METHOD DEVELOPMENT AND VALIDATION FOR THE QUANTITATION OF TRIFLURIDINE IN HUMAN PLASMA BY USING LC-MS/MS TECHNIQUE

The main objective of the current study was to develop a highly sensitive LC-MS/MS (liquid chromatography-tandem mass spectrometry) technique for the quantification of trifluridine (TFD) in human plasma. Chromatography was processed on Phenomenex-RP-C18 (5 µm, 250 mm × 4.6 mm) column comprising acetonitrile, methanol and 5 mM ammonium formate (45:40:15, v/v) as a mobile phase, infused at a flow of 0.8 ml/min. TFD and internal standard ?-thymidine (TMD) were separated from 150 ml plasma utilizing Phenomenex cartridges. Quantification of the [M+H] +ion of analyte was done with MRM (multiple reaction monitoring) modes in LC-MS/MS utilizing electro-spray ionization. The parent and product ions transitions were monitored atm/z 297-181 and m/z² 41–42 for TFD and TMD respectively. The linearity was processed in the concentration range of 5.0–2000.0 ng/ml with regression coefficient (r²) = 0.999. Method intra and inter-batch precision findings were <3.33% and intra and inter-batch accuracy findings at three different concentrations were within 97.34% to 103.74%. Matrix effects were calculated by determining the %CV values for High and Low QC samples and was found to be 1.65% and 0.73% respectively. The % change for all the re-injected QC solutions was ≤7.45. The method was specific, accurate, precise and stable for more time and the technique was successfully applied for the quantification of trifluridine in human plasma samples.