水体中苯并芘检测试剂盒研制及效果评价

陈超超; 邵辉峰; 王吕; 陈雪岚

doi:10.11847/zgggws2016-32-02-33

水体中苯并芘检测试剂盒研制及效果评价

Development and evaluation of an enzyme-linked immunosorbent assay kit for determination of benzo(a)pyrene in water

摘要

摘要: 目的研制间接竞争酶联免疫吸附法(ELISA)检测苯并芘试剂盒。方法采用棋盘滴定法确定抗原抗体的最佳用量;采用间接ELISA分析法确定苯并芘检测的最适条件,包括pH值、盐离子浓度及甲醇浓度。结果抗原最佳包被浓度为10 μg/mL,单克隆抗体稀释倍数为 1:5000;在最优pH值为7.4、盐离子浓度为0.01 mol/L及甲醇浓度为10%的条件下,检测方法在5~50 ng/mL范围内线性相关,线性方程为y=-28.105 Ln(x)+123.66,R²=0.9937,IC₅₀为13.74 ng/mL,苯并芘的最低检出限为3.3 ng/mL;在水样中的加标回收率为94.8%~112.3%;变异系数为4.38%~9.72%。结论该方法适用于水体中苯并芘的检测,可用于大批量样品的快速筛查。

Abstract: Objective To develop an indirect competitive enzyme-linked immunosorbent assay kit for the determination of benzo(a)pyrene(BaP).Methods The optimum concentration of antigen and monoclonal antibody were determined with chessboard titration.The optimum conditions,including pH,saltion and methanol concentration,were separately measured with indirect competitive enzyme-linked immunosorbent assay(icELISA).Results The optimum coating concentration of antigen was 10 μg/mL,while the optimum dilution ratio of antibody was 5 000.The established icELISA for the detection of BaP exhibited a linear range of 5 to 50 ng/mL,a sensitivity of 3.30 ng/mL,and a half inhibitory concentration(IC₅₀)of 13.74 ng/mL in the conditions of pH 7.4,0.01 mol/L of saltion concentration and 10%of methanol.The average recovery was 105%in water samples.Conclusion These results indicate that the icELISA kit developed could be applied in rapid screening detection of BaP in water samples.

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