摘要：建立高效液相色谱串联质谱法测定饲料中苯乙醇胺A的分析方法。样品用80%（V∶V）乙腈水溶液提取，经PRiME HLB净化后，采用高效液相色谱串联质谱法测定，外标法定量。以01%甲酸水溶液和甲醇作为流动相，用Hypersil GOLD C18色谱柱 （19 μm，21 mm×100 mm）进行梯度洗脱，采用电喷雾离子源（ESI+）、正离子扫描多反应监测（MRM）模式进行定性和定量分析。结果表明：苯乙醇胺A在50～2000 ng· mL-1的范围内线性关系良好（r=09992）。在100、200、500 μg· kg-1添加水平下的回收率为822%～915%，相对标准偏差（n=6）为68%～85%，检出限（以信噪比>3计）为01 μg·kg-1，定量限（以信噪比>10计）为03 μg·kg-1。该方法精密度好，灵敏度高，可用于大批量快速准确测定饲料中的苯乙醇胺A，有效提高工作效率。

关键词：PRiME HLB;高效液相色谱串联质谱法;苯乙醇胺A;饲料

中图分类号：O 658文献标志码：A文章编号：0253-2301（2021）02-0024-08

DOI： 1013651/jcnkifjnykj202102005

Determination of Phenylethanolamine A in Feed Based on High Performance Liquid

ChromatographyTandem Mass Spectrometry After Purified by PRiME HLB

CHEN Qihuang

（Xiamen Inspection and Test Center of Agricultural Products Quality and Safety， Xiamen， Fujian 361009， China）

Abstract： A analytical method for the determination of phenylethanolamine A in feed by high performance liquid chromatographytandem mass spectrometry was established. The samples were extracted with 80% （V∶V） acetonitrile aqueous solution. After purified by PRiME HLB， the samples were determined by high performance liquid chromatographytandem mass spectrometry and quantified by external standard method. Then， the gradient elution was performed through Hypersil GOLD C18 chromatographic column （1.9 μm ， 2.1 mm×100 mm） by using 0.1% formic acid aqueous solution and methanol as the mobile phase. And the qualitative and quantitative analysis was carried out by using the modes of electrospray ionization （ESI+） and positiveion scanning multiple reaction monitoring （MRM）. The results showed that phenylethanolamine A had a good linear relationship in the range of 5.0～200.0 ng·mL-1 （r=0.9992）. The recoveries were 82.2% to 91.5% at the supplemented levels of 10.0， 20.0 and 50.0 μg·kg-1， the relative standard deviations （n=6） were 6.8% to 8.5%， the limits of detection （the SNR being greater than 3） were 0.1 μg·kg-1， and the limits of quantification （the SNR being greater than 10） were 0.3 μg·kg-1. The method had good precision and high sensitivity， and could be used for the rapid and accurate determination of phenylethanolamine A in large quantities， which would effectively improve the work efficiency.

Key words： PRiME HLB; High performance liquid chromatography tandem mass spectrometry（HPLCMS/MS）; Phenylethanolamine A; Feed

苯乙醇胺A，又稱克伦巴胺，英文名称Phenylethanolamine A，CAS：1346746-81-3，分子式C19H24N2O4，分子量34417。据报道，苯乙醇胺A被非法添加到饲料中以促进动物生长，减少脂肪沉积，提高瘦肉率，但其会在动物组织中残留，食用含有苯乙醇胺A残留的动物源性产品，对人体健康具有潜在危害＼[1＼]。为加强饲料及养殖环节质量安全监管，保障饲料及畜产品质量安全，我国农业部于2010年12月27日发布中华人民共和国农业部第1519号公告，禁止在饲料和动物饮水中使用苯乙醇胺A等物质。因此，加强对苯乙醇胺A的残留检测具有非常重要意义。

目前，苯乙醇胺A的检测方法主要有胶体金快速检测法（CGIA）＼[2＼]、酶联免疫吸附测定法（ELISA）＼[3-4＼]、高效液相色谱法（HPLC）＼[1，5-6＼]和高效液相色谱串联质谱法（HPLCMS/MS）＼[7-12＼]。HPLCMS/MS能准确定量确证，方法检出限低、灵敏度高，前处理大多采用固相萃取净化技术，但固相萃取小柱需要活化平衡等步骤， 过程耗时长，消耗溶剂量大，前处理比较复杂烦琐，又有流速控制要求，操作不当易造成损失，降低回收率。Oa……