Gas chromatography–mass spectrometry and high-performance thin-layer chromatography quantifications of some physiologically active secondary metabolites in Averrhoa carambola L. fruits

Abstract

The extraction of crude drugs by using different solvents provides polarity-based fractions containing specific types of secondary metabolites. Averrhoa carambola L. fruits were extracted and fractionated, and petroleum ether extract was processed by fatty acid methyl ester (FAME) technique for characterization by gas chromatography–mass spectrometry (GC–MS) analysis. The remaining part was extracted with methanol for high-performance thin-layer chromatography (HPTLC) analysis, for simultaneous quantitative determination of gallic acid, protocatechuic acid, and quercetin in methanolic fractions. Petroleum ether and methanol fractions were found to be the best for the highest possible recovery of target analytes. The chromatographic elutions of FAME compounds generated from ether extract were evaluated by GC–MS profiling. Ten fatty acid compounds were separated with the highest quantity of oleic acid methyl ester (42.88%). On other hand, polar fraction was processed by HPTLC profiling. For achieving good separation, a mobile phase of toluene–ethyl acetate–formic acid (5:4:1, v/v) was used. The densitometric determination was carried out at 310 nm in reflection–absorption mode. The calibration curves were linear in the range of 100–600 ng per spot for gallic acid, protocatechuic acid, and quercetin. During the analysis, the dried raw material from A. carambola L. fruits showed the presence of gallic acid (0.96%), protocatechuic acid (0.05%), and quercetin (0.40%). The proposed method is simple, precise, specific, and accurate. The statistical analysis of the data obtained proves that the method is reproducible and selective and can be used for the routine analysis of the reported phenolic compounds in crude drug and extracts. The results indicated that the methanolic extracts of the plant contained a considerable amount of bioactive compounds. The presence of phytochemicals especially phenolics and flavonoids explains its use in various diseases. It may be concluded that the results obtained from the quantitative evaluation of quercetin by HPTLC fingerprinting could be useful in its authentication, the quality control of the drug, and in ensuring therapeutic efficacy.