Development and Validation of a High-Performance Liquid Chromatographic Method for the Simultaneous Quantification of Marker Constituents in Cheonwangbosimdan

Abstract

A high-performance liquid chromatography–photodiode array detector method was established for the simultaneous determination of 7 components in Cheonwangbosimdan extract. The components were 5-hydroxymethyl-2-furaldehyde (1), coptisine (2), berberine (3), nodakenin (4), harpagoside (5), cinnamic acid (6), and β-asarone (7). All analytes were separated by gradient elution using two mobile phases on a Gemini C₁₈ column and maintained at 40°C. The flow rate was 1.0 mL/min and the injection volume was 10 μL. Calibration curves of the 7 compounds showed good linearity with correlation coefficients (r²) ≥ 0.9996. The limits of detection and quantification of the 7 analytes were 0.01–0.04 and 0.03–0.12 μg/mL, respectively. The recoveries of the 7 marker constituents were 97.6–104.2% with relative standard deviations (RSD) of less than 2.2%. The RSD values of intra- and interday precision were 0.11–1.78 and 0.19–1.92%, respectively. Among the 7 biomarker compounds, the major compounds of Cheonwangbosimdan were berberine and coptisine, which originated from Coptis japonica. The results indicate that the developed analytical method is suitable for quality control use.