A sort of validated Bioanalytical method developed for the estimation of Etoposide and Cisplatin in rat plasma by using two different advanced liquid chromatographic techniques like HPLC and UPLC and its application in bioequivalence studies

Abstract

An easy, quick, precise, active and reproducible reverse method high performance liquid chromatography (RP-HPLC), as well as Ultra pressure liquid chromatography (UPLC) unique technique, was developed for the bioanalytical method of Etoposide and Cisplatin with Oxaplatin as an internal standard. Chromatographic analysis was performed using HPLC was Waters Alliance e-2695, and UPLC was Waters Acquity UPLC by using x-bridge phenyl 150x4.6mm, 3.5µ column and therefore the mobile phase containing 0.1% triethylamine and acetonitrile at a ratio out of 60:40 v/v. The flow rate was 1 ml/min, and analytes were detected at 283 nm employing a photodiode array detector at ambient temperature in both liquid chromatographic systems. The proposed biological method was proved in both HPLC and UPLC with USFDA guidelines. USP tailing is 1.1, 1.02 for etoposide and 1.16, 1.05 for Cisplatin in HPLC and UPLC. USP plate count is 4794, 3884 for Etoposide and 9289, 14487 for Cisplatin. The calibration curve under accumulation set of 5-100 ng/ml of etoposide and 10-200 ng/ml of Cisplatin in both HPLC and UPLC. The accuracy of low, middle and high-level quality control samples are taken in 50%, 100% and 150% levels. Stability study was administered altogether conditions are bench top, wet extract and auto sampler, freeze-thaw, short term stability and in long term stability. This is the advanced technique established to the straightforward, economical, suitable, precise, accurate and stable method for the analysis of Etoposide and Cisplatin and study of its stability and pharmacokinetic studies using rat plasma.