KLT (TLC) & PC
KLT (TLC) & PC
(PC)
Bentuk paling sederhana dari kromatografi
Prinsip:
Sampel diteteskan pada kertas saring dengan mikropipet, selanjutnya
ujung kertas dicelupkan pada pelarut yang sesuai (jangan sampai
mengenai sampel).
Solven akan merambat ke atas dengan daya kapiler dan komponen sampel
akan bergerak naik dengan kecepatan yang berbeda sesuai dengan tingkat
retensinya pada kertas. Masing-masing komponen dapat dilihat dengan
mereaksikan kertas (menyemprot) dengan pereaksi yang dapat
membentuk warna.
Tingkat pemisahan
komponen dalam
sampel dinyatakan
dengan harga Rf
Each component is
characterized by its Rf
value, the ratio of a and b:
Rf = a/b: 0 ~ 1
A B U C D
Stationary Phase: Alumina
O OH OH OH OH
Al Al Al Al Al
O O O O O O
Acidic: -Al-OH
Neutral: -Al-OH + -Al-O-
Basic: -Al-O-
Stationary Phase: Silica (SiO2)
OH
OH
OH
OH
OH Si
Si O
Si O
Si O O
Si O O
O O
O O
O Si Si
O O
Si
Si O O O
Si O O
O
O O
O
Si
Si O
O
O O
O
Reverse phase chromatography
Silica is alkylated with long chain hydrocarbon groups, using 18
carbons long (ODS). This is usually referred to as C-18 silica.
CH3
CH2
CH3 17 CH3
Si
CH2 SiCH3)3 CH3
17 CH3
SiCH3)3
SiCH3)3
O
Si O
CH3 O
O
O Si
Si O
Si O
Si O O
Si O O
O O
O O
O Si
Si O
Si O
Si O O
Si O O
O O
O O
O
Si
Si O
O
O O
O
THIN LAYER CHROMATOGRAPHY
Distance B
migrated = 2.0 cm 3.0 cm Rf (D) = 4.0 cm = 0.80
5.0 cm
Distance C
migrated = 0.8 cm
0.8 cm Rf (U1) = 3.0 cm = 0.60
Origen
x x x x x 5.0 cm
A B U C D
0.8 cm
Rf (U2) = = 0.16
5.0 cm
The Rf is defined as the distance the center of the spot moved divided
by the distance the solvent front moved (both measured from the origin)
THIN LAYER CHROMATOGRAPHY
Calculation of Rf’s
2.0 cm
Rf (A) = = 0.40
5.0 cm
Solvent Front
Distance B
migrated = 2.0 cm 3.0 cm Rf (D) = 4.0 cm = 0.80
5.0 cm
Distance C
migrated = 0.8 cm
0.8 cm Rf (U1) = 3.0 cm = 0.60
Origen
x x x x x 5.0 cm
A B U C D
0.8 cm
Rf (U2) = = 0.16
5.0 cm
The Rf is defined as the distance the center of the spot moved divided
by the distance the solvent front moved (both measured from the origin)
THIN LAYER CHROMATOGRAPHY – Rf’s
Absorption of Solutes
The adsorption strength of compounds increases with increasing polarity of
functional groups, as shown below:
-CH=CH2, -X, -OR, -CHO, -CO2R, -NR2, -NH2, -OH, -CONR2, -CO2H.
(weakly adsorbed) (strongly adsorbed)
(nonpolar) (more polar)