DETERMINATION OF TRAZODONE IN HUMAN PLASMA BY REVERSED-PHASE LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY WITH ELECTROSPRAY IONISATION

Abstract

Objective: The development and validation of LC-MS/MS method for quantification of Trazodone (a serotonin antagonist and reuptake inhibitor (SARI), which is a second generation antidepressant compound belonging to the class of phenyl piperazine) in human plasma is described.

Methods: The method involves protein precipitation (extraction) using Trazodone d6 as an internal standard (IS). Chromatographic separation is achieved on Zorbax eclipse XDB C8 150×4.6 mm, 5 μm column with a mobile phase consisting of 2 mM ammonium formate (pH 3.0) and methanol (30:70 v/v) at a flow rate of 1.0 mL / min and the total run time was 2.5 minute. Detection was carried out by AB Sciex API 3200 tandem mass spectrometer using positive electro-spray ionization mode by multiple reactions monitoring method at m/z 372.00→176.10 and 378.20→182.10 for Trazodone and Trazodone d6 (ISTD) respectively with dwell times of 300 msecs for each of the transitions.

Results: The standard curve was linear from 5.203 ng / mL to 3025.166 ng / mL with goodness of fit (r²) greater than 0.990 observed during the method validation batches. This assay allows quantification of Trazodone at a concentration as low as 5 ng / mL in human plasma. The observed mean recovery was 88% for the drug.

Conclusions: The method described here is found to be simple, cost effective and suitable for the use in bioequivalence and bioavailability studies.