Determination of Itraconazole in Bulk and Human Plasma by High Performance Liquid Chromatographic-Electrospray Ionization-Tandem Mass Spectrometry

AbstractA fast, sensitive and selective high performance liquid chromatography-electron spray ionization-tandem mass spectrometry method (HPLC-ESI-MS/MS) was produced and validated for the determination of itraconazole (ITC) in bulk and human plasma on an intersil, ODS-3, C18 column using a mobile phase consisting of acetonitrile: ammonium formate (80:20, v/v)(pH 3.5) with flow rate 0.5 ml/min. Detection was carried out on a triple quadrupole tandem mass spectrometer in multiple reaction monitoring (MRM) mode utilizing m/z 705.3 → m/z 392.3 for ITC and m/z 559.3 → m/z 440.3 for atorvastatin as an internal standard (IS). Different parameters were discussed, e.g: type of columns, mobile phase pH, mobile phase composition, rate of fragmentation, and rate of collision. .The mass detection was performed using electrospray ionization in the positive mode (ESI +). The method was linear over the concentration range 4- 1000 ng/ml for ITC with r2 = 0.99 . The limit of detection and limit of quantification were 1.17...