Simple and Rapid HPLC Method for the Determination of Alfuzosin in Human Plasma

Ashok K. Shakya, Tawfiq A. Arafat, Ahmad Abuawaad, Hamza Al-Hroub, Munther Melhim


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 containing 25 % v/v acetonitrile and 75 % v/v water (containing 1ml/L triethylamine as peak modifier, pH adjusted to 2.5 with orthophosphoric acid). Chromatographic separation was achieved on a BDS Hypersil-C18 column (50 mm × 4.6 mm i.d., particle size 5μm) 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 nm 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 ± 5oC. The absolutes extraction recoveries of drug from plasma was ≥ 98%. This validated method is sensitive and simple with between-batch precision of ≤ 3%. The method can be used for bioequivalence and pharmacokinetic studies of alfuzosin formulations.


Alfuzosin, Assay; HPLC; Plasma

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