A new sensitive and specific liquid chromatography-tandem mass spectrometry methodology was needed for the estimation of alpelisib in plasma samples. Drug components were extracted by liquid-liquid extraction technique utilizing ethyl acetate. Liquid chromatographic system was processed with Zorbax C18 (50 × 4.6 mm, 5 µ) reverse phase analytical column with isocratic mobile solvent system consisting of acetonitrile and 0.1% formic acid in the ratio of 90:10 (%V/V). Analytes were detected on mass system equipped with triple quadrupole system and electrospray ionization, functioning in multiple reaction monitoring, with the transitions of m/z 442.15 → 70.06, m/z 409.14 → 391.13 for alpelisib, dapagliflozin, respectively, in the positive ionization mode. Method has given more than 90.0% recovery values and accuracy values were present in between −4.32% and 4.37% of relative error. All the relative standard deviation findings were less than 4.21%. Alpelisib pharmacokinetic parameters were determined from the curve obtained by taking time on X-axis and concentration of plasma on Y-axis. Alpelisib has mean Tmax of 2.0 ± 0.21 and AUC0 →α , AUC0t and mean Cmax, for test formulation is 3,624.21 ± 315, 3,075.20 ± 316, and 269 ± 12.17, respectively.
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