Original Articles: 2012 Vol: 4 Issue: 6
Development, validation and Comparative statistical Evaluation of HPLC and HPTLC method for determination of Alfuzosin in Human plasma
Rapid, precise , accurate, simple, selective, and sensitive HPLC and HPTLC methods for the determination of Alfuzosin in human plasma have been developed. The method utilizes simple protein precipitation as the sample preparation method in both the techniques. HPLC method was developed using HiQ sil C8 HS column, with mobile phase containing mixture of Acetonitrile: Sodium acetate buffer(0.04M) containing n-hexane sulphonic acid salt (0.005mM) (pH 4.0, adjusted with glacial aceteic acid) (55:45 v/v),at the flow rate of 1ml/min and detection was performed at 244nm. Retention times for Alfuzosin (Alfu) and the internal standard (IS) were 6.7 and 4.32 min, respectively. The calibration curve was linear ( r2>0.99) through the range of 25-45ng/ml. The mean recovery was found to be 96.94% for Alfuzosin. The HPTLC separation was carried out on the Aluminium plates precoated with silica gel 60 F254 using Toluene: Methanol: Triethylamine (7:3:0.2%v/v/v)as mobile phase and scanned at 244nm with camag TLC scanner. Quantification was achieved with HPTLC, over the concentration range of 1000 to 1800 ng/band with mean recovery of 97.07% for Alfuzosin. Retention factor (Rf) for Alfuzosin and the internal standard were 0.59 and 0.28 respectively.