BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF LEVOCETIRIZINE IN BLOOD PLASMA BY USING RP-HPLC

A rapid, selective, precise and sensitive reverse phase high-performance liquid chromatography method was developed for the quantitative estimation of Levocetirizine Dihydrochloride (LD) in human plasma and pharmaceutical dosage form. Extraction of drug from plasma was done by employing optimized liquid-liquid extraction procedure. The sample was analyzed using acetonitrile: methanol: 20mM ammonium acetate buffer pH-5 (25:55:20 % v/v/v) as mobile phase. Chromatographic separation was achieved on Thermo C-18 column (4.6 x 250mm, 5μ particle size) as stationary phase using isocratic elution (at a flow rate of 1 ml/min). The peak was detected using UV-PDA detector set at 232 nm and the total time for a chromatographic separation was 8 min. The calibration curve obtained was linear (r2= 0.9998) over the concentration range of 2-10 μg/ml. Method was validated for precision, robustness and recovery. The limit of detection and limit of quantitation was 0.0057 and 0.174 µg/ml respectively. There was no significant difference between the amount of drug spiked in plasma and the amount recovered and plasma did not interfere in estimation. Thus, the proposed method is suitable for the analysis of LD in tablet dosage forms and human plasma. Keywords: RP-HPLC, Levocetirizine Dihydrochloride, Human plasma, Liquid-liquid extractio