High‐performance Liquid Chromatographic Ultraviolet Detection of Nilotinib in Human Plasma from Patients with Chronic Myelogenous Leukemia, and Comparison with Liquid Chromatography?Tandem Mass Spectrometry

摘要

Background: A method for determiningnilotinib concentration in human plasma isproposed using high-performance liquidchromatography and ultraviolet detection.Materials & Methods: Nilotinib and theinternal standard dasatinib were separatedusing a mobile phase of 0.5%Na2PO4H2O (pH 2.5)-acetonitrile-methanol(55:25:20, v/v/v) on a Capcell Pak C18MG II column (250 9 4.6 mm) at a flowrate of 1.0 ml/min, and ultraviolet measurementat 250 nm. Results: The calibrationcurve exhibited linearity over thenilotinib concentration range of 50–2,500 ng/ml at 250 nm, with relative standarddeviations (n = 5) of 7.1%, 2.5%,and 2.9% for 250, 1,500, and 2,500 ng/ml,respectively. The detection limit for nilotinibwas 5 ng/ml due to three blank determinations(q = 3). Conclusion: Thismethod was successfully applied toassaying nilotinib in human plasma samplesfrom patients with chronic myelogenousleukemia. In addition, we comparedthe results with those measured by liquidchromatography tandem mass spectrometry(LC-MS/MS) at BML, Inc. (a commerciallaboratory). A strong correlation wasobserved between the nilotinib concentrationsmeasured by our high-performanceliquid chromatographic method and thoseobtained by LC/MS-MS (r2 = 0.988,P 0.01).