Development and Validation of High-Performance Liquid Chromatography for Simultaneous Determination of Seven Bioactive Compounds in Flos Lonicerae japonicae

Abstract

A selective high-performance liquid chromatographic method for simultaneous determination of seven bioactive compounds in Flos Lonicerae japonicae was developed and validated. In this paper, the seven components were separated on a Waters Symmetry C₁₈ column (250 mm × 4.6 mm, 5 μm) and detected by a diode array detector. The mobile phase was composed of (A) aqueous formic acid (0.5 %, v/v) and (B) acetonitrile using a gradient elution. Analytes were performed at 30 ºC with a flow rate of 1 mL min^-1 and the detection wavelength was set at 238 nm. All calibration curves showed good linear regression (r² ³ 0.9991) within tested ranges. Overall intra-day and inter-day variations (RSDs) were less than 2.13 % and the average recoveries of seven analytes varied from 97.41 to 103.10 % with RSDs ranging from 0.87 to 2.81 %. The proposed method would be sensitive enough and reliable for comprehensive quality control of Flos Lonicerae japonicae and its related processed drugs or preparations.