Liquid chromatography–tandem mass spectrometry method for determination of five antidepressants and four atypical antipsychotics and their main metabolites in human serum

The rapid and simple ultra performance liquid chromatography–tandem mass spectrometry method was developed and validated for simultaneous determination parent drugs: sertraline, fluoxetine, citalopram, paroxetine, venlafaxine, clozapine, olanzapine, quetiapine, risperidone, and their active and nonactive metabolites N-desmethylsertraline, norfluoxetine, desmethylcitalopram, didemethylcitalopram, N-desmethylvenlafaxine, O-desmethylvenlafaxine, N-desmethylclozapine, N-desmethylolanzapine, 2-hydroxyolanzapine and 9-hydroxyrisperidone in human serum. Precipitation of serum proteins was performed with a precipitation reagent consisting of 0.05% solution of ZnSO4·7H2O in acetonitrile/methanol (40:60, v/v). Alprenolol was used as an internal standard. Chromatographic separation was carried out on a BEH C18 column using gradient elution mobile phase A (2 mmol/L ammonium acetate, 0.1% formic acid in 5% acetonitrile, v/v/v) and B (2 mmol/L ammonium acetate, 0.1% formic acid in 95% acetonitrile, v/v/v). Electrospray in positive mode was used for ionization. Detection was performed on a triple–quadrupole tandem mass spectrometer by multiple reaction monitoring. Analysis time was 5 min. Drugs were separated into three groups with low, medium and high levels. Correlation coefficients of calibration curves were in the range 0.995–1.000. Coefficients of variation were 4.2–9.5% for intra-assay and 3.0–11.9% for inter-assay. Recoveries were 87.1–110% for intra-assay and 88.1–108.2% for inter-assay. The method was fully validated and can be successfully applied for routine analyses.

► The determination of nine antipsychotics in human serum.
► simultaneous measurement of parent drugs and main metabolites.
► This method can be used for fenotyping.
► Method was fully validated and can be successfully applied for routine analyses.