A simple and sensitive LC-MS/MS method for the quantitation of sertraline and N-desmethylsertraline in human plasma.

A simple and sensitive LC-MS/MS method for the simultaneous quantification of the second-generation antidepressant sertraline, and its active metabolite, N-desmethylsertraline, in human plasma was developed and validated. The analytes were extracted from 200 µL human plasma using a simple protein precipitation method. A gradient elution mode of water and 0.1% formic acid and acetonitrile was used for chromatographic separation on a Poroshell EC-C18 column (3 × 100 mm, 2.7 µm) at a flow rate of 0.450 mL/min. MS/MS analysis was performed in positive ionization mode using the transitions of m/z 306.1 → 159.1, 309.1 → 275.2, 292.1 → 159.1, and 296.2 → 279.0 for sertraline, sertraline-d3, N-desmethylsertraline, and N-desmethylsertraline-d4, respectively. The calibration curves for sertraline and N-desmethylsertraline in human plasma ranged from 2.50-320 ng/mL and 10.0-1280 ng/mL, respectively, with correlation coefficients (r) of ≥ 0.9992. The intra- and inter-assay precisions for both analytes at four concentrations (LLOQ, QCL, QCM, and QCH) ranged between 2.2% and 12.2%, respectively, while their accuracies ranged between 92.0 and 111.7%, with the exception of LLOQ, which ranged between 84.3 and 106.0%. The mean percentage recovery and process efficiency at three concentrations (QCL, QCM, and QCH) was 94.2 and 87.9% for sertraline, and 95.7 and 95.2% for N-desmethylsertraline, respectively. Both analytes were stable in plasma at room temperature for 2 h, at -80 °C for 28 days and through three freeze/thaw cycles. This method was successfully applied to a clinical study investigating the use of sertraline during pregnancy.