Mortimer M, Kasemets K, Heinlaan M, Kurvet I, Kahru A
Laboratory of Molecular Genetics, National Institute of Chemical Physics and Biophysics, Akadeemia tee 23, Tallinn, Estonia.
Toxicol In Vitro. 2008 Aug;22(5):1412-7. doi: 10.1016/j.tiv.2008.02.011. Epub 2008 Feb 26.
Despite of the growing production and use of nanoparticles (NPs) in various applications, current regulations, including EC new chemical policy REACH, fail to address the environmental, health, and safety risks posed by NPs. This paper shows that kinetic Vibrio fischeri luminescence inhibition test--Flash Assay--that up to now was mainly used for toxicity analysis of solid and colored environmental samples (e.g. sediments, soil suspensions), is a powerful tool for screening the toxic properties of NPs. To demonstrate that Flash Assay (initially designed for a tube luminometer) can also be adapted to a microplate format for high throughput toxicity screening of NPs, altogether 11 chemicals were comparatively analyzed. The studied chemicals included bulk and nanosized CuO and ZnO, polyethylenimine (PEI) and polyamidoamine dendrimer generations 2 and 5 (PAMAM G2 and G5). The results showed that EC50 values of 30-min Flash Assay in tube and microplate formats were practically similar and correlated very well (log-logR2=0.98), classifying all analyzed chemicals, except nano CuO (that was more toxic in cuvette format), analogously when compared to the risk phrases of the EC Directive 93/67/EEC for ranking toxicity of chemicals for aquatic organisms. The 30-min EC50 values of nanoscale organic cationic polymers (PEI and dendrimers) ranged from 215 to 775 mg/l. Thirty-minute EC50 values of metal oxides varied largely, ranging from approximately 4 mg/l (bulk and nano ZnO) to approximately 100 mg/l (nano CuO) and approximately 4000 mg/l (bulk CuO). Thus, considering an excellent correlation between both formats, 96-well microplate Flash Assay can be successfully used for high throughput evaluation of harmful properties of chemicals (including organic and inorganic NPs) to bacteria.
尽管纳米颗粒（NPs）在各种应用中的产量和使用量不断增加，但包括欧盟新化学政策REACH在内的现行法规未能解决纳米颗粒所带来的环境、健康和安全风险。本文表明，动力学费氏弧菌发光抑制试验——快速检测法——迄今为止主要用于固体和有色环境样品（如沉积物、土壤悬浮液）的毒性分析，是筛选纳米颗粒毒性特性的有力工具。为了证明快速检测法（最初设计用于管式发光计）也可适用于微孔板形式以进行纳米颗粒的高通量毒性筛选，共对11种化学品进行了比较分析。所研究的化学品包括块状和纳米级的氧化铜和氧化锌、聚乙烯亚胺（PEI）以及第2代和第5代聚酰胺胺树枝状聚合物（PAMAM G2和G5）。结果表明，30分钟管式和微孔板形式的快速检测法的半数有效浓度（EC50）值实际上相似且相关性非常好（对数-对数R2 = 0.98），与欧盟指令93/67/EEC用于对水生生物化学品毒性分级的风险短语相比，除纳米氧化铜（在比色皿形式中毒性更大）外，所有分析化学品的分类类似。纳米级有机阳离子聚合物（PEI和树枝状聚合物）的30分钟EC50值范围为215至775毫克/升。金属氧化物的30分钟EC50值差异很大，范围从约4毫克/升（块状和纳米氧化锌）到约100毫克/升（纳米氧化铜）和约4000毫克/升（块状氧化铜）。因此，考虑到两种形式之间的良好相关性，96孔微孔板快速检测法可成功用于高通量评估化学品（包括有机和无机纳米颗粒）对细菌的有害特性。
