环境水样中六价铬酶联免疫检测方法的建立
evelopment of enzyme-linked immunoassay for hexavalent chromium detection in environmental water samples
以抗Cr3+-EDTA单克隆抗体为基础，建立优化的间接竞争ELISA方法和样品前处理方法，用于环境水样中Cr6+的检测。分别配制不同pH及不同离子强度的PBS保温液，优化筛选ELSIA的最佳反应pH值与离子强度。前处理过程中，通过调节样品pH，沉淀去除水样原有Cr3+；优化各项还原条件使Cr6+还原为Cr3+，调节pH进行ELISA检测，通过检测出的Cr3+的量计算原有Cr6+的含量。结果表明，当ELISA 保温液pH为7.4，离子强度为0.15M时，ELISA的灵敏度（IC50）最佳，为5.9 ng/mL，检测限为0.35 ng/mL，线性范围是0.35~50 ng/mL。当还原剂为60 μg/mL NaHSO3，还原pH为3，反应时间为15 min时样品前处理效率最高。以结合最优样品前处理方法的ELISA检测实际水样并与ICP检测结果比较，得到两种方法检测结果数据的相关系数达0.9887，表明本研究建立的ELISA方法检测结果可靠。加标实验结果显示该ELSIA方法变异系数小于12%，说明该方法具有良好的重复性和精密度，可用于环境水样六价铬离子的监测。
n optimal indirect competitive ELISA was developed using monoclonal antibody against Cr3+-EDTA. The pretreatment procedure was also established. Reaction conditions, such as pH and ionic strength, were studied to optimize the immunoassay. In pretreatment procedure, the Cr3+ existent in samples was precipitated by adjusting pH and the reducing conditions of Cr6+ were researched completely, the concentration of Cr6+ in samples were calculated as Cr3+ detected by ELISA. Results showed that the ELISA had the most sensitive to soluble Cr3+-EDTA when pH was 7.4 and ionic strength was 0.15 M, the IC50 and limit of detection was 5.9ng/mL, 0.35ng/mL, respectively, and the liner range was 0.35~50ng/mL. The collected supernatant containing Cr6+ was mixed with 60mg/L of NaHSO3 solution, the Cr6+ was reduced to Cr3+ at pH 3.0 and reacted for 15 min. Finally, ELISA was used to detect the concentration of Cr6+ in samples. The correlation coefficient between the ELISA and ICP was 0.9887 and the coefficient of variation was below 12% indicated that the immunoassay was reliable and precise, it was suitable for detect water samples.
宋建勋、唐勇、欧子强、邹军辉、钟辉
环境污染、环境污染防治
六价铬ELISA环境水样
hexavalent chromiumELISAwater samples
宋建勋,唐勇,欧子强,邹军辉,钟辉.环境水样中六价铬酶联免疫检测方法的建立[EB/OL].(2012-03-14)[2025-08-18].http://www.paper.edu.cn/releasepaper/content/201203-454.点此复制
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