Thin Layer Chromaographic Analysis Essay

Chromatographic analysis: Thin Layer Chromatography175
Procedure
Thin layer chromatography is used for the separation, identification and quality control of drug. It also determines small amount of impurities or adulterants if present.
Apparatus
Glass plates of uniform thickness, 15-20 cm long and wide, spreader, chromatographic chamber of glass with a tightly fitting lid having suitable size to accommodate the glass plates and capillary, ultraviolet light source emitting short 254 nm and long 366 nm wavelengths.
Preparation of samples
The extract of Bhrungaraj samples were prepared by taking 1.0 g of drug and 1-10 ml of solvent, extract was stirred for about 30 minutes and filtered.
Preparation of Thin layers in plates
The plates were cleaned, rinsed thoroughly until the water

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Procedure
Preparation of Test Solution
The sample was prepared by adding 100 mg of powdered in 10 mL of methanol, soaked for 15 min. Centrifugation was done and the filtrates or supernatants were used as the sample solution. Out of this filtrate 50 μL of samples were dissolved in 1 mL of toluene and were used as the sample solution.
Preparation of the standard solutions
Wedelolactone, Ecliptaalbasaponin-I and Ecliptaalbasaponin-II were used as marker compounds; these markers were dissolved in methanol at a concentration of 1 mg/ml. The extracts of Bhrungaraj prepared were shaken and sonicated in methanol at a concentration of 10 mg/ml.
Sample application and plate layout
Samples were applied as narrow bands 8-9 mm above the lower edge of the plate. The sample application developing points were marked with a pencil before the development of chromatography.
Preconditioning of the plate
After the sample application the plate kept standing in a closed chamber containing saturated solution of magnesium chloride for preconditioning before keeping in the mobile phase.
Preparation of the developing chamber and development of the