A High-Performance Liquid Chromatography:Chemiluminescence Method for Potential Determination of Vardenafil in Dietary Supplement

A flow method of high-performance liquid chromatography (HPLC) seperation and chemiluminescence (CL) detection for sensitive vardenafil analysis in dietary supplements was developed. The vardenafil separation was achieved on a C18 column at 30°C using ethanol-H(3)PO(4) and ethylenediaminetetraacetic acid disodium salt (Na(2)EDTA) aqueous solution (25 : 75, v/v%) as mobile phase. The followed continuous CL detection was conducted based on the strong CL enhancement by the presence of vardenafil to luminol-K(3)Fe(CN)(6) reaction in alkaline medium. At the flow rate of 0.8 mL/min, the vardenafil retention time (t(R)) was 6.4 min. Factors that affected the HPLC resolution and CL detection were studied and optimized. The calibration curve obtained for vardenafil standard was linear in concentration range of 8.0 × 10(−7) ~ 1.0 × 10(−4) mol/L. The relative standard deviations (RSD) of intraday and interday precision were less than 3.5%. The proposed method was applied to the vardenafil determination in oral liquid, wine, and capsule samples.