超高效液相色谱-串联质谱法测定发酵乳制品中米酵菌酸和异米酵菌酸  

Determination of Bongkrekic Acid and Iso-bongkrekic Acid in Fermented Dairy Products by UPLC-MS/MS

作　　者：李文廷[1] 叶沛 刘玲 陶蓉蓉 张瑞雨[1] 师真[1] 蒋孟圆[3] LI Wen-ting;YE Pei;LIU Ling;TAO Rong-rong;ZHANG Rui-yu;SHI Zhen;JIANG Meng-yuan(Kunming Center for Disease Control and Prevention,Kunming 650228,China;School of Public Health,Kunming Medical University,Kunming 650500,China;Key Laboratory of Chemistry in Ethnic Medicinal Resources,State Ethnic Affairs Commission and Ministry of Education,Yunnan Minzu University,Kunming 650504,China)

机构地区：[1]昆明市疾病预防控制中心,云南昆明650228 [2]昆明医科大学公共卫生学院,云南昆明650500 [3]云南民族大学民族药资源化学国家民族事务委员会-教育部重点实验室,云南昆明650504

出　　处：《分析测试学报》2025年第2期326-333,共8页Journal of Instrumental Analysis

基　　金：云南省基础研究计划项目(202401AT070040);昆明市科技计划项目(昆科计字2024-1-NS-0034);昆明市卫生科技人才培养项目(2022-SW(带头)-16)。

摘　　要：该研究采用液液萃取技术结合超高效液相色谱-串联质谱法对发酵乳制品中的米酵菌酸和异米酵菌酸两种细菌毒素进行监测分析,以防控细菌毒素中毒事件的发生。以发酵乳制品为基质,通过考察提取试剂、净化剂和盐析剂以及仪器参数对两种细菌毒素测定效果的影响,确定前处理条件为采用90%丙酮-1%甲酸溶液涡旋提取,高速离心浓缩,经0.22μm滤膜过滤后进行检测。在最佳条件下,米酵菌酸和异米酵菌酸在1~200 ng/mL范围内线性关系良好,相关系数(r)均大于0.999,检出限分别为0.075、0.11μg/kg,在3个加标水平下的回收率为90.8%~106%,相对标准偏差为0.80%~6.1%。该方法相较于其他方法更高效便捷,且无需前处理净化操作,展现了更佳的提取率、更高的灵敏度及更强的准确性,适用于发酵乳制品尤其是批量样品中米酵菌酸和异米酵菌酸的测定。Bacterial toxins produced by fermented food are very harmful,and the establishment of efficient and reliable detection methods can ensure food safety and protect human health.Liquid-liq⁃uid extraction combined with ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)was used to monitor and analyze two bacterial toxins bongkrekic acid and iso-bong⁃krekic acid in fermented dairy products,so as to prevent and control the occurrence of bacterial toxin poisoning.In this study,fermented dairy products were used as the substrate,and the effects of ex⁃traction reagents,purification agents and salting agents in sample pretreatment and instrument pa⁃rameters on the determination of two bacterial toxins were investigated.The final treatment conditions were as follows:90%acetone-1%formic acid solution was used for vortex extraction,after high-speed centrifugation and concentration,it was filtered by 0.22μm filter membrane and detected by instrument.Under the optimum conditions,the linear relationship of the two bacterial toxins was good in the range of 1-200 ng/mL,with the correlation coefficient(r)more than 0.999.The limits of detection for bongkrekic acid and iso-bongkrekic acid were 0.075μg/kg and 0.11μg/kg,respective⁃ly.The recoveries were 90.8%-106%and the relative standard deviations were 0.80%-6.1%.Com⁃pared with other methods,this method is more efficient and convenient,without pre-treatment and purification operation,and shows better extraction rate,higher sensitivity and higher accuracy.It is suitable for the determination and analysis of bongkrekic acid and iso-bongkrekic acid in fermented dairy products,especially in batch samples.

关 键 词：发酵乳制品 米酵菌酸 异米酵菌酸 超高效液相色谱-串联质谱 细菌毒素 

分 类 号：O657.7[理学—分析化学] TS207.3[理学—化学]