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As noted above, particle counting may not be sufficient to evaluate potential risks of exposure to nanoparticles For each type of nanomaterial, the surface area of individual particles can significantly affect the activity of the material toward biological tissues [44]

Development of a real-time instrument that can monitor exposure as opposed to a single parameter represents an important advance toward ensuring safe working environments for the engineered nanoparticle industry. The electrical aerosol detector (EAD) measures a unique aerosol parameter called aerosol diameter concentration, or total aerosol length . This measurement (reported as mm/cm3) represents a number concentration multiplied by average diameter, and thus is directly related to surface area The aerosol diameter concentration, when complemented by CPC data for particle number, can be used to calculate the average particle size Continuous measurements of aerosol diameter concentration with the EAD correlate well with the surface area of deposited particles and are believed to provide a better estimate of actual inhalation exposure than either the mass or number concentration of particles could. EAD has been used in ambient air studies at the St. Louis Supersite [45] .

5.3 SAMPLING AND ANALYSIS OF WATERS AND SOILS FOR NANOPARTICLES

The second subset of the challenge to develop instruments to assess exposure to engineered nanomaterials is "to develop instruments that can track the release, concentration and transformation of engineered nanoparticles in water systems" [41] . Measurements of natural nanoparticles in environmental waters have been reported by a variety of techniques, but there are few reports at this time of field studies designed to detect engineered nanoparticles

A separation technique called field-flow fractionation (FFF) separates nanoparticles from larger particles, permitting their direct analysis or collection for detailed characterization

As shown in Figure 5 .4, field-flow fractionation is similar to the chromatographic separations typical of environmental analyses for organic contaminants The water sample passes through a thin flow channel designed so that the flow will be laminar, that is, not turbulent and faster in the center of the column than at the walls The bottom side of the channel is a membrane that will allow water through but not the particles of interest A second force is applied perpendicular to the channel flow to generate a cross-flow All particles in the sample will be pushed downward toward the membranes, but smaller particles will diffuse upward toward the center of the channel to a greater degree and will be in the faster stream lines of the channel flow The smaller particles in the sample will exit from the channel before the larger particles

Once separated, natural particles in the nanometer size range can be directly introduced into an inductively coupled mass spectrometer [46] for elemental analysis, collected for ESEM analysis [47], and coupled to a light scattering instrument for particle size measurements [48] . Field-flow fractionation was applied for size distribution analysis of trace concentrations of iron oxi/hydroxide colloids being considered as potential carriers for the radionuclide migration from a nuclear waste repository [49]

Top of Channel

Cross Flow

FIGURE 5.4 Flow field fractionation. (From Postnova Analytics, Inc. With permission .)

Cross Flow

FIGURE 5.4 Flow field fractionation. (From Postnova Analytics, Inc. With permission .)

Field flow fractionation also has been applied to the analysis of nanoparticles in soil and sediment systems Engineered zinc nanoparticles have been separated from larger soil particles through the preparation of suspensions that are then shaken and allowed to settle gravitationally. The supernatant, containing the less than 1-pm particle fraction, was then separated by field-flow fractionation for further analysis [50]

Analyses of engineered nanoparticles directly in soil or sediment matrices by SEM or TEM imaging techniques is possible when the material has some unique property, such as fluorescence or light absorption, or contains a rare metal or unique organic compound [51] Nanoparticles of natural origin are ubiquitous, and the detection of engineered particles against background using these techniques, which are at best time-consuming and costly, is not likely to be a practical means of routine environmental assessments

5.4 NANOTECHNOLOGY MEASUREMENT RESEARCH AND FUTURE DIRECTIONS

Both within the United States and internationally, private and governmental organizations have recognized the need for improved analytical tools for nanotechnology The American Society for Testing and Materials (ASTM) Committee E56 on Nano-technology was formed in 2005 to develop standards and guidance for nanotechnol-ogy and nanomaterials. The first ASTM standard, published in July 2007, precisely defines the language for nanotechnology [52] This should allow more consistent and effective technical communication within the diverse fields involved in nanotechnol-ogy and with the public .

In late 2005, the International Organization for Standardization (ISO) established a new technical committee, ISO T/C229 Nanotechnologies, with three working groups The United States is represented on the Measurement and Characterization Workgroup, WG2 . The Draft Business Plan for T/C229 [53] details the high-priority needs and strategies for this group A limited number of standards relating to nanoparticle measurements have been published, with several more in progress ISO/TR 27628:2007 contains guidelines on characterizing occupational nanoaerosol exposures, with a discussion of applicable measurement terms . Specific information is provided on methods for bulk aerosol characterization and single-particle analysis . Other standards currently near completion include N 270 TS: Terminology and definitions for carbon nanomaterials; N 271 TS: Format for reporting the engineered nanomaterials content of products; and N 272 TR: Guide to nanoparticles measurement methods and their limitations

5.4.1 United States 5.4.1.1 NIOSH

In the United States, the National Institute for Occupational Safety and Health (NIOSH) established the Nanotechnology Research Center (NTRC) in 2004 to coordinate and facilitate research on the impact of nanotechnology in the workplace . The NTRC recognized that in order to evaluate risks, accurate measurement data would

TABLE 5.2

NIOSH Goals for Nanoparticle Measurement

1 Evaluating methods of measuring mass of respirable particles in the air and determining if this measurement can be used to measure nanomaterials

2 Developing and field-testing practical methods to accurately measure airborne nanomaterials in the workplace

3 Developing testing and evaluation systems to compare and validate sampling instruments

TABLE 5.3

NIOSH Research Agenda

NIOSH Nanoparticle Research Strategic Plan and Timeline Fiscal Year (Measurement and Analysis Programs)

2005 Surveillance Phase I: Identify and gather baseline information . Develop techniques for online surface area measurement.

2006 Conduct measurement studies of nanoparticles in the workplace . Analyses of filter efficiency for nanomaterials .

2007 Evaluate surface area-mass metric results . Establish a suite of instruments and protocols for nanomaterial measurements Conduct measurement studies of nanoparticles in the workplace . Further development of online and offline nanoparticle measurement methods

2008-2009 Develop performance results for nanoparticle measurement instruments and methods .

Complete evaluation of viable and practical workplace sampling devices and methods for nanoparticles (affordable, portable, effective) . Quantification of systemic nanoparticle concentrations in laboratory animals after pulmonary exposure to nanospheres and nanofibers be needed . Measurement method development objectives, as listed in Table 5 . 2, were accordingly included as critical topics for their strategic workplace goals .

The NTRC established several partnerships with other agencies, including U. S . EPA, NIST, the Department of Defense (DOD) and the Department of Energy (DOE), and ASTM . In addition, the NTRC is partnering with and in some instances supporting research at academic institutions, instrument manufacturers, and private industry. Table 5 . 3 summarizes method analysis studies included in the research program planned for the period 2005 through 2009 [54] .

Accomplishments and publications for 17 completed and ongoing research programs are listed in the 2007 NIOSH report entitled "Progress toward Safe Nanotechnology in the Workplace" [55] . "Project 1, Generation and Characterization of Occupationally Relevant Airborne Nanoparticles," includes numerous accomplishments relevant to the characterization and workplace measurement of carbon nanotubes and TiO2 . Project 11, Nanoparticles in the Workplace, is designed to develop partnerships with industry, academia, and other government agencies for research and development of monitoring instrumentation and protocols Project 13, The Measurement and Control of Workplace Nanoparticles, will provide a basis for

TABLE 5.4

U.S. Government Supported Research on Nanotechnology Environmental Measurements

Project Title Sponsor

Biological Fate and Electron Microscopy Detection of EPA

Nanoparticles during Wastewater Treatment Development of Detection Techniques and Diagnostics for DOE Airborne Carbon Nanotubes

Anticipated End Year

2010

2007

Fate and Transport of Carbon Nanotubes in Unsaturated and EPA 2008

Saturated Soils

Identifying and Regulating Environmental Impacts of NSF 2007

Nanomaterials

Monitoring and Characterizing Airborne Carbon Nanotube NIOSH 2008

Particles

New Instruments for Real-Time, High Resolution NSF 2007

Characterization of Nanoparticles in the Environment understanding how nanoparticles are released in the workplace and how they can be monitored and exposure controlled . Research Project 14 is an exposure study of TiO2 in manufacturing and end-user facilities using a variety of monitoring techniques .

5.4.1.2 U.S. Government-Sponsored Research

The Project on Emerging Nanotechnologies of the Woodrow Wilson International Center for Scholars in Arlington, Virginia, is developing an inventory of government-sponsored research into the environmental, health, and safety implications of nanotechnology [56] . While the intent of this inventory is to include research projects on an international basis, the current listing is dominated by projects supported by U S agencies Included in these are several funded by the U S EPA, the Department of Energy (DOE), the National Science Foundation (NSF), and NIOSH that should provide information on environmental measurements Current projects included in this inventory that are of particular relevance to environmental measurements include those listed in Table 5 4

5.4.1.3 National Institute of Standards and Technology (NIST)

In early 2006, the National Institute of Standards and Technology (NIST) launched a new state-of-the-art Center for Nanoscale Science and Technology (CNST) [57] . The CNST is specifically dedicated to developing the measurement methods and tools needed to support all phases of the nanotechnology industry While the Center's focus is not specifically toward environmental methods or analyses, measurement advances for discovery, development, and manufacturing of nanoparticles should have applicability in monitoring their presence and effects in the environment

NIST has responsibility for the development and supply of standard reference materials for analyses of various chemicals and materials Academic laboratories as well as commercial facilities use these standard reference materials to verify the accuracy of their data NIST currently provides certified polystyrene spheres for nanoparticle size analyses . NIST [57] also reports a recent development of a prototype atomic "ruler" for calibrating dimensional measurements of 100 nm and below. This ruler will be able to document the accuracy of scanning electron or atomic force microscopy data NIST now is transferring the technology to a commercial standards supplier

5.4.2 European Union

The European Union (EU) launched its largest ever funding program for research and technological development on January 1, 2007. The EU Member States have earmarked a total of €3 . 5 billion (approx . U. S . $4 .5 billion) for funding nanotechnology related research over the period 2007 to 2013 . The program calls for proposals for a wide range of activities related to the risk assessment of nanomaterials [58] First among five areas where proposals are invited is NMP-2007-1.3-1, "Specific, easy to use portable devices for measurement and analysis ." Based on the belief that workplace exposure is the area of greatest concern, the objective of this work will be "to develop and validate affordable, portable, adequate sampling and measurement equipment for monitoring working environments (i e , quantification and characterization of airborne nanoparticles in particular) "

German governmental agencies, including the Federal Institute for Occupational Safety and Health (BAuA), the Federal Institute for Risk Assessment (BfR), and the Federal Environment Agency (UBA), have jointly developed a nanotechnology research strategy and are currently conducting a limited number of projects [59] . Two of these are designed to test instrumentation or modify and validate existing measurement methods to be applicable for workplace measurements of nanoparticles

5.4.3 Asia-pacific

The Industrial Technology Research Institute (ITRI) of Chinese Taipei is undertaking an international project awarded by the Asia-Pacific Economic Cooperation (APEC) Industrial Science and Technology Working Group (IST WG) to assist in the establishment of the Technological Cooperative Framework on Nanoscale Analytical and Measurement Methods among APEC economies This cooperative project was formed to create an avenue for sharing advances in nanometer analytical measurement methods and to promote the best available technology to meet the needs for nanoscale standards [60] . The United States is one of six nations participating in this project

As part of the project, the NanoTechnology Research Center (also using the acronym NTRC) of ITRI is organizing an interlaboratory comparison study on nanopar-ticle characterization The aim of the comparison is to establish the effectiveness and comparability of different measurement methods across different laboratories on nanometer-scale particles This multi-year program will include a series of measurement challenges using standardized material, as shown in Table 5 5

The results for 2005 interlaboratory comparison measurements of size and diameter have been published [60] The NTRC provided samples of polystyrene

TABLE 5.5

APEC Nanometer Measurement Interlaboratory Comparison Studies for Nanoparticles

Year Characteristic

2005 Size/diameter

2006 Surface area, refractive index, light absorption/reflection

2007 Number and distribution, conductivity, dispersion spheres with diameters of 30, 50, and 100 nm to participating laboratories . The study generated a total of 32 data sets from 15 participating laboratories . Particle size measurements were made using DLS, SEM, TEM, and SPM instrumentation . The results for the 30-nm particles were satisfactory from all measurements, while two measurements for the 50-nm particles and four for the 100-nm sample fell outside three standard deviations of the mean Measurements taken by TEM for the 30-nm and 100-nm particles were significantly below the expected diameters and below the results from the other techniques . Samples were distributed for the 2006 studies, and 16 laboratories have reported results, but these have not been made publicly available at the time of writing

5.5 Summary

Reliable and accurate measurements of nanoparticle physical and chemical properties have been recognized as critical elements required for meaningful assessments of impact and risk . While numerous technologies do exist, many challenges to measuring engineered nanoparticles in the environment have yet to be addressed The combination of their small size and the range of attributes that may factor into their activity requires a complex matrix of complementary analyses and methods, for many critical parameters have yet to be devised for nanoparticles in environmental settings Environmental analyses of nanoparticles are far from routine or readily available at this point, but the increased interest and focus of governmental agencies and research organizations allows for optimism

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