ALA VIJAY POSTER
Poster Title: APPLICATION OF ENANTIOSELECTIVE TECHNIQUE FOR SIMULTANEOUS ESTIMATION OF SIBUTRAMINE AND ITS ACTIVE METABOLITES IN HUMAN PLASMA BY LIQUID CHROMATOGRAPHY-ELECTROSPRAY IONIZATION TANDEM MASS SPECTROMETRY
Author(s) Name and Affiliation:
Vijay Gokarn, Bhalerao Khairnar, Sasikumar Menon
Institute for Advanced Training & Research in Interdisciplinary Sciences
(Therapeutic Drug Monitoring Laboratory) &
Ramnarain Ruia College,
A new, automated, rapid, and sensitive chiral liquid chromatography – tandem mass spectrometric method is developed and validated to separate and quantify R & S (±) enantiomers of sibutramine and its active metabolites desmethyl sibutramine (M1) and di-desmethyl sibutramine (M2) using propranolol as the internal standard in human plasma samples for pharmacological studies. The method involves protein precipitation followed by solid phase extraction from plasma. The enontiomeric separation of sibutramine (±) and its active metabolites (±) was performed using HPLC on Chiral AGPTM column (100 mm x 4.0 mm, id.) 5µm particle size, analyzed by mass spectrometry in the multiple reaction monitoring mode. The method has an LOQ of 250 pg/mL for all analytes with a chromatographic run time of 12.0 min. The method is validated with a linear range of 250 pg/mL - 40000 pg/mL for sibutramine and its active metabolite M1 and M2, with good correlation coefficient (r = 0.9990). The method is sensitive and specific having intra- and inter- assay precision at three different levels covering the entire linearity range. This method validated as per the ICH guidelines significantly reduces analysis time and this automated method has been successfully applied for bioequivalence study.