本文選題：牛奶 + 6種抗菌藥物�。� 參考：《昆明醫(yī)科大學》2017年碩士論文

【摘要】：[目的]畜牧業(yè)中,抗菌藥物被廣泛用于奶牛各種疾病的治療,造成乳制品外源性抗菌藥物的存在,對人體健康造成很大危害。為了對牛奶中痕量多種抗菌藥物獸藥殘留進行快速檢測分析,建立并優(yōu)化一種快速分析方法尤為重要。本研究建立了牛奶中6種抗菌藥物獸藥殘留的超高效液相-串聯(lián)質(zhì)譜(UPLC-QDa)檢測方法。該方法可以滿足日常牛奶中常用獸類抗菌藥物的痕量分析要求。[方法]首先,對超高效液相色譜和QDa質(zhì)譜條件進行優(yōu)化。其次,優(yōu)化了樣品的前處理方法,具體步驟為:準確稱取5.0 g牛奶樣品于50 mL離心管中,加入5 mL PBS緩沖溶液(pH值為7.0),渦旋混勻1.5 min,加入7.0 mL乙腈,1.0 g氯化鈉,2.0 g無水硫酸鈉,30℃水浴超聲5 min,渦旋混勻3 min,5500 r/min高速離心6 min,取上清液于10 mL具刻度玻璃管中。離心后的牛奶樣品再次加入3.0 mL乙腈,重復上述過程(無需再加入氯化鈉和無水硫酸鈉),合并上清液,30℃下氮氣吹至近干。加水5mL稀釋溶解,待上樣。先將HLB固相萃取小柱用6 mL甲醇,6 mL水預處理,將提取液上樣,6 mL水淋洗,抽干小柱中淋洗液,10mL洗脫液(甲醇-乙酸乙酯7/3,V/V)進行洗脫,控制流速1～2mL/min,使其中的液體完全流出。收集洗脫液于10mL具刻度玻璃管,30℃下氮氣吹干。1mL甲醇定容,渦旋1min,過0.22 μm濾膜,待上機。最后,運用所建立的方法對市售牛奶進行檢測。[結(jié)果]6種混合標準樣品在10～250 μg/L范圍內(nèi)線性關(guān)系良好;以鮮牛奶空白基質(zhì)進行檢出限(根據(jù)S/N=3計算)和定量限(根據(jù)S/N=10計算)研究,檢出限范圍為1.5～6 μg/kg;定量限范圍為5～20 μg/kg。分別以定量限,定量限5倍和定量限10倍作為三個添加水平,每個添加水平做5次平行試驗,所有目標化合物的平均回收率范圍為72.1%～95.0%;相對標準偏差范圍為1.31%～13.8%。[結(jié)論]本研究建立了牛奶中3種硝基咪唑類、2種大環(huán)內(nèi)酯類、1種林可酰胺類的6種抗菌藥物獸藥殘留的超高效液相-串聯(lián)質(zhì)譜(UPLC-QDa)檢測方法。優(yōu)化了樣品的前處理條件,方法操作簡單,靈敏度高。
[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，

本文編號：1789794