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基于磁珠的快速免疫分析测定法用于灵敏检测双氯芬酸。

A rapid magnetic bead-based immunoassay for sensitive determination of diclofenac.

机构信息

Department of Analytical Chemistry; Reference Materials, Bundesanstalt für Materialforschung und -prüfung (BAM), 12489, Berlin, Germany.

Department of Chemistry, Humboldt-Universität zu Berlin, 12489, Berlin, Germany.

出版信息

Anal Bioanal Chem. 2022 Feb;414(4):1563-1573. doi: 10.1007/s00216-021-03778-7. Epub 2021 Nov 20.

Abstract

Increasing contamination of environmental waters with pharmaceuticals represents an emerging threat for the drinking water quality and safety. In this regard, fast and reliable analytical methods are required to allow quick countermeasures in case of contamination. Here, we report the development of a magnetic bead-based immunoassay (MBBA) for the fast and cost-effective determination of the analgesic diclofenac (DCF) in water samples, based on diclofenac-coupled magnetic beads and a robust monoclonal anti-DCF antibody. A novel synthetic strategy for preparation of the beads resulted in an assay that enabled for the determination of diclofenac with a significantly lower limit of detection (400 ng/L) than the respective enzyme-linked immunosorbent assay (ELISA). With shorter incubation times and only one manual washing step required, the assay demands for remarkably shorter time to result (< 45 min) and less equipment than ELISA. Evaluation of assay precision and accuracy with a series of spiked water samples yielded results with low to moderate intra- and inter-assay variations and in good agreement with LC-MS/MS reference analysis. The assay principle can be transferred to other, e.g., microfluidic, formats, as well as applied to other analytes and may replace ELISA as the standard immunochemical method.

摘要

环境水中药物的污染不断增加,对饮用水的质量和安全构成了新的威胁。在这方面,需要快速、可靠的分析方法,以便在污染发生时迅速采取对策。在此,我们报告了一种基于磁珠的免疫测定法(MBBA)的开发,该方法可快速、经济有效地测定水样中的镇痛药双氯芬酸(DCF),该方法基于双氯芬酸偶联磁珠和一种强大的单克隆抗 DCF 抗体。一种新的合成策略用于制备珠子,使得可以用比相应的酶联免疫吸附测定(ELISA)明显更低的检测限(400ng/L)来测定双氯芬酸。由于孵育时间更短,且仅需进行一次手动洗涤步骤,因此与 ELISA 相比,该测定法所需的时间更短(<45 分钟),且所需设备更少。用一系列加标水样评估测定的精密度和准确性,结果显示内和间测定的变异度低至中度,与 LC-MS/MS 参考分析结果一致。该测定原理可转移到其他例如微流控的格式,以及应用于其他分析物,并可能取代 ELISA 成为标准免疫化学方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/45a0/8761716/3b309591d014/216_2021_3778_Fig1_HTML.jpg

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