| 全自动免疫亲和在线净化/高效液相色谱法快速高通量测定饲料中黄曲霉毒素 |
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| 基金项目:粮食公益性行业科研专项(201313005) |
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| 中文摘要:建立了全自动免疫亲和在线净化/高效液相色谱快速高通量测定饲料中黄曲霉毒素(Aflatoxins，AFT)的分析方法。饲料样品经乙腈-水(80∶20，体积比)提取，3 g/L Triton X-100水溶液10倍稀释后，用自动进样器注入RIDA-CREST在线固相萃取系统并流经黄曲霉毒素免疫亲和小柱，以甲醇-水(45∶55，体积比)为流动相，流速为1.0 mL/min，C18色谱柱(150 mm×3.5 mm，5 μm)分离，光化学衍生，荧光检测器测定。根据3倍信噪比的峰响应值，确定黄曲霉毒素B1，B2，G1，G2的检出限分别为0.08，0.05，0.18，0.08 μg/kg，分别在1~100，0.24~24，0.56~56，0.24~24 μg/kg 范围内呈线性相关，相关系数(r2)分别为0.999 4，0.999 7，0.999 8 和0.999 8；AFT在猪饲料、鸡饲料、宠物饲料和饲料原料4类样品中的加标回收率为72.6%~103%，相对标准偏差为2.5%~4.9%。该方法一次装柱可检测60个样品，液相色谱分析一个样品总的运行时间为15 min，所以1 d可检测70~80个样品，满足饲料中黄曲霉毒素快速高通量准确定量检测的需要。 |
| 中文关键词:饲料 黄曲霉毒素 在线免疫亲和柱净化 高效液相色谱 |
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| Rapid and High Throughput Analysis of Aflatoxins in Feed Matrix Using On-line Immunoaffinity Column Clean-up and High Performance Liquid Chromatography |
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| Abstract:A rapid analytical method using on-line immunoaffinity column clean-up and high performance liquid chromatography(HPLC) was developed for the determination of aflatoxins(B1，B2，G1，G2) in feed samples.The samples were extracted with acetonitrile-water solution(80∶20,by volumn).The extraction solution was diluted by 10 times using 3 g/L triton X-100 water solution,then injected into the RIDA-CREST on-line immunoaffinity column clean-up system,and washed out with methanol-water(45∶55,by volume) as mobile phase at a flow rate of 1.0 mL/min.Aflatoxins were analyzed by HPLC/FLD with post-column photochemical derivatizaton.The detection limits of AFB1,AFB2,AFG1 and AFG2 were 0.08,0.05,0.18,0.08 μg/kg,respectively.The linear ranges for AFB1,AFB2,AFG1 and AFG2 were 1-100,0.24-24,0.56-56,0.24-24 μg/kg,with their correlation coefficients(r2) of 0.999 4,0.999 7,0.999 8 and 0.999 8,respectively.The recoveries for four kinds of feed samples spiked with aflatoxins were in the range of 72.6%-103%,with relative standard deviations of 2.5%-4.9%.Each immunoaffinity colomn could be used for 60 samples cleanup,and the total running time for each HPLC analysis is 15 min,thus 70-80 samples could be detected per day.This method could satisfy the demands for rapid and accurate analysis of aflatoxin in feed. |
| Key Words:feed aflatoxins on-line immunoaffinity column clean-up high performance liquid chromatography(HPLC) |
| 引用本文：李丽，黎睿，谢刚,叶金，王松雪.全自动免疫亲和在线净化/高效液相色谱法快速高通量测定饲料中黄曲霉毒素[J].分析测试学报,2017,36(6):800-804. |
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