离子液体—分散液液微萃取富集动物源性食品中抗生素类药物的研究及应用

Antibiotics are widely used to prevent and/or treat diseases in animals, as well as added in the feed to promote the growth of animals in livestock and aquaculture operations. If people often eat foodstuffs of animal origin that containing antibiotics, their health will be harmed. Therefore, it is very important to determine antibiotics residues in foodstuffs of animal origin. In this research, dispersive liquid-liquid microextraction (DLLME) based on room temperature ionic liquids (RTILs or ILs) which have high extraction efficient for antibiotics as extraction solvent will be used to concentrate antibiotics in foodstuffs of animal origin which have the complex matrices. The technology avoids the shortcomings of solid phase extraction (SPE) technology. For example, SPE often suffers from the consumption of appreciable amount of hazardous organic solvents at the elution step, the plugging of cartridge and high cost. Compared with the traditional organic solvent, ILs are less toxic, friendly to environment and have better extractability by designing them. Several important parameters influencing the extraction efficiency of IL-DLLME will be tested and the extraction mechanism will be discussed. A novel method termed as IL-DLLME combining with high performance liquid chromatography (HPLC) or high performance liquid chromatography-mass spectrometry (HPLC-MS) will be developed for the determination of antibiotics in foodstuffs of animal origin. This method is sensitive, easy to operate, fast, friendly to environment and economic. The method will be applied in determination of antibiotics in foodstuffs of animal origin which are sampled from the markets. The results will provide data support about antibiotics residues in foodstuffs of animal origin and then avoid antibiotics from being misused by human beings.

抗生素被大量用于动物的防病、治病及添加于饲料中以提高饲料利用率和促进动物生长。人如果长期食用含抗生素的动物源性食品，人的健康将受到危害。因此测定动物源性食品中的抗生素就显得很重要。本研究拟筛选出对抗生素具有高萃取效率的室温离子液体（RTILs,ILs），结合分散液液微萃取（DLLME）技术分离富集基质复杂的动物源性食品中的抗生素。该技术避免了固相萃取法的缺点，如需使用有毒有害的有机溶剂来洗脱、易堵塞、成本较高等。ILs作为萃取剂，与传统有机溶剂相比，具有毒性小、更环保、可对ILs进行设计获得更高萃取效率等优点。同时研究各种因素对萃取效率的影响，初步探讨萃取的机理。将该技术与液相色谱（或液相色谱－质谱）联用，建立一种灵敏、简单、快速、环保及经济的测定动物源性食品中抗生素的新方法。并把此方法应用于市场上动物源性食品的测定，为动物源性食品中抗生素的残留提供一定的数据支持，避免人类滥用抗生素。

抗生素对治疗感染性疾病发挥了巨大作用，在水产养殖业和畜牧业中被广泛的应用。残留在动物源性食品中的抗生素对人类健康有潜在的危害。目前分离富集动物源性食品中的抗生素主要采用液液萃取和固相萃取法相结合。但是固相萃取法仍需要一些有毒有害的有机溶剂来洗脱、柱子容易堵塞、 操作费时，而且一次性的固相萃取柱成本高。因此本课题主要以绿色环保试剂离子液体作为萃取溶剂，采用分散液液微萃取技术分离富集动物源性食品中的抗生素。.本课题先建立了一种基于离子液体（IL）的分散液液微萃取（DLLME）-高效液相色谱法（HPLC）测定蜂蜜中11种磺胺类抗生素的分析方法。最佳的萃取条件：以50 μL的1-己基-3-甲基咪唑六氟磷酸盐（[C6MIM][PF6]）作为萃取剂，溶液pH=3，不加盐。方法的检出限0.006～0.015 mg/kg（S/N=3）；对各种蜂蜜进行2个浓度（0.1mg/kg 、0.5mg/kg）的加标，得回收率为85.3%～97.4%，相对标准偏差（RSD）为5.2～7.3%（n=5）。该方法简单、快速、环保、准确，可用于蜂蜜中磺胺类抗生素的测定。另外，采用以离子液体为萃取剂的分散液液微萃取（DLLME）技术，结合柱前荧光衍生高效液相色谱荧光法建立了同时测定鱼肉中8种磺胺类抗生素的分析方法。优化出的最佳实验条件为：鱼肉样品先用0.1 mol/L盐酸溶液振荡提取20 min后，再用40 μL[C6MIM][PF6]作萃取剂、0.1 mL丙酮作分散剂，对pH = 4且不含NaCl的鱼肉样品预处理液进行分散液液微萃取，以60 μL 0.02%荧光胺丙酮溶液为柱前衍生剂，衍生化反应6 h后用HPLC测定。LOD为0.1~1 μg/kg (S/N=3)。对鱼肉样品进行测定，发现草鱼和金昌鱼中可测出磺胺甲噻二唑，在3个浓度水平（10、100、500 μg/kg）进行加标回收率实验，得回收率为84.4%~95.0%， RSD为4.1%~7.2%。该方法灵敏、简单、环保、经济，可用于测定鱼肉中8种磺胺类抗生素。根据要求，本项目发表论文（含接收待刊）6篇，其中1篇SCI收录，3篇中文核心期刊论文，1篇国际会议论文，有望被EI检索收录，另外还有部分成果正在整理成论文发表。