SIMULTANEOUS DETERMINATION OF CLOZAPINE, NORCLOZAPINE, CLOZAPINE-N-OXIDE IN SERUM AND ORGANS

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DOI: https://doi.org/None
Issue: 
5
Year: 
2016

O.L. Romanova, E.S. Stepanova, S.S. Barsegyan, PhD; D.V. Sundukov, MD; V.V. Chistyakov, PhD Peoples’ Friendship University of Russia; 6, Miklukho-Maklay St., Moscow 117198, Russian Federation

Introduction. Clozapine has been reported to be much more commonly used in criminal activity in recent years. Procedures that can simultaneously measure the levels of clozapine and its metabolites in rat plasma and liver, kidney, heart, and lung homogenates by high performance liquid chromatography in combination with mass spectrometry (HPLC-MS/MS) have not been devised. Objective. To devise and validate a procedure for the simultaneous determination of clozapine and its metabolites in the serum and visceral organ homogenates to be used in toxicokinetic examinations. Material and methods. Clozapine, norclozapine, clozapine-N-oxide, and amitriptyline (Sigma Aldrich) were used. Clozapine and its metabolites were extracted from rat plasma via precipitation with methanol (serum) and acetonitrile (organ homogenates). Their analysis was carried out using the Aligent Technologies 6430 Triple Quad HPLC/MS system. Results. Conditions were selected for the mass-spectrometric detection of the study objects. MS/MS detection to achieve maximum sensitivity was optimized for quantitative analysis. Selectivity, reproducibility, and accuracy, detection limit, and extraction were assessed when validating the procedure devised. Conclusion. The developed and validated procedure was simple, rapid, and sensitive in simultaneously determining clozapine and its metabolites in bioobjects by liquid chromatography in combination with tandem mass spectrometry. It may be used to study the toxicokinetics of clozapine.
Keywords: 
clozapine
norclozapine
clozapine-N-oxide
determination
high performance liquid chromatography
mass spectrometry
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