Development and Validation of LC-MS/MS Method for the Quantification of Chiral Separated R-Bicalutamide in Human Plasma
- 1 Alkem Research Center, Navi Mumbai, India
- 2 Chebrolu Hanumaiah Institute of Pharmaceutical Sciences, Guntur, India
- 3 Chebrolu Hanumaiah Institute of Pharmaceutical Sciences, Guntur, India
- 4 Vignan Pharmacy College, Guntur, India
- 5 GVK Biosciences, Hyderabad, India
Abstract
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 v/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 quantifica tion of R-bicalutamide in human plasma was developed and validated can be used in the bioequivalence study of this drug.
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