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食品中农药残留的高通量微生物检测方法研究

Detection of pesticides residues in food with high-throughput method

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【作者】 李亮林曦薛早早胡文坚胡瑜杜季梅刘佳明

【Author】 LI Liang1,LIN Xi2,XUE Zao-zao1,HU Wem-jian1,HU Yu1,DU Ji-mei3,LIU Jia-ming1(1.School of Environment al Science and Public Health,Wenzhou Medical College,Wenzhou 325035,China;2.Wenzhouy Environmental Protection Design & Science Institute,Wenzhou 325035,China;3.School of Medical Laboratory Science,Wenzhou Medical College,Wenzhou 325035,China)

【机构】 温州医学院环境与公共卫生学院温州市环境保护设计科学研究院温州医学院检验医学院

【摘要】 目的甲胺磷的微生物高通量定量分析法,并在待检蔬菜样品进行初步的探索。方法以短双歧杆菌LJM-006作为测试菌,选择浓度为0、0.25、0.5、1.0、2.0和4.0 mg/L的甲胺磷标准品溶液20μL加入所述检测管内,37℃厌氧培养8~10 h,分别测定反应管和阴性对照反应管的A500值,B值为样品反应管的A500值,B0值为阴性对照反应管的A500值;以B/B0为纵坐标Y,甲胺磷浓度的对数值为横坐标X,绘制标准曲线,建立甲胺磷含量与B/B0的线性回归方程;以此方法进行模拟食品样品中所含甲胺磷检测,进行准确度、精密度分析。结果双歧杆菌菌株LJM-006对甲胺磷敏感性高,与其他有机磷农药的交叉反应率均小于2%;在0.01~100 mg/L浓度范围内B/B0与甲胺磷浓度对数值的线性关系良好,甲胺磷检测的标准曲线为Y=-24.68X+66.72,R2为0.9902,IC50=0.25 mg/L,检测下限达到0.1 mg/L,回收率为78.5%~105.7%,批内变异系数≤12%,批间变异系数≤8.2%。结论该方法具有操作简便,具备一定灵敏度和特异性等优点,可作为一种食品中甲胺磷残留筛检方法。

【Abstract】 Objective To establish microbiological high-throughput method for Methamidophos residue detection,and conduct a preliminary evaluation of the method with vegetables as the test samples.Method Bifidobacterium breve LJM-006 was used as the test microorganism.Each 20 μL of standard methamidophos solutions at the concentrations of 0 mg/L,0.25 mg/L,0.5 mg/L,1 mg/L,2 mg/L,and 4 mg/L was added to the testing tubes,respectively,then the tubes were cultured at 37℃ in anaerobic condition for 8 to 10 hours.The A500 values of the sample reaction tubes(B) and the negative control reaction tubes(B0) were detected to get a standard curve using B/B0 as the vertical ordinate and the logarithm of Methamidophos concentration as the horizontal coordinate;The linear regression equation of Methamidophos content and B/B0 was established to detect the methamidophos contained in simulated food samples;Accuracy and precision analyses were performed.Result The Bifidobacterium breve LJM-006 showed high sensitivity to methanmidophos and a very low cross reaction rate with other organic phosphorus pesticides(<2%).In concentration range of 0.01~100 mg/L B/B0 and methamidophos concentration had a good linear relation.A standard curve was obtained and the regression equation was Y=-24.68X+66.72,and R2 was 0.9902.The 50% inhibition value(IC50) was 0.25 mg/L.The low detection limit was 0.1 mg/L;the recovery rate was 78.5%~105.7%;Intraassay variation coefficient was less than 12%,while interassay variation coefficient less than 8.2%.Conclusion The method is easy to operate,with certain sensitivity and specificity,and can be used as a screening method of Methamidophos Residue in food.

【基金】 浙江省科技厅公益性技术应用研究计划项目(2010C33091);浙江省大学生科技创新活动计划项目(2010R413040)
  • 【文献出处】 中国微生态学杂志 ,Chinese Journal of Microecology , 编辑部邮箱 ,2012年04期
  • 【分类号】TS207.53
  • 【被引频次】8
  • 【下载频次】242
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