Abstract
A simple, sensitive, Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) method was developed for the determination of Trimipramine maleate in human plasma using Opipramol dihydrochloride as internal standard (ISTD). The analyte and ISTD were isolated from plasma by protein precipitation method using methanol. The chromatographic separation was carried out on ZORBAX ECLIPSE XDB- C18 column (4.6 x 150 mm, 5 µm). The elution was done with isocratic mobile phase composed of 5mM ammonium formate with 0.1% formic acid and methanol in the ratio of 25:75(v/v) at a flow rate of 1.0 ml/min. Trimipramine and Opipramol were detected with proton adducts at m/z 295.20→100.10 and m/z 364.30→171.20 in multiple reaction monitoring (MRM) positive mode respectively. The retention time of Trimipramine and ISTD were found to be 1.67min and 1.48min respectively. The calibration curves were linear over concentration range of 0.1 to 100.1 ng/ml. The lower limit of quantification was found to be 0.1 ng/ml. The method was validated by EMEA (European medicines agency) guidelines for linearity, accuracy & precision, matrix factor, recovery, selectivity and stability. The validation parameters were found to be well within the acceptance limit.
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