DEVELOPMENT OF A LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY METHOD FOR THE DETERMINATION OF INDIRUBIN-5-NITRO-3′-MONOXIME, A NOVEL CDK INHIBITOR, IN RAT PLASMA

• Published in: Journal of liquid chromatography & related technologies 2012-09, Vol.35 (15), p.2175-2187
• Main Authors: Kim, Hyun Mi, Kim, Yong-Chul, Kim, Jae Il, Choi, Yong Seok, Kim, Yoon Gyoon
• Format: Article
• Language: English
• Subjects: Analysis; Biological and medical sciences; cyclin-dependent kinase inhibitor; General pharmacology; indirubin-5-nitro-3′-monoxime; LC-MS/MS; Medical sciences; pharmacokinetics; Pharmacology. Drug treatments; plasma; rat
• ISSN: 1082-6076; 1520-572X
• DOI: 10.1080/10826076.2011.629389

A simple and rapid liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantitation of indirubin-5-nitro-3′-monoxime (5-NIO) in rat plasma using 5-chloro-5′-chloro-indirubinoxime as the internal standard. The sample preparation involved protein precipitation using acetonitrile. Separation was achieved on an Xterra column C 18 (2.1 mm × 50 mm, i.d. 5 µm) using a mixture of 0.1% formic acid-water/acetonitrile (18:82, v/v) as the mobile phase at a flow rate of 300 µL/min. Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization and multiple reaction monitoring (MRM) in negative ion mode. The total chromatographic run time was 2.5 min, and the calibration curves of 5-NIO were linear (correlation coefficient range: 0.9989-0.9999) over the concentration range of 0.2-2000 ng/mL in rat plasma. Also, the detection limit of the method was 0.05 ng/mL and it showed good accuracy and precision within the concentration range of 0.45-376 ng/mL. This validated method was successfully applied to estimating the pharmacokinetic parameters of 5-NIO following intravenous administration of 5-NIO to rats.