Bioanalytical HPLC method of Piper Betle L. for quantifying phenolic compound, water-soluble vitamin, and essential oil in five different solvent extracts

Abstract

A reversed-phase high-performance liquid chromatography with diode-array detection (HPLC-DAD) was developed and validated to estimate the phenolic acids (gallic acid, caffeic acid, syringic acid, p-coumaric acid, sinapic acid, and ferrulic acid), flavonoids (catechin rutin, myricetin, quercetin, apigenin, and kaempferol), ascorbic acid, and eugenol. The chromatogram condition was set in suitable wavelength 272 nm and run flow rate 0.7 μl/minutes using HPLC Agilent Technologies 1260 Infinity, a reversed-phase Zorbax SB-C18 column (3.5 μm particle size, i.d. 4.6 mm × 250 mm) with the mobile phase solution (1:9, HPLC-grade acetonitrile:1% acetic acid). The linearity, precision, limit of detection, limit of detection, and accuracy were R 2 > 0.9907, relative standard deviation < 1%, 0.005 μg/ml, 0.015 μg/ ml, and 96%-102%, respectively. As a result, all the selective compounds were successfully separated, identified, and quantified. The enormous contents were found in quercetin and eugenol, expressing crude content (mean, 5.989 mg/g) and residue content (mean, 1.934 mg/g) for quercetin, while crude content (mean, 3.209 mg/g) and residue content (mean, 0.184 mg/g) for eugenol. Consequently, this method could be applied, repeated, and developed for the later observation, especially in commercially inclination of Piper betle analysis.