蜡样芽胞杆菌呕吐毒素引发的米面制品食物中毒快速确证方法研究  

Rapid confirmation method of food poisoning caused by Bacillus cereus cereulide in rice and flour products

作　　者：崔霞[1] 王莉莉[1] 刘平[1] 段佳丽[1] 赵榕[1] 范赛[1] Cui Xia;Wang Lili;Liu Ping;Duan Jiali;Zhao Rong;Fan Sai(Beijing Center for Disease Prevention and Control,Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning,Beijing 100013,China)

机构地区：[1]北京市疾病预防控制中心食物中毒诊断溯源技术北京市重点实验室,北京100013

出　　处：《卫生研究》2023年第4期573-578,共6页Journal of Hygiene Research

基　　金：高层次公共卫生技术人才建设项目(No.学科骨干-01-003)。

摘　　要：目的建立快速测定米面制品中蜡样芽胞杆菌呕吐毒素的超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS),为蜡样芽胞杆菌呕吐毒素引起的食物中毒提供应急处置措施。方法单一基质米面样品用乙腈水溶液提取,以盐析分层,离心过膜后直接上机检测;炒米饭、炒面等复杂基质样本用乙腈水溶液提取,经HLB-P小柱净化,采用ACQUITY UPLC Peptide BEH C18300?色谱柱(2.1 mm×100 mm,1.7μm)分离后于电喷雾正离子模式下,采用多反应监测扫描(multiple reaction monitoring,MRM)进行测定,以溶剂标准曲线法定量。结果蜡样芽胞杆菌呕吐毒素在蒸米饭、馒头、面条等阴性样品中加标水平在0.5、1.0和5.0μg/kg下回收率为87.4%~98.3%,精密度为1.4%~4.2%(n=6)。在炒米饭、炒面等阴性样本中加标水平在1.0、2.0、5.0μg/kg下,回收率为89.5%~99.3%,精密度为1.1%~7.5%(n=6)。目标化合物的检出限为0.2~0.3μg/kg,定量限为0.5~1.0μg/kg。对北京某地引起食物中毒的实际样品进行了检测,中毒样品中蜡样芽胞杆菌呕吐毒素含量为1287~7398μg/kg,2份原料凉面中蜡样芽胞杆菌呕吐毒素含量分别为0.4和9.4μg/kg。结论该方法操作简便、快速,灵敏度和准确度高,实现了蜡样芽胞杆菌引发食物中毒的快速处置。OBJECTIVE To develop a method for rapid determination of Bacillus cereus cereulide in rice and flour products by ultra-performance liquid chromatography-tandem mass spectrometry,providing emergency measures for food poisoning caused by cereulide.METHODS Single rice and flour samples were extracted with acetonitrile aqueous solution,salting out,after centrifuged and filmed,the organic phase was directly determined.The complex matrix samples fried rice and noodles were extracted with acetonitrile aqueous solution,cleaned up with HLB column,a ACQUITY UPLC Peptide BEH C_(18)300?column(2.1 mm×100 mm,1.7μm)was used for liquid chromatography separation,multiple reaction monitoring(MRM)mode was used for detection in electrospray ionization with positive ion mode,and quantified by the solvent standard curve method.RESULTS At the spiked level of 0.5,1.0 and 5.0μg/kg,the recoveries of cereulide in negative steamed rice,steamed bread and noodles samples were 87.4%-98.3%,and the relative standard deviations were 1.4%-4.2%.At the spiking levels of 1.0,2.0 and 5.0μg/kg,the recoveries of cereulide in the negative samples such as fried steamed rice and fried noodles were 89.5%-99.3%with the relative standard deviations of 1.1%-7.5%(n=6).The detection limit of cereulide was 0.2-0.3μg/kg,and the quantification limit was 0.5-1.0μg/kg.The established method was applied to the detection of the actual samples causing food poisoning in a certain place in Beijing.The content of cereulide in poisoned food samples was 1287-7398μg/kg,the content of cereulide in two raw materials cold noodles was 0.4 and 9.4μg/kg.CONCLUSION The method is simple,rapid,high sensitivity and accurate,and can realize the rapid treatment of food poisoning caused by cereulide.

关 键 词：蜡样芽胞杆菌 呕吐毒素 食物中毒 液相色谱-串联质谱 

分 类 号：R155.5[医药卫生—营养与食品卫生学] TS207.3[医药卫生—公共卫生与预防医学]