UPLC-MS/MS法检测人体毛发中右美沙芬及其代谢物去甲右美沙芬

Detection of dextromethorphan and dextrorphan in hair by UPLC-MS/MS

目的:建立一种准确、快速检测毛发中右美沙芬及其代谢物去甲右美沙芬的超高效液相色谱-串联质谱(UPLC-MS/MS)方法。方法:用含内标双苯戊二氨酯(SKF525A)的甲醇溶液提取含有右美沙芬及其代谢物的毛发,经0.22μm微孔有机滤膜过滤,UPLC-MS/MS检测。采用ACQUITY UPLC HSS T3超高液相色谱柱(100 mm×2.1 mm,1.8μm),以0.2%甲酸(10 mmol·L^(-1)甲酸铵)和乙腈为流动相进行梯度洗脱,流速0.3 mL·min-1,柱温为室温。采用电喷雾离子源,正离子多反应模式检测。结果:右美沙芬和去甲右美沙芬质量浓度均在1~100 ng·mL^(-1)范围内线性响应良好,线性方程分别为Y=1.34949X-0.02080(r=0.9988)和Y=0.77510X-0.01387(r=0.9991),检测限和定量限均分别为0.01 ng·mL^(-1)和0.025 ng·mL^(-1),平均回收率在97.0%~104.8%,日内精密度和日间精密度分别在1.5%~3.9%和2.1%~5.5%。将本方法应用于实际案例,结果在6例右美沙芬滥用者毛发中均检测出右美沙芬及其代谢物去甲右美沙芬。结论:该方法操作简便,检测灵敏度高,可用于毛发中右美沙芬及其代谢物去甲右美沙芬的检测。

Objective:To establish a rapid and accurate ultra performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)method for the detection of dextromethorphan and one metabolite dextrorphan in hair.Methods:The hair sample containing dextromethorphan and dextrorphan were extracted with methanol containing internal standard proadifen hydrochloride(SKF525A).The extract was filtered with 0.22μm organic filter membrane and detected by UPLC-MS/MS.All components were separated by an ACQUITY UPLC HSS T3 column(100 mm×2.1 mm,1.8μm),using a gradient elution procedure consisting of 0.2%formic acid(10 mmol·L^(-1)ammonium formate)and acetonitrile,at a flow rate of 0.3 mL·min-1,and the column temperature was room temperature.Positive electrospray ionization was performed using multiple reaction monitoring mode(MRM).Results:The linear relationships of dextromethorphan and dextrorphan were good in the range of 1-100 ng·mL^(-1).The linear equations were Y=1.34949X-0.02080(r=0.9988)and Y=0.77510X-0.01387(r=0.9991),respectively.The detection limit and quantitation limit were 0.010 ng·mL^(-1)and 0.025 ng·mL^(-1)respectively.Their recoveries ranged from 97.0%-104.8%.The intra-day RSD and inter-day RSD were 1.5%-3.9%and 2.1%-5.5%,respectively.The method was applied to cases,and the results showed that dextromethorphan and dextrorphan were detected in the hair of 6 abusers.Conclusion:This method is simple and sensitive enough to be applied to detect dextromethorphan and one metabolite dextrorphan in hair.