THE ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THE SIMULTANEOUS ESTIMATION OF FLOUXETINE & OLANZAPINE BY USING RP- HPLC

A new method was established for simultaneous estimation of Fluoxetine and Olanzapine by RP-HPLC method. The chromatographic conditions were successfully developed for the separation of Fluoxetine and Olanzapine by using Agilent C18 column (4.6×150mm)5µ, flow rate was 1ml/min, mobile phase ratio was (60:40 v/v) methanol: Phosphate buffer pH 3.0, detection wavelength was 256 nm. The instrument used was WATERS HPLC Auto Sampler, Separation module 2695, photo diode array detector 996, Empower-software version-2. The retention times were found to be 2.327 mins and 4.342 mins. The % purity of Fluoxetine and Olanzapine was found to be 99.84% and 100.14% respectively. The system suitability parameters for Fluoxetine and Olanzapine such as theoretical plates and tailing factor were found to be 2937, 1.3 and 2300 and 1.3, the resolution was found to be 4.6. The analytical method was validated according to ICH guidelines (ICH, Q2 (R1)). The linearity study of Fluoxetine and Olanzapine was found in concentration range of 50µg-250µg and 10µg-50µg and correlation coefficient (r2) was found to be 0.999 and 0.999, % recovery was found to be 100.07% and 100.06%, %RSD for repeatability was 0.3 and 0.39, % RSD for intermediate precision was 0.1 and 0.16 respectively. The precision study was precision, robustness and repeatability. LOD value was 3.041 and 3.08 and LOQ value was 9.79 and 10.37 respectively. Keywords: Fluoxetine, Olanzapine, methanol and Phosphate buffer