AJAC  Vol.4 No.2 , February 2013
Development and Validation of LC-MS/MS Method for the Quantification of Chiral Separated R-Bicalutamide in Human Plasma
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-1with a 100 μL plasma sample. The concentrations of eight working standards showed linearity between 20 to 3200 ng·mL-1(r2≥ 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.
Cite this paper: N. Ramarao, S. Vidyadhara, R. Sasidhar, B. Deepti and R. Yadav, "Development and Validation of LC-MS/MS Method for the Quantification of Chiral Separated R-Bicalutamide in Human Plasma," American Journal of Analytical Chemistry, Vol. 4 No. 2, 2013, pp. 63-76. doi: 10.4236/ajac.2013.42009.

[1]   B. J. Furr, “The Development of Casodex (Bicalutamide): Preclinical Studies,” European Urology, Vol. 29, Suppl. l 2, 1996, pp. 83-95.

[2]   S. Lee, Y. J. Chung, B. H. Kim, J. H. Shim and S. H. Yoon, “Comparative Pharmacokinetic Evaluation of Two Formulations of Bicalutamide 50-mg Tablets: An Open Label, Randomized-Sequence, Single-Dose, Two-Period Crossover Study in Healthy Korean Male Volunteers,” Clinical Therapeutics, Vol. 31, No. 12, 2009, pp. 3000 3008. doi:10.1016/j.clinthera.2009.12.004

[3]   R. NageswaraRao, A. NarasaRaju and D. Nagaraju, “An Improved and Validated LC Method for Resolution of Bicalutamide Enantiomers Using Amylase Tris-(3,5-Ime thylphenylcarbamate) as a Chiral Stationary Phase. Journal of Pharmaceutical and Biomedical Analysis, Vol. 42, No. 3, 2006, pp. 347-353. doi:10.1016/j.jpba.2006.04.014

[4]   D. McKillop, G. W. Boyle, I. D. Cockshott, D. C. Jones, P. J. Phillips, et al. “Metabolism and Enantioselective Pharmacokinetics of Casodex in Man,” Xenobiotica, Vol. 23, No. 11, 1993, pp. 1241-1253. doi:10.3109/00498259309059435

[5]   F. Boccardo, A. Rubagotti, G. Conti, D. Potenzoni, A. Manganelli and D. Del Monaco, “Exploratory Study of Drug Plasma Levels during Bicalutamide 150 mg Therapy Co-Administered with Tamoxifen or Anastrozole for Prophylaxis of Gynecomastia and Breast Pain in Men with Prostate Cancer,” Cancer Chemotherapy and Pharmacology, Vol. 56, No. 4, 2005, pp. 415-420. doi:10.1007/s00280-005-1016-1

[6]   US Food and Drug Administration, “Guidance for Indus try. Bioavailability and Bioequivalence Studies for Orally Administered Drug Products-General Considerations,” 2009.

[7]   “ICH Text on Validation of Analytical Procedures, Q2A (R1),” International Conference on Harmonization, IF PMA, Geneva, 1994.

[8]   Anonymous, “Guide for Industry, Bioanalytical Method Validation,” US Department of Health and Human Services, Food and Drug Administration (FDA), Silver Spring, 2001.