UPLC-MS快速测定5种中药中米酵菌酸的含量及病原菌的鉴定  

Rapid determination of bongkrekic acid content in 5 Chinese herbs by UPLC-MS and the identification of pathogenic bacteria

作　　者：朱仁愿 陈婷 续艳丽 白雯静 丁辉 ZHU Renyuan;CHEN Ting;XU Yanli;BAI Wenjing;DING Hui(Gansu Provincial Engineering Research Center for Monitoring Exogenous Pollutants in Planted Chinese Medicinal Materials,Key Laboratory for Monitoring Pesticide and Veterinary Drug Residues in Food under National Market Supervision,Lanzhou Institute of Food and Drug Control,Lanzhou,Gansu,730050 P.R.China)

机构地区：[1]兰州市食品药品检验检测研究院国家市场监管食品中农药兽药残留监控重点实验室甘肃省种植中药材外源性污染物监测工程研究中心,甘肃兰州730050

出　　处：《华西药学杂志》2024年第6期705-709,共5页West China Journal of Pharmaceutical Sciences

基　　金：国家市场监督管理总局科技计划项目(2023MK120);甘肃省科技重大专项-企业创新联合体项目(22ZD6FA021-3);甘肃省药品监督管理局科研项目(2022GSMPA0067);兰州市科技计划项目(2023-3-29)。

摘　　要：目的采用超高效液相色谱-四极杆-静电场轨道阱高分辨质谱联用(UPLC-MS)快速筛查5种中药(六神曲、建曲、灵芝、猪苓、海藻)中米酵菌酸的含量。方法样品水浸泡后,用1%乙酸乙腈振荡涡旋提取,经分散固相萃取净化,氮吹干后用乙腈溶液复溶,采用UPLC-MS电喷雾负离子同时进行一级、二级全扫描,得到母离子和碎片离子精确质量数,构建质谱数据库实现快速定性分析,同时用外标法定量,并采用微生物病原菌鉴定进行验证。结果5种中药的米酵菌酸基质对照液在30~200μg·L^(-1)线性关系良好,r≥0.9917,检出限为10~20μg·kg^(-1),定量限为30~50μg·kg^(-1),平均回收率为72.9%~118.0%,RSD为1.70%~7.40%(n=6);样品中均未检出米酵菌酸。病原菌鉴定出与唐菖蒲伯克霍尔德氏菌菌落特征相似的肺炎克雷伯菌、阴沟肠杆菌复合菌和洋葱伯克霍尔德菌。结论所用方法基质干扰小、样品分析快,与传统微生物检验方法的结果基本一致,且检验时长缩短。OBJECTIVE To rapid determine the bongkrekic acid content in 5 Chinese herbs including Liushenqu,Jianqu,Ganoderma,Polyporus,and Sargassum by UPLC-MS.METHODS The samples were soaked in water and extracted with 1%glacial acetic acid acetonitrile solution with oscillating vortex,purified by dispersed solid phase extraction,nitrogen blow-dried and redissolved with acetonitrile solution.The primary and secondary high-precision full scanning was carried out simultaneously by electrospray negative ion of UPLC-MS at the same time,and the exact mass numbers of parent ions and fragment ions were obtained.A mass spectrum database was constructed to achieve rapid qualitative analysis;quantitative analysis was performed by external standard method;and microbial pathogen identification was used for verification.RESULTS The matrix standard liquid of 5 Chinese herbs had a good linear relationship in the concentration range of 30-200μg·L^(-1),with correlation coefficient r≥0.9917,the limit of detection of 10-20μg·kg^(-1),and the limit of quantification of 30-50μg·kg^(-1).The average recoveries were 72.9%-118.0%,and RSD was 1.70%-7.40%(n=6).No bongkrekic acid was detected in the samples.Bacteria such as Klebsiella pneumoniae,Enterobacter cloacae complex and Burkholderia cepacia were identified with similar colony characteristics to Burkholderia gladiolus.CONCLUSION This method demonstrates low matrix interference and fast sample analysis.Compared with the traditional method,the experimental results are basically the same,and the test time is greatly shortened.

关 键 词：六神曲 建曲 灵芝 猪苓 海藻 米酵菌酸 超高效液相色谱-四极杆-静电场轨道阱高分辨质谱联用 病原菌鉴定 

分 类 号：R917[医药卫生—药物分析学]