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.

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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.

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This document provides guidance and requirements for the determination of the mean (spherical) equivalent diameter of nano-objects (i.e. particles, droplets or bubbles) dispersed in liquids using the static multiple light scattering (SMLS) technique. The technique is applicable to a wide range of materials and does not require dilution of concentrated samples.

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This document specifies requirements and recommendations for the identification of measurands to characterize nano-objects and their agglomerates and aggregates, and to assess specific properties relevant to the performance of materials that contain them. It provides recommendations for relevant measurement.

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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.

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This document specifies characteristics to be measured of magnetic beads in suspension and powder forms for nucleic acid extraction applications. This document deals with magnetic beads that contain a substantial amount of magnetic nanoparticles (which can be superparamagnetic). Potential applicable measurement methods are listed for the individual characteristics. Specifically, this document lists critical characteristics of magnetic beads and suspensions, and additional characteristics to describe the magnetic beads and the suspension for nucleic acid extraction. Health, safety and environmental aspects of magnetic beads are not within the scope of this document.

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This document specifies the antimicrobial performance assessment method of textiles containing manufactured (metals/metal oxides) nanomaterials (TCMNMs). The textiles in this document include fabric, yarn and fibre in which manufactured nanomaterials are used during production or finishing process. Further, this document also specifies protocols to determine the quantity of nanomaterials released from textile following washing and/or exposure to artificial human body sweat. This document only covers the antibacterial, antifungal, and the anti-odour performance assessment method of TCMNMs. This document does not cover textiles that have therapeutic application as well as environment, health and safety (EHS) issues related to TCMNMs. Further, it does not cover the release of nanomaterials from TCMNMs as a result of aging, dry attrition and abrasion, although it is considered as an effective factor in releasing nanomaterials.

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This document describes methods for the measurement of particle size distributions for cellulose nanocrystals using atomic force microscopy and transmission electron microscopy. The document provides a protocol for the reproducible dispersion of the material using ultrasonication, as assessed using dynamic light scattering. Sample preparation for microscopy, image acquisition and data analysis are included.

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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.

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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.

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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.

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This document specifies methods of determining nanoparticle size and shape distributions by acquiring and evaluating scanning electron microscope images and by obtaining and reporting accurate results. NOTE 1 This document applies to particles with a lower size limit that depends on the required uncertainty and on the suitable performance of the SEM, which is to be proven first -according to the requirements described in this document. NOTE 2 This document applies also to SEM-based size and shape measurements of larger than nanoscale particles.

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  • Standard
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This document provides guidance for sample preparation, data acquisition by transmission electron microscopy, data processing, and three-dimensional image reconstruction to measure size and shape parameters of nano-objects on rod-shaped supports. The method is applicable to samples dispersed on or within an electron-transparent rod-shaped support.

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This document describes methods for the quantification of nano-object release from powders as a result of treatment, ranging from handling to high energy dispersion, by measuring aerosols liberated after a defined aerosolization procedure. Particle number concentration and size distribution of the aerosol are measured and the mass concentration is derived. This document provides information on factors to be considered when selecting among the available methods for powder sampling and treatment procedures and specifies minimum requirements for test sample preparation, test protocol development, measuring particle release and reporting data. In order to characterize the full size range of particles generated, the measurement of nano-objects as well as agglomerates and aggregates is adressed in this document. This document does not include the characterization of particle sizes within the powder. Tribological methods are excluded where direct mechanical friction is applied to grind or abrade the material.

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This document specifies characteristics to be measured and measurement methods for clay nanoplate samples in powder and suspension forms used for gas-barrier films. In addition, measurement protocols for the individual characteristics are described. This document does not deal with characteristics of post-manufacturing modification of clay nanoplates. This document does not cover considerations specific to health and safety issues either during manufacturing or use.

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This document defines terms related to the characterization of nano-objects in the field of nanotechnologies. It is intended to facilitate communication between organizations and individuals in research, industry and other interested parties and those who interact with them.

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IEC TR 63258:2021 is a Technical Report focused on the practical protocol of ellipsometry to evaluate the thickness of nanoscale films. This document does not include any specification of the ellipsometers, but suggests how to minimize the data variation to improve data reproducibility.

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This document specifies characteristics to be measured of individualized cellulose nanofibril (iCNF) samples in suspension and powder forms and their measurement methods. In addition, it provides sample preparation, measurement and data analysis procedures. This document does not apply to the characterization of iCNFs that have been modified after they are manufactured.

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This document specifies the sequence of methods for characterizing the structural properties of graphene, bilayer graphene and graphene nanoplatelets from powders and liquid dispersions using a range of measurement techniques typically after the isolation of individual flakes on a substrate. The properties covered are the number of layers/thickness, the lateral flake size, the level of disorder, layer alignment and the specific surface area. Suggested measurement protocols, sample preparation routines and data analysis for the characterization of graphene from powders and dispersions are given.

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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.

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This document specifies characteristics to be measured of nanostructured porous alumina in powder form as catalyst support for vehicle exhaust emission control and their relevant measurement methods. It includes critical characteristics that are required to be measured and additional characteristics that are recommended to be measured, based upon agreement between the interested parties. Measurement methods for each characteristic are recommended. This document is applicable to nanostructured porous alumina for gasoline-powered cars. It does not apply to characteristics specific for health, the environment and safety issues.

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This document defines terms related to nanomanufacturing processes in the field of nanotechnologies. All the process terms in this document are relevant to nanomanufacturing, however, many of the listed processes are not exclusively relevant to the nanoscale. Terms that are not exclusive are noted within the definitions. Depending on controllable conditions, such processes can result in material features at the nanoscale or, alternatively, at larger scales. There are many other terms that name tools, components, materials, systems control methods or metrology methods associated with nanomanufacturing that are beyond the scope of this document. Terms and definitions from other parts of the ISO/TS 80004 series are reproduced in Clause 3 for context and better understanding.

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This document defines terms related to carbon nano-objects in the field of nanotechnologies. It is intended to facilitate communication between organizations' and individuals' research, industry and other interested parties and those who interact with them. Additional terms and definitions for graphene and two-dimensional materials (2D) materials are provided in ISO/TS 80004-13. Related carbon nanoscale materials are given in Annex A.

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This document specifies the fundamental characteristics to be measured of a sample of engineered nanoparticles in powder form to determine the size, the chemical content and the surface area. This document also specifies measurement methods for determining each of the characteristics. It is intended to facilitate communication among consumers, regulators and industries with the necessary characteristics. It excludes characteristics that pertain to specific industrial applications of nanoparticles in powder form and detailed measurement protocols, as well as characteristics related to health, safety and environmental issues.

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This document specifies how to capture, measure and analyse transmission electron microscopy images to obtain particle size and shape distributions in the nanoscale. This document broadly is applicable to nano-objects as well as to particles with sizes larger than 100 nm. The exact working range of the method depends on the required uncertainty and on the performance of the transmission electron microscope. These elements can be evaluated according to the requirements described in this document.

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This document specifies characteristics including barrier properties to be measured of polymeric nanocomposite films used for improving food packaging. The barrier properties cover gas (oxygen), water vapour transmission and UV-Vis light transparency. This document also describes the relevant measurement methods. This document addresses neither safety and health issues related to the food packaging nor environmental aspects.

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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.

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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.

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This document specifies the characteristics to be measured of nano-object-assembled layers on electrodes by means of a solution process and of nano-objects constituting the layers for electrochemical applications such as nano-biosensor or diagnosis applications. It also provides measurement methods for determining the characteristics. It does not apply to: — the requirements of nanostructures by top-down nanomanufacturing; — the subsequent coating of materials such as biomaterials onto nano-object-assembled layers; — specific health and safety requirements during manufacturing; — the experimental conditions of electrochemical sensing; — the packaging, labelling, expiratory dates and transport of nano-object-enhanced electrochemical electrodes.

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This document gives guidelines for the characterization of carbon nanotube (CNT)-containing samples by thermogravimetric analysis (TGA), performed in either an inert or oxidizing environment. Guidance is provided on the purity assessment of the CNT samples through a quantitative measure of the types of carbon species present as well as the non-carbon impurities (e.g. metal catalyst particles) within the material. In addition, this technique provides a qualitative assessment of the thermal stability and homogeneity of the CNT-containing sample. Additional characterization techniques are required to confirm the presence of specific types of CNT and to verify the composition of the metallic impurities present.

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This document specifies the characteristics to be measured of air filter media containing polymeric nanofibres on the surfaces of a substrate. It also describes measurement methods for determining the individual characteristics. This document does not cover characteristics specific for health and safety issues. NOTE The properties/performances assessment of air filter media related to applications requires the use of relevant published standards. The physical properties and performances of filtration media, such as pressure drop, and the particle removal efficiency of air filter media are measured with test methods suitable for specific applications.

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This document specifies the characteristics to be measured of suspensions containing multi-walled carbon nanotubes (CNT suspensions). It includes the essential and additional characteristics of the CNT suspension, and the corresponding measurement methods. Characteristics specific to health, environmental and safety issues are excluded from this document. WARNING — The use of this document can involve hazardous materials, operations and equipment. It does not purport to address all of the safety or environmental problems associated with its use. The execution of this document is entrusted to appropriately qualified and experienced people.

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This document gives guidelines for the characterization of single-wall carbon nanotubes (SWCNTs) using near infrared (NIR) photoluminescence (PL) spectroscopy. It provides a measurement method for the determination of the chiral indices of the semi-conducting SWCNTs in a sample and their relative integrated PL intensities. The method can be expanded to estimate the relative mass concentrations of semi-conducting SWCNTs in a sample from their measured integrated PL intensities and knowledge of their PL cross-sections.

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This document specifies a method for the measurement of the photocatalytic activity (PCA) of nanoparticles (NPs), suspended in an aqueous environment in physiologically relevant conditions, by measuring the ultraviolet (UV)-induced nicotine adenine dinucleotide hydrate (NADH) oxidation. The measurement is intended to assess the potential for the photo-toxicity of nanomaterials. The method is also applicable to NP aggregates and agglomerates.

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This document specifies characteristics to be measured of layered clay nanomaterials in powder form and chemically modified ones, and describes their relevant measurement methods. This document does not deal with health, safety and environmental issues.

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This document specifies a method for the characterization of evolved gas components in single-wall carbon nanotube (SWCNT) samples using evolved gas analysis/gas chromatograph mass spectrometry (EGA/GCMS). NOTE Some difference could appear between qualitative and quantitative results of emitted gas and gas content in the sample due to the heating and the possible presence of catalysts.

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This document specifies the characteristics of magnetic nanosuspensions to be measured and lists measurement methods for measuring these characteristics. This is a generic document and does not deal with any particular application.

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This document provides guidance for the specification of characteristics and relevant measurement methods for silver nanoparticles in powder or colloidal forms that are intended for antibacterial applications in nanotechnology. This document is intended to aid the producer in providing the physicochemical characteristics of silver nanoparticles that have an antibacterial effect to the buyer. This document does not cover considerations specific to health and safety issues either during manufacturing or use.

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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.

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This document provides guidelines to quantify and identify air concentration (number of particles/cm3) of particles of carbon black and/or amorphous silica by size in air samples collected in a mixed dust industrial manufacturing environment. The method is defined for air samples collected with an electrical low pressure cascade impactor (ELPCI). on a 25 mm polycarbonate substrate. The method is suitable for sampling in manufacturing environments where there are a variety of particle types contributing to the overall atmosphere. This method is applicable only to environments with chemically and physically distinct particles contributing to aerosols or when confounders can be controlled (e.g. diesel sources). Other sampling methods can also be suitable, though this document is limited to describing methods associated with the electrical low pressure cascade impactor. Samples collected with the electrical low pressure cascade impactor are analyzed via TEM and EDS to for particle morphology and elemental composition, respectively, to permit identification of particles by type. This information is then used, in conjunction with particle concentration by size range, as determined by the electrical low pressure cascade impactor, to determine concentration of the materials of interest by size.

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This document provides a matrix which links key properties of graphene and related two-dimensional (2D) materials to commercially available measurement techniques. The matrix includes measurement techniques to characterize chemical, physical, electrical, optical, thermal and mechanical properties of graphene and related 2D materials.

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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.

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This document describes health and safety practices in occupational settings relevant to nanotechnologies. This document focuses on the occupational manufacture and use of manufactured nano-objects, and their aggregates and agglomerates greater than 100 nm (NOAAs). It does not address health and safety issues or practices associated with NOAAs generated by natural processes, hot processes and other standard operations which unintentionally generate NOAAs, or potential consumer exposures or uses, though some of the information in this document can be relevant to those areas.

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This document specifies development of representative test materials consisting of nano-objects in dry powder form, to enable test method development and improve comparability of data for nanotechnology applications. It includes the physico-chemical properties (specifically, size and shape, specific surface area, crystal structure, and bulk chemical composition) that are required to be measured and reported with the representative test material.

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This document provides an overview of approaches of sample preparation (i.e. pre-treatment and size-fractionation) for analytical measurements applied to surface and drinking water, potentially containing relevant amounts and types of metal and metal oxide nano-objects, including collection from source and storage of samples, pre-concentration of analytes, and their fractionation.

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This document identifies parameters and conditions, as part of an integrated measurement system, necessary to develop and validate methods for the application of asymmetrical-flow and centrifugal field-flow fractionation to the analysis of nano-objects and their aggregates and agglomerates dispersed in aqueous media. In addition to constituent fractionation, analysis can include size, size distribution, concentration and material identification using one or more suitable detectors. General guidelines and procedures are provided for application, and minimal reporting requirements necessary to reproduce a method and to convey critical aspects are specified.

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ISO 19007:2018 specifies a method for evaluating the effects of nano-objects and their aggregates and agglomerates (NOAA) on cellular viability using the MTS assay. The assay design includes performance requirements and control experiments to identify and manage variability in the assay results. ISO 19007:2018 is applicable to the use of a 96-well plate.

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    32 pages
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ISO/TS 13278:2017 provides methods for the determination of residual elements other than carbon in samples of single-wall carbon nanotubes (SWCNTs) and multiwall carbon nanotubes (MWCNTs) using inductively coupled plasma mass spectrometry (ICP-MS). The purpose of this document is to provide optimized digestion and preparation procedures for SWCNT and MWCNT samples in order to enable accurate and quantitative determinations of elemental impurities using ICP-MS.

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ISO/TS 20787:2017 specifies a test method, aiming to maximize repeatability and reliability of testing, to determine whether MNMs are toxic to aquatic organisms, specifically Artemia sp. nauplius. ISO/TS 20787:2017 is intended to be used by ecotoxicological laboratories that are capable in the hatching and culturing of Artemia sp. and the evaluation of toxicity of nanomaterials using Artemia sp. nauplius. This method uses Artemia sp. nauplii in a simulated environment, artificial seawater, to assess effects of nanomaterials. ISO/TS 20787:2017 is applicable to MNMs that consist of nano-objects such as nanoparticles, nanopowders, nanofibres, nanotubes, nanowires, as well as aggregates and agglomerates of such MNMs.

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    15 pages
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ISO/TR 19057:2017 reviews the use and application of acellular in vitro tests and methodologies implemented in the assessment of the biodurability of nanomaterials and their ligands in simulated biological and environmental media. ISO/TR 19057:2017 is intended to focus more on acellular in vitro methodologies implemented to assess biodurability and, therefore, excludes the general review of relevant literature on in vitro cellular or animal biodurability tests.

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