Development and Validation of LC-MS/MS Method for the Quantification of Chiral Separated R-Bicalutamide in Human Plasma

N. T. Ramarao, S. Vidyadhara, R. L. C. Sasidhar, B. Deepti, R. Surendra Yadav
2013 American Journal of Analytical Chemistry  
A simple, rapid, specific and precise liquid chromatography-tandem mass spectrophotometric (LC-MS/MS) method was developed and validated for the quantification of chiral separated R-bicalutamide from S-bicalutamide, in human plasma. Topiramate (TPM) was used as internal standard, added to plasma sample prior to extraction using t-butyl methyl ether (TBME). Chromatographic separation was achieved on CHIRALPAK AD-RH column (150 mm × 4.6 mm, 5 μm) with acteonitrile: 0.1% formic acid Buffer (50:50
more » ... /v) as an isocratic mobile phase with a flow rate of 1.0 mL·min −1 . Quantitation was performed by transition of 429.0 → 255.0 (m/z) for R-bicalutamide and 338.1 → 77.8 (m/z) for topiramate. The lower limit of quantitation was 20 ng·mL −1 with a 100 μL plasma sample. The concentrations of eight working standards showed linearity between 20 to 3200 ng·mL −1 (r 2 ≥ 0.9990). Chromatographic separation was achieved within 6 min, compared to the 15 min of previous methods. The average extraction recoveries of 3 quality control concentrations were 98.56% for R-bicalutamide and 92.42% for topiramate. The coefficient of variation was ≤15% for intra-and inter-batch assays. Therefore a rapid, specific and sensitive LC-MS/MS method for the quantification of R-bicalutamide in human plasma was developed and validated can be used in the bioequivalence study of this drug. N. T. RAMARAO ET AL. Data Collection and Integration The data were gathered and processed with Analyst version 1.5 data collection and integration software on a Microsoft windows compatible computer. Bioanalytical Method Validation ICH guidelines and USFDA guidelines were followed for
doi:10.4236/ajac.2013.42009 fatcat:ah5bdihnmnamthsdwomfq5xbrm