comparison of PC and TLC

1. PRESENTED BY
THUSHARA C
1ST YEAR MPHARM
DEPT. OF PHARMACY PRACTICE
COMPARISION OF THIN LAYER
CHROMATOGRAPHY AND PAPER
CHROMATOGRAPHY :

2. PAPER CHROMATOGRAPHY TLC
PRINCIPLE Principle of separation is partition Principle of separation is adsorption
Stationary phase Whatman filter papers of
different grades like No.1, No.2,
No.3, No.4, No.20, No.40, No.42
etc are used
Silica gel coated on a plate
Silica gel H ( Silica gel with out
binder )
Silica gel G ( Silica gel + CaSO4
Silica GF (Silica gel + binder +
fluorescent indicator)
Alumina, Cellulose powder,
Kieselguhr G
Mobile phase Pure solvents, buffer solutions or
mixture of solvents
Examples- Hydrophilic mobile
phase
Isopropanol: ammonia:water
Methanol : water
N-butanol : glacial acetic acid :
water
Hydrophobic mobile phases
dimethyl ether: cyclohexane
kerosene : isopropanol
pyridine, pet. ether, carbon
tetrachloride, acetone, water,
glycerol, ethanol, benzene

3. Development
techniques
Ascending , descending, circular or
radial
One dimensional
development,
Two dimensional
development,
Horizontal
development,
Multiple development
Visualising agents Non specific methods ( Physical methods)
E.g. iodine chamber method,
UV chamber for fluorescent
compounds – at 254 or at 365nm.
Specific methods (Chemical methods) or
Spraying method - examples, Ferric
chloride,Ninhydrin in acetone
Dragendroff’s reagents3,5 dinitro
benzoic acid , Phenolic comp. &
tannins,Amino acids,Alkaloids
Cardiac glycosides
a) Non specific
methods
Iodine chamber method
Sulphuric acid spray
reagent
Same as PC

4. Quantitative
estimation
1. Direct technique:
• Comparison of visible spots
• Photo densitometry
• Radiotracer Method
2. Indirect method
3. Rf value
4. Rx value
1. Direct technique
2. Indirect method
3. Rf value
4. Rx value
Application  Separation of mixtures of
drugs,
 Separation of carbohydrates,
vitamins, antibiotics,
proteins, etc.
 Identification of drugs
 Identification of impurities
 Analysis of metabolites of
drugs in blood , urine ….
Purity of sample
»Examination of reaction
»Identification of
compounds
»Biochemical analysis
»In pharmaceutical industry
»Separation of
multicomponent
pharmaceutical formulations
»In food and cosmetic
industry

5.  COMPARISION OF THIN LAYER CHROMATOGRAPHY
AND PAPER CHROMATOGRAPHY :
THIN LAYER CHROMATOGRAPHY PAPER CHROMATOGRAPHY
The principle of separation is adsorption. The principle of separation is partition
More amount of substance is required. Less amount of substance is required.
Less time consuming i.e. 15—45 minutes More time consuming i.e. 2—3 hours
TLC plates can be heated in an oven for a
long time
Paper cannot be heated in an oven for a long
time.
Separation is more sharp Separation is less sharp.
Thin layer have physical strength. Ascending
techniques are preferred for this type
Due to lack of physical strength in paper,
descending techniques are used.
In TLC, chromatoplates are prepared by
coating thin layers of silica gel or alumina on
glass plates.
Here paper is generally used whatman filter
paper. Silica gel or alumina cannot be used
because they cannot be made into sheets.
in TLC, corrosive reagents may be coated on
glass plates.
Here, corrosive reagents destroy the paper.
Sensitivity of detection of the fraction on
plate is high. The spots are less diffused
Less sensitivity of detection. Spots easily
diffused
TLC can be evaluated under UV. Paper chromatography cannot be evaluated
under UV.

6. Thank you