Determination of Major Polyphenolic Components in Euphoria longana Lam. by Validated High Performance Thin-Layer Chromatography Method and Direct Analysis in Real Time Mass Spectrometry

Abstract

In the current study, a simple, sensitive, precise, and specific method has been developed and validated for the determination of four major polyphenolic compounds in Euphoria longana Lam. seeds based on high performance thin-layer chromatography (HPTLC) and confirmed by direct analysis in real time mass spectrometry (DART-MS). The chromatographic separation was accomplished on Merck HPTLC aluminum plates precoated with silica gel G60 F254 (20 cm × 10 cm) with 250 μm thickness as stationary phase using a mobile phase composed of n-butanol: water: methanol: formic acid (7.59: 1.27: 0.13: 1.01, v/v/v/v). Densitometric measurement was performed in the absorbance mode at 280 nm. The compounds were resolved satisfactorily with Rf values of 0.40 ± 0.01, 0.57 ± 0.02, 0.69 ± 0.01, and 0.79 ± 0.01 for corilagin, ellagic acid, epicatechin, and gallic acid, respectively. The method developed was validated with acceptable linearity (r2>0.995), sensitivity, precision (RSD ≤ 1.60%), robustness, and recovery (RSD ≤ 1.77%). The proposed method was successfully applied for the determination of active components for comprehensive quality control of E. longana products. Furthermore, peak identities of compounds from each band were confirmed by direct analysis in real time mass spectrometry.