Objective:To establish a robust ultra-high performance liquid chromatography coupled to mass spectrometry(UHPLC-MS/MS)method for the simultaneous determination of imidazole compounds and their enantiomers in blood,uri...Objective:To establish a robust ultra-high performance liquid chromatography coupled to mass spectrometry(UHPLC-MS/MS)method for the simultaneous determination of imidazole compounds and their enantiomers in blood,urine,and hair.Materials and Methods:Blood and urine samples were subjected to protein precipitation with acetonitrile,whereas hair samples were pulverized after washing and extracted with methanol,followed by ultrasonication and centrifugation.The supernatant was filtered through a 0.22μm membrane for analysis.Enantiomeric separation was achieved on a Phenomenex Lux^(■)Cellulose-3 chiral column using an isocratic mobile phase of 5 mmol/L ammonium acetate with 0.1%formic acid(60%)and acetonitrile(40%)at a flow rate of 0.5 mL/min.Detection was performed in positive electrospray ionization mode with multiple reaction monitoring.The method was validated according to international forensic toxicology standards and applied to authentic case samples.Results:The method demonstrated excellent selectivity and linearity(R^(2)>0.996).Calibration ranges were 5–3000 ng/mL for blood and urine and 0.05–30 ng/mg for hair.Limits of detection were 1 ng/mL for blood and urine and 0.02 ng/mg for hair,with corresponding limits of quantification(LLOQ)of 5 ng/mL and 0.05 ng/mg.Accuracy ranged from 86.00%to 114.58%,recoveries exceeded 86.75%,and matrix effects were within 0.01%–14.82%,all meeting forensic validation criteria.Application to 22 authentic hair samples revealed widespread exposure to etomidate,with the R-isomers of metomidate,propoxate,and isopropoxate detected more frequently and at higher concentrations than their S-counterparts.Conclusion:The established UHPLC-MS/MS method provides high sensitivity,selectivity,and reproducibility,with a wide linear range and straightforward sample preparation.Its successful application to multiple biological matrices underscores its potential as a powerful tool for the screening,quantification,and source tracing of imidazole compounds and their enantiomers in forensic toxicology and drug abuse monitoring.展开更多
文摘Objective:To establish a robust ultra-high performance liquid chromatography coupled to mass spectrometry(UHPLC-MS/MS)method for the simultaneous determination of imidazole compounds and their enantiomers in blood,urine,and hair.Materials and Methods:Blood and urine samples were subjected to protein precipitation with acetonitrile,whereas hair samples were pulverized after washing and extracted with methanol,followed by ultrasonication and centrifugation.The supernatant was filtered through a 0.22μm membrane for analysis.Enantiomeric separation was achieved on a Phenomenex Lux^(■)Cellulose-3 chiral column using an isocratic mobile phase of 5 mmol/L ammonium acetate with 0.1%formic acid(60%)and acetonitrile(40%)at a flow rate of 0.5 mL/min.Detection was performed in positive electrospray ionization mode with multiple reaction monitoring.The method was validated according to international forensic toxicology standards and applied to authentic case samples.Results:The method demonstrated excellent selectivity and linearity(R^(2)>0.996).Calibration ranges were 5–3000 ng/mL for blood and urine and 0.05–30 ng/mg for hair.Limits of detection were 1 ng/mL for blood and urine and 0.02 ng/mg for hair,with corresponding limits of quantification(LLOQ)of 5 ng/mL and 0.05 ng/mg.Accuracy ranged from 86.00%to 114.58%,recoveries exceeded 86.75%,and matrix effects were within 0.01%–14.82%,all meeting forensic validation criteria.Application to 22 authentic hair samples revealed widespread exposure to etomidate,with the R-isomers of metomidate,propoxate,and isopropoxate detected more frequently and at higher concentrations than their S-counterparts.Conclusion:The established UHPLC-MS/MS method provides high sensitivity,selectivity,and reproducibility,with a wide linear range and straightforward sample preparation.Its successful application to multiple biological matrices underscores its potential as a powerful tool for the screening,quantification,and source tracing of imidazole compounds and their enantiomers in forensic toxicology and drug abuse monitoring.
