UPLC-QDa同时检测牛奶中多种抗菌药物兽药残留的快速分析方法研究

发布时间：2018-04-23 01:16

  本文选题：牛奶 + 6种抗菌药物　； 参考：《昆明医科大学》2017年硕士论文

【摘要】：[目的]畜牧业中,抗菌药物被广泛用于奶牛各种疾病的治疗,造成乳制品外源性抗菌药物的存在,对人体健康造成很大危害。为了对牛奶中痕量多种抗菌药物兽药残留进行快速检测分析,建立并优化一种快速分析方法尤为重要。本研究建立了牛奶中6种抗菌药物兽药残留的超高效液相-串联质谱(UPLC-QDa)检测方法。该方法可以满足日常牛奶中常用兽类抗菌药物的痕量分析要求。[方法]首先,对超高效液相色谱和QDa质谱条件进行优化。其次,优化了样品的前处理方法,具体步骤为:准确称取5.0 g牛奶样品于50 mL离心管中,加入5 mL PBS缓冲溶液(pH值为7.0),涡旋混匀1.5 min,加入7.0 mL乙腈,1.0 g氯化钠,2.0 g无水硫酸钠,30℃水浴超声5 min,涡旋混匀3 min,5500 r/min高速离心6 min,取上清液于10 mL具刻度玻璃管中。离心后的牛奶样品再次加入3.0 mL乙腈,重复上述过程(无需再加入氯化钠和无水硫酸钠),合并上清液,30℃下氮气吹至近干。加水5mL稀释溶解,待上样。先将HLB固相萃取小柱用6 mL甲醇,6 mL水预处理,将提取液上样,6 mL水淋洗,抽干小柱中淋洗液,10mL洗脱液(甲醇-乙酸乙酯7/3,V/V)进行洗脱,控制流速1～2mL/min,使其中的液体完全流出。收集洗脱液于10mL具刻度玻璃管,30℃下氮气吹干。1mL甲醇定容,涡旋1min,过0.22 μm滤膜,待上机。最后,运用所建立的方法对市售牛奶进行检测。[结果]6种混合标准样品在10～250 μg/L范围内线性关系良好;以鲜牛奶空白基质进行检出限(根据S/N=3计算)和定量限(根据S/N=10计算)研究,检出限范围为1.5～6 μg/kg;定量限范围为5～20 μg/kg。分别以定量限,定量限5倍和定量限10倍作为三个添加水平,每个添加水平做5次平行试验,所有目标化合物的平均回收率范围为72.1%～95.0%;相对标准偏差范围为1.31%～13.8%。[结论]本研究建立了牛奶中3种硝基咪唑类、2种大环内酯类、1种林可酰胺类的6种抗菌药物兽药残留的超高效液相-串联质谱(UPLC-QDa)检测方法。优化了样品的前处理条件,方法操作简单,灵敏度高。
[Abstract]:[objective] in animal husbandry, antimicrobial agents are widely used in the treatment of dairy cow diseases, resulting in the existence of exogenous antibiotics in dairy products and great harm to human health. It is very important to establish and optimize a rapid method for the determination and analysis of veterinary drug residues of trace antibiotics in milk. A high performance liquid phase tandem mass spectrometry (UPLC-QDa) method was developed for the determination of veterinary drug residues in milk. The method can meet the requirements of trace analysis of veterinary antimicrobial agents in daily milk. [methods] first, the conditions of ultra-high performance liquid chromatography and QDa mass spectrometry were optimized. Secondly, the pretreatment method of the sample was optimized. The specific steps were as follows: accurately weighing 5.0 g milk sample in 50 mL centrifuge tube, The pH value of 5 mL PBS buffer solution was 7.0 min, vortex mixing was 1.5 min, adding 7.0 mL acetonitrile 1.0 g sodium chloride 2.0 g sodium chloride 2.0 g anhydrous sodium sulfate for 5 min, vortex mixing for 3 min and 5 500 r/min high speed centrifugation for 6 min. The supernatant was extracted from 10 mL glass tube. After centrifugation, 3.0 mL acetonitrile was added again to the milk sample. The above process was repeated (no further addition of sodium chloride and anhydrous sodium sulfate), and the supernatant was combined with nitrogen at 30 鈩，

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