Simultaneous determination of paclitaxel-stearate(PTX-SA)and its bioactive me-tabolite paclitaxel(PTX)by RP-HPLC method / 药学实践杂志

ABSTRACT

Objective To establish a reversed phase high performance liquid chromatography method which couldb be ap-plied to determine the content of paclitaxel stearate(PTX-SA)and its bioactive metabolites paclitaxel(PTX)simultaneously. Methods Diazepam was chosen to be the internal standard and liquid-liquid extraction method was used for the preparation of sample solution with methyl tert-butyl ether.The two compounds were separated on Agilent Eclipse plus C18column (4.6 mm×250 mm,5 μm)with gradient elution by using mobile phase of acetonitrile(A)-methanol(B)-water(C),the detection wavelength was 227 nm and the column temperature was 30 ℃.The flow rate was 1 ml/min and the injection volume was 20 μl.Results The standard curve exhibited good linearity within the concentration of PTX-SA from 0.5-250 μg/ml(r=0.997 6)and PTX from 0.05-25 μg/ml(r=0.997 9),meanwhile the LOQ of PTX-SA and PTX were 0.15 and 0.05 μg/ml, respectively.The recoveries were both larger than 90% with RSD less than 5%.Moreover,the RSD of intra-day and inter-day assays were both less than 5%.Conclusion The method was simple,effective,stable and accurate which could be suitable for the determination of PTX-SA and PTX in pharmacokinetic study.