Tests using algae and/or cyanobacteria, invertebrates (crustaceans) and fish form the basic elements of an ecotoxicological assessment in a number of regulations, in particular for classification of a substance as hazardous or not to the aquatic environment according to the Globally Harmonised System of Classification and Labelling of Chemicals (GHS-CLP) (GHS, 2022) and the REACH regulation (Registration, Evaluation, Authorisation and Restriction of Chemicals, EC, 2006). Standardised test guidelines (TGs) of the Organisation for Economic Co-operation and Development (OECD) are available to address the regulatory relevant endpoints of growth inhibition in algae and cyanobacteria (TG 201), acute toxicity to invertebrates (TG 202), and acute toxicity in fish (TG 203). Applying these existing OECD TGs for testing two dimensional (2D) graphene nanoforms may require more attention, additional considerations and/or adaptations of the protocols, because graphene materials are often problematic to test due to their unique attributes. In this review a critical analysis of all existing studies and approaches to testing used has been performed in order to comment on the current state of the science on testing and the overall ecotoxicity of 2D graphene materials. Focusing on the specific tests and available guidance's, a complete evaluation of aquatic toxicity testing for hazard classification of 2D graphene materials, as well as the use of alternative tests in an integrated approach to testing and assessment, has been made. This information is essential to ensure future assessments generate meaningful data that will fulfil regulatory requirements for the safe use of this “wonder” material.

使用藻类和/或蓝藻、无脊椎动物（甲壳类动物）和鱼类进行的测试构成了一些法规中生态毒理学评估的基本要素，特别是根据全球协调制度将物质分类为对水生环境有害或不有害的物质化学品分类和标签 (GHS-CLP)（GHS，2022 年）和 REACH 法规（化学品注册、评估、授权和限制，EC，2006 年）。经济合作与发展组织 (OECD) 的标准化测试指南 (TG) 可用于解决藻类和蓝藻生长抑制 (TG 201)、对无脊椎动物的急性毒性 (TG 202) 和急性鱼类毒性 (TG 203)。将这些现有的经合组织 TG 用于测试二维 (2D) 石墨烯纳米形式可能需要更多的关注、额外的考虑和/或协议的改编，因为石墨烯材料由于其独特的属性而经常难以测试。在这篇综述中，对所有现有研究和测试方法进行了批判性分析，以评论测试科学的现状和二维石墨烯材料的整体生态毒性。着眼于特定的测试和可用的指南，对二维石墨烯材料危险分类的水生毒性测试进行了完整的评估，以及在测试和评估的综合方法中使用替代测试。