TITLE:
Development and Validation of LC-MS/MS Method for the Quantification of Chiral Separated R-Bicalutamide in Human Plasma
AUTHORS:
N. T. Ramarao, S. Vidyadhara, R. L. C. Sasidhar, B. Deepti, R. Surendra Yadav
KEYWORDS:
LC-MS/MS; R-Bicalutamide; Topiramate; Validation; Human Plasma
JOURNAL NAME:
American Journal of Analytical Chemistry,
Vol.4 No.2,
February
17,
2013
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.