Validated RP-HPLC method for quantification of doxazosin in human plasma: Application in a bioequivalence study

The aim of the present study was to validate a simple, sensitive, HPLC method of analysis of doxazosin in human plasma with fluorescence detection.

The validated method employed one-step direct protein precipitation with acetonitrile. Chromatographic separation was attained using a reverse-phase 250mm×4.6mm 5μ Hypersil® BDS C 18 column and the mobile phase consisted of 10mm sodium dihydrogen phosphate dihydrate (pH=3.0) and acetonitrile at a ratio of (65:35 v/v). The method was evaluated in terms of linearity, precision, accuracy, selectivity and stability as per standard guidelines. The total run time was about 4.5min which make this method suitable for high throughput analyses. This method was applied to the bioequivalence study of two doxazosin tablets in healthy human volunteers.

Good linear response was achieved over the range of 5.0-200ng/mL. The observed within- and between-day assay precision ranged from 0.64% to 14.73%; accuracy varied between 94.11% and 105%. The 90% confidence intervals for the ratio Cmax, and AUC 0-∞ of the test product over those of reference were within the acceptable range (0.8-1.25) for bioequivalence.

The developed method was simple and could be applied to therapeutic drug monitoring of doxazosin.