DEVELOPMENT AND VALIDATION OF BIOANALYTICAL METHOD FOR SIMULTANEOUS ESTIMATION OF TENOFOVIR DISOPROXIL FUMARATE, LAMIVUDINE AND EFAVIRENZ IN FORMULATION BY ADVANCED DERIVATIVE UV-SPECTROSCOPY

A selective and sensitive derivative spectrophotometry method was developed to detect and quantify tenofovirdisoproxilfumarate, lamivudine and efavirenz in rat plasma. After liquid-liquid extraction the analyte were analysed. Mixture of methanol and phosphate buffer pH.5.0 (60:50 v/v) was used as solvent. The method was based on the measurement of absorbance at zero crossing points of three drugs 260.8nm, 271nm, and 247.6 nm respectively. The method was validated according to USFDA guidelines over the range of 100-3200 ng/mL for tenofovirdisoproxilfumarate and lamivudine and 200-6400ng/mL for efavirenz, with correlation coefficient values (R2) 0.999 for tenofovirdisoproxilfumarate and efavirenz, and 0.997 for lamivudine. The absolute recovery for analyte achieved from spiked plasma sample was consistent and reproducible. The % CV reported for precision and accuracy determinations across four validation runs (LLOQ, LQC, MQC and HQC) were less than 15. LLOQ was found to be 100ng/mL for tenofovirdisoproxilfumarate and lamivudine and 200ng/mL for efavirenz. Keywords: TenofovirDisoproxilFumarate, Lamivudine, Efavirenz, Derivative Spectrophotometry, BioanalyticalMethod Validation, USFDA guidelines.