Beruflich Dokumente
Kultur Dokumente
Chromatography
Distribution Coefficient
Definition:
Concentration of component A in stationary phase
Concentration of component A in mobile phase
Kinds of Chromatography
S ample
(A+B+C)
Eluant (eluate)
Si - O - H
Silica Gel
HEXANE
Si - OH
CH 3
OH
CH 3
C-CH 3
CH 3
CH 3 - C
CH 3
CH 3
Water-Soluble Vitamins
1.
Niacinamide
2.
Pyridoxine
H3C
N
HO
CONH 2
3.
H 3C
H 3C
CH 2OH
Riboflavin
CH 2OH
HOCH
HOCH
HOCH
CH 2
N
N
4. Thiamin
O
NH
N
O
CH 2OH
H3C
N
N
NH 2
CH 2
S
N
CH 2CH 2OH
Cl
CH 3
Water-Soluble Vitamins
2
3
Inject
1
10
15
20
Liquid-Liquid Chromatography
ODPN (oxydipropionylnitrile)
Normal Phase LLC
Reverse Phase LLC
The stationary solid surface is coated with a 2nd liquid (the Stationary Phase)
which is immiscible in the solvent (Mobile) phase.
Partitioning of the sample between 2 phases delays or retains some components
more than others to effect separation.
Types of Chromatography
LIQUID
MOBILE PHASE
Liquid-Liquid
Chromatography (Partition)
FORMAT
STATIONARY PHASE
Normal Phase
Liquid-Solid
Chromatography (Adsorption)
Solid
Liquid
Reverse Phase
Normal Phase
Reverse Phase
Ion-Exchange Chromatography
SO 3- Na +
pH2
-
SO 3
Na
H3N
COOH
Ion-exchange Resin
SO 3
H 3N
Na
COO
pH4.5
Mobile Phase
H3 N
SO 3 Na+
COOH
+
Na
SO 3
OH
H3 N
COOH
Exchange Resin
-
SO 3 H3N+
COOH
SO3
pH3.5
OH
H 3 N+
+
Na
COO
OH = H 2 O
Na
SO 3
H3 N
+
-
COO
OH = H 2 O
SO 3Na+
pH4.5
Gel-Permeation Chromatography
Solvents
Polar Solvents
Water > Methanol > Acetonitrile > Ethanol >
Oxydipropionitrile
Non-polar Solvents
N-Decane > N-Hexane > N-Pentane >
Cyclohexane
LC Mode
Positional isomers
LSC or LLC
LSC or LLC
LSC or LLC
Ionizable Species
IEC
LLC
GCC
Detector
1.
Ultraviolet Detector
200-400nm
254 nm
2.
Retention Time
Time required for the sample to travel from the injection port through
the column to the detector.
Response
D
10
15
Retention Time
20
25
Selectivity
Ratio of Net Retention Time of 2 components.
(Distribution Coefficient)
X2
X1
X0
X0
Selectivity
Selectivity
Response
X
X1
X0
3
Retention Time
Resolution Equation
V2 - V
R=
1/2(W + W2)
1
Response
V2
V1
W2
W1
W2
Volumes
Resolution
X
Theoretical plates (N) = 16 ( )2
Y
2
4
V2
V1
V0
W2
W1
W3
W4
V3