A Sensitive Bioanalytical Method Development And Validation Of Pazopanib In Human Plasma By LC-ESI-MS/MS

A simple, sensitive and specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of Pazopanib in human plasma using Verapamil as an internal standard (IS). Chromatographic separation was performed on Water X Bridge c18 2.1x100 column with an isocratic mobile phase composed of acetonitrile and 0.2% formic acid in2mM Ammonium formate in the ratio of (70:30 v/v), at a flow-rate of 0.250mL/min. Pazopanib and verapamil were detected with parent ions at m/z 438.15 to 357.14 and the daughter mass was found to be 353.55 to 342.13 in multiple reaction monitoring (MRM) positive mode respectively. The protein precipitation method was used to extract the drug and IS. The method was validated over a linear concentration range of 1.0-1000.0 ng/mL with a correlation coefficient (r2) >= 0.9994. This method demonstrated Intra and inter-day Precision within 0.5 to 3.0 and 0.6 to 3.3 % and Accuracy within 95.55 to 106.32 and 94.62 to 112.6%. Pazopanib was found to be stable throughout Long-term stability studies, benchtop, and postoperative stability studies.