Validated Method to Determine Quetiapine and Norquetiapine in Microsomal Matrix by LC MS–MS: Implication in Quetiapine Metabolism

摘要

A sensitive and selective LC–MS/MS method for the quantification of the atypical antipsychotic agent quetiapine and its metabolite norquetiapine (N-desalkyl quetiapine) was developed and validated. Following the protein precipitation technique, the analytes were separated using a reversed phase column with gradient elution. The compounds were ionized in the electrospray positive ionization (ESI?) ion source tandem MS detection in multiple reaction monitoring (MRM) mode. Calibration curves were generated by plotting the peak area ratio of quetiapine and norquetiapine to the IS clozapine for each calibration concentration. The method provides a linear response from a quantitation range of 2.3–452.9 nM (0.9–173.7 ng/mL) and 2.7–543.0 nM (1.0–200.0 ng/mL) for quetiapine and norquetiapine, respectively. Regression analysis showed a correlation coefficient greater than 0.999 and 0.991 for quetiapine and norquetiapine, respectively. To evaluate the metabolism of quetiapine by the cytochrome P450 in microsomes, the method has been subsequently employed. LC–MS/MS procedure has been carried out to determine increasing concentrations of both drugs in microsomal matrix obtained by a pool of mammalian liver microsomes BD UltraPool~(TM) Human Liver Microsomes (HLM 150).