CC PHNG PHP C BN CA K THUT SC K

Chromatography
Mc ch:
-Tch mt hn hp phc tp gm nhiu loi hp cht
ra ring thnh tng loi n cht.
C ch:
- Da vo tnh i lc khc nhau ca nhng loi hp
cht i vi mt h thng (h thng ny thng
gm hai pha: mt pha ng v mt pha tnh).
CC PHNG PHP C BN:
SC K BN MNG -THIN-LAYER CHROMATGRAPHY (TLC)

SC K KH GAS CHROMATOGRAPHY (GC)

SC K LNG LIQUID CHROMATOGRAPHY (LC)

SC K LNG HIU NNG CAO- HIGH PERFORMANCE
LIQUID CHROMATOGRAPHY (HPLC)
SC K LP MNG
THIN-LAYER CHROMATGRAPHY (TLC)
Nguyn liu c s dng lm cht hp
thu (thng l silica gel) c trng
thnh mt lp mng, ph u ln mt tm
knh, nhm hoc tm nha
Pha ng thng l mt h cht lng thng gi l dung
mi gii ly (eluent, eluant).
Trong TLC, hay s dng thng s Rf (retardation factor, retention factor )
V tr ca mu cht c quan st bng
nh sng UV hoc hin hnh bng thuc
th
u im:
- Ch cn mt lng nh mu phn tch
- Thi gian trin khai sc k tng i nhanh
- C th tin hnh i chng cng lc nhiu mu

Khuyt im:
- Ch nh tnh, khng nh lng.
- Thng ch phn tch ch khng phn lp.

Mt s tc nhn nh hng n gi tr Rf:
1.1 - S phn cc ca pha tnh (silica gel).
1.2 - S phn cc ca pha ng (dung mi).
1.3 - S tng tc gia hp cht cn phn tch vi
pha tnh v pha ng.

1.1- nh hng ca pha tnh - Silica gel:
Silica gel thng: l polymer ba chiu, gm nhng
n v SiO2.H2O
Silica gel pha o (reversed phase):
*) i vi silica gel thng:
Nhng hp cht phn cc ( -OH, -NH2, -COOH.)
b gi cht, b gii ly chm.
Nhng hp cht km phn cc b gii ly nhanh
hn.

*) i vi silica gel pha o th ngc li
1.2 nh hng ca pha ng - Dung mi gii ly:
- Kh nng ha tan: dung mi di chuyn n u th
mu cht b li ko theo y.
- Hin tng thay th ch (s cnh tranh ch): phn
t dung mi cnh tranh y mu cht phn tch ri xa
pha tnh.
** S la chn dung mi phn ln vn da vo kinh
nghim c nhn.

1.3 Hin hnh sau khi gii ly
Cc cht c mu c th quan st c bng mt
thng.
Bng nh snh t ngoi ( 254nm, 366 nm).
Bng thuc th ha hc:
+ Hi Iod.
+ H2SO4 10% trong etanol.
+ Ninhydrin (amin, amino acid)

1.4 Cc cng dng ca sc k lp mng
Gi tr Rf ca mt hp cht
Kim tra 2 hp cht c ging nhau ?
Tm hiu s b tnh cht ca mu kho st ( tinh
khit ?, bao nhiu cht ?, phn cc?).
Theo di phn ng.
Chun b cho vic sc k ct.