Abstract

SENSITIVE AND RAPID HIGH PERFORMANCE LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY METHOD (LC/MS/MS) FOR THE ESTIMATION OF CLOFARABINE IN RABBIT PLASMA

Dr. V. Kiran Kumar, Dr. VVSS. Appala Raju, Dr. Macharla Venkata Ramana, Dr. T. Shyam. Principal, K. N. Rajanikanth and *Dr. N. Appala Raju

Abstract

A simple, sensitive and fast throughput liquid chromatography tandem mass spectrometry (LC-MS/MS) method has been developed for the simultaneous estimation of Clofarabine in rabbit plasma. The method involved Liquid-Liquid Extraction of the analyte from rabbit plasma. The chromatographic separation was achieved on a Sunfire C18, (150×4.6mm and 5μm particle size) analytical column using isocratic mobile phase, mobile phase was found in acetonitrile: water: formic acid (75:25:0.1) % v/v/v respectively at a flow-rate of 1.0 mL/min with 90% flow splitting. The parent→product ion transitions m/z 541.9 → (443.9 + 461.9), for Clofarabine and 299.250>109.050 for internal standard respectively were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring (MRM) positive ion mode. The method was validated over the concentration range of 0.092-16.937 ng/mL for clofarabine. The mean recovery values for analyte from spiked plasma samples were reproducible. The method was rugged and rapid with a total run time of 4.0 minutes.

Keywords: Clofarabine, LC-MS/MS, pharmacokinetics, plasma, quantitation.