PEER-REVIEWED ABSTRACT PRESENTED AT THE 2014 ANNUAL MEETING OF THE INTERNATIONAL ASSOCIATION OF FORENSIC TOXICOLOGISTS
PTD2 DEVELOPMENT AND VALIDATION OF AN LCMS/MS APPROACH FOR IDENTIFICATION AND QUANTIFICATION OF NEUROLEPTICS IN HUMAN WHOLE BLOOD, BLOOD PLASMA, AND SERUM Deborah Montenarh1,2, Markus Hopf1, Peter H. Schmidt1 and Andreas H. Ewald1 1Institute of Legal Medicine, Saarland University, 66421, Homburg, Saar, Germany 2Kantonsspital Aarau, Institut of Legal Medicine, Forensic Toxicology, 5000 Aarau, Switzerland
Email: [email protected]
Aims: An LC-MS/MS multi-analyte approach based on a simple liquid-liquid-extraction (LLE, according to Maurer et al., JCB, 2002) was developed and validated for fast target screening and quantification of 33 neuroleptics in whole blood, plasma, and serum. The method is based on a corresponding approach for quantification of benzodiazepines, z-drugs, and antidepressants (Montenarh et al., ABC, 2014 a, b).
Methods: Whole blood, plasma, or serum (500 μL each) were extracted twice at pH 7.4 and at pH 10 with ether/ethyl acetate (1:1). Separation, identification, and quantification were performed with a Shimadzu Prominence LC 20 system coupled to an AB Sciex 3200 Q- TRAP(ESI+). The method was validated according to GTFCh guidelines. For accuracy and precision, full calibration was performed with ranges from subtherapeutic to toxic concentrations.
Results: Selectivity problems could not be observed, but matrix effects from 47 – 169 % for fluphenazine, flupenthixol, fluspirilene, hydroxy-risperidone, perazine, promethazine, pipamperone, prothipendyl, olanzapine, ziprasidone, and zotepine in all samples. For quality control low, recovery ranged from 32-112 %, process efficiency from 31-132 % and for quality control high recovery from 42-142 %, process efficiency from 29-154 %. Accuracy and precision were within the accepted range of }15% ( }20% near LOQ) except for bromperidol, clozapine, flupenthixol, fluspirilene, moperone in all biosamples, risperidone in whole blood and serum, quetiapine and zotepine in plasma and serum, perphenazine, olanzapine, and ziprasidone in serum. Analytes were stable (bench top) for 15-20 h, except for aripiprazole, olanzapine, asenapine, droperidol, perazine, thioridazine, ziprasidone (stable for 5-10 h). The LLOQs were the lowest calibrator concentrations (lowest therapeutic concentrations) and ranged from 0.001-0.1 mg/L.
Conclusion: The presented LC-MS/MS approach as part of a universal multi-analyte concept was applicable for selective detection as well as accurate and precise quantification of 27 neuroleptics in whole blood, 26 in plasma, and 21 in serum.
Keywords: LC-MS/MS, neuroleptics, validation
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