BIOANALYTICAL METHOD DEVELOPMENT & VALIDATION OF FULVESTRANT IN RAT PLASMA BY USING LIQUID CHROMATOGRAPHY AND TANDEM MASS SPECTROMETRY AS PER USFDA GUIDELINES
A rapid, specific and sensitive high-performance liquid chromatography and electro spray tandem mass spectrometry (LC-MS/MS) method was developed and validated for estimation of Fulvestrant in Rat plasma using liquid-liquid extraction. The separation and quantification of Fulvestrant were achieved by reverse-phase chromatography on a YMC Pack ODS- AM column (150mm x 4.6mm x 5µm) with isocratic elution at a flow rate of 1mL/min using mobile phase consisting of 2mM Ammonium acetate in water and Methanol(10:90). Fulvestrant-D3 was used as internal standard. A heated electro spray ionization interface in negative polarity mode was used. The volume of plasma about 100 µl was used. The method was validated over the concentration range of 0.322- 35.823ng/ml. The within-batch and between-batch accuracy and precision were found to be within acceptance criteria at all QC levels. The recovery was found to be 85.94% and 96.67% for Fulvestrant and Fulvestrant-D3 respectively. The matrix effect from rat plasma was found to be within the acceptance criteria at all QC levels. The chromatographic run time was 5 min.