Ex vivo conversion of prodrug prothionamide to its metabolite prothionamide sulfoxide with different extraction techniques and their estimation in human plasma by LC-MS/MS.

Vikas Trivedi, Vivek Upadhyay, Gaurang Shah, Manish Yadav, Pranav S Shrivastav, Mallika Sanyal

Chemistry Department, Kadi Sarva Vishwavidyalaya, Sarva Vidyalaya Campus, Sector 15/23, Gandhinagar 382015, India.

Bioanalysis 2013 Jan

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The present work reports an ex vivo conversion study of prothionamide to its active metabolite prothionamide sulfoxide in human plasma during sample preparation by three conventional extraction techniques. The chromatography was done on a Hypersil™ Gold C18 (50 × 2.1 mm, 3.0 µm) column using 0.1% acetic acid and acetonitrile (20:80, v/v) as the mobile phase. Quantitation of the analytes was done by MS/MS in the positive ionization mode. The method was validated over a wide concentration range of 30 to 6000 ng/ml for prothionamide and 50 to 10,000 ng/ml for prothionamide sulfoxide. The recovery for both the analytes was greater than 89%. Stability was extensively validated under different storage conditions. The extraction protocol was optimized using acetonitrile as protein precipitant for their simultaneous determination in human plasma by LC-MS/MS. The method was applied to a bioequivalence study of 250 mg prothionamide tablet formulation in 14 healthy Indian subjects.

Citation

Vikas Trivedi, Vivek Upadhyay, Gaurang Shah, Manish Yadav, Pranav S Shrivastav, Mallika Sanyal. Ex vivo conversion of prodrug prothionamide to its metabolite prothionamide sulfoxide with different extraction techniques and their estimation in human plasma by LC-MS/MS. Bioanalysis. 2013 Jan;5(2):185-200