• Thin Layer Chromatography (TLC) studies were carried out for various extracts to confirm the presence of different phytoconstituents in these extracts.

• TLC is a mode of liquid chromatography, in which, the extract is applied as a small spot or band at the origin of thin sorbent layer supported on a glass / plastic / metal plate.

• The mobile phase migrates through the stationary phase by capillary action.

• The separation of solutes takes place due to their differential adsorption / partition coefficient with respect to both mobile and stationary phases.

• Each separated component has same migration time but different migration distance.

• The mobile phase consists of a single solvent or a mixture of solvents.

• Although, a number of sorbents like silica gel, cellulose, polyamide, alumina, chemically modified silica gel etc. are used, Silica gel (type 60) is most commonly used.

• Handmade plates are prepared by using techniques like, pouring, dipping or spraying.

• Now-a-days, readymade precoated plates are also available. The plates need to be activated at 110⁰C for 1 h. This removes water / moisture loosely bound to silica gel surface

The Retardation Factor (Rf) is calculated using following formula,

The Details of TLC are as follows: -

Adsorbent : Silica gel GF 254 (activated)

Thickness : 0.4 mm

Plate size : 10 x 20 cm

Activation temp. : 110ºC for 1 hr

Volume of spot : 20 µl

Solvent system : As per drug

Procedure: -

• A thick layer of silica gel GF-254 was coated on the square shaped plate and activated at 110⁰C for one hour. The sample was applied on the plate.

• The plate was developed in a saturated chamber having desired solvent system.

• After developing the plate was dried and if the band gives fluorescence then it can be easily scraped.

• Otherwise a small portion of the band was sprayed with detecting agent by taking care to avoid the exposure of remaining plate to spray reagent.

• Then the band is scraped by measuring the height of sprayed band.