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. 2011:2011:982186.
doi: 10.1155/2011/982186. Epub 2011 Jan 20.

A high-performance liquid chromatography:chemiluminescence method for potential determination of vardenafil in dietary supplement

Youjun Di  1 , Min Zhao, Yingchun Nie, Fei Wang, Jiagen Lv
Affiliations

A high-performance liquid chromatography:chemiluminescence method for potential determination of vardenafil in dietary supplement

Youjun Di et al. J Autom Methods Manag Chem. 2011.

Abstract

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.

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Figures

Figure 1
Figure 1
Structure of vardenafil.
Figure 2
Figure 2
Schematic diagram of the proposed HPLC-CL integration for vardenafil analysis. a: luminol+Na2EDTA+NaOH; b: K3Fe(CN)6; PMT: photomultiplier tube; NHV: negative high voltage generator (−400 V); PC: computer; T1 and T2: three-way mixer 1 and 2.
Figure 3
Figure 3
Vardenafil chromatograms with (A) and without (B) H3PO4 in mobile phase. Mobile phase contained 25% ethanol and 2.0 × 10−4 mol/L Na2EDTA.
Figure 4
Figure 4
Original (O) and 2.0 × 10−5 mol/L vardenafil standard spiked (S) oral liquid, wine, and capsule chromatograms under the optimum conditions.
See this image and copyright information in PMC

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