ABSTRACT
A simple, sensitive and precise HPLC method for the quantitation of alfuzosin in human plasma has been developed and validated. Commercially available atenolol was used as an internal standard. After liquid-liquid extraction, alfuzosin and atenolol [I.S.] in human plasma were analyzed using mobile phase containing25% v/v acetonitrile and 75% v/v water [containing Iml/L triethylamine as peak modifier, pH adjusted to 2.5 with orthophosphoric acid]. Chromatographic separation was achieved on a BDS Hypersil-C18 column [50mm x 4.6 mm i.d., particle size 5 micro m] using isocratic elution at [flow rate 0.5 mL/min.] The peak was detected using a fluorescence detector programmed for 0.2 min at Ex 222 mm and Em 300 nm for atenolol and 2-4 min at Ex 265 nm and Em 380 nm for alfuzosin, and the total time for a chromatographic separation was 4 min. The validated quantitation ranges of this method were 0.1-25 ng/ml with coefficients of variation between 1.6 - 4.8%. Mean recoveries were 98.0 +/- 0.9%. The within and between batch precision was 2.74 - 3.28% and 2.65 - 2.77%, respectively. The within and between batch relative error [bias] were -7.1 - [-0.3]% and -1.60 - 2.46%, respectively. Stability of alfuzosin in plasma was >/= 94.9%, with no evidence of degradation during sample processing and 30 days storage in a deep freezer at -70 +/- 5°C. The absolutes extraction recoveries of drug from plasma was >/= 98%. This validated method is sensitive and simple with between-batch precision of