03-12-2012, 12:30 PM
SCREENING OF IAA BY TLC METHOD
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Regent Preparation
Salkowsk’s Reagent – the salkowsk’s reagent is consist of 50ML,35% perchloric acid and 1Mi of 0.5 M FeCl3
Solvent Preparation
The solvent was prepared by taking methanol Glacial acetic acid and water in the ratio 120:30:50 and made up to 200Ml
Seperation of Auxin by TLC
Method
The bacterial filtrate is applied separately to the plates at correct distances micropette was used and small spots of bacteria as culture were made. The developing solvents (methanol, glacial acetic acid, water) in the ratio (120:3050) were prepared and poured into the tank .Allow it to stand. A cover plate over the top of the tank assure that the atmosphere within the tank. Tank was saturated with solvent solution. The glass plate is kept in such a way that solvent lies below the sample spot. Then the tank was covered. The separation of components. Occurs as the solvent moves upward once the solvent reaches the top of the plate. The plate was removed fro the tank and the solvent front was marked. Then allow it to dry and the plate was sprayed with salkowsky reagent the plates were dried in a hot air oven at 105oC far 5-8 min the RF value of Auxin can be calculated using the formula.
RF =