A Rapid and Sensitive Bio Analytical RP-HPLC Method for Detection of Docetaxel: Development and Validation

Abstract:

Objective: A rapid and selective high-performance liquid chromatographic (HPLC) method has been developed and validated for Docetaxel anhydrous (DTX) using Ketoconanzole (KCZ) as an internal standard in biological fluids. Methods: The analyte was extracted from human plasma by liquid-liquid extraction method using acetonitrile. The analysis was carried out on Licrosphere IV, C8 column (LC–GC Chromatography Solutions Pvt. Ltd, Mumbai, and India) (4.6×250 mm) with isocratic elution using a mobile phase composition of 0.2% triethylamine (pH 6.4 with orthophosphoric acid) (45%) and Acetonitrile (55%). The flow rate was 1.5ml/min with absorbance wavelength of 230 nm. Results: The developed method was found to be linear over the range of 100- 2500 ng/mL with correlation coefficients of about 0.999. The limit of quantification for docetaxel was found to be 100 ng/mL. The limit of detection was found to be 50 ng/ mL. The intra- and inter-day precision was determined to be <5% and the accuracy was 99–105%. The overall recovery for docetaxel was about 89.0% -91.0%. The total analysis time was only 10.0 min. The method was successfully applied for determination of DTX in rat plasma, to establish the pharmacokinetic profile of a marketed formulation. Conclusion: A simple, sensitive, rapid, accurate and prudent RP-HPLC method was developed, validated and applied for the Docetaxel anhydrous.

Key words: Docetaxel anhydrous, Ketoconazole, Bio-analytical method, HPLC, Pharmacokinetic study.