A VALIDATED LCMS METHOD FOR THE ANALYSIS OF ISOPROTERENOL – A β ADRENORECEPTOR AGONIST IN SPIKED HUMAN PLASMA

A simple, sensitive, and rapid Liquid Chromatography-Mass Spectroscopy method was developed and validated for the quantification of Isoproterenol in human plasma using Dobutamine as an internal standard. The method utilized simple liquid-liquid extraction using a mixture of diethyl ether and dichloromethane for the sample preparation involved prior to LCMS analysis. The analytes were chromatographed on Prontosil ODS C18 Column with isocratic elution using methanol, Acetonitrile and Tri ethyl-amine in the ratio of 60:25:15 (v/v) at pH 6.3 as the mobile phase at a flow rate of 0.9 mL/min and the UV detector response of the column eluents were recorded at a wavelength of 248 nm. Quantification was performed in multiple-reaction-monitoring mode with the ion transitions m/z 212.19 → 135.21 for Isoproterenol, m/z 302.19 → 107.05 for Dobutamine. Good linearity was obtained in the range of 0.50–300 ng/mL (r² = 0.999). The method was fully validated with accuracy, precision, matrix effects, recovery and stability. The results of the stability study confirm that the method was found to be stable. The validated data have met the acceptance criteria in FDA guidelines. This study could be readily applied in therapeutic drug monitoring of Isoproterenol in patients receiving such drug combinations.