng mt cht kh hoc cht lng (pha ng).
Trong nhng nm 1930, hang loat cac nha hoa hoc a c vinh danh bng giai
thng Nobel v Hoa hoc cac hp cht thin nhin nh vao nhng thanh cng trong
vic s dung ky thut sc ky ct hp phu:
- 1937, P. KARRER (Thuy Si) v hoa hoc Carotenoid c bit la vitamin A va
vitamin B.
- 1938 R.KUHN (c) hoa hoc Carotenoid va Vitamin.
- 1939 L.RIZICKA (Thuy Si) hoa hoc Polymetylen va Terpenoid.
- 1940 MARTIN va SYNGE (Anh) bt u nghin cu v sc ky phn b trn lp
silica gel va v sau phat trin sang sc ky giy. Hai ng c giai thng Nobel nm
1952 v sc ky phn b.
- 1948 TISLIUS (Thuy in) c giai Nobel v ky thut in di va sc ky hp
phu.
4

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

2. nh ngha sc ky
- inh nghia cua Mikhail S. Tsvett (1906): sc ky l mt phng php tch trong
cc cu t ca mt hn hp c tch trn mt ct hp th t trong mt h thng
ang chy.
- nh ngha ca IUPAC (1993): sc ky l mt phng php tch trong cc cu
t c tch c phn b gia hai pha, mt trong hai pha l pha tnh ng yn cn
pha kia chuyn ng theo mt hng xc nh. Trong sc ky ct, pha tnh c gi
trong mt ct ngn v pha ng c cho chuyn ng qua ct bi p sut hoc do
trng lc.
- Hin nay: sc k l qu trnh tch lin tc tng vi phn hn hp cc cht do s
phn b khng ng u ca chng gia pha tnh v pha ng i xuyn qua pha tnh.
3. Cc nguyn tc c ban cua ky thut sc k
Sc k l mt phng php vt l tch ring cc thnh phn trong mt hn
hp bng cch phn chia chng thnh 2 pha: pha ng v pha tnh.

Pha tnh

Pha ng

(Stationar phase)

(Mobile phase)

Lc khi u

Cc thnh phn ca hn hp
phn b khc nhau vo hai
pha

Trng thi cn bng, cc

thnh phn ca hn hp
phn b gia hai pha theo
mt t l nht nh

Nhn xt:Hp cht ( ) c i

lc mnh hn i vi pha tnh

H s phn chia l s phn b th d: ca 2 loi hp cht ( ) v ( ) trong mt h thng

gm c hai pha: pha ng v pha tnh
i vi cc cht ring bit trong hn hp, ty theo kh nng hp ph v kh
nng ha tan ca n i vi dung mi ra ct c ly ra ln lt trc hoc sau.
5

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Sc k hp ph c thc hin trn mt ng tr v mt hoa hoc i vi cac cht

trong qua trinh sc ky, ct co th la: thy tinh, kim loai, nha thng ng gi l ct
vi cht hp ph ng vai tr pha tnh, dung mi ra ct ng vai tr pha ng chy
qua cht hp ph .
Cht hp ph trong sc k ct thng dng l oxid nhm, silica gel, CaCO3, than
hot tnh, polyamid, cac loai co gn nhom ion, Cc cht ny phi c tiu chun
ha.
Dung mi dng c th l mt hoc hn hp nhiu loi dung mi c t l thch
hp. Quy trinh ra giai noi chung la s dung dung mi co tinh cht hng v mt phia
hoc tng hoc giam v tinh cht. Vi cc cht hp ph pha thun c in, dung mi
s dng c phn cc tng dn.
Vic tch hai hp cht no ra ring c t kt qu tt hay khng l ty thuc
vo h s phn chia (partition coeffiicient). Bt k mt hp cht no khi c t vo
mt h thng gm c 2 pha (th d: hai pha lng-lng hoc rn-lng), lc t n trng
thi cn bng, hp cht s phn b vo mi pha vi mt t l nng c nh, t l
ny thay i ty vo cc tnh cht ng hc ca cc hp cht v ca c hai pha
H s phn chia K c biu din nh sau:
K=

Cs No
ng ocu
a ca
c hp cha
t trong phatnh
=
Cm
No
ng ocu
a hp cha
t trong pha o
ng

Mi hp cht s c i lc ring ca n i vi hai pha, v th s c tng tc

mnh/yu khc nhau i vi pha tnh. H qu l mi hp cht s di chuyn ngang qua
pha tnh vi mt vn tc khc nhau, nh vy k thut sc k c th tch ring cc loi
hp cht.
4. Phn loai sc k
4.1. Phn loai theo ban cht hai pha s dung
Ty thuc vo bn cht ca pha tnh v pha ng, ngi ta phn bit mt s k
thut sc k khc nhau.
- Pha tnh: c th l cht rn hoc cht lng. Pha tnh tch ring cc hp cht
trong mt hn hp no l nh vo tnh cht hp ph ca n.
+ Pha tnh l cht rn: thng l alumina hoc silica gel c x l, c th
c np nn vo trong mt ct hoc c trng thnh mt lp mng, ph ln trn b
mt mt tm king, tm nhm hoc tm nha.

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

+ Pha tnh l cht lng: c th l mt cht lng c tm ln b mt mt cht

mang rn hoc mt chui dy cacbon di c gn ln trn cht mang rn.
- Pha ng: c th l cht lng hoc cht kh
+ Pha ng l cht kh: v d trong k thut sc k kh
+ Pha ng l cht lng: v d trong k thut sc k giy, sc k lp mng, sc
k ct.
4.2. Phn loai theo ban cht tng tac
Sc k phn chia:
* Pha ng: cht lng hoc cht kh (trong sc k kh)
* Pha tnh l cht lng, l mt lp cht lng vi chiu di tht mng, cht
lng ny c ni ha hc ln b mt ca nhng ht rn, nhuyn mn, c tr.
Sc k hp ph:
* Pha ng l cht lng hoc cht kh.
* Pha tnh l cht rn, l nhng ht rn nhuyn mn, c tnh tr, c nhi
trong mt ci ng. Nhng ht rn tr ny cng ging nh nhng ht rn trong sc k
phn chia, nhng khng c ph cht lng bn ngoi, bn thn ht rn l pha tnh.

Tiu biu ca sc k hp ph. Pha tnh rn l nhng ht rn, nhuyn, tr

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Sc k trao i ion:

Tiu biu ca sc k trao i ion

* Pha ng l ch c th l cht lng
* Pha tnh l cht rn, l nhng ht hnh cu rt nh, c cu to ha hc gi l
polyme, nn c gi l cc ht nha. B mt ca cc ht mang nhm chc dng
ion.
Sc k lc gel:
* Pha ng ch c th l cht lng
* Pha tnh l cht rn, l nhng ht hnh cu bng polyme, trn b mt ca cc
ht c nhiu l rng. Mi loi nha c mt kch c nht nh, bit trc.

Tiu biu ca sc k lc gel

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

4.3. Phn loai theo cu hinh

Sc k giy v sc k lp mng:
Phng php sc k lp mng bao gm pha tnh l mt lp mng cc cht hp
ph, thng l silica gel, aluminium oxide, hoc cellulose c ph trn mt mt
phng cht tr. Pha ng bao gm dung dch cn phn tch c ha tan trong mt
dung mi thch hp v c ht ln bn sc k bi mao dn, tch dung dch th
nghim da trn tnh phn cc ca cc thnh phn trong dung dch.
Ch phn tch ca sc k giy ch yu l phn b, trong pha tnh (thng l
nc) c thm trn mt t giy thm c bit gi l giy sc k. Nh cc xoang
rng trong si cellulose ca t giy sc k khc nhau, phn bit theo thm dung
mi v mc dy mng ca giy, vi cc m hiu ty thuc vo hng sn xut. Khi
tin hnh sc k cn chn loi giy thch hp.
Sc k ct
Sc k ct h c in l tn gi ch loi sc k s dng mt ng hnh tr,
c t dng ng, vi u trn h v u di c gn mt kha, dng c ny ging
nh ci buret nh phn trong phng th nghim.
C th ni sc k ct l mt dng ca sc k giy hoc sc k lp mng nhng
y pha tnh c nhi vo ct, nh vy c th trin khai mt cch lin tc vi nhiu
h dung mi khc nhau t phn cc yu n phn cc mnh.
Trong sc k ct pha tnh l cht rn c nhi thnh ct. Ty theo tnh cht
ca cht dng lm ct m s tch trong ct xy ra ch yu theo c ch hp ph (ct
hp ph) hoc theo c ch phn b (ct phn b).
Sc k ct c tin hnh iu kin p sut
kh quyn. Pha tnh thng l nhng ht c kch thc
tng i ln (50-150m), c np trong mt ct
bng thy tinh. Mu cht cn phn tch c t pha
trn u pha tnh, c mt lp bng thy tinh t ln trn
b mt khng b xo trn lp mt. Dung mi gii ly
c a ra v hng trong nhng l nh pha di
ct, ri em c quay ui dung mi, dng sc l lp
mng theo di qu trnh gii ly.

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Trong sc k ct vi pha tnh l silica gel loi thng th hp cht khng hoc
km phn cc c gii ly ra khi ct trc, hp cht phn cc c gii ly ra sau.
Cn trong sc k ct vi pha tnh l silica gel pha o th nhng hp cht phn cc s
gii ly ra khi trc v nhng cht phn cc km s gii ly sau.

10

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Chng 2: SC K CT H
1. DNG C - HA CHT
1.1. Ct
Ct l nhng ng hnh tr bng thy tinh di
30-70 cm, ng knh 1-5 cm, u di c mt
vi thy tinh v 1 kha iu chnh tc
chy.
Kch c ca ct ty thuc vo s lng mu
cht cn phn tch
- Trng lng cht hp ph phi ln hn 2550 ln trng lng mu cn sc k. Tuy nhin
vi nhng hn hp cc hp cht kh tch ring
th cn s dng s lng cht hp ph nhiu hn
(ln hn 100-200 ln), cn vi cc hn hp d tch th c th s dng lng cht hp
ph t hn.
- T l gia chiu cao cht hp ph v ng knh trong ca ct vo khong
10:1
Mun bit lng cht hp ph c ph hp vi ct th cho cht hp ph kh vo
ct quan st.
1.2. Cc loi pha tnh dng nhi ct
1.2.1. Silica gel
* Silica gel pha thng: c ch to bng cch thy gii silicat natri (cho tc
dng vi acid sulfuric) thnh polysilisic, tip theo l ngng t v polyme ha t
cc ch tiu vt l cn thit nh c cc ht vi kch c, th tch l rng trn b mt,
din tch b mt, nh yu cu.

11

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Silica gel
Ht silica gel s dng cho sc ky c in c ng knh ht trung bnh khong
40-200m, cc l rng c ng knh trung bnh khong 40-300A, din tch b mt
khong 100-800 m2/g.
Cc v tr hot ng trn b mt ca ht silica gel l cc nhm silanol, mi nhm
cch nhau 5A. y l nhng tm rt hot ng c th to ni hydrogen mnh vi
nhng hp cht c sc k. V th, khi sc k ct vi ct nhi bng silica gel, nhng
hp cht phn cc (c mang nhm chc -OH, -NH 2, -COOH...) c kh nng to ni
hydro mnh, b silica gel gi cht li trong ct v b gii ly ra chm hn so vi nhng
cht khc c tnh km phn cc nh alkan, terpen (l nhng hp cht khng cha
nhng nhm chc c th to nn ni hydrogen) t b silica gel gi li, s ra khi ct
sm.
- Mun iu chnh hot tnh b mt ca silica gel ch cn thm hoc loi b
nc. Khi silica gel hp ph nc, cc phn t nc s che khut nhng v tr hot
ng trn b mt ca ht silica gel lm ht b gim hot tnh; mun silica gel hot tnh
tr li, ch cn un nng loi b nc. y l qu trnh thun nghch, mun lm
gim hot tnh silica gel th ch cn cho thm nc vo. Tuy nhin, khi un nng
khong 400-500C, qu trnh thun nghch bin mt, silica gel bi mt vnh vin hot
tnh b mt, do hai nhm silanol k bn mt mt phn t nc, to thnh ni eter,
khng cn tnh sc k.

M t b mt ht silica gel

- Hot tnh ca silica gel l kh nng hp ph nc, c o bi s Brockmann.

Silica gel c s Brockmann cng ln th b mt cng km hot tnh.

12

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

- C th hiu chnh tnh ca silica gel bng cch cho silica gel kt hp vi
nhng hp cht khc nh base, cc dung dch m mc pH xc nh. Cng c th
thm nitrat bc lm gia tng kh nng tch cc hp cht, nht l alken.
Tuy nhin cng cn nh l vi mt hp cht no ang b silica gel gi li
trong ct, gii ly cht ra khi ct c hay khng cng cn ty vo vic s dng
dung mi gii ly c phn cc mnh hay yu. Dung mi no c th to ni hydrogen
mnh s l dung mi thch hp gii ly cc hp cht phn cc mnh ra khi ct silica
gel. Hn na, mun ui ht cc hp cht phn cc th d cc flavonoid, triterpen
glycosid nn dng 1-2% acid axetic trong metanol.

* Silica gel ch ha: c iu ch bng cch cho cc nhm chc silanol ca

silica tc dng vi nhiu loi clorur silil khc nhau to thnh nhng loi cht hp
ph mi, vi cc c tnh vt l i khc, c gi l silica gel-to ni. C th ch ha
thnh silica gel-to ni dng cho pha thng hoc pha o.
- Silica gel to ni dng cho pha o: Cho silica gel tc dng vi
clorodimetylalkylsilan R-Si(CH3)2-Cl to silica gel mi c tnh khng phn cc.
Dy alkyl R thng l C-1, C-2, C-4, C-6, C-8, v C-18. Tuy nhin, thng hay ch
to dy C-8 v C-18. Silica gel pha o c i lc mnh vi cc hp cht km phn cc
v gi cht cc hp cht ny li trong ct. Pha ng thng l nc, sau thm vo
cc dung mi hu c lm gim phn cc ca dung mi gii ly, nh th cht phn
cc s gii ly ra khi ct trc, v nhng cht km phn cc s ra sau.

13

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Cu trc b mt silica gel pha o

- Silica gel to ni dng cho pha thng: iu ch tng t nh silica gel to
ni dng cho pha o vi R thng l: phenyl, cyano, amino, diol,u im ca n
l bn i vi tt c cc loi dung mi k c cc dung mi c tnh phn cc mnh nn
c th s dng dung mi phn cc mnh gii ly ra khi ct tt c cc hp cht c
tnh phn cc mnh, trong khi silica gel thng s gi rt mnh cc hp cht c
tnh phn cc, nhiu khi hp cht b dnh lun trong ct, khng th gii ly ra khi ct.

Cu trc v tn gi mt s loi silica gel-to ni dng cho pha thng

- Silica gel to ni dng cho sc k th tnh:
Hot tnh sinh hc ca nhng hp cht th tnh c trng ty theo ha hc lp
th ca chng. C i phn c hot tnh cha bnh trong khi i phn cn li khng c
c tnh , m li c khi cn c hi, v th nht thit phi tch ring chng. Nhng
hai i phn c c tnh vt l ging nhau, cng ha tan vo cng mt loi dung
mi no , nn khng th s dng sc k tch chng c, cho d s dng silica
gel pha thng hay pha o, ct thng hay ct hiu nng cao HPLC.
C nhiu loi silica gel thng phm s dng cho sc k th tnh vi c ch lu
gi cht khc nhau, mt s nhng cht s dng silica gel lm cht nn.
Bng 1: Phn loi mt vi pha tnh th tnh
Loi pha th tnh
Pirkle
Polymer hnh xon c - th tnh
(polysaccarid)
Cyclodextrin

eter

(Crown ether)
Enzym

gi

vng

C ch bt gi cht ca pha tnh

Tng tc qua ba im
Cc ni k nc (hydrophobic bond)
Tng tc gia ch-khch (tng tc gia cht
tan i vi l rng c tnh th tnh ca ch l

cyclodextrin)
nh i lc th tnh
14

Tch chit v phn lp hp cht thin nhin

(Immobilised)
Phc amino acid kim loi
1.2.2. Alumina

Phng php sc k ct

Phc xuyn lp th phn

Alumina l oxid aluminium Al2O3, c iu ch qui trnh tng t nh silica

gel nhng trn nguyn liu l hydroxid aluminium, hn th na c th iu chnh
pH ca dung dch phn ng sn xut ra ht alumina vi b mt c tnh acid, tnh
kim hay trung tnh.
- Alumina base: cha khong 0,1-0,5% NaOH bm trn mt alumina dng
natri aluminat, c pH 10.
- Alumina trung tnh: pH t 6,5-7
- Alumina axit: pH 4

Alumina
Nhit lc hot ha trong qu trnh iu ch l c im khc nhau gia
alumina v silica gel. Mun c alumina hot tnh mnh, cn phi un nng alumina
400-450oC trong 12-16 gi. Mun gim hot tnh alumina, thm nc vo.
1.2.3. Kieselguhr-Celite
Kieselguhr v Celite l loi t st diatomit. Nguyn liu ny c cc l rng
ln, din tch b mt ln v c tnh hp ph rt yu. Kh nng hp ph cn c th gim
hn na bng cch cho tc dng vi dung dch HCl 3%, ra sch clor v sy kh
80oC. Vi cc tnh cht trn, n c s dng lm pha tnh trong sc k phn chia.
1.2.4. Gel
Gel l tn gi chung cho cc loi pha tnh c iu ch t tinh bt, agar
(polysaccarid) hoc polyacrylamid, trong cc chui dy di c ni mng ngang
to thnh mng khng gian 3 chiu.
15

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

1.3. Dung mi:

1.3.1. Cc dung mi thng dng cho sc k ct: hexan, benzene, chloroform,
aceton, etanol, methanol, butanol, nc.
Th t phn cc tng dn: Ete du ha < hexan < xiclohexan < CCl 4,<
benzen < toluen < diclometan < CHCl 3 < dietyl ete < etyl axetat < axeton < pyridin <
propanol < etanol < nc < axit axetic
1.3.2. Cch chn dung mi thch hp
Trc khi trin khai sc k ct, nht thit phi s dng sc k lp mng d
tm h dung mi gii ly cho ph hp, vi cc bc tun t nh sau:
Bc 1: Mu cn sc k c ha tan hon ton trong dung mi ph hp, vi
nng 10mg/ml gi l dung dch mu (A).
Bc 2: Chun b 4-6 tm bn mng 2,5 x 10 cm. Chm ln nhng tm bn ny
mi tm 1 chm khong 2-5l dung dch (A).
Bc 3: Mi bn mng c trin khai vi 1 loi dung mi gii ly khc nhau,
k hin hnh cc vt trn bn bng n UV hoc bng cc thuc th. Vi n dung
mi s d dng thy c dung mi no l ph hp. T kt qu , c gng tm mt
hn hp dung mi, trong mt dung mi km phn cc v mt dung mi phn cc.
Bc 4:
- Vi hn hp mu cht l kt qu ca phn ng tng hp hu c, ta chn h
dung mi c th y hp cht cn quan tm ln v tr trn bn vi Rf = 0,2-0,3.
- Vi mu cao th chit xut t cy c, chn dung mi gii ly u tin l dung
mi c th y vt t phn cc nht ca cao chit ln v tr bn vi Rf = 0,5 v chn
dung mi chm dt sc k ct l dung mi c th y vt phn cc nht ca cao chit
ln v tr bn vi Rf = 0,2.
Sau khi chn c h dung mi ph hp, c th p dng h dung mi ny cho
sc k ct. Gii ly trc tin bng dung mi khng phn cc v tng dn tnh phn cc
cho dung mi gii ly.
Lu :
- Phi s dng pha tnh ca sc k lp mng v sc k ct ging nhau.
- Phi chnh t l dung mi gii ly ct sao cho c tnh km phn cc 1 t so vi
h dung mi chn bng sc k lp mng.
1.4. Mu sc k
16

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Mu th trc khi phn tch phi c loi tp cht bng phng php thch
hp. Mu th c th 2 dng:
- Dng dung dch kh m c: ha tan mu trong dung mi khng qu phn
cc so vi h dung mi pha ng. VD: Pha ng l benzen-EtOAc th tt nht l ha
tan mu trong benzene, nu mu khng tan trong benzene th dng dung mi phn cc
hn benzene mt cht.
- Dng bt kh: ha tan mu trong dung mi nh etyl axetat hoc methanol,
thm vo silica gel ht ln va , c quay hn hp ui ht dung mi thu c
mu dng bt mn ti xp.

Mu dng dung dch m c

Mu dng bt kh

2. K THUT TIN HNH

2.1. Chun b ct: Yu cu l cht rn lm ct phi phn tn ng u mi im
trong ct thnh mt khi ng nht.
- Ra ct tht sch, trng vi nc ct v sy kh.
- Cho bng gn vo y ct (c th cho thm mt lp ct mn sch). Kp ct thng
ng trn gi.
- Cho cht hp ph vo ct thng c gi l nhi ct. C 2 cch nhi ct: nhi
ct t v nhi ct kh.
+ Nhi ct t: Cht hp ph c np vo ct dng st, c chun b nh
sau:
Trong 1 becher c cha sn dung mi, cho cht hp ph
tng lng nh v u n vo becher, va cho va khuy u.
t ln u ct mt phu lc ui di, rt hn hp st vo
ct, m nh kha bn di ct cho dung mi chy qua, hng
dung mi vo mt becher trng, dung mi ny c s dng li
rt ln u ct.

17

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Rt cht st vo ct cho n khi ht s lng, va rt va dng 1 thanh cao su

g nh vo bn ngoi thnh ct cht hp ph nn u trong ct.
Sau khi np xong, cho dung mi chy ra v rt tr li u ct vi ln vic
np ct c cht ch cho n khi thy cht hp ph trong ct c dng ng nht v
mt thong cht hp ph u ct phi nm ngang. Khng c u ct b kh,
ngha l dung mi phi lun ph trn phn u ct.
- Nhi ct kh: Cho dung mi loi km phn cc nht vo khong 2/3 chiu
cao ct.
Cho cht hp ph dng bt kh vo thng trong ct qua phu lc ui di, u
n, mi ln mt lng nh, va cho vo va g nh thnh ct. Khi lp cht hp ph
t c chiu cao khong 2 cm trong ct th m nh kha cho dung mi chy ra,
hng dung mi v 1 becher trng, dung mi c s dng li rt tr li ln u ct.
Sau khi np xong, cho dung mi chy qua cht hp ph vi ln n khi thy
cht hp ph trong ct c dng ng nht.
Lu : Qu trnh nhi ct phi thc hin mt cch cn thn, nu quan st thy
c nhiu bt kh hoc c nhng bt thng trong ct th phi x ht ct v np li t
u.

Ct sau khi c np silica gel

2.2. Np mu cn tch ln ct sc k
2.2.1. Np mu dng dung dch
- M kha cho dung mi chy ra khi ct h mc dung mi trong ct xung
sao cho va st vi mt thong ca cht hp ph trong ct.

18

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

- ng kha li, dng pipette ht dung dch mu cht v np vo u ct. Mun

np mu t u ca pipette gn st vi mt thong ca cht hp ph trong ct, va
bp va ry pipette dc quanh thnh trong ca ct, cho dung dch cht chy ra dc
theo thnh trong ca ct chm xung b mt ca cht hp ph .
- M kha bn di cho dung mi chy ra khi ct, lm
cho dung dch mu c thm ht vo cht hp ph trn u
ct.
- Dng pipette cho mt lng nh dung mi mi ln u
ct, dng dung mi ny ra sch dung dch mu cht dnh
trn thnh ct. Tip tc m kha cho dung mi chy qua. Lp
li vi ln nh th dung dch mu thm su vo phn cht
hp ph phn u ct v phn dung mi trn u ct tr nn
trong sut.
- Cho mt lp ct (hoc bng gn) dy khong 3-6 mm t nh ln trn mt
thong ca cht hp ph bo v mt ct.
Sau khi np mu xong cho dung mi vo y ct bt u qu trnh gii ly.

Ct sau khi np mu
2.2.2. Np mu dng bt kh:
Nu mu cht khng tan trong dung mi loi dung mi la chn bt u qu
trnh sc k ct, v y l loi dung mi km phn cc thay v phi ho tan mu trong
dung mi phn cc c th nh hng n qu trnh gii ly, c th np mu kh.

19

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Trong mt bnh cu dng c quay, mu cn sc k (Xg) c ho tan trong

dung mi nh etylaxetat hoc metanol (50Xg) cho thm vo silica gel c ht ln
(10Xg). Hn hp ny c c quay chn khng n khi c bt silica gel kh, by gi
mu sc k c tm u ln b mt ca nhng ht silica gel.
t mu bt kh ny ln trn u ct, dng mt t dung mi (loi chn bt
u qu trnh sc k ct) thm t phn bt silica gel. Cho mt lp ct dy khong 3-6
mm t nh ln trn mt thong ca cht hp ph bo v b mt. Cui cng cho
dung mi vo y ct bt u qu trnh gii ly.
2.3. Gii ly cht ra khi ct
2.3.1. Cc phng php gii ly
- Gii ly nh vo trng lc: cc ht gel np ct phi c kch thc ln hn
60m. Nu s dng nhng ht gel c kch thc nh hn, dung mi ra khi ct rt
chm v cn phi nh n mt lc c th ra khi ct.
- Gii ly s dng lc y: dng my bm to p lc y dung mi i ngang
qua pha tnh. S dng ht gel c kch thc 40-63 m v ct sc k phi c thnh dy
vi lp bao li che chn bn ngoi trnh khi s dng p sut cao, ct c th b n
v.

Hai loi dng c gip y dung mi vo u ct cho sc k chp nhong

- Gii ly s dng lc ht: dng my bm ht to chn khng u ra ca ct.
K thut ny cng cho kt qu tng t nh trn, nhng vic thu ly dung mi gii ly
c phn km thun tin v phi ngng to chn khng, tho thit b rt dung mi ra.
Tuy vy k thut ny an ton hn.

20

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Gii ly s dng lc ht
Nh vy, cc k thut sc k khc nhau l vic s dng kich thc khc nhau
ca ht gel lam pha tnh v vic s dng p lc gii ly dung mi ra khi ct.
Ct p sut thp, s dng ct lm bng thu tinh: thng s dng ht c kch c
40 200 m, s dng p sut bnh thng ca bu kh quyn.
Ct p sut trung bnh, s dng ct lm bng thu tinh dy: thng s dng ht
c kch thc c 25 40 m; thc hin p sut 75 600 psi (tc 5 40 bar).
HPLC s dng ct bng thp khng r: ht np c kch c 3 10 m; thc hin
p sut 500 3000 psi.
2.3.2. Dung mi gii ly v k thut tng dn tnh phn cc cho dung mi gii ly
- Gii ly s dng dung mi n nng : ch s dng n dung mi hoc hn
hp dung mi nhng trong hn hp t l gia cc thnh phn khng thay i gii ly
cho n khi vic tch cht hon tt.
- Gii ly s dng dung mi c nng tng theo kiu bc thang: Vic s dng
mt loi dung mi s ch gii ly ra khi ct mt s cu t nht nh v mt s cu t
khc c tnh phn cc hn vn cn nm u ct. V th mun ui chng ra khi ct,
phi dng mt dung mi c i lc mnh hn.
Mun tng tnh phn cc cho bt k mt dung mi no, nn cho thm t t mi
ln vi phn trm mt dung mi mi c tnh phn cc hn vo dung mi c s
dng. Khng nn tng tnh phn cc nhanh, t ngt s lm gy ct.
V d: ang gii ly vi hexan, mun chuyn sang benzen, s pha benzen vo
hexan theo t l 1%, 2%, 3%, 5%, 10%, 50%, 100% benzen.
Bng 2: Cc dung mi thng dng trong sc k ct, c sp xp theo ch s phn
cc tng dn

21

Tch chit v phn lp hp cht thin nhin

Dung mi

Nhit si
(oC)

Phng php sc k ct

Hng s
in mi
25oC

Ch s

nht

phn cc

(mN.S.m-2)

tan
trong nc
(%w/w)

Pentan

36

1,8

0,0

0,23

0,004

Hexan

69

1,9

0,0

0,33

0,001

Heptan

98

0,0

0,39

0,0003

Ciclohexan

81

2,0

0,2

1,00

0,01

Tetraclorur

77

2,2

1,6

0,97

0,08

Toluen

111

2,38

2,4

0,59

0,51

Xylen

139

2,5

0,61

0,018

Benzen

80

2,3

2,7

0,65

0,18

Diethyl ether

35

4,34

2,8

0,32

6,89

Diclorometan

41

8,9

3,1

0,44

1,6

Isopropanol

82

18,3

3,9

2,3

100

n-Buthanol

118

3,9

2,98

7,81

Tetrahydrofuran

65

7,58

4,0

0,55

100

n-Propanol

92

20,1

4,0

2,27

100

Acetate butyl

125

4,0

0,73

0,43

Chloroform

61

4,87

4,1

0,57

0,815

Acetate ethyl

77

6,0

4,4

0,45

8,7

Methyl ethyl

80

4,7

0,45

24

Dioxan

101

2,2

4,8

1,54

100

Aceton

56

20,7

5,1

0,32

100

Methanol

65

33,6

5,1

0,6

100

Ethanol

78

24,3

5,2

1,2

100

Acetonitryl

82

37,5

5,8

0,37

100

Acid acetic

118

6,2

6,2

1,26

100

Dimethyl

189

4,7

7,2

2,0

100

Nc
100
78,5
9,0
1,0
100
- Gii ly s dng dung mi vi nng tng dn tuyn tnh: Dung mi gii ly
c phn cc tng dn u.
2.3.3. Vn tc gii ly
22

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Vn tc chy ca dung mi gii ly khng c qu nhanh (s khng kp cn

bng vi cht hp ph) cng khng c qu chm hoc b cho ngng li mt thi
gian v lc cc dy cht tan s khuch tn hoc tri di theo mi hng lm xu qu
trnh tch.
Thng thng trong a s sc k ct, vn tc gii ly khong 5-50 git/pht hoc
1-2 cm/pht.
V d: Chit dch t l xanh Dung mi s dng l dung mi no? Vn tc chy
ca dung mi?
Ly 10g l rau mun cn ti (khong 30 l) ct nh cho vo ci s (b gn l),
trn thm 2g bt CaCO3 trung ha dch axit ca t bo. Nghin cc l n khi thnh
mt th ng nht. Cho 10 -15ml aceton 80% hoc cn etylic 90% vo ci s v khuy
u, 5 pht ri lc bng bng v bng phu thu tinh, dung dch thu c trn vi
2ml toluen ri chit bng phu chit, ly phn trn c dch mu. Tin hnh sc k
ct.
Sau khi np silica gel vo buret t chiu cao ct khong 10-20cm, bt ng
glucoz 20-30cm, bt canxi cacbonat np khong 15-20cm.
Dung mi s dng l cc h: ete du ho-aceton; toluen-aceton; toluen-cn
etylic; aceton-HCl c; NH4NO3 4M - NH3 4M; CH3COOH 1,5M - CH3COONH4
1,5M.
Vn tc chy ca dung mi gii ly khong 15- 20 git/pht.
2.3.4. Theo di qu trnh gii ly ct
- Vi mu c mu: quan st bng mt thng ta thy cc dy lp c mu sc
khc nhau ang tch xa nhau ra. Theo di cc dy mu v hng chng khi c gii ly
khi ct.

Cc cht c mu sc khc nhau tch xa nhau trong ct

23

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

S dng sc k bn mng kim tra mi dy mu c mt hay nhiu cht

- Vi cc mu khng mu: hng dung dch gii ly trong nhng h bi c th tch
nh nhau v nh s th t. Dung dch trong nhng h bi hng c s c sc k
lp mng trn cng mt bn mng. Nhng l no c sc k bn mng ging nhau s
c gom chung li vi nhau thnh mt phn on. ui dung mi p sut km cc
phn on ny s cho cao ca cc phn on . Ch ngng ct khi thu c lng cao
cc phn on bng 70-80% trng lng mu np vo u ct.
2.3.5. Ghi nhn kt qu sc k
- Phn on thu c l mt cht tng i sch tuy vn c cha mt t tp
cht: khi trn bn mng c mt vt chnh chim 80%, ngoi ra cn mt s vt m.
- Phn on thu c l hn hp nhiu cht: khi trn bn mng c nhiu vt,
cc vt c m nh nhau.
- kim tra xem mt hp cht c tinh khit hay khng ta s dng sc k lp
mng ln lt vi 3 h dung mi gii ly hon ton khc hn nhau (khng dng mt h
dung mi vi 3 t l khc nhau). Nu cht ch cho hin ra mt vt duy nht th c th
kt lun l cht tinh khit. Nu cht khng tinh khit th s cho thy bn cnh vt chnh
c thm vt khc. Mun kho st r hn th dng HPLC.
2.4. Xc nh cu trc ha hc ca mt hp cht
Ch nn tm cch xc nh cu trc ha hc ca mt hp cht tinh khit sau khi
kt tinh nhiu ln trong dung mi ph hp ( tinh khit 95%).
Kt qu kho st cu trc ha hc ca mt hp cht theo cc chi tit sau:
- M t hnh dng, mu sc v mi ca sn phm.
- Sc k lp mng trong h dung mi g, thuc hin hnh g, gi tr Rf ca vt,
hnh chp bn mng.
- im nng chy.
- Nng lc trin quang.
- Khi ph.
24

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

- Phn tch nguyn t.

- Ph hng ngoi IR.
- Ph t ngoi: max (ch o khi hp cht c cha ni i lin hp)
- Ph 1H NMR.
- Ph 13C-NMR.
Sau khi bin lun cc tn hiu ca cc loi ph, so snh vi cc s liu trong ti
liu s xut cu trc ha hc ca hp cht c lp c. Cu trc ny phi p ng
tt c cc ch tiu ha l nu trn.
3. U, NHC IM V NG DNG CA SC K CT H
3.1. u im
- C th s dng trong c hai lnh vc phn tch v iu ch.
- Sc k ct khng nhng dng xc nh s lng cc thnh phn ca hn
hp m n cn c th c dng tch v tinh ch nhng thnh phn phn tch.
- Pha tnh v cc dng c th nghim r tin, d kim; c th trin khai vi mt
lng ln mu cht.
3.2. Nhc im
- Khi chy ct sc k i hi s theo di lin tc duy tr mc dung mi v
nu khng s d gy xut hin cc bt kh lm tc ct, nt ct, bin dng ng i ca
dung mi.
- Qu trnh chy ct mt thi gian.
- Qu trnh tch chm v hiu qu thp so vi sc k lng cao p.
3.3. ng dng
- Phn lp cc alkaloid, khng sinh, hormon tng trng steroid, hormon sinh
dc v cc hp cht lin quan.
- Tch cc cht lipid, cc hp cht cha nitro t cc hydrocarbon, cc hp cht
vng thm t hn hp mch thng vng thm.
- Ra gii v tch cc thuc tr su.
- Lm tinh khit dc liu.
- Phn tch cc vitamin.
- Kh mu du, cht bo v sp bng phng php thm lc.

25

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Chng 3: MT VI K THUT SC K CT KHC

1. Sc k ct kh (Dry column chromatography DCC)
- Cht hp ph: s dng alumina hoc silica gel loi dng cho sc k lp
mng, c th trn thm cht ch th pht hunh quang. Liu lng s dng: 1g mu
cht cn 300g cht hp ph kh.
- M t h thng: s dng mt ng Celophan c ng knh 2-5cm c dng
thng ng. y ng c hn li, to dng hnh tr v u y ny c t mt lp
bng thy tinh. y ng c soi thng nhiu l nh bng cy kim may.

Chun b ng sc k
- Np cht hp ph vo ng: cht hp ph c np kh vo ng. u tin cho
cht vo 1/3 ct, g nh ng ln mt gh hay mt bn nn cht cho cht ch, tip tc
nhi c 1 ng thng cng c th cm trn tay (trong sut qu trnh nhi, u trn
c h).

Np cht hp ph vo ng
- Np mu vo u ct:
26

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

+ Np mu dng t: mu c ha tan trong mt lng ti thiu dung mi,

dng ng nh git t dung dch mu ph ln trn u ct v yn mt lc cho dung
mi bay i.
+ Np mu dng kh: ha tan mu trong mt dung mi d bay hi nh dietyl
ete hoc diclorometan vo mt becher, cho vo mt lng cht hp ph loi dng cho
sc k ct vi trng lng gp nm ln lng mu, trn u ri c quay thu hi dung
mi s c c bt rn kh. Ph lp bt ny ln trn u ct nhi lc ny. Sau cng
ph mt lp ct (hoc bng gn) ln trn u ct bo v mt thong ca ct.

Np mu vo ct
- Trin khai sc k: ng c dng ng v dung mi c cho chy t trn
u ng xung di cui ng nh vo trng lc. Cho dung mi vo u ct, gi sao
cho c mt lp dung mi dy 3-5cm trn u ct.

Trin khai sc k
- Hin hnh sc k: t ng tr nm ngang trn mt tm king v quan st
bng n UV, nh du cc on. Ly ng ra ngoi v dng li dao lam ct ng
thnh nhng khc thch hp. Dng dung mi trch cht trong mi on.
27

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Ct ng tr trch cht
u v nhc im ca phng php sc k ct kh
u im:
- Phng php sc k ct kh l phng php bt cu gia phng php sc
k lp mng (TLC) v sc k ct iu ch c in. p lc chi ph v thit b t hn
so vi phng php sc k lng iu ch.
- Cng nh cc phng php sc k ct ni chung, DCC dng c c trong
phn tch v iu ch.
- DCC nhanh, d dng v hiu qu cao trong vic tch hoc tinh ch cc cht
vi s lng ln.
- Ging nh TLC, DCC l mt phng php "khng ra gii" s dng ng
nilon nh l vt h tr v c th dng UV-254 nhn bit. Cng nh cc loi sc k
ct, cc ct vi hu ht mi kch c cha y pha tnh trong sc k ct kh c th
dng tch, hoc lm sch mt lng rt ln cc cht.
Nhc im:
- thit lp ct i hi k thut, s kho lo v cn thi gian.
- i hi s ch lin tc trong sut qu trnh tin hnh.
ng dng: Cng nh cc phng php sc k ct ni chung, sc k ct kh
dng c c trong phn tch v iu ch. Sc k ct kh c dng xc nh s
lng thnh phn cht trong hn hp, phn tch v lm tinh khit nhng thnh phn
ny dng cho nhng phn tch sau . Sc k ct kh p dng thnh cng cho
nhng cht nh thuc nhum, alkaloids, v nhng hp cht d vng khc, mc d c
th tch bng cc loi ct sc k khc nhng s gp nhiu kh khn ng k. Lipid
cng c th c tch bng sc k ct kh.
2. Sc k ct nhanh (Flash column chromatography-FCC)
- Cht hp ph: s dng silica gel c c ht 40-63m cho kt qu tt nht.
- M t h thng: H thng gm ct sc k bng thy tinh nh h thng sc k
28

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

c in, u ct c rp ni vi mt h thng bm nn c p lc trung bnh, p lc

c th iu chnh. Hi nn t trn u ct y dung mi xuyn xung lp cht hp ph
nhanh hn.

H thng sc k ct nhanh
- Lng mu v kch thc ct: nn s dng ct ngn, ng knh khong 110 cm v chiu di khng qu 40 cm.
Bng 3: Mt s thng s cho sc k ct nhanh
Lng mu cho vo (mg)

Th tch hng

ng knh

Th tch gii ly

ct (mm)

(ml)

Rf 0,2

Rf 0,1

10

100

100

40

20

200

400

160

10

30

400

900

360

20

40

600

1600

600

30

50

1000

2500

1000

50

mt l
(ml)

- Vn tc gii ly: mc dung mi h xung trong ng vi tc 5 cm/pht.

u v nhc im ca phng php sc k ct nhanh
u im:
- Nh kh nn y dung mi gii ly nn thi gian trin khai nhanh hn sc k
ct c in (thi gian trin khai nh hn 20 pht). Cho php phn lp mu 0.01-10g
29

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

trong10-15 pht. C th tch rt tt hn hp c khong 4 cht.

- Thit b n gin, r tin so vi HPLC.
Nhc im:
- phn gii trung bnh (ln hn 0.15).
- Khng tch c hn hp phc tp hay tch ring cht cn phn lp m ch
thu c cc phn on cha cht cn phn lp.
- thc hin cn phi c my bm nn kh v m tb phn gm nhng ct v
nhng np vn kn chuyn dng, c kch thc t nh n ln. Nhng b phn ny
tng i t tin nu so vi sc k c in.
ng dng: Sc k ct nhanh c s dng tch cc phn on n gin t
hn hp phc tp. Sau a cc phn on ny ln sc k ct c in tch thnh
cc phn on tinh khit.
3. Sc k nhanh - ct kh (Dry-column flash chromatography; Vacuum liquid
chromatography VLC)
- Dng c: Phu lc xp bng thy tinh, bnh tam gic, h thng to chn
khng nh bng vi nc.

H thng sc k nhanh ct kh
- Np cht hp ph vo ct:
H thng gm: phu lc xp gn trn bnh tam gic, bnh ny ni vi mt my
bm ht to chn khng.
Cho my bm chn khng hot ng, cho tng mung silica gel kh (loi c
ht 15-40 m) vo phu n tn y. Dng nt thy tinh c y bng phng nn
silica gel xung y phu to thnh mt khi rn ng nht c b mt phng.

30

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

Lu : lp silica gel ch dy ti a 5 cm (gia tng theo ng knh, khng

tng theo chiu cao).
Bng 4: Mi lin quan gia lng mu v kch thc phu trong sc k nhanh-ct
kh
Lng mu

ng knh trong

ca phu (cm)
< 100 mg
0,5 1,0
0,5 1,0 g
2,0 3,0
1,0 10,0 g
5
10,0 20,0 g
10
- Cn bng ct sc k:

Chiu cao

Phn on dung

Lp hp ph (cm)
4
4
5
5

dch gii ly (ml)

10 15
15 20
30 50
50

+ t mt lp bng gn dy ln b mt pha tnh bo v lp mt.

+ Cho mt lng ln dung mi (loi dung mi t phn cc nht) ln u ct,
cho h thng to p sut km hot ng, dung mi c ht xung i qua ct v c
hng vo bnh tam gic. Lng dung mi ny c s dng cho li ln u ct.
Thc hin vi ln n khi thy cht hp ph trong ct ng nht.
- Np mu ln u ct: tho ming bng gn bo v b mt
+ Np mu t: ct c ht kh tng i, tt my p sut km. Ha tan
mu trong dung mi t phn cc sau ph u ln khp b mt silica gel. Gn my p
sut km.
+ Np mu kh: cho lng mu kh ln u ct, g nh v tri u mu thnh
mt lp mng.
Sau khi np mu ln u ct, ly lp bng gn ph li ln trn mt ct.
- Gii ly: Cho my bm to p sut km hot ng. Rt mt th tch dung mi
nht nh ln b mt phu (bng 4), dung mi c ht xung bnh tam gic bn di.
Ch cho phu kh tng i, ngt p sut, rt phn dung dch va gii ly vo cc l c
nh s th t. Tip tc cho dung mi gii ly vo phu v lp li cc thao tc nh trn.
Mi ln phi ht kh phu ri mi cho dung mi mi. Dung mi gii ly c s dng
ln lt t khng phn cc n phn cc.
u v nhc im ca phng php sc k nhanh-ct kh
u im:
- Dng c d tm.
- Thi gian sc k nhanh nh lc ht bn di ht dung mi gii ly.

31

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

- Thun tin cho ngi thao tc: qu trnh thc hin c th b gin on m
khng b nh hng.
Nhc im:
- Khng tch ngay thnh cht tinh khit c (tr nhng hn hp n gin hay
cc vt cch xa nhau).
- Phi c dng c thch hp (phu lc xp; h thng ht).
ng dng:
- Tch hn hp thnh vi phn on c phn cc khc nhau, mi phn on
gm t cht hn d dng phn lp tip bng sc k ct c in.
- Hot ng n gin nn c ng dng rng trong lnh vc hp cht thin
nhin. Thch hp i vi dch chit dc liu th.
- Gip nh hng trong th nghim sinh hc mt cch tp trung v t lng ph.

32

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

KT LUN
Sc k l mt k thut c ng dng rng ri trong nhiu lnh vc trn th
gii. Vit Nam tip cn k thut ny vo nhng nm ca thp nin 80. V iu kin
cuc sng ngy cng cao nn i hi phi c nhng cng ngh k thut hin i
p ng nhu cu.
Ngy nay, phng php sc k l phng php ch o phn lp cc hp
cht t nhin cng nh tng hp. N c nhng ng gp to ln trong qu trnh
phn lp cc dc cht t dc liu. Qua chng minh c dc tnh v gip bo
ch ra cc thuc c hot cht t dc liu vi tinh khit cao gp phn lm tng
hiu qu iu tr cng nh gim cc tc dng ph. Ngoi ra, k thut sc k cng c
vai tr ln trong cng tc kim tra, gim st cht lng hng ho p ng nhu cu
cuc sng nh: thc phm, dc phm, ho cht
V vy, vi tng mc ch s dng m ta c th chn mt phng php sc k
ph hp phn tch mu tng ng./.

33

Tch chit v phn lp hp cht thin nhin

Phng php sc k ct

TI LIU THAM KHO

1. Nguyn Vn n, Nguyn Vit Tu (1986), Phng php nghin cu ha hc
cy thuc, Nh xut bn Y hc chi nhnh Thnh ph H Ch Minh.
2. Nguyn Ngc Hnh (2002), Gio trnh tch chit v c lp hp cht hu c, Vin
cng ngh Ha hc Thnh ph H Ch Minh.
3. Nguyn Kim Phi Phng (2007), Phng php c lp hp cht hu c, Nh xut
bn i hc Quc gia Thnh ph H Ch Minh.
4. http://www.sorbtech.com/chromatography/dry-column-chromatography-dcc/
5. http://www.saiadsorbents.com/dcc.htm
6. http://www.wfu.edu/chem/courses/organic/CC/index.htm
7. http://orgchem.colorado.edu/Technique/Procedures/Columnchrom/Procedure.html
8. http://www.chemguide.co.uk/analysis/chromatography/column.html
9. http://www.chemistryviews.org/details/education/2040151/Tips_and_Tricks_ for
_ the_Lab_ Column_Packing.html

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