通过式固相萃取-超高效液相色谱串联质谱法快速测定中兽药制剂中4种硝基咪唑类药物

Rapid and simultaneous determination of 4 nitroimidazoles in traditional Chinese veterinary medicine by pass-through solid phase extraction purification and ultra performance liquid chromatography tandem mass spectrometry

目的建立通过式固相萃取-超高效液相色谱串联质谱法(ultra performance liquid chromatographytandem mass spectrometry,UPLC-MS/MS)同时快速测定中兽药制剂中4种硝基咪唑类药物的方法。方法样品(2.0 g)经乙醇超声提取,采用PRiME HLB固相萃取柱净化,以0.1%甲酸溶液和乙腈为流动相,ACQUITY BEH C18色谱柱分离,电喷雾正离子扫描,多反应监测模式进行液相色谱串联质谱检测。结果4种目标化合物在1~100 ng/mL范围内线性良好,相关系数r^2>0.999。在3个加标水平(25、50和250μg/kg)下,4种化合物的平均回收率范围为70.0%~93.8%,相对标准偏差范围为1.2%~13.0%。方法的检出限为10μg/kg;定量限为25μg/kg。结论该方法操作简便、灵敏度高,可满足中兽药制剂中硝基咪唑类药物的监测需求。

Objective To establish a method for the rapid and simultaneous determination of 4 nitroimidazoles in traditional Chinese veterinary medicine by pass-through solid phase extraction(SPE)purification with ultra performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS).Methods The samples(2.0 g)were extracted by ethanol,and then cleaned up with an Oasis PRiME HLB pass-through SPE column.The analytes were separated on a C18 reversed-phase column with 0.1%(V/V)formic acid aqueous solution and acetonitrile as mobile phases and finally analyzed using electrospray ionization in positive ion mode(ESI+)with multiple reaction monitoring(MRM)for determination by LC-MS/MS.Results The linear ranges of the 4 compounds were from 1 to100 ng/mL with the correlation coefficients(r2)more than 0.999.The average recoveries and relative standard deviations were 70.0%-93.8%and 1.2%-13.0%respectively at 3 spiked levels(25,50 and 250μg/kg).The limits of detection and the limits of quantification were 10μg/kg and 25μg/kg respectively for the 4 nitroimidazoles.Conclusion This method is simple and sensitive,and can meet the monitoring needs of nitroimidazoles in traditional Chinese veterinary medicine.