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Studies of portable immunochromatographic methods for analysis of pesticide residues.

机译：便携式免疫色谱法分析农药残留的研究。

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摘要

This work details the design, development and characterization of a portable immunoextraction method coupled to reversed-phase liquid chromatography for the analysis of herbicide residues. The method gave low limits of detection (LOD), excellent precision and reproducibility. The method was optimized to give a total analysis time of 10 min for the three model s-triazine compounds at a sample throughput of less than 5 min. The lower LOD was dependent upon sample size, and for a 2.00 mL sample was 0.3 μg/L at a S/N = 2. Good stability of the system was observed with within-day precisions of 16% and day-to-day precisions of 16%. This work also explored theoretical aspects of the coupling of these two methods. Assays and equations were developed for examining thermodynamic and kinetic properties of immunoaffinity columns.;A three step assay was used to determine thermodynamic and kinetic properties of antibody columns. The three steps were: (1) an initial application step in which analyze at various concentrations saturated the affinity column, (2) an elution step in which a step gradient elution buffer was applied, and (3) a regeneration step in which the antibodies were returned to the initial buffer conditions. This method allowed calculation of association equilibrium constants as well as association and dissociation rate constants for the antibody-antigen interaction in less than fifteen minutes per assay. Accurate on-column estimations of the thermodynamic and kinetic parameters of interest were shown.;Behavior of analytes at the interface of the antibody column and reversed-phase liquid chromatography column was investigated. An equation describing the application profile of analyte to the reversed-phase column was coupled with partitioning equations describing analyte behavior in the reversed-phase column. Comparison of various parameters illustrated the tendency of peaks to give skewed shapes with decreasing dissociation from the antibody column. Retention of analyte on the precolumn under aqueous conditions dictated the length of time required to elute the analyte, but did not have much effect on the overall peak shape.
机译：这项工作详细介绍了便携式免疫提取方法与反相液相色谱联用用于除草剂残留分析的设计，开发和表征。该方法检测限低（LOD），精密度和重现性极佳。对方法进行了优化，以使三种模型s-三嗪化合物的总分析时间为10分钟，而样品通量少于5分钟。较低的LOD取决于样品量，对于2.00 mL样品，S / N = 2时为0.3μg/ L。日间精度为16％，日精度为系统观察到的系统稳定性良好。占16％。这项工作还探讨了这两种方法耦合的理论方面。建立了检测免疫亲和柱的热力学和动力学特性的方法和方程。采用三步法测定抗体柱的热力学和动力学特性。这三个步骤是：（1）初始应用步骤，其中在各种浓度下分析亲和柱均已饱和;（2）洗脱步骤，其中应用了逐步梯度洗脱缓冲液;（3）再生步骤，其中了抗体返回到初始缓冲条件。该方法允许在每次测定少于十五分钟的时间内计算抗体-抗原相互作用的缔合平衡常数以及缔合和解离速率常数。显示了感兴趣的热力学和动力学参数的准确的柱上估计。;研究了抗体柱和反相液相色谱柱界面处分析物的行为。将描述分析物在反相色谱柱上的应用曲线的方程式与描述分析物在反相色谱柱中的行为的分配方程式结合在一起。各种参数的比较说明了随着与抗体色谱柱的解离减少，峰出现倾斜形状的趋势。在水性条件下，分析物在预柱上的保留时间决定了洗脱分析物所需的时间，但对总体峰形没有太大影响。

著录项

  • 作者

    Nelson, Mary Anne.;

  • 作者单位

    The University of Nebraska - Lincoln.;

  • 授予单位 The University of Nebraska - Lincoln.;
  • 学科 Chemistry Analytical.
  • 学位 Ph.D.
  • 年度 2003
  • 页码 276 p.
  • 总页数 276
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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