ISO/TC 229/WG 3 - Health, Safety and Environmental Aspects of Nanotechnologies

This document describes the characteristics of working suspensions of nano-objects to be considered when conducting in vitro assays to evaluate inherent nano-object toxicity. In addition, the document identifies applicable measurement methods for these characteristics. This document is applicable to nano-objects, and their aggregates and agglomerates greater than 100 nm. This document intends to help clarify whether observed toxic effects come from tested nano-objects themselves or from uncontrolled sources.

This document specifies a method for assessing the peroxidase-like activity of metal and metal oxide nanoparticles by spectrophotometry. This document can serve as a reference for the measurements of peroxidase-like activities in other types of nanoparticles.

This document provides a reliable and repeatable method for simultaneous assessment of both exposure and toxicity of manufactured nano-objects (MNOs) using Tetrahymena sp. The ingested, internalized material (MNOs) indicates aquatic exposure. This document is intended to be used by all the centers working with nano(eco)toxicity of MNOs and capable of culturing of Tetrahymena sp. The method uses Tetrahymena sp. to assess exposure and effects of MNOs. In addition, the test can be used by centers (laboratories) interested in investigating the biological interaction of MNOs with living cells. This method is applicable to nano-objects such as nanoparticles, nanofibres of certain size (in a µm size range), nanoplates, as well as their aggregates and agglomerates.

This document reviews characterization of CNT and CNF aerosols for inhalation exposure studies. The document also provides useful information on appropriate characterization of CNT and CNF, which is required to evaluate and understand the inhalation toxicity of CNT and CNF aerosols. This document neither provides guidance on aerosol characterization for other carbon nanomaterials, nor provides guidance for characterization of carbon nanotube and nanofibre aerosols in the workplace or ambient air.

This document describes a method for high throughput evaluation of cytotoxic response of 3D model cells exposed to NPs without optical interference. The method in this document is intended to be used in biological testing laboratories that are competent in the culture and growth of cells and the evaluation of cytotoxicity of NPs using 3D-model cells. This method applies to materials that consist of nano-objects such as nanoparticles, nanopowders, nanofibres, nanotubes, and nanowires, as well as aggregates and agglomerates of these materials.

This document describes a near-infrared optical absorption method to estimate the in vitro cellular uptake of carbon nanomaterials including both internalized and/or tightly adhered to the cell membrane from liquid dispersions. This is a simple method to screen carbon nanomaterials uptake; additional analysis using a different technique can be required if quantification is desired.

This document describes a methodology of a label free and real-time detection for non-invasive monitoring of cell-based assays to assess toxicity of nanomaterials to eukaryotic and prokaryotic cells.

This document reviews and evaluates the utility of available methods to assess material released from commercial polymer composites in support of product use and safety decisions, and describes what revised or additional methods are needed. The document is not focused on describing methods per se; rather the goal is to describe information that is appropriate for consideration in the selection of methods to support decision-making.

This document specifies measurement protocols and test conditions to determine alterations to protein secondary structure induced by their interaction with nanomaterials using ultraviolet circular dichroism (UV-CD) spectroscopy. This document does not apply to the characterization of conformational changes of disordered proteins.

This document specifies a method for rapidly assessing nanomaterial toxicity (fish early life stage, 0 HPF to 120 HPF). It includes information on the importance of acellular chorion removal, detailed chorion removal procedures, and a complete protocol for the toxicity assessment of nanomaterials using dechorionated zebrafish embryos. The focus of this document is on testing nanomaterial toxicity.

This document collates information regarding the systems available for exposure and assessment of nano-objects and their aggregates and agglomerates (NOAA) for in vitro air exposure studies. It provides an overview of the various exposure systems and in vitro cell systems used to perform in vitro studies that simulate an inhalation toxicology study design.

This document describes the background and principles for toxicokinetic studies relevant for nanomaterials. Annex A shows the definitions for terminology with respect to toxicokinetics as used in OECD TG 417:2010.

This document provides some considerations for the collection of environmental samples to be analysed for manufactured NOAA, considerations to distinguish manufactured NOAA from background levels of naturally occurring nanoscale particles of the same composition, and preparation procedures to aid in the quantification of manufactured NM in environmental matrices.

ISO/TS 12901-2:2014 describes the use of a control banding approach for controlling the risks associated with occupational exposures to nano-objects, and their aggregates and agglomerates greater than 100 nm (NOAA), even if knowledge regarding their toxicity and quantitative exposure estimations is limited or lacking. The ultimate purpose of control banding is to control exposure in order to prevent any possible adverse effects on workers' health. The control banding tool described here is specifically designed for inhalation control. Some guidance for skin and eye protection is given in ISO/TS 12901‑1. ISO/TS 12901-2:2014 is focused on intentionally produced nano-objects such as nanoparticles, nanopowders, nanofibres, nanotubes, nanowires, as well as of aggregates and agglomerates of the same. As used in ISO/TS 12901-2:2014, the term "NOAA" applies to such components, whether in their original form or incorporated in materials or preparations from which they could be released during their lifecycle. ISO/TS 12901-2:2014 is intended to help businesses and others, including research organizations engaged in the manufacturing, processing or handling of NOAA, by providing an easy-to-understand, pragmatic approach for the control of occupational exposures.

ISO/TS 12901:2012 provides guidance on occupational health and safety measures relating to engineered nanomaterials, including the use of engineering controls and appropriate personal protective equipment, guidance on dealing with spills and accidental releases, and guidance on appropriate handling of these materials during disposal. ISO/TS 12901:2012 is intended for use by competent personnel, such as health and safety managers, production managers, environmental managers, industrial/occupational hygienists and others with responsibility for the safe operation of facilities engaged in production, handling, processing and disposal of engineered nanomaterials. ISO/TS 12901:2012 is applicable to engineered materials that consist of nano-objects such as nanoparticles, nanofibres, nanotubes and nanowires, as well as aggregates and agglomerates of these materials (NOAA).

ISO/TR 13121:2011 describes a process for identifying, evaluating, addressing, making decisions about, and communicating the potential risks of developing and using manufactured nanomaterials, in order to protect the health and safety of the public, consumers, workers and the environment. ISO/TR 13121:2011 offers guidance on the information needed to make sound risk evaluations and risk management decisions, as well as how to manage in the face of incomplete or uncertain information by using reasonable assumptions and appropriate risk management practices. Further, ISO/TR 13121:2011 includes methods to update assumptions, decisions, and practices as new information becomes available, and on how to communicate information and decisions to stakeholders. ISO/TR 13121:2011 suggests methods organizations can use to be transparent and accountable in how they manage nanomaterials. It describes a process of organizing, documenting, and communicating what information organizations have about nanomaterials.

ISO 10801:2010 gives requirements and recommendations for generating metal nanoparticles as aerosols suitable for inhalation toxicity testing by the evaporation/condensation method. Its application is limited to metals such as gold and silver which have been proven to generate nanoparticles suitable for inhalation toxicity testing using the technique specified.

ISO 10808:2010 specifies requirements for, and gives guidance on, the characterization of airborne nanoparticles in inhalation exposure chambers for the purpose of inhalation toxicity studies in terms of particle mass, size distribution, number concentration and composition.

ISO 29701:2010 describes the application of a test using Limulus amebocyte lysate (LAL) reagent for the evaluation of nanomaterials intended for cell-based in vitro biological test systems. The test is suitable for use with nanomaterial samples dispersed in aqueous media, e.g. water, serum or reaction medium, and to such media incubated with nanomaterials for an appropriate duration at 37 °C. ISO 29701:2010 is restricted to test samples for in vitro systems, but the methods can also be adapted to nanomaterials to be administered to animals by parenteral routes.