两种同时检测茶叶中赭曲霉毒素A与桔霉素方法的比对  

Comparison of 2 methods for the simultaneous detection of ochratoxin A and citrinin in tea

作　　者：王亚青 陈欢 滕建文[1] 夏宁[1] 韦保耀[1] 黄丽[1] WANG Ya-Qing;CHEN Huan;TENG Jian-Wen;XIA Ning;WEI Bao-Yao;HUANG Li(Institute of Light lndustry and Food Engineernig,Guangxi University,Nanning 530000,China)

机构地区：[1]广西大学轻工与食品工程学院,南宁530000

出　　处：《食品安全质量检测学报》2023年第21期71-80,共10页Journal of Food Safety and Quality

基　　金：广西重点研发计划项目(桂科AB21220068);2021—2025年国家现代农业产业技术体系广西牛羊产业创新团队专项(nycytxgxcxtd-21-09-06)。

摘　　要：目的对比两种同时检测茶叶中赭曲霉毒素A(ochratoxin A,OTA)与桔霉素(citrinin,CIT)的方法。方法考察不同提取方法、溶剂、料液比的提取效果,比较适配体结合超滤法与免疫亲和柱纯化效果;对比高效液相色谱-荧光检测器(high performance liquid chromatography-fluorescence detector,HPLC-FLD)与超高效液相色谱-三重四极杆质谱法(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)测定OTA与CIT的检出限、精密度、准确度等差异,评价方法的适用性。结果采用70%甲醇超声与振荡的提取方法,在料液比为1:3的条件下,OTA、CIT的提取率最佳,回收率分别为99.49%的92.77%;经适配体结合超滤纯化后,CIT与OTA的回收率为82.07%~92.95%,与免疫亲和柱纯化方法效果类似;UPLC-MS/MS测定CIT和OTA的基质效应(20.0%、-20.0%)小于HPLC-FLD测定CIT和OTA的基质效应(48.8%、50.3%),二者测定CIT与OTA均具有良好线性度,相关性系数均不小于0.9989;HPLC-FLD测定CIT与OTA的检出限分别为0.09μg/kg、0.12μg/kg;UPLC-MS/MS测定CIT和OTA的检出限分别为0.12μg/kg、0.09μg/kg,两种方法的加标回收率分别为84.20%~100.70%与89.22%~97.88%,相对标准偏差分别为1.30%~4.31%与0.74%~4.04%。结论适配体结合超滤纯化方法操作简单,成本低廉,免疫亲和柱纯化方法成本较高且低浓度下的准确度高;HPLC-FLD和UPLC-MS/MS均适用于茶叶基质中CIT与OTA的提取与检测,但后者基质效应更低,定性更准确。Objective To compare 2 methods for the simultaneous detection of ochratoxin A(OTA)and citrinin(CIT)in tea.Methods The effects of different extraction methods,solvents,and sample-to-solvent ratios on extraction efficiency were investigated,the binding of molecularly imprinted superfiltration was compared with immunoaffinity column purification.The limits of detection,precision,accuracy and other differences between high performance liquid chromatography-fluorescence detector(HPLC-FLD)and ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)in determining OTA and CIT were evaluated to assess the applicability of the detection methods.Results When the ratio of solid to liquid was 1:3,the best extraction rates of OTA and CIT were obtained by 70%methanol ultrasonic and oscillating extraction method,and the recoveries were 99.49%and 92.77%,respectively.After purification by aptamer-binding ultrafiltration,the recoveries of CIT and OTA were 82.07%‒92.95%,which were similar to the results obtained using immunoaffinity column purification alone.The matrix effects of UPLC-MS/MS for quantitation of CIT and OTA(20.0%,‒20.0%)were lower than HPLC-FLD(48.8%,50.3%).Both methods showed good linearity for CIT and OTA,with correlation coefficients not less than 0.9989.The limits of detection for CIT and OTA by HPLC-FLD were 0.09μg/kg and 0.12μg/kg,respectively.The limits of detection for CIT and OTA by UPLC-MS/MS were 0.12μg/kg and 0.09μg/kg,respectively.Spike recoveries ranged from 84.20%‒100.70%with relative standard deviations of 1.30%‒4.31%for HPLC-FLD and 89.22%‒97.88%with relative standard deviations of 0.74%‒4.04%for UPLC-MS/MS.Conclusion Aptamer combined with ultrafiltration purification method is simple and cost-effective,immunoaffinity column purification is more costly and has high accuracy at low concentrations.Both HPLC-FLD and UPLC-MS/MS are suitable for the extraction and detection of CIT and OTA in tea matrix,but the latter has lower matrix effect and more accurate characteriza

关 键 词：茶叶 赭曲霉毒素A 桔霉素 适配体 超滤法 

分 类 号：TS272.7[农业科学—茶叶生产加工]