超高效液相色谱-高分辨质谱法同时测定猪肉样品中5种巴比妥类药物

Simultaneous determination of 5 kinds of barbiturates in pork by ultra performance liquid chromatography-high resolution mass spectrometry

目的建立超高效液相色谱-高分辨质谱法(ultra performance liquid chromatography-high resolution mass spectrometry,UPLC-HRMS)同时测定猪肉中5种巴比妥类药物(巴比妥、苯巴比妥、司可巴比妥、异戊巴比妥以及硫喷妥钠)的分析方法。方法猪肉样品采用超声辅助甲醇提取、离心、净化后,经C18色谱柱(100 mm×2.1 mm,1.9μm)分离,以0.1%甲酸水和甲醇作为流动相,梯度洗脱。质谱采用全扫描/数据依赖的二级扫描模式(full scan data-dependent MS/MS acquisition mode,Full MS/dd-MS2),外标法定量。结果在5~100 ng/mL质量浓度范围内,标准曲线的相关系数均大于等于0.9908。5种巴比妥类药物的检出限(limits of detection,LODs)(S/N≥3)范围为0.17~0.64µg/kg,添加浓度水平为5、10和25µg/kg时,平均回收率在63.05%~90.34%之间,相对标准偏差(relative standard deviations,RSDs)范围为3.65%~11.22%。结论本方法灵敏度和准确性高、重现性好,可对猪肉样品中5种巴比妥类药物进行定性和定量分析。

Objective To establish an analytical method for the simultaneous determination of 5 kinds of barbiturates(barbital,phenobarbital,scobarbital,secobarbital,and pentobarbital sodium)in pork by ultra performance liquid chromatography-high resolution mass spectrometry(UPLC-HRMS).Methods Samples were extracted with methanol under ultrasonic condition,centrifuged by centrifugal machine and cleaned up,separated on C18 chromatographic column(100 mm×2.1 mm,1.9μm).Gradient elution was carried out with water(containing 0.1%formic acid)and methyl alcohol as mobile phase.The high resolution mass spectrometry data was simultaneously acquired in full scan data-dependent MS/MS acquisition mode(Full MS/dd-MS2),and external standard calibration curves were used for quantification.Results The correlation coefficient of the standard curve was greater than or equal to 0.9908 within the mass concentration range of 5–100 ng/mL.The limits of detection(LODs)of 5 kinds of barbiturates were in the range of 0.17–0.64µg/kg.The average recoveries of barbiturates were 63.05%–90.34%at spiked levels of 5.0,10.0 and 25.0µg/kg,the relative standard deviations(RSDs)of the method were between 3.65%–11.22%.Conclusion This method has high sensitivity,accuracy,and good reproducibility,and can be used for qualitative and quantitative analysis of 5 kinds of barbiturates in pork samples.