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氯化钙对花生中黄曲霉毒素B1提取及其快速检测结果的影响
引用本文:李敏,陈冉,李培武,张奇,张文,王督,李冉.氯化钙对花生中黄曲霉毒素B1提取及其快速检测结果的影响[J].中国油料,2014(1):117-121.
作者姓名:李敏  陈冉  李培武  张奇  张文  王督  李冉
作者单位:[1]中国农业科学院油料作物研究所,湖北武汉430062 [2]农业部油料作物生物学与遗传育种重点实验室,湖北武汉430062 [3]农业部生物毒素检测重点实验室,湖北武汉430062 [4]农业部油料产品质量安全风险评估实验室武汉,湖北武汉430062 [5]农业部油料及制品质量监督检验测试中心,湖北武汉430062
基金项目:国家科技支撑计划(2012BABl9809);农业行业科技专项(201303088);吉林省世界银行贷款项目(2011-Z57)
摘    要:以氯化钙为蛋白沉淀剂、黄曲霉毒素B1为靶标,探究了70%甲醇水(V/V)提取液中不同含量氯化钙对花生中黄曲霉毒素提取及其免疫检测结果的影响;采用样品加标回收的方法,研究比较了五种不同含量氯化钙对胶体金免疫层析试纸条和ELISA检测结果的影响。研究结果表明,样品经含1%CaCl2(m/V)的甲醇水处理后,用胶体金免疫层析试纸条检测时,试纸条T线颜色梯度变化,不同浓度加标回收率等参数均得到明显改善,前处理效果显著;选取20份花生实际样品,经过这种前处理之后,进行胶体金免疫层析试纸条和ELISA检测,同时将其检测结果与免疫亲和层析高效液相色谱法(IAC-HPLC)进行比对,胶体金免疫层析试纸条与IAC-HPLC检测结果符合率为90%,间接竞争ELISA与IAC-HPLC检测结果相关系数达0.996,表明本研究所改进的样品前处理方法能提高免疫检测准确度,具有很好的应用性。

关 键 词:花生  黄曲霉毒素B1  氯化钙  胶体金免疫层析  ELISA

Effect of calcium chloride on extraction and rapid detection of AFB1 in peanut
Authors:LI Min  CHEN Ran  LI Pei - Wll  ZHANG Qi  ZHANG Wen  WANG Du  LI Ran
Affiliation:1. Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, China; 2. Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan 430062, China ; 3. Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, China; 4. Laboratory of Risk Assessment for Oilseeds Products ( Wuhan) , Ministry of Agriculture, Wuhan 430062, China ; 5. Quality Inspection and Test Center for Oilseeds Products, Ministry of Agriculture, Wuhan 430062, China)
Abstract:In order to establish a pretreatment method of samples to match the immune analysis, dltIerent quantity of calcium chloride were added in methanol aqueous solution, their effects on extraction and rapid detection of AFB1 to medium in peanut was studied. Samples were detected by IC strip test and ELISA after adding AFB1 to matrixes of peanut samples. The results showed that methanol aqueous solution with 1% CaCl2 was most suitable for pretreatment of peanuts. Samples extracted by methanol aqueous solution with 1% CaCl2 were determined by IC strip test and ELISA. Meanwhile, we compared the results with that obtained by HPLC method. The coincidence rate of results obtained by both IC strip test and HPLC method was 90% ; the correlation coefficient of results ob- tained by both indirect competitive ELISA and HPLC method was 0. 996. The results demonstrated this method could accurately measure the content of AFB1 in peanut and provide technical support for further development of pretreatment method for aflatoxin detection.
Keywords:Peanut  AFB1  Calcium chloride  Colloidal gold immunochromatography  ELISA
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