Simultaneous Quantification of Eight Marker Compounds in Yongdamsagan-Tang Using a High-Performance Liquid Chromatography Equipped with Photodiode Array Detector.

Abstract

Yongdamsagan-tang (YDSGT) has been used clinically for the treatment of acute- and chronic-urethritis, cystitis, orchitis and hypertension in Korea. In this study, a powerful method based on high-performance liquid chromatography (HPLC) with photodiode array (PDA) detection was established and validated for the quantitative analysis of eight components: chlorogenic acid, gentiopicroside, liquiritin apioside, liquiritin, nodakenin, baicalin, wogonoside and glycyrrhizin in YDSGT extract. The compounds were separated with a Gemini C18 analytical column (column temperature: 40°C; mobile phase: 0.1% (v/v) aqueous trifluoroacetic acid (A) and acetonitrile (B); flow rate: 1.0 mL/min; injection volume: 10 μL). The PDA detector scanned the range 190-800 nm and the marker compounds were monitored at 254, 275, 325 and 335 nm. The correlation coefficients of all compounds were 1.000 and the results showed excellent linearity. The lower limits of detection and quantification of the analytes were 0.01-0.09 μg/mL and 0.03-0.28 μg/mL, respectively. The extraction recoveries of the marker compounds were 98.13-103.86%, with relative standard deviation values not exceeding 2.10%. The precision of intra- and inter-day measurements were 0.09-1.78% and 0.12-2.09%, respectively. The content of the eight marker compounds in the freeze-dried YDSGT extract were 1.41-23.71 mg/g.