3本研究建立了鱼油和猪肉中万古霉素残留液相色谱-串联质谱（LC-MS/MS）的检测方法，优化了鱼油与猪肉中万古霉素的提取、纯化方法。两者均用外标法定量。鱼油用有机溶剂进行溶解，然后使用酸性水溶剂进行提取。通过C18固相萃取柱净化，最后用0.1%甲酸定容， C18柱分离、质谱进行分析。通过回收率的比较，选择鱼油中最佳的万古霉素提取方式。猪肉使用酸性水溶液进行蛋白质的沉淀、万古霉素的提取，C18固相萃取柱净化，最后用0.1%甲酸定容，C18柱分离、质谱进行分析。通过回收率的比较，选择猪肉中最佳的万古霉素提取方法。结果表明，鱼油用5mL石油醚、0.1mol/L 盐酸提取，并通过固相萃取柱纯化，其回收率为74.386%～82.195%，相对偏差<10%；猪肉用2%高氯酸离心过滤，并通过MCX纯化，其回收率为71.08%～78.379%，相对偏差<6%。结果表明，该方法能有效去除杂质干扰，方法可靠，灵敏度更高，能满足鱼油和猪肉中万古霉素糖肽类抗生素残留量的检测。
目 录
Abstract3
Key words3
前言3
1材料与方法5
1.1材料与仪器 5
1.2实验方法 5
1.2.1猪肉中万古霉素的提取5
1.2.2猪肉中万古霉素的净化5
1.2.3基质曲线的制备5
1.2.4鱼油中万古霉素的提取5
1.2.5鱼油中万古霉素的净化5
1.2.6色谱条件6
1.2.7质谱条件6
2结果与分析6
2.1提取条件的优化6
2.1.1猪肉中万古霉素提取条件的优化6
2.1.2 鱼油中万古霉素提取条件的优化7
2.2净化条件的优化7
2.3猪肉中万古霉素的检测方法8
2.4鱼油中万古霉素的检测方法9
3讨论 10
致谢11
参考文献11
液相色谱串联质谱法测定鱼油和猪肉中万古霉素残留
动物医学院专业学生 李梦雅
指导教师 苏娟
Determination of Vancomycin in *景先生毕设|www.jxszl.com +Q: ^351916072^ 
fish oil and pork by LCMS/MS
Student majoring in in Veterinary Mengya Li
Tutor Juan Su
Abstract：A method was established for determination of vancomycin residues in fish oil and pork by liquid chromatography tandem mass spectrometry (LCMS/MS). The sample was supersonically extracted and purified. Both of them were quantified by external standard method. Fish oil was dissolved in organic solvents and extracted with acid water. C18 solid phase extraction column was used to purify, and constant volume with 0.1% formic acid. C18 column separation and mass spectrometry were used for analysis. Through the comparison of the recovery rate, the optimum extraction method of vancomycin in fish oil was selected. Pork uses acidic aqueous solution to precipitate protein and extract vancomycin. It is purified by C18 solid phase extraction column. The final content is determined by 0.1% formic acid and analyzed by C18 column separation and mass spectrometry. Through the comparison of the recovery rate, the optimum method of extracting vancomycin from pork was selected. The results showed that fish oil with 5mL petroleum ether, 0.1mol/L hydrochloric acid extraction, and after purification of MCX, the recovery rate is 74.386%~82.195%, the relative deviation of <10%; with 2% perchloric acid centrifugal filtration of pork, and after purification of MCX, the recovery rate is 71.08%~78.379%, RSD<6%.
Keywords: Fish oil; pork; vancomycin;LCMS/MS
前言
万古霉素（Vancomycin）是一种糖肽类大分子抗生素，其性质为【1】游离碱为无色结晶,等电点PH值为5；常用其盐酸盐,白色固体状,易溶于水；不耐高温,需要低温保存。其治疗作用有：抑制或杀灭葡萄球菌（包括耐青霉素和耐新青霉素株）、难辨梭状芽胞杆菌等，如心内膜炎、败血症、伪膜性肠炎。它是治疗耐甲氧西林金黄色葡萄球菌( MRSA) 感染的一线药物，虽然对厌氧菌和革兰氏阴性细菌无效，但是在其他抗生素对病菌无效时会被使用，从而常作为超级细菌的最后一道防线【2】。
已有研究揭示万古霉素的杀菌机理为【3】：1.直接与细胞壁肽聚糖前体五肽侧链末端D丙胺酰D丙氨酸结合，阻断肽聚糖多聚酶的转肽作用，影响细菌细胞壁肽聚糖前体的交叉联结，使细胞壁不能形成三维空间结构，从而发挥杀菌效果。2.万古霉素也能够起到损伤细菌细胞膜和抑制细菌RNA合成的作用。

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