High-performance thin-layer chromatographic analysis for the simultaneous quantification of four phenolic compounds in green, red, and black fruits of<i>Trapa natans</i>var.<i>bispinosa</i>Roxb. (singhara)

Abstract

The fruits of Trapa natans var. bispinosa Roxb. (singhara) of nutraceutical importance are commercially consumed in food commodities. It is nutritious while having several biological activities. In the present study, a simple, rapid, cost-effective, and sensitive highperformance thin-layer chromatography (HPTLC) method was developed for the simultaneous determination of four phenolic compounds viz. gallic (phenolic acid), caffeic acid (hydroxycinnamic acid), quercetin, and kaempferol (flavonols) in seeds and pericarp of green-, red-, and black-colored singhara fruits. The method is economical in terms of the time taken and the amount of solvent used for analysis. Simultaneous separation and quantification of compounds were achieved on HPTLC precoated silica gel 60 F254 aluminum plates using mobile phase of toluene-ethyl acetate-formic acid (13:11:2). Densitometric determination was carried out at λmax 282 nm. The calibration curves were linear ranging between 0.996 and 0.999; the limits of detection and quantification ranged between 86.8–135.0 ng μL−1 and 263.2–409.2 ng μL−1 and recovery ranged between 96.4% and 98.5%. The validated method was successively used to analyze the above compounds in fruit parts of singhara. The amount of phenolic compounds ranged from 0.04% kaempferol (green pericarp) to 2.11% gallic acid (red pericarp). The developed method may be used in the quality control and standardization of plant extracts as well as herbal drugs and formulations having polyphenols. As this study reveals the presence of specific phenolic compounds in green, red, and black pericarp of singhara, the agrowaste shall be a good source for isolation of the above compounds for industrial use.