间接竞争酶联免疫吸附法快速筛检莲子中黄曲霉毒素B1的污染水平(1)
[摘要]建立间接竞争酶联免疫吸附法(ic-ELISA)快速筛检“药食同源”中药-莲子中黄曲霉毒素B1(AFB1)的污染水平，并采用超快速液相串联质谱(UFLC-MS/MS)进行结果确证。通过基质匹配，AFB1在0.171～7.25 μg·L-1线性关系良好(R2 > 0.978)，半数抑制浓度(IC50)为1.29 μg·L-1，加样回收率为74.73% ～ 126.9%，RSD<5%，检测限(IC10)为0.128 μg·L-1。将ic-ELSIA法应用于20批莲子中AFB1的筛查，结果显示：于30 ℃，30%湿度条件下贮藏一年的1～15号样品中AFB1污染水平为1.19～115.3 μg·kg-1，40%样品中AFB1含量超过限量标准(5 μg·kg-1);新购买的16～20号样品中AFB1污染率为40%。以上结果经液质联用技术确证，通过计算ic-ELISA和液质测定结果之间的皮尔森相关系数可知，二者测定结果的匹配度达0.997。建立的ic-ELISA分析方法操作简便、灵敏、结果准确，可用于莲子中AFB1的高通量现场快速检测。
, http://www.100md.com
[关键词]莲子;黄曲霉毒素B1;快速筛检;间接竞争-酶联免疫吸附法;液质联用法
Contamination level of aflatoxin B1 in lotus seeds rapid screening
by indirect competitive ELISA method
CHU Xian-feng1，2， DOU Xiao-wen2， KONG Wei-jun2， YANG Mei-hua2*，ZHAO Chong3， ZHAO Ming1*， OUYANG Zhen1
(1.School of Pharmacy， Jiangsu University， Zhenjiang 212013， China;
2. Institute of Medicinal Plant Development Chinese Academy of Medical Science，Peking Union Medical College，Beijing 100193， China; 3. Beijing Hapten and Protein Biomedical Institute， Beijing 102202， China)
, http://www.100md.com
[Abstract]A simple and cost-effective indirect competitive enzyme-linked immune sorbent assay(ic-ELISA) was developed to rapidly screen the content of aflatoxin B1(AFB1) in lotus seeds， and the results were confirmed by ultra-fast liquid chromatography-tandem mass spectrometry(UFLC-MS/MS). Matrix-matched calibration expressed a good linearity ranging from 0.171 to 7.25 μg·L-1 for AFB1 with R2>0.978. The medium inhibitory concentration(IC50) for AFB1 was 1.29 μg·L-1， the recovery for AFB1 was 74.73% to 126.9% with RSD<5%， and the limit of detection(IC10) was 0.128 μg·L-1. The developed ic-ELSIA method was applied to rapid analysis of AFB1 in 20 lotus seeds samples and the results indicated that the contents of AFB1 in samples 1-15 were in the range of 1.19-115.3 μg·kg-1 and in 40% of the samples exceeded the legal limit(5 μg·kg-1)， while the contamination rate of AFB1 in samples 16-20 was 40%. Pearson correlation coefficient(r) reached 0.997 for AFB1 content in the samples detected by ic-ELSIA and UFLC-MS/MS methods. The results proved that the developed ic-ELISA method is simple， sensitive and reliable， and can be used for rapid and high-throughput screening of AFB1 in lotus seeds.
, http://www.100md.com
[Key words]Lotus seeds; AFB1; rapid screening; ic-ELISA; UFLC-MS/MS
doi：10.4268/cjcmm20150424
真菌毒素(mycotoxin)是由产毒真菌在适宜的环境条件下产生的有毒次生代谢产物[1]，食品、农作物及饲料、中药材[2]等植物源性产品储存不当极易造成真菌毒素污染[3]。目前已发现约400种真菌毒素[4-5]，其中，黄曲霉毒素类(aflatoxins，AFs)较为常见，而黄曲霉毒素B1(AFB1)分布最广，且毒性是氰化钾的10倍，具有严重的肝毒性和致癌、致畸、致突变等作用[6-7]，常作为黄曲霉毒素的主要检测指标[8-9]。, http://www.100md.com(褚先锋 豆小文 孔维军 杨美华 赵翀 赵明 欧阳臻)
, http://www.100md.com
[关键词]莲子;黄曲霉毒素B1;快速筛检;间接竞争-酶联免疫吸附法;液质联用法
Contamination level of aflatoxin B1 in lotus seeds rapid screening
by indirect competitive ELISA method
CHU Xian-feng1，2， DOU Xiao-wen2， KONG Wei-jun2， YANG Mei-hua2*，ZHAO Chong3， ZHAO Ming1*， OUYANG Zhen1
(1.School of Pharmacy， Jiangsu University， Zhenjiang 212013， China;
2. Institute of Medicinal Plant Development Chinese Academy of Medical Science，Peking Union Medical College，Beijing 100193， China; 3. Beijing Hapten and Protein Biomedical Institute， Beijing 102202， China)
, http://www.100md.com
[Abstract]A simple and cost-effective indirect competitive enzyme-linked immune sorbent assay(ic-ELISA) was developed to rapidly screen the content of aflatoxin B1(AFB1) in lotus seeds， and the results were confirmed by ultra-fast liquid chromatography-tandem mass spectrometry(UFLC-MS/MS). Matrix-matched calibration expressed a good linearity ranging from 0.171 to 7.25 μg·L-1 for AFB1 with R2>0.978. The medium inhibitory concentration(IC50) for AFB1 was 1.29 μg·L-1， the recovery for AFB1 was 74.73% to 126.9% with RSD<5%， and the limit of detection(IC10) was 0.128 μg·L-1. The developed ic-ELSIA method was applied to rapid analysis of AFB1 in 20 lotus seeds samples and the results indicated that the contents of AFB1 in samples 1-15 were in the range of 1.19-115.3 μg·kg-1 and in 40% of the samples exceeded the legal limit(5 μg·kg-1)， while the contamination rate of AFB1 in samples 16-20 was 40%. Pearson correlation coefficient(r) reached 0.997 for AFB1 content in the samples detected by ic-ELSIA and UFLC-MS/MS methods. The results proved that the developed ic-ELISA method is simple， sensitive and reliable， and can be used for rapid and high-throughput screening of AFB1 in lotus seeds.
, http://www.100md.com
[Key words]Lotus seeds; AFB1; rapid screening; ic-ELISA; UFLC-MS/MS
doi：10.4268/cjcmm20150424
真菌毒素(mycotoxin)是由产毒真菌在适宜的环境条件下产生的有毒次生代谢产物[1]，食品、农作物及饲料、中药材[2]等植物源性产品储存不当极易造成真菌毒素污染[3]。目前已发现约400种真菌毒素[4-5]，其中，黄曲霉毒素类(aflatoxins，AFs)较为常见，而黄曲霉毒素B1(AFB1)分布最广，且毒性是氰化钾的10倍，具有严重的肝毒性和致癌、致畸、致突变等作用[6-7]，常作为黄曲霉毒素的主要检测指标[8-9]。, http://www.100md.com(褚先锋 豆小文 孔维军 杨美华 赵翀 赵明 欧阳臻)