高效液相色谱-串联质谱法测定动物尿液中23种β-受体激动剂

Determination of 23 kinds of β-receptor agonists in animal urine by high performance liquid chromatography-tandem mass spectrometry

目的建立高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)快速检测动物尿液中23种β-受体激动剂残留的分析方法。方法样品经酸解释放结合态药物,混合型阳离子固相萃取柱富集净化,5%氨水甲醇洗脱,浓缩。采用Aglient EC C18(100 mm×3.0 mm,2.7μm)为色谱柱,乙腈和5 mmol/L乙酸铵-0.2%甲酸溶液作为流动相进行梯度洗脱,质谱采用多反应监测模式(multiple reaction monitoring,MRM)对β-受体激动剂的定量离子和定性离子进行监测。结果本方法在10 min内完成23种兴奋剂的有效分离。在猪尿液、牛尿液、羊尿液中0.2、0.4和2μg/L添加水平下,23种β-受体激动剂的平均回收率为65.7%~112.6%,相对标准偏差≤16.5%(n=6),方法定量限为0.2μg/L,检出限为0.1μg/L。结论该方法高效、准确,适合测定动物尿液中23种β-受体激动剂药物残留,并为相关标准制修订提供参考。

Objective To establish an analytical method for rapid detection of 23 kinds ofβ-receptor agonists residues in animal urine by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Methods The sample was treated with acid to interpret the bound drug,enriched and purified by a mixed cation solid-phase extraction column,followed by elution with 5%ammonia and methanol,and concentrate.Aglient EC C18(100 mm×3.0 mm,2.7μm)was used as the chromatographic column,and acetonitrile and 5 mmol/L ammonium acetate-0.2%formic acid solution were used as the mobile phases for gradient elution.The quantitative and qualitative ions of β-receptor agonists were monitored by multiple reaction monitoring(MRM)using mass spectrum.Results This method completed the effective separation of 23 kinds of agonists within 10 minutes.At the addition levels of 0.2,0.4 and 2μg/L in pig,cow and goat urine,the average recoveries of the 23 kinds of β-receptor agonists were 65.7%‒112.6%,the relative standard deviations were less than or equal to 16.5%(n=6),the limit of quantitation was 0.2μg/L and the limit of detection was 0.1μg/L.Conclusion This method is efficient,accurate and suitable for the determination of 23 kinds of β-receptor agonists drug residues in animal urine,and provides reference for the revision of relevant standards.