GCMS analysis of partially purified chloroform sub fractions of methanol extract of drymaria cordata (Linn) and their effects on mitochondrial membrane permeability transition pore

Abstract

The plant drymaria cordata is folklorically used in the treatment of uterine fibroids. Investigating its phytoconstituents and their effects on mitochondrial permeability transition (mPT) pore is of pharmacological importance. The aim of this study was to identify the possible phytochemical compounds present in the most potent sub-fraction of the plant and test them on mPT pore. The methanol extract of drymaria cordata was successively partitioned between n-hexane, chloroform, ethyl acetate and methanol solvent using Vacuum Liquid Chromatography (VLC) technique to obtain n-hexane, chloroform, ethyl acetate and methanol fractions. The chloroform fraction was partitioned into five different subfractions using VLC. The subfractions were tested for their potency in the induction of mPT pore opening. The subfraction D which was the most potent with respect to mPT pore opening was loaded on a column and eluted with n-hexane and chloroform varying the solvent systems in order of increasing polarity. The flow rate was 2.5ml/min. Thin Layer Chromatography (TLC) plates were used to monitor samples with similar components. Samples with similar Rf values were pooled together. For further resolution, samples which showed the same single band on the TLC plates were further subjected to purification using micro-column chromatography. The eluents were also monitored using the TLC plates. The partially purified samples were subjected to GC-MS analysis and certain phytochemicals including hexadecanoic acid methyl ester, hexadecanoic acid ethyl ester, hexadecanoic acid, 9-Octadecenoic acid, Oxalic acid 2-ethylhexyl isobutyl ester, 3-(p-Fluorobenzoyl)-propionic acid and Bis(2-ethylhexyl) phthalate were identified. The presence of these major compounds might be responsible for the pharmacological properties ascribed to the fraction. Also, these could give an insight into developing chemopreventive and therapeutic approaches to various diseases in human associated with dysregulated apoptosis.