ESTIMATION OF TERIFLUNOMIDE ALONG WITH CONCOMITANT DRUGS IN DIFFERENT BIOLOGICAL MATRICES USING LC-MS/MS

A simple, rapid and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) assay method has been developed and fully validated for the quantification of Teriflunomide (TFM), in different biological matrices like human plasma, rat plasma, rabbit plasma and dog plasma and as well as their serums. The analytes were extracted from 200 mu L aliquots of each plasma/serum via protein precipitation using acetonitrile. The extracted samples were chromatographed on a XBridge C18 column by using acetonitrile and 5 mM ammonium formate (pH 9.0) as the mobile phase in gradient mode at a flow rate of 1.0 mL/min. The calibration curves obtained were linear (r(2) = 0.98) over the concentration range of 1.0-4000 ng/mL. Detection was carried out on a triple quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) with an electro spray ionization (ESI) mode using the respective m/z 268.8 -> 82 for TFM and 281 -> 86 for IS. The method was validated interms of specificity, accuracy, precision and limit of quantification. The results of the intra-and inter-day precision and accuracy studies were well within the acceptable limits. The analyte was found to be stable up to 72 hrs in the matrices. Along with concomitant drug analysis the method is precise and sensitive enough for its intended purpose and can be applied to clinical samples