CC PHUONG PHP CO BN CA KY THUAT SAC K

Chromatography
Mc dch:
-Tch mt hn hp phc tap gm nhiu Ioai hp cht
ra ring thnh tng Ioai don cht.
Co ch:
- Da vo tnh i Ic khc nhau ca nhng Ioai hp
cht d di vi mt h thng (h thng ny thung
gm hai pha: mt pha dng v mt pha tnh).
CC PHUONG PHP CO BAN:
W SAC K BAN MONG -THIN-LAYER CHROMATGRAPHY (TLC)
W SAC K KHI - GAS CHROMATOGRAPHY (GC)
W SAC K LONG - LIQUID CHROMATOGRAPHY (LC)
W SAC K LONG HIJU NNG CAO- HIGH PERFORMANCE
LIQUID CHROMATOGRAPHY (HPLC)
SAC K LP MNG
THIN-LAYER CHROMATGRAPHY (TLC)
Nguyn Iiu duc s dng Im cht hp
thu (thung I siIica geI) duc trng
thnh mt Ip mng, ph du In mt tm
knh, nhm hoc tm nha
Pha dng thung I mt h cht Ing thung gi I dung
mi gii Iy (eIuent, eIuant).
Trong TLC, hay s dng thng s Rf (retardation factor, retention factor )
' tr ca mu cht duc quan st bng
nh sng U' hoc hin hnh bng thuc
th
Uu dim:
- Chi cn mt Iung nh mu d phn tch
- Thi gian trin khai sc k tuong di nhanh
- C th tin hnh di chng cng Ic nhiu mu
Khuyt dim:
- Chi dnh tnh, khng dnh Iung.
- Thung chi d phn tch ch khng phn Ip.
Mt s tc nhn nh hung dn gi tr Rf:
1.1 - S phn cc ca pha tnh (siIica geI).
1.2 - S phn cc ca pha dng (dung mi).
1.3 - S tuong tc gia hp cht cn phn tch vi
pha tnh v pha dng.
1.1- nh hung ca pha tnh - SiIica geI:
SiIica geI thung: I poIymer ba chiu, gm nhng
don v SiO2.H2O
SiIica geI pha do (reversed phase):
) i vi siIica geI thung:
Nhng hp cht phn cc ( -OH, -NH2, -COOH..)
b gi cht, b gii Iy chm.
Nhng hp cht km phn cc b gii Iy nhanh
hon.
) i vi siIica geI pha do th nguc Iai
1.2 - nh hung ca pha dng - Dung mi gii Iy:
- Kh nng ha tan: dung mi di chuyn dn du th
mu cht b Ii ko theo dy.
- Hin tung thay th ch (s canh tranh ch): phn
t dung mi canh tranh dy 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 Iy
Cc cht c mu c th quan st duc bng mt
thung.
Bng nh snh t ngoai ( 254nm, 366 nm).
Bng thuc th ha hc:
+ Hoi Iod.
+ H2SO4 10% trong etanoI.
+ Ninhydrin (amin, amino acid).
TAO THNH ESTER????
1.4 Cc cng dng ca sc k Ip mng
Gi tr Rf ca mt hp cht
Kim tra 2 hp cht c ging nhau ?
Tm hiu so 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.????( Ktra sau khi ra
khoi cot????)