均匀设计在ELISA分析方法条件优化实验中的应用
The Application of Uniform Design for the Optimization of ELISA Method
-
摘要: 以对硫磷为对象,通过与常规方法相比较,对均匀设计法在ELISA分析方法条件优化实验中应用的适用性进行了研究。采用常规方法对对硫磷ELISA分析方法条件优化的结果为:甲醇体积分数为5％,离子强度0.50 mol/L和pH 7.40,此时建立的ELISA方法IC50值为 89.17 ng/mL,IC20值为 12.49 ng/mL。而采用均匀设计法对对硫磷ELISA分析方法条件进行优化,并对实验结果进行多元二项式统计回归,预测了最佳条件组合的取值,并进行了ELISA实验验证,结果表明:当甲醇体积分数为10％、离子强度为2.00 mol/L、pH为 5.00时,建立的ELISA方法的IC50值为9.96 ng/mL,IC20值为1.21 ng/mL。将两种方法应用于5种性质不同的环境土壤中对硫磷的检测,结果表明,采用均匀设计法进行ELISA分析方法条件优化实验快速简便,能够筛选得到最优条件组合,建立的ELISA方法具有更好的广谱性,能够更广泛地应用于基质差异较大的同类样品检测。Abstract: The application of uniform design(UD) for the optimization of ELISA method for determining parathion in soil was investigated and compared with the regular optimization method.Firstly,the regular method for the optimization of ELISA showed the optimal conditions were 5% methanol,ionic strength 0.50 mol/L and pH 7.40.Under these conditions,the method showed the IC50 and IC20 were 89.17 ng/mL and 12.49 ng/mL,respectively.Further,the uniform design was introduced into the optimization of ELISA and the results were computed by statistical method.Then the forecasting model was brought out and the optimized conditions were recommended.The results showed the optimal conditions were with 10% methanol,ionic strength 2.00 mol/L and at pH of 5.00.The IC50 and IC20 of this UD optimized method were 9.96 ng/mL and 1.21 ng/mL,respectively.Both two optimized methods were applied for the residue determination of parathion in five types of soils.The results showed the application of uniform design for the optimization of ELISA method was quick and easy compared with the regular method . Moreover ,the optimized conditions from the uniform design method contributed to a more broad-specific method,which could be applied for parathion determination in complex samples with different matrix effects.
下载: