Extraction and RP-HPLC determination of taxol in rat plasma,cell culture and quality control samples

摘要

A rapid,sensitive,selective and validated reverse phase high-performance liquid chromatography(RP-HPLC)method for the estimation of paclitaxel in micro-sample of rat plasma and in culture of cancer cells was performed in this study.The mobile phase consisted of an optimized mixture of methanol:water:trifluroacetic acid(80:20:0.1,v/v/v).Column elution at a flow rate of 1 mL/minute with UV detection at 225 nm at room temperature was used.The RP-HPLC method was successfully applied for the determination of paclitaxel in plasma samples and in culture of cancer cells with nano-quantity of estimation.The validation studies were performed in accordance with the International Conference on Harmonization(ICH)guidelines.The intra-and inter-day precision showed that the coefficients of variation ranged from 1.07%to 4.27%at different levels of concentrations.To the best of our knowledge,this study also reported for the first time the optimization of different solvents for effective extraction of paclitaxel wherein tert.-butyl methyl ether(TBME):diethyl ether(DEE)in 50:50 v/v composition was found most efficient with extraction efficiency ranging between 77.99%and 91.74%and between 76.14 and 93.66%in the plasma and cell culture,respectively.This proposed method was successfully applied to study the pharmacokinetics of paclitaxel and the influence of verapamil and all-trans retinoic acid(atRA)on paclitaxel pharmacokinetics in rat models.This proposed method might emerge as a valuable aid in the laboratory monitoring of paclitaxel in a variety of in vitro as well as in vivo scenarios.