Identification and characterization of Ethyl gallate from Ethyl acetate fraction of Phyllanthus emblica fruit, and invitro free radical scavenging activity

Phyllanthus emblica fruit has a rich source of polyphenolic compounds and is widely used due to its medicinal properties. A portion of the ethyl acetate fraction (EAF) from 70% methanol extract (70% ME) showed greater 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) scavenging activities. Moreover, EAF has more polyphenol content than 70% ME. The EAF was subjected to a dry silica gel column eluted successively with a mixture of chloroform: ethyl acetate: formic acid (2.8: 2.4: 0.4ml) as a mobile phase solvent system, and to afford various fractions. EAF and isolated fraction-1 of ethyl gallate (EG) showed a unique pattern of EG standard chromatogram in thin layer chromatography (TLC) analysis. EAF and isolated fraction-1 real time (R.T) of the peaks were unique compared with those of the standard chromatogram in high-performance liquid chromatography (HPLC) analysis. This chromatogram fraction was collected from isolated fraction-1 at the same R.T time by multiple HPLC analysis. The collected fraction from HPLC was injected into Mass spectroscopy (MS) and it showed a molecular ion at m/z 197 corresponding to C9H10O5. The substance was identified as EG based on the results mentioned above.