Simultaneous Determination of 12 Antidepressants in Human Blood by UHPLC-MS/MS
Objective To establish an analytical method for the simultaneous determination of 12 antidepressants including moclobemide and bupropion in human blood by ultra high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS).Methods Blood samples were purified through protein precipitation with acetonitrile that contained internal standard(proadifen),then the samples were injected into the UHPLC-MS/MS.The separation was performed on an ACQUITY UPLC HSS T3 column(2.1 mm×100 mm,1.8 µm)using gradient elution with a flow rate of 0.35 mL/min.The mobile phase consisted of water solution and acetonitrile solution that both contained 2 mmol/mL ammonium formate and 0.1%formic acid.The analytes were detected using multiple reaction monitoring mode.The electrospray ionization source was performed in positive mode.The internal standard method was used for quantification.Results The correlation coefficients of the 12 antidepressants were better than 0.999 in the corresponding concentration ranges,and the limits of detection ranged from 0.05 ng/mL to 0.2 ng/mL,the limits of quantification ranged from 0.5 ng/mL to 1 ng/mL.At different spiked concentrations(low,medium and high),the accuracy ranged from-7.1%to 11.7%,the intra-day precision ranged from 0.1%to 11.5%,the inter-day precision ranged from 1.9%to 9.4%,the extraction recoveries ranged from 85.7%to 113.5%,and the matrix effect ranged from-18.2%to 10.2%.The stability of each component at different levels of concentration and under different environment conditions ranged from-9.30%to 11.30%.All of the above results meet the requirements for biological sample analysis and forensic toxicology analysis.Conclusion The method is selective,reproducible and precise.The simultaneous determination of 12 antidepressants in human blood can be applied in forensic toxicology practice and it will be of great use in the analysis of the cause of death in unnatural death cases.
forensic toxicologyultra high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)antidepressanthuman blood
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