My Compound is Leaf powdered Methanolic extract which is showing good phenolic compounds upon dipping in FeCl3. -My problem is phenolic compounds are not eluting with any common solvent system.
Try with Dichloromethane and Methanol:
DCM:MeOH, 100%, 98%, 95%, 90%, 85%, and 80%
Dear Sandesh,
Phenolic compounds elutes at lower Rf range . So you will need proper mobile phase. You can use Mobile Phase: Ethyl acetate: Formic acid: glacial acetic acid: Water (99:12:11:27 v/v/v)
Try this method:
Sample (3 g) was extracted with aqueous methanol. It was kept for 24 hr in order to get phenolic extract. The extract evaporated under vacuum pump to yield a sticky residue. Using a capillary tube, 3 drops of aqueous methanolic that they line up with the notches etched for each plant species. The developing solvent which is a mixture of Toluene - Formic acid - Ethyl acetate – H2O (25: 5.5: 5.5: 18.8) was prepared. Enough volume of the developing solvent was poured in a jar so as to reach 1 cm deep in the bottom and apiece of filter paper was placed into the jar to saturate the atmosphere with the solvent. Then the plate was dipped carefully in the developing solvent and the chromatogram was allowed to develop.
Plates were removed from the chamber and the highest solvent level on them was traced with pencil. Also the spots were traced while held under a UV lamp. Then the spots were marked with a pencil. The Rf values were then calculated for each spot.
I will follow your procedure
Thank you sir Faiq H S Hussain , Raju Ratan Wadekar , Ikram Madani Ahmed for your valuable answers.
Ikram Madani Ahmed sir i tried DCM : methanol and EtAc : formic acid : Acetic acid : Water with different ratio but they didn't shown any proper separation.
so i feel my methanolic extract may contain Gallic acid , Ellagic acid etc...
can you help to separate heavy phenolic molecule in leaf extract.
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