液相色谱—串联质谱法测定牛奶中粘杆菌素和多粘菌素B残留量.docVIP

液相色谱—串联质谱法测定牛奶中粘杆菌素和多粘菌素B残留量 　　摘要 建立了同时测定牛奶中粘杆菌素和多粘菌素B素残留量的液相色谱-串联质谱（LC-MS/MS）的方法。样品通过5%高氯酸溶液沉淀蛋白，经乙醚除脂，Strata-X固相萃取柱净化，浓缩后经LC-MS/MS电喷雾正离子多反应检测模式（ESI+MRM）检测。结果显示，粘杆菌素和多粘菌素B在25～1 000 ng/mL范围内均具有良好的线性关系，相关系数不低于0.997 2，加标回收率为71.5%～98.1%，相对标准偏差为3.8%～9.5%。试验证明该方法前处理简单，快速，稳定性好，适用于大量样品中粘菌素和多粘菌素B残留的同时检测。 　　关键词 液相色谱-串联质谱法；残留量；粘杆菌素；多粘菌素B 　　中图分类号 R155.5+7 文献标识码 A 文章编号 1007-5739（2013）22-0260-02 　　Determination of Colistin and Polymycin B Residues in Milk by Liquid Chromatography-Tandem Mass Spetrometry 　　LIANG Xue-qi LIU Hong * HE Feng-qin 　　（Zhongshan Agricultural Products Quality Supervision Inspection Institute in Guangdong Province，Zhongshan Guangdong 528400） 　　Abstract A method was established to simultaneously detect colistinin and polymycin B in milk by liquid chromatography tandem mass spetrometry（LC-MS/MS）.The samples，protein was precipitated with 5% perchloric acid and the fat was removed with diethyl ether，the other impurities were removed with Strata-X solid phase extraction cartridge，after concentration，theanalvtes were detected by LC-MS/MS（ESI）with multiple reaction monitoring（MRM）.The results indicated that the calibration curves were in good linearity between the peak area and the concentration of 25～1 000 ng/mL for colistinin and polymycin B with correlation cofficients not less than 0.997 2，the spiked recoveries ranged from 71.5%～98.1% with reative standard deviations of 3.8%～9.5%.The proposed method was simple and rapid，the stability was good and it was suitable for the simultaneous quantitative detection of colistinin and polymycin B in milk. 　　Key words LC-MS/MS；residues；colistinin；polymycin B 　　粘杆菌素和多粘菌素B是常用的多肽类抗生素并广泛使用于兽药。多粘菌素系由多粘芽孢杆菌的培养液中提取获得的碱性环状多肽类抗生素，主要有A、B、C、D、E 5种成分，粘杆菌素和多粘菌素B为兽医临床应用最广泛，其中粘杆菌素是由A和B组分构成[1-3]。 　　粘杆菌素和多粘菌素B对革兰氏阴性菌感染，例如：绿脓杆菌、大肠杆菌、沙门氏菌等有较强的抗菌作用，还被广泛用于治疗家禽、猪、牛等肠道呼吸道、泌尿道系统感染，特别是对牛乳腺炎等疾病有较好的疗效；作为饲料添加剂同时对畜禽有明显的促进生长作用[1，4]。多肽类抗生素具有肾及神经系统毒性，过度使用在动物体内大量残留，通过肉、蛋、奶等动物源性食品传递给人类，其产生的毒性副作用对人体产生直接的危害。如果长期暴露，还会增加细菌抗原性。现今社会随着抗生素的广泛使用，滥用和忽略休