Abstract

A new, simple and sensitive high-performance liquid chromatography-diode array detector (HPLC-DAD) method was developed for the simultaneous determination of eight phenolic components in Phyllanthus simplex Retz. The chemical profile of the eight components, including gallic acid, caffeic acid, quercetin, ferulic acid, luteolin-7-O-glucoside, 8,9-epoxy brevifolin, brevifolin and globulariacitrin were acquired by using high-performance liquid chromatography-diode array detector coupled to an electrospray ionization mass spectrometer (HPLC-DAD-ESI/MS). The optimal condition of separation was achieved on a ZORBAX Extend C₁₈ column (5 μm, 250×4.6 mm i.d.) with a gradient elution of methanol and 1% acetic acid at a flow rate of 1.0 ml/min. Under ultra violent (UV) detection at 275 nm, the recoveries of the analytes were in the range of 99.2 to 99.8% with a RSD of 0.41 to 1.50%. The limits of detection and quantification for the analytes  ranged from 55 to 140 ng/ml and from 155 to 320 ng/ml, respectively. The intra-day RSD% ranged from 0.32 to 0.82% and the inter-day RSD% were not higher than 5.0%. The proposed method is suitable for quantitative and qualitative determination of the eight bioactive phenolic components in P. simplex Retz.

Key words: Phyllanthus simplex Retz, phenolic components, high-performance liquid chromatography-diode array detector coupled to an electrospray ionization mass spectrometer (HPLC-DAD-ESI/MS).