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使用测试生物组和光学氧呼吸测定法对化学和环境样品进行毒理学分析。

Toxicological profiling of chemical and environmental samples using panels of test organisms and optical oxygen respirometry.

作者信息

Zitova Alice, O'Mahony Fiach C, Cross Maud, Davenport John, Papkovsky Dmitri B

机构信息

Biochemistry Department and ABCRF, University College Cork, Cavanagh Pharmacy Building, Cork, Ireland.

出版信息

Environ Toxicol. 2009 Apr;24(2):116-27. doi: 10.1002/tox.20387.

Abstract

A simple and versatile methodology for high throughput toxicological assessment of chemical and environmental samples is presented. It uses panels of test organisms ranging from prokaryotic (E. coli, V. fischeri) and eukaryotic (Jurkat) cells to invertebrate (Artemia salina) and vertebrate (Danio rerio) organisms, to analyze alterations in their oxygen consumption by optical oxygen respirometry. All the assays are carried out in a convenient microtiter plate format using commercial reagents (phosphorescent oxygen probe, microplates) and detection on a standard fluorescent plate reader. Simple experimental set-up and mix-and-measure procedure allow parallel assessment of up to 96 samples (or assay points) in 2 h, easy generation of dose- and time-dependent responses, and EC(50) values. The methodology was demonstrated with several different classes of chemicals including heavy metal ions, PAHs, pesticides, their mixtures, and also validated with complex environmental samples such as wastewater from a wastewater treatment plant. It has been shown to provide high sensitivity, sample throughput and information content, flexibility and general robustness. It allows ranking and profiling of samples, compares favorably with alternative methods such as MicroTox and mortality tests with animal models, and is well suited for large-scale monitoring programs such as CWA and WFD.

摘要

本文介绍了一种用于化学和环境样品高通量毒理学评估的简单通用方法。该方法使用一系列测试生物,从原核生物(大肠杆菌、费氏弧菌)和真核生物( Jurkat细胞)到无脊椎动物(卤虫)和脊椎动物(斑马鱼),通过光学氧呼吸测定法分析它们耗氧量的变化。所有测定均采用方便的微孔板形式,使用商业试剂(磷光氧探针、微孔板),并在标准荧光酶标仪上进行检测。简单的实验设置和混合测量程序允许在2小时内并行评估多达96个样品(或测定点),轻松生成剂量和时间依赖性响应以及半数有效浓度(EC50)值。该方法已通过包括重金属离子、多环芳烃、农药及其混合物在内的几种不同类型的化学品进行了验证,还通过诸如污水处理厂废水等复杂环境样品进行了验证。结果表明,该方法具有高灵敏度、样品通量和信息含量、灵活性和总体稳健性。它可以对样品进行排序和分析,与诸如MicroTox和动物模型死亡率测试等替代方法相比具有优势,非常适合诸如《清洁水法》和《水框架指令》等大规模监测计划。

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