摘要
建立了全自动柱固相萃取超高效液相色谱法测定粮油中黄曲霉毒素的分析方法.样品经84/16(V/V)的乙腈/水涡旋振荡提取20 min,7000 r/min,离心5 min,0.1%PBST水溶液10倍稀释后,稀释液置于全自动固相萃取仪样品架上,选取柱本底未检出,柱回收在90%以上的黄曲霉毒素免疫亲和柱进行全自动免疫亲和净化富集,上样速度为5 mL/min,2 mL甲醇洗脱,50℃下氮气吹干,1 mL流动相复溶上机检测.结果表明:方法的加标回收率为85%~104%,日内相对标准偏差为1.8%~3.0%,日间精密度为1.8%~4.8%.测定三种基体质控物质的结果均在标示值范围内,测定结果准确可靠.通过配对t检验,仪器间无显著台间差,多次重复验证无仪器残留,方法稳定性优于手动法.利用本方法对样品中黄曲霉毒素B1进行了测量不确定度评定,不确定度的主要来源是标准储备液、标准曲线配制和回收率.本方法能够最大限度地减少实验人员的工作量,提高工作效率,还可避免因人员操作导致的结果偏差,适用于大量粮油中黄曲霉毒素的精准测定.
A high throughput automated immunoaffinity cleaning up analytical method for aflatoxin in cereals and oils was developed.The aflatoxin in samples were extracted by using 84/16 acetonitrile-water mixture by vortex for 20 min.0.1%PBST water solution was used to dilute the extraction for 10 times.After being centrifuged with 7000 rpm for five minutes,the supernatant was collected for cleaning up by automated solid phase extraction with the aflatoxin immunoaffinity columns.The performance of the columns was checked,no background was observed and the recovery was above 90%.Sample loading speed was set at 5 mL/min,columns were eluted by 2 mL methanol and the eluant was dried up under nitrogen atmosphere at 50℃.The solid residue was dissolved with 1 mL mobile phase and ready for analysis.As the results shown,the spiking recoveries of automated method were 85%~104%,with within-day precision RSD of 1.8%~3.0%and inter-day precision RSD of 1.8%~4.8%obtained.The testing results of three different matrix reference materials were within the assigned range.This method was accurate and reliable,and with no differences between machines and blank residue observed during the whole progress.Compared to manual method,a better stability was gained.The uncertainty of analysis for aflatoxin B 1 in sample was evaluated.The main sources of uncertainty were aflatoxin B 1 standard solution,the course of standard curve preparation and recovery.The automated method could be used to analyze the aflatoxins in cereals and oil samples to avoid the errors caused by human,save the liberty and improve the efficiency using batch processing.
作者
李丽
吴宇
王海波
叶金
轩志宏
刘洪美
王松雪
张峰
何建洪
Li Li;Wu Yu;Wang Haibo;Ye Jin;Xuan Zhihong;Liu Hongmei;Wang Songxue;Zhang Feng;He Jianhong(Academy of National Food and Strategic Reserves Administration,Beijing100037;Guangxi-ASEAN Food Inspection and Testing Center,Nanning530021;Raykol Instrument Co.Ltd.,Xiamen361006)
出处
《中国粮油学报》
EI
CAS
CSCD
北大核心
2020年第7期157-164,共8页
Journal of the Chinese Cereals and Oils Association
基金
“十三五”国家重点研发计划(2017YFC1601300)
中国科协青年人才托举工程(2018QNRC001)。
关键词
全自动固相萃取
超高效液相色谱仪
黄曲霉毒素
免疫亲和柱
不确定度评定
automated immunoaffinity system
high performance liquid chromatography
Aflatoxin Immunoaffinity Column
uncertainty evaluation
