alexa Determination of sildenafil citrate and its main metabolite by sample stacking with polarity switching using micellar electrokinetic chromatography.
Pharmaceutical Sciences

Pharmaceutical Sciences

Pharmaceutica Analytica Acta

Author(s): Berzas Nevado JJ, Flores JR, Pealvo GC, Rodrguez FN

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Abstract Micellar electrokinetic capillary chromatography (MEKC) coupled with sample stacking and polarity switching was investigated for the determination of Viagra (sildenafil citrate, SC) and its metabolite (UK-103,320, UK) in human serum in the concentration range of clinical interest. Human serum samples spiked with SC and UK were eluted with methanol from a C18 cartridge, the extract was evaporated and regenerated in a solution that contained 1 mM phosphate buffer (pH 12.3) and 20\% methanol. The MEKC separation was performed using an injection time of 275 s, a polarity switching time of 93 s, a phosphate buffer, (pH 12.3, 15 mM) containing 25 mM sodium dodecyl sulfate as separation electrolyte and a fused-silica capillary. The analysis takes about 6 min and gives satisfactory inter-day precision with respect to migration times and linear responses over the 80-900 ng/ml concentration range investigated for SC and UK. Intra-day RSDs (n=4 graphs) for the slopes of the calibration graphs were 4.86\% for SC and 3.50\% for UK. Inter-day RSDs for the slopes were 4.37\% for SC and 5.39\% for UK. Detection limits (S/N=3) were about 17 ng/ml for both compounds in human serum. A 1-ml volume of blood serum was necessary to do this determination.
This article was published in J Chromatogr A and referenced in Pharmaceutica Analytica Acta

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