Many scientific organizations or task forces have strongly recommended
that toxicologists adequately characterize physicochemical properties of
the nanoparticle-types that are being evaluated for hazard testing.
However, too often this recommendation becomes a “laundry list” of
physicochemical characteristics and does not have adequate
prioritization.  As a consequence, in order to adequately describe the
physical characteristics of the nanoparticle-type being evaluated, I
would recommend that toxicologists characterize the following
(prioritized) physicochemical properties prior to conducting hazard
studies with nanoparticle-types:

Particle size and size distribution (wet state) and surface area (dry
state) in the relevant media being utilized – depending upon the route
of exposure;

Crystal structure/crystallinity;

Aggregation status in the relevant media;

Composition/surface coatings;

Surface reactivity;

Method of nanomaterial synthesis and/or preparation including
post-synthetic modifications (e.g., neutralization of ultrafine TiO2
particle-types);

Purity of sample;

This represents a focused approach concomitant with a minimum,
standardized assessment of physicochemical properties that should be
investigated prior to the development of toxicity testing with
nanoparticles.

Additional points to consider for material characterization:

Depending upon the exposure scenario, it may be appropriate to conduct
assessments of nanoparticle characteristics in both the wet state (in
the relevant media) and the dry state.  Therefore a BET surface area
measurement may not suffice – but should be carried out in parallel
with different types of measurements such as cryo TEM (dry state) and
DLS (dynamic light scattering - particle sizing in the wet phase).

The chemical composition of the nanoparticle core vs. shell (hard or
soft) should be evaluated.

In this regard, the surface chemistry could be specific for each
nanoparticle-type.

Below, I have listed 5 recently-published nanoparticle toxicity studies
from our Laboratory concomitant with the specific physicochemical
characteristics that were identified from each of the studies.
Accordingly, we have concluded that characteristics of nanoparticle
surface reactivity appeared to be the best predictors of in vivo
pulmonary toxicity in rats.  However, it should be pointed out that
surface reactivity evaluations are particle-type specific (e.g. Vitamin
C assay for TiO2 particles; erythrocyte hemolysis for silica particles).
 Therefore, the same tests for surface reactivity cannot be utilized for
each nanoparticle-type.

A Listing of Material Characterization Methods utilized in the 5 studies
listed below

“Pulmonary Instillation Studies with Nanoscale TiO2 Rods and Dots in
Rats: Toxicity is not dependent upon Particle Size and Surface Area”.
Warheit DB, Webb TR, Sayes CM, Colvin VL, and Reed KL. .  Toxicol Sci.
91: 227-236, 2006

Material characterization employed in this study:

synthesis method

crystal structure

particle size

surface area

composition/surface coating

aggregation status

cryo TEM

crystallinity

purity (TGA)

“Pulmonary bioassay studies with nanoscale and fine quartz particles
in rats: Toxicity is not dependent upon particle size but on surface
characteristics”. Warheit DB, Webb TR, Colvin VL, Reed KL and Sayes
CM.  Toxicol Sci. 95:270-280, 2007;

Material characterization employed in this study:

synthesis method

crystal structure/crystallinity (XRD)

median particle size - particle size (range)

purity (% Fe content)– ICP-AES

surface area

TEM

aggregation status

purity

surface reactivity (erythrocyte hemolysis)

reactive oxygen species (ESR)

“Pulmonary Toxicity Study in Rats with Three Forms of ultrafine-TiO2
Particles:  Differential Responses related to Surface Properties”.
Warheit DB, Webb TR, Reed KL, Frerichs S, and Sayes CM. Toxicology
230:90-104, 2007;

Material characterization employed in this study:

crystal phase

median particle size and size distribution in water and PBS

pH in water and PBS

surface area (BET)

TEM

aggregation status

chemical (surface) reactivity – (Vitamin C assay)

surface coatings/composition, purity

Assessing toxicity of fine and nanoparticles: Comparing in vitro
measurements to in vivo pulmonary toxicity profiles Sayes CM, Reed KL,
and Warheit DB.  Toxicol Sci, 97:163-180, 2007.

Particle-types utilized in this study:

Fine-sized carbonyl iron

Fine-sized crystalline silica

Fine-sized amorphous silica

Nano ZnO

Fine ZnO

Particle characterizations conducted both in the “dry state” and
“wet state”

Material characterization employed in this study:

Particle characterization in the dry state

particle size

surface area

density

calculated size in dry state (based on surface area determinations)

crystallinity

purity

Particle characterization in the wet state

particle size  in solutions – PBS, culture media, water

average aggregated size in solutions

% distribution

surface charge

aggregation status

Conversion and comparisons of in vitro and in vivo doses for dosimetric
comparisons

Comparative Pulmonary Toxicity Assessments of C60 Water Suspensions in
Rats: Few Differences in Fullerene Toxicity In Vivo in Contrast to In
Vitro Profiles.    Sayes CM, Marchione AA, Reed KL, and Warheit DB. Nano
Lett. 2007 Aug;7(8):2399-2406.

Material characterization employed in this study:

particle size and size distribution

surface charge

crystallinity

TEM

Composition

oxidative radical activity (ESR measurements)

surface reactivity (erythrocyte hemolytic potential)

Nanoscale manufacturing and processing

	The number of manufacturing and processing methods for generating
nanoscale material continues to grow and become more sophisticated.  The
two primary areas for this discussion include physical reduction methods
(milling) and engineering methods (e.g., particle stabilization, vapor
deposition, self assembly).

Question 9: What are the common processes used to manufacture nanoscale
materials?

Dr. BelBruno:

The list in the paragraph above covers the general “bottom up” and
“top down’ methods.

Dr. Canady:

A list was developed in the ICON workshop last year at NIH – I don’t
have the full list, but it is extensive.

Dr. Cassell:

Solution based methods are quite common and include a number of
approaches (self-assembly, colloidal preparation), chemical vapor
deposition is also used extensively.  All techniques conventionally used
in organic and inorganic synthesis are also adapted for nanoscale
materials manufacture.

Dr. Lahann:

Organic materials: self-assembly

Inorganic materials: CVD

Polymers: staged synthesis, self-assembly, suspension polymerization,
grafting, electrified jetting

Composites: similar to polymers

Dr. Murashov:

The draft ISO TC229 Technical Report on “Health and Safety Practices
in Occupational Settings Relevant to Nanotechnologies” lists the
following categories of major processes used to manufacture nanoscale
materials:

Gas phase processes such as flame pyrolysis, high temperature
evaporation and plasma synthesis;

Vapor deposition;

Liquid phase methods: colloidal, self-assembly, sol-gel;

Supercritical fluid;

Electro-spinning for polymer nanofiber synthesis;

Mechanical processes including grinding, milling and alloying;

Bioreactors.

Dr. Murday:

Since NS, by definition, have large surface to volume ratios, quality
manufacturing processes must utilize a controlled environment.

For large scale bulk NS the processes are largely either a variant on
colloid chemistry or gas phase nucleation and growth.  Fumed silica and
carbon black are examples of the latter – both can be NS and have been
made in volume for decades.

Dr. Small:

Common methods would include: lithography, vacuum and spray coating,
CVD, Vapor deposition, Plasma vaporization, laser ablation,
arc-discharge, gas-phase catalytic growth of CNT from carbon monoxide,
and Sol gel.  There are of course many “variations” on these methods
some of which are proprietary.  In the case of the more complex
materials these would probably start with one of the above and then have
proprietary additional steps.

Dr. Warheit:

No specific comments provided.

Question 10: How are processes used to produce specific characteristics
or properties?

Dr. BelBruno:

The size of synthesized particles can be controlled through the
chemistry, either by limiting reagents or by the specific conditions of
the chemistry (heat, solvent, reaction mechanism, etc).  To the extent
that properties reflect size (and surface area), then the properties are
controlled by the chemistry.  Of course, that relationship
(size-properties) is not yet well-defined.  Similarly, self assembly can
be chemically controlled and the surface characteristics, therefore,
influenced by the process.  Finally, chemistry can be used to modify or
functionalize the particles.  We know that functionalized nanotubes, for
example, have different properties from the naked nanotubes.

Dr. Canady:

I don’t know.

Dr. Cassell:

Self assembly for instance can be used to control nanoparticle diameter.
 Polymerization techniques (stoichiometric control, temperature,
pressure, etc.) can be used to control engineered polymer systems.
Thermal/chemical treatment can be used to increase surface area or
surface chemistry.  There are a wide variety of techniques borrowed from
conventional disciplines that are used in the manipulation of nanoscale
materials.

Dr. Lahann:

By exactly controlling chemical composition and structure of
nanoparticles/nanoobjects, a specific characteristic or property can be
achieved.  For instance, in the case of CdSe nanocrystals, the particle
diameter defines important optical properties and developing processes,
which can generate monodisperse particles with defined diameters, will
give a unique function.

Dr. Murashov:

This depends on the process, materials and application.

Dr. Murday:

After the formation of most NS it will be necessary to tailor the
surface to make the NS compatible with further processing.  The tailored
surfaces can be thought of as a shell.  There may be several shells as
we progress toward “smart” NS.

As an example, the first shell of a semiconductor QD is designed to trap
the electron-hole pair in the core, a second shell might have a
therapeutic function and a third shell a recognition function.

Processes used to produce the shells will likely vary considerably, but
would again be mostly variants on colloid/gas phases processing.

Dr. Small:

No specific comments provided.

Dr. Warheit:

No specific comments provided.

Question 11: Which methods reduce particle size but do not result in
property changes?  Which methods reduce particle size and result in
property changes?

Dr. BelBruno:

See response to question 10.

Dr. Canady:

First, the question should be about change not reduction.  Size going up
from 50 to 200nm will cause different disposition effects in the body,
so increase in size in the nanoscale range is relevant as well.

But I don’t understand this question.  Size is a property that has
relevance in biology on its own.  If “property” as a function of
size means lack of biological or environmental impact, then how could
you know that properties are not changed without testing?  How could we
know the answer to this question unless the answer is that all methods
to change size will result in some unknown likelihood of a change in
biological or environmental response?  This is a smaller set of the
question of whether you can assume that any modification will have no
effect (by limiting the modification to “size change” rather than
“any modification”).

Dr. Cassell:

This depends on the system under question.  Cutting a carbon nanotube in
half reduces its particle size, but does not greatly effect other
properties.  Reducing gold particle diameters dramatically alter melting
point for instance.

Dr. Lahann:

If a material’s change in particle size results in fundamental
property changes, the outcome is more a function of the material itself
and less of the method used to reduce the size.  However, the process
used to reduce the size of a material may determine polydispersity and
may still be influencing properties.

Dr. Murashov:

If the reduction in size results in changes in properties of interest,
then it does not matter how the reduction in size was achieved.  If
properties of interest also depend on surface characteristics, then the
production method can affect these properties and additional steps such
as surface passivation might be required (see also answer to question
7).

Dr. Murday:

The question is not meaningful – all methods may/may not change
properties.

Processes without a controlled environment are likely to lead to wider
variation in surface composition and thereby different behaviors.  For
instance, the technique of ball milling larger particles into smaller
frequently is inadequately controlled; compacted powder studies have
suffered from inability to reproduce results.

To control size, processes growing NS will need to have a nucleation
event separate from the growth step.  If nucleation and growth proceed
in parallel, there is an inevitable distribution in NS size.

Dr. Small:

Many of the production methods are bottom up so this question does not
apply to those. In general the property change results for the nanoscale
characteristics of the materials not on how they are produced.  Not sure
what this question refers to.

Dr. Warheit:

No specific comments provided.

Impurities

	Impurity content is a growing area of interest in nanotechnology due to
improved performance observed in some cases (e.g., solar cells and
semiconductors) and deleterious effects observed in others (e.g.,
quantum dot quantum computers).  The confounding effects that impurities
have with respect to toxicological endpoints are also being studied
(National Nanotechnology Initiative 2006 Environmental Health and Safety
research report).

Question 12: How important are impurity identity and impurity levels to
the understanding and characterization of nanoscale materials?

Dr. BelBruno:

Impurity level and identity are important factors, since the impurities
are also nanometer scale and their own properties add to the difficulty
in assessing the impact of the target particles.  Again using nanotubes
as an example, the catalytic metals that are sometimes incorporated in
the final product need to be identified, quantified and, often, removed.
 This has been accomplished in a number of different ways.  One example
is provided by Y. Wang, et al, Chemical Physics Letters (2006), 432,
205.

Dr. Canady:

Impurity identity is critical and not clear at this point.  It could be
defined relative to the set of nanoscale properties of interest which
would also be the specification of the material, or of the nanoscale
particle.  Something could be an impurity if it has slight topographic
variation from the optimal and conversely, something may not be an
impurity if it has dramatically differing composition but similar enough
structure so that the effect does not depend on composition.

Impurity could also be defined based on unwanted side effects of a
variation in nanoscale material even if the nanoscale material that is
“impure” (in the sense of its unwanted side effects) is “pure”
with respect to its intended effects.

Impurity could also just be a solvent or carryover from the
manufacturing in the typical sense, such as the PAHs in CNTs.

The need to measure impurity level is a function of effect and would be
case-dependent.

Dr. Cassell:

Extremely important.  I would say that impurities become more relevant
as you scale your system to smaller dimensions.  Impurities could be
even more important in the context of toxicological effects.

Dr. Lahann:

Depending on the application, the effects can either be hugely important
or can be neglected. However, impurities always introduce an additional
source for structural variability into a nanomaterial.

Dr. Murashov:

There is a need to clarify what is meant by “impurity”.  Term
“impurity” when it is applied to nanomaterials is more similar to
molecular chemicals (e. g. organic polymers) than to macroscopic
materials with a high degree of ionic or metallic bonding (e.g. SiO2,
Si, NaCl).  Specifically, target nanomaterials can be contaminated by
other nanomaterials and chemicals (example is a raw carbon nanotube
material composed of a mixture of carbon nanotubes and nanoscale metal
catalyst particles).  Nanomaterials can be also “contaminated” by
atomic/molecular species covalently bonded to nanomaterials.  If such a
“defect” or “contamination” results in significant changes of
nanomaterial properties, it could be considered a new nanomaterial
rather than “contaminated” or “defective”, just as replacement
of an atom in a molecule for a different chemical element creates a new
molecule/chemical.  Presently, in many instances nanomaterials are
produced as mixtures of nanoparticles with a range of particle sizes,
shapes and chemistries, which is, to some degree, similar to a mixture
of oligomers obtained during polymer synthesis.

The importance of “impurity” or “impurity” level (or the amount
of other molecules and nanomaterials in the mixture) for a particular
application will be strongly dependent on that application.  The
importance of “impurity” or “impurity” level on risk of a target
nanomaterial will be strongly dependent on risk-related properties of
the “impurity” and the target nanomaterial.

Dr. Murday:

Impurity identify and level can be very important, i.e., doping of a
semiconductor NS.  The NS size is sufficiently small that a single
dopant atom will have dramatic impact on its electrical properties.
Doping at the nanoscale might be better accomplished by injecting charge
through surface bond compensation.

CNT manufacture provides an excellent case study in quality control (or
lack thereof).  Figure A shows the results of an analysis of eight
different nominal CNT samples – the worst has only 40% carbon (as a
fraction of C, H, O; all other elements were excluded from the
analysis), and that carbon might not be NT.

Dr. Small:

This is a critical area when characterizing these materials. For
example, the catalyst in NT production is nanosized metal (Fe, Ni)
particles. Some studies indicate that these may be more toxic than the
NTs.  The ability to determine the amount and the physical-chemical
character of the impurities contained in ENM relates directly to
understanding and evaluating exposure and risk.

Dr. Warheit:

No specific comments provided.

Question 13: Are there routine purification procedures that can
effectively control or remove impurities, when desirable, for certain
classes of nanoscale materials?

Dr. BelBruno:

For nanotubes, the response is yes (see Q12).  Removing impurities from
inorganic materials is more difficult because procedures could also
affect the desired particles.

Dr. Canady:

I don’t know.

Dr. Cassell:

Yes.  There are many techniques that are borrowed from those known in a
number of disciplines (chemistry, materials science, biology) and used
in nanoscale materials manipulation.  They are too numerous to list
here, but include techniques such as annealing, chemical treatment,
separation, chromatography, sedimentation, plasma treatment.

Dr. Lahann:

Processes to remove impurities from nanoscale materials are often
available.

Dr. Murashov:

The following broad classes of purification processes can be
distinguished:

chemical modification to reduce heterogeneity;

dissolution of ionic, molecular and particulate species from target
nanomaterials (e.g. acid-washing has been reported as a means of
removing metal catalytic particles from raw carbon nanotube material);

mobility sizing using difference in mobility of nanomaterials of
different sizes, masses, charges and shapes in gas and liquid phases,
across membranes and subject to force-fields.

Dr. Murday:

Chemistry provides a rich source of purification processes that can be
exploited for NS.  One limitation could be difficulty in sustaining
solubility/suspension in fluid media.

Dr. Small:

Acid washing for CNT along with other steps has been used for
purification of CNTs.  There are several lab-based methods these are not
routine.

Dr. Warheit:

No specific comments provided.

Obtaining characterization data

	Determining identity, quantifying the nanoscale particle range, and
measuring physical-chemical properties for that identity and particle
range are essential to the characterization of nanoscale materials.
Because of the challenges associated with size, shape, surface
characteristics, and possibly other aspects of nanoscale materials, an
evaluation of existing measurement techniques is critical to nanoscale
material characterization.  The National Nanotechnology Initiative
report stated that “...Accurate and useful measurement techniques are
also important because agglomerated nano materials may either retain or
lose their emergent properties - or take on new properties - thus
affecting the potential biological response.”

Question 14: Are validated methods available for the different
categories of nanoscale materials?

Dr. BelBruno:

Particles are easily sized using commercially available equipment.  This
measurement is a bit more difficult (larger uncertainty) for coated
nanoparticles.  Force and electron microscopy may also be used to
measure the size of particles.  These are essentially universal, but
tedious, methods.  X-ray spectroscopic techniques can be used to
identify metallic impurities (in many electron microscopes).  Surface
area measurement is also a well-developed technique. (see Q4)

Dr. Canady:

No specific comments provided.

Dr. Cassell:

I am unaware of validated methods for nanoscale materials in particular,
but there may be methods (e.g. ISO) that are inclusive of nanoscale
materials.

Dr. Lahann:

Some validated methods are available, but further efforts are needed.

Dr. Murashov:

There are no validated methods specific to engineered nanoparticles
(NIOSH 2006).  There are validated methods for materials that exist in
the nanometer scale, such as welding fume, carbon black; but these
methods do not discriminate by size.  There are some methods that have
been validated for the larger (bulk) form of nanomaterials that may be
applied to the nano form; for instance dust sampling with a cyclone
(ISO, 2007); sampling for a specific element with analysis by NIOSH 7300
(NIOSH, 1994); fiber sampling and analysis by Transmission Electron
Microscopy (TEM).  These methods have not been validated for nanoscale
material, but there is potential for reapplication and they can provide
useful information for evaluating occupational exposures with respect to
particle size, mass, surface area, number concentration, composition,
and surface (NIOSH, 2006).  The challenge is demonstrating that the data
generated by the measurement (the metric) was created by the engineered
nanoparticle rather than by incidental nanoparticles of similar chemical
composition.  Variations of the sampling process for mixtures of
nanoscale and fine TiO2 are being tested (NIOSH, 2005).  One of the
challenges of that method is collecting enough material for the first
mass-based step in the analysis.  Often the electron microscopy analysis
is used to verify that the material collected has the expected elemental
composition and is in the nanometer size range though there is always
some degree of agglomeration.

Dr. Murday:

MS and light scattering are the closest to validated.

NIST and NCL are working this issue – EPA should partner.

Dr. Small:

There are several methods under development be few in publication.  One
I am aware of is IEEE P1650 standard test method of meas. of electrical
properties of CNTs adopted in 2005.

Dr. Warheit:

No specific comments provided.

Question 15: Are there techniques that can be universally applied?

Dr. BelBruno:

See response to question 14.

Dr. Canady:

No specific comments provided.

Dr. Cassell:

I would think so, yes.  Especially the more generic particle
characterization techniques (surface area, particle size determination,
etc.) that have been utilized in other materials characterization work.

Dr. Lahann:

Particle size and shapes can be characterized using Transmission
electron microscopy, scanning electron microscopy and scanning probe
microscopy independent of the materials composition.

Dr. Murashov:

The collection and analysis of an air sample by TEM is an example of a
universal technique which can be applied across several types of
nanomaterials.

Dr. Murday:

In a broad context, there are no techniques that provide all the
necessary information, all the time.  NS can be found in free form and
embedded in matrices.  The NS itself is likely to have
composition/structure that varies with distance from its center.

Light scattering is likely to be useful for size/shape measurement.  It
might be useful/important to extend the photon energy in the far UV to
make the wavelength more comparable to the NS size (however, this may
invoke a problem for the optics).

Mass Spectrometry has been developed to handle very large molecules –
100s of KDa.  Fragmentation patterns can illuminate substructures.  This
technique could prove very useful in probing core/shell NS.

MS and light scattering require multiple copies of a NS and yield
ensemble averaged results.  Techniques to probe individual NS properties
are also very useful.  Some variant on HRSEM with energy dispersive
analysis for composition, coupled with AFM, is likely to be the most
versatile.

SERS might be developed with enough sensitivity to analyze one or a few
NS for a vibrational approach to composition.

Dr. Small:

Not that I am aware of.  Perhaps some of the microscopy methods could be
used to characterize multiple classes.

Dr. Warheit:

No specific comments provided.

Question 16: For small quantities of materials, are there sampling,
handling, and collection techniques as well as sample integrity,
accuracy and precision QA/QC methodologies available?

Dr. BelBruno:

Many nanoparticles are available from commercial suppliers (quantum
dots, fullerenes, nanotubes, inorganic cluster molecules), with
specified purity levels and chemical composition.  Clearly, commercial
suppliers can arrange QA procedures and we should look to these methods
as a means of introducing standards.

For example, from American Elements
(http://www.americanelements.com/fenp.html): “Iron (Fe) Nanoparticles,
nanodots or nanopowder are spherical or faceted high surface area metal
nanostructure particles. Nanoscale Iron Particles are typically 20-40
nanometers (nm) with specific surface area (SSA) in the 30 - 50 m2/g
range and also available in with an average particle size of 100 nm
range with a specific surface area of approximately 7 m2/g. Nano Iron
Particles are also available in Ultra high purity and high purity and
coated and dispersed forms.”

Dr. Canady:

No specific comments provided.

Dr. Cassell:

I would say that there may be generic techniques published in the
technical literature, but these are probably not standardized by a
leading organization.  Quality control procedures for nanoscale
materials are largely dependent upon the organization that handles the
materials and most of these are based upon the needs of that particular
organization.

Dr. Lahann:

Some specialized procedures are available, but further efforts are
needed.

Dr. Murashov:

At this point, the greatest QA/QC challenge is the lack of methods
specific for nanoscale particles and the lack of reference materials
that accurately represent the material being evaluated.

Dr. Murday:

The traditional handling of chemicals provides proven precedents.  But
they will have to be adapted to the broader range of size/properties
incumbent in NS.

Testing on NS shows (at least in some circumstances) that they can be
collected effectively on filters – van der Wall forces or surface
charges attract to the filter material; NS are small enough that fluid
drag forces are small and don’t displace them from the filter.

Dr. Small:

No.  I think best-practices methods (handling of ultrafines) are used
although there are several efforts by government agencies and industry
to develop safe handling practices for ENMs that would apply to
“small” quantities.

Dr. Warheit:

No specific comments provided.

Question 17: What is the status of standardization efforts? Are these
efforts focused on broadly applicable characterization methods or
category-specific methods?

Dr. BelBruno:

Mostly category specific, as the response to Q19 indicates.

Dr. Canady:

No specific comments provided.

Dr. Cassell:

NIST is active in generating standards for single walled carbon
nanotubes see:    HYPERLINK
"http://polymers.nist.gov/Nanotube3/Agenda.pdf"
http://polymers.nist.gov/Nanotube3/Agenda.pdf .  There are a variety of
techniques described for characterizing single walled carbon nanotubes.

Dr. Lahann:

Depending on the field and material, standardization efforts vary and
further activities are needed.

Dr. Murashov:

Standardization efforts are focused on both broadly applicable
characterization methods and category-specific methods. For example, the
ISO TC 229 (Nanotechnologies), Working Group 3 (Health, Safety and the
Environment) is working on five projects:

Technical Report “Health and Safety Practices in Occupational Settings
Relevant to Nanotechnologies” – broadly applicable characterization
methods;

International Standard “Endotoxin test on nanomaterial samples for in
vitro systems” – broadly applicable;

International Standard “Generation of silver nanoparticles for
inhalation toxicity testing” – specific to silver nanoparticles;

International Standard “Monitoring silver nanoparticles in inhalation
exposure chambers for inhalation toxicity testing” - specific to
silver nanoparticles;

Technical Report “Guidance on physico-chemical characterization of
engineered nanoscale materials for toxicologic assessment” - broadly
applicable.

The ISO TC 229 Working Group 2 (Metrology and Instrumentation) is
working on a number of standards for characterizing carbon nanotubes.

Dr. Murday:

No specific comments provided.

Dr. Small:

Physical standards: Standardization efforts are in the process of
ramping up both from an activity as well as a funding view point.
Government, industry, and academia, in the US as well as other areas
like Europe are in the process of defining and measuring candidate
materials. Efforts are focused on both category-specific i.e. CNT and on
broader definitions such as metal oxides or on schemes used for
classifications of materials like toxicity, commercial impact, and
environmental impact.

Documentary Standards (methods): As with the physical standards these
efforts are also increasing although development of “standard methods,
in most cases, requires the availability of well characterized physical
standards.

Dr. Warheit:

No specific comments provided.

Question 18: What alternative or innovative methods or technologies can
be applied to nanomaterial analysis?

Dr. BelBruno:

No specific comments provided.

Dr. Canady:

No specific comments provided.

Dr. Cassell:

Atomic force microscopy along with some of the latest scanning probe
microscopy techniques could be useful for nanomaterials analysis.  I’m
unsure if standards for characterizing with these techniques have been
set.

Dr. Lahann:

Transmission electron microscopy, scanning electron microscopy and
scanning probe microscopy, fluorescence microscopy, X-ray microscopy.

Dr. Murashov:

Some examples of alternative or innovative methods or technologies that
could be applied to nanomaterial analysis as part of safety and health
assessments are:

A number of instruments providing on-line size-resolved chemical
speciation of aerosols, such as aerosol mass-spectrometry, have been
developed.  These techniques can provide compositional information on
single particles or ensembles of size-selected particles in real time.

Size distribution and number concentration of nanoparticles in liquid
phase can be characterized using light scattering, laser diffraction,
size exclusion chromatography, acoustic techniques and field flow
fractionation (Powers et al, 2006), while space-resolved spectroscopic
techniques can be useful in obtaining information about chemical
composition and structure of nanoparticles.

Dr. Murday:

ASTM interlab standards not yet in place.

Tip characterization for proximal probes is a major challenge.

Absence of standard materials another factor.

Lab on a chip and variants on microarrays might offer new approaches
that could be readily adapted to field use.

To the extent that NS impact on living systems is paramount, cell based
microarrays (being developed for drug candidate screening) might provide
a “canary on a chip.”

Dr. Small:

There are many novel technologies, e.g. microscopy-based, as well as
others, which are applicable to nanoscale regimes.  Is the EPA looking
for methods that are likely to be widely used (cheap and fast) or for
very specific research areas?  Many of these methods are currently
difficult and expensive to make and are confined to laboratory studies
some may eventually become routine others will not.  Examples of some
techniques include: super-resolution optical, 3D chemical tomography,
TGA on chip, size-exclusion chromatography.

Dr. Warheit:

No specific comments provided.

Modeling

Empirical modeling can be a useful approach to predict physical-chemical
properties when experimental data are not known or ascertainable.  The
initial problem with modeling is that, to accurately predict property
endpoints for a given category of substances, there must be some
experimental data available in the tool’s database for at least some
representative substances in that category.  For newly discovered or
studied materials, the minimum but necessary quantity and type of
experimental data often is not available to sufficiently populate a
tool’s database and allow accurate prediction by the tool.  Some
estimation methods have been developed for specific property endpoints,
but many others are lacking.

Question 19: Are there models that are currently used to obtain property
data for nanoscale materials?  For which properties and which nanoscale
materials?

Dr. BelBruno:

I believe that we need to have a long-term view for this.  Simulations
provide agreement with already established data, but are not predictive.
 Quantum mechanical calculations provide a basis for the emission in
quantum dots.  However, structure function type relationships of the
type used in the pharmaceutical industry do not exist.

Dr. Canady:

I don’t know.

Dr. Cassell:

There are some atomistic approaches that have been used to predict
properties of nanoscale materials and some of these models have been
extended to molecular dynamics and mesoscale modeling approaches.  I am
not an expert in the field, but I am aware of these techniques.  The
properties modeled are typically physical in nature such as mechanical,
thermal or electrical in nature.  Primarily these approaches are
utilized to understand transport phenomena.

Dr. Lahann:

A series of different approaches is currently under development
depending on specific materials categories.

Dr. Murashov:

Only few risk-related models exist that are currently used to obtain
property data for nanoscale materials.  Examples are:

Several models currently used to obtain property data for nanoscale
materials are models based on aerodynamic properties of nanoparticles
such as penetration through filter media, coagulation and deposition in
the lung (Lee and Liu, 1982; Maynard and Kuempel, 2005; ICRP, 1994).

Research data indicate that health hazard properties of poorly soluble,
low-toxicity nanoparticles can be modeled using their total surface area
(Maynard and Kuempel, 2005; Duffin et al., 2007).

Dr. Murday:

It is possible to extrapolate from smaller (atomistic) and larger
(continuum) scale models, but caution must be exercised.

First principle models of CNT were able to predict accurately many of
their properties well before the experimental community could measure
them (see Figure C).  However, CNT is a special case of NS – CNT is
all C, has only on type of bond, and has a high degree of symmetry.

Dr. Small:

No specific comments provided.

Dr. Warheit:

No specific comments provided.

Question 20:  Has any validation work been conducted that compares
predicted values with measured data?  For which properties and which
nanoscale materials?

Dr. BelBruno:

See response to question 19.

Dr. Canady:

NCL has published standard methods through ASTM for some assays that
have validation data.

Dr. Cassell:

Individual carbon nanotube based thermal conductivity has been predicted
by theory and subsequently validated using experimental approach.  Prof.
Arun Majumdar’s group at UC Berkeley performed these elegant
experiments.

Dr. Lahann:

Some of the models have been validated.  A good example is the work of
Nick Kotov and Sharon Glotzer at University of Michigan that
experimentally validated the formation of nanoparticle sheets (Science
2006).  More work is needed.

Dr. Murashov:

Validation work has been conducted for data related to penetration of
filter media by nanoscale particles (see for example, Rengasamy et al.,
2007).

Dr. Murday:

CNT model predictions have been verified by experiment (see Figure C).

QD model predictions have been verified by experiment.

Fluid flow in nanopores has been examined by Navier Stokes (continuum)
and molecular dynamics approaches to understand the modeling
limitations.  Results have been partially validated by experiment.

Dr. Small:

No specific comments provided.

Dr. Warheit:

No specific comments provided.

Question 21: Are there current significant characterization needs for
which the NMSP should investigate model development?

Dr. BelBruno:

See response to question 19.

Dr. Canady:

Yes – but an entire workshop or similar effort should be devoted to
developing ideas and projects for modeling.  The most immediate one may
be developing a modeling approach that allows updating of the
information needs relative to the decision needs.  As more is learned
about the materials and the properties that are most relevant to
decisions there will need to be a process for adjusting the information
requirements.

Dr. Cassell:

I’m not sure if there are aggregation/agglomeration models that could
be used for various types of nanomaterials, but I see this as being a
valuable method to augment characterization approaches.  Any predictive
tool that is useful in a contextual setting as opposed to idealized
models would be extremely useful.

Dr. Lahann:

Specifically in the area of correlation between nanoparticle properties,
such as size, shape, composition, and biological properties, such as
cell uptake or cytotoxicity, there would be a great potential for model
developments.

Dr. Murashov:

Examples of current significant characterization needs for which the
NMSP should investigate model development are:

Risk assessment models for nanoscale materials are needed.
Specifically, models to extrapolate toxicity data from in vitro to in
vivo; from in vivo to human; external exposure levels to internal dose
should be further developed.

There is a need to develop models correlating biomarkers of exposure
with specific nanomaterials.

There is also a need to determine if certain physico-chemical properties
of nanoparticles are predictive of biological activity.

Dr. Murday:

Chemical reactivity – partner with catalysis efforts.

Toxicology – partner with NIH, NIOSH, NIST, NCL.

Fate and effect in soils – a topic more closely aligned with EPA than
other agencies.

Dr. Small:

No specific comments provided.

Dr. Warheit:

No specific comments provided.

Supporting Figures

Figures provided by Dr. Murday:

Figure   SEQ Figure \* ALPHABETIC  A

FIELD	PROPERTY	SCALE LENGTH

ELECTRONIC	ELECTRON WAVELENGTH

INELASTIC MEAN FREE PATH

TUNNELING	10

1

1	--

--

--	100nm

100nm

10nm

MAGNETIC	DOMAIN WALL

EXCHANGE ENERGY

SPIN-FLIP SCATTERING LENGTH	10

0.1

1	--

--

--	100 nm

1nm

100nm

OPTIC	QUANTUM WELL

EVANESCENT WAVE DECAY LENGTH

METALLIC SKIN DEPTHS	1

10

10	--

--

--	100nm

100nm

100nm

SUPERCONDUCTIVITY	COOPER PAIR COHERENCE LENGTH

MEISSNER PENETRATION DEPTH	0.1

1	--

--	100nm

100nm

MECHANICS	DISLOCATION INTERACTION

GRAIN BOUNDRIES	1

1	--

--	1000nm

10nm

NUCLEATION/

GROWTH	DEFECT

SURFACE CORRUGATION	0.1

1	--

--	10nm

10nm

CATALYSIS	LOCALIZED BONDING ORBITALS

SURFACE TOPOLOGY	0.01

1	--

--	0.1nm

10nm

SUPRAMOLECULES	PRIMARY STRUCTURE

SECONDARY STRUCTURE

TERTIARY STRUCTURE	0.1

1

10	--

--

--	1nm

10nm

1000nm

IMMUNOLOGY	MOLECULAR RECOGNITION	1	--	10nm

Figure   SEQ Figure \* ALPHABETIC  B : Characteristic Lengths in Solid
State Science Models

Figure   SEQ Figure \* ALPHABETIC  C

Supporting References

References provided by Dr. Murashov:

Duffin, R., Tran, L., Brown, D., Stone, V., and Donaldson, K. (2007).
Proinflammogenic effects of low-toxicity and metal nanoparticles in vivo
and in vitro: highlighting the Role of Particle Surface Area and Surface
Reactivity, Inhal. Tox. 19(10), 849-856.

ICRP, Human respiratory tract model for radiological protection, 1994,
ICRP publication 66.

ISO, Workplace Atmospheres – Ultrafine, nanoparticle and
nano-structured aerosols – Inhalation exposure characterization and
assessment, ISO/TR 27628:2007, 2007.

Jevprasesphant, R., Penny, J., Jalal, R., Attwood, D., MckEown, N. B.,
and D’Emanuele, A. (2003). The influence of surface modification on
the cytotoxicity of PAMAM dendrimers, Int. J. Pharm. 252, 263-266.

Lee, K. W. and Liu, B. Y. H. (1982). Theoretical study of aerosol
filtration by fibrous filters, Aerosol Sci. Technol. 1(2), 147-162.

Maynard, A. D. and Kuempel, E. D. (2005). Airborne nanostructured
particles and occupational health. J. Nanopart. Res. 7, 587-614.

Mecke, A., Uppuluri, S., Sassanella, T. M., Lee, D. K., Ramamoorthy, A.,
Baker, J. R., Jr., Orr, B. G., and Banaszak Holl, M. M. (2004). Direct
observation of lipid bilayer disruption by poly(amidoamine) dendrimers.
Chem. Phys. Lipids 132, 3-14.

NIOSH (1994). NIOSH Manual of Analytical Methods (NMAM®), 4th ed. DHHS
(NIOSH) Publication 94-113 (August, 1994), Schlecht, P.C. & O'Connor,
P.F., Eds., 1994. Available on line at   HYPERLINK
"http://www.cdc.gov/niosh/nmam/chaps.html"
www.cdc.gov/niosh/nmam/chaps.html .

NIOSH (2005). Evaluation of Health Hazard and Recommendations for
Occupational Exposure to Titanium Dioxide. Available on line at
HYPERLINK "www.cdc.gov/niosh/review/public/TIo2/"
www.cdc.gov/niosh/review/public/TIo2/ .

NIOSH (2006). Approaches to Safe Nanotechnology: An Information Exchange
with NIOSH. Available online at:   HYPERLINK
"http://www.cdc.gov/niosh/topics/nanotech/safenano/"
http://www.cdc.gov/niosh/topics/nanotech/safenano/ .

Powers, K. W., Brown, S. C., Krishna, V. B., Wasdo, S. C., Moudgil, B.
M., and Roberts, S. M. (2006). Research Strategies for Safety Evaluation
of Nanomaterials. Part VI. Characterization of Nanoscale Particles for
Toxicological Evaluation. Tox. Sci. 90(2), 296-303.

Rengasamy, S., Verbofsky, R., King, W. P., and Shaffer, R. E. (2007).
Nanoparticle Penetration through NIOSH-approved N95 Filtering-facepiece
Respirators, J. Int. Soc. Resp. Protec. 24, 49-59.

Sayes, C. M., Fortner, J. D., Guo, W., Lyon, D., Boyd, A. M., Ausman, K.
D., Tao, Y. J., Sitharaman, B., Wilson, L. J., Hughes, J. B., West, J.
L., and Colvin, V. L. (2004). The differential cytotoxicity of
water-soluble fullerenes, Nano Lett. 4 (10), 1881-1887.

Shvedova, A.A., Sager, T., Murray, A.R., Kisin, E., Porter, D.W.,
Leonard, S., Schwegler-Berry, D., Robinson, V.A., and Castranova, V.
(2007). Critical issues in the evaluation of possible adverse pulmonary
effects resulting from airborne nanoparticles.  In: Nanotoxicology –
Characterization, Dosing and Health Effects. N.A. Monteiro-Riviere and
C.L. Tran (eds), Informa Healthcare, NY, Chap14, pp 225-236.

Singh, R., Pantarotto, D., Lacerda, L., Pastorin, G., Klumpp, C., Prato,
M., Bianco, A., Kostarelos, K. (2006). Tissue biodistribution and blood
clearance rates of intravenously administered carbon nanotube
radiotracers, Proc. Nat. Acad. Sci. 103 (9), 3357-3362.

Wexler, A. S., and Johnston, M. V. (2001). Real-time single-particle
analysis. In Aerosol Measurement: Principles, Techniques and
Applications, Baron, P. A. and Willeke, K. eds., John Wiley & Sons, New
York, pp. 365-385.

References provided by Dr. Warheit:

References for Nanoparticle Material Characterization Strategies for
Toxicity Studies

International Life Sciences Institute – Panel – “Principles for
characterizing the potential human health effects from exposure to
nanomaterials: elements of a screening strategy” - Oberdorster G,
Maynard A, Donaldson K, Castranova V, Fitzpatrick J, Ausman K, Carter J,
Karn B, Kreyling W, Lai D, Olin S, Monteiro-Rivere N, Warheit DB, and
Yang H.  Particle and Fibre Toxicology 2005, 2:8.

“Physicochemical properties that may be important in understanding the
toxic effects of test materials include particle size and size
distribution, agglomeration state, shape, crystal structure, chemical
composition, surface area, surface chemistry, surface charge, and
porosity”.

Testing strategies to establish the safety of nanomaterials:
Conclusions of an ECETOC workshop.  Inhal. Toxicol., 19: 631-643, 2007.
Warheit DB, Borm PJA, Hennes C, and Lademann J.  Inhal. Toxicol., 19:
631-643, 2007.

“In summary, it was concluded that nanoparticle composition,
dissolution, surface area and characteristics, size, size distribution,
and shape are parameters needed for any target organ toxicity
assessment.  Depending on the type of toxicological study undertaken,
other physicochemical parameters would also be required

 http://www.particleand fibretoxicology.com/content/2/1/8

 http://www.ansi.org/standards_activities/standards_boards_panels/nsp/ov
erview.aspx?menuid=3

    HYPERLINK
"http://www.iso.org/iso/en/stdsdevelopment/tc/tclist/TechnicalCommitteeD
etailPage.TechnicalCommitteeDetail"
http://www.iso.org/iso/en/stdsdevelopment/tc/tclist/TechnicalCommitteeDe
tailPage.TechnicalCommitteeDetail ? COMMID=5932

    HYPERLINK
"http://publicaa.ansi.org/sites/apdl/Documents/Forms/AllItems.aspx"
http://publicaa.ansi.org/sites/apdl/Documents/Forms/AllItems.aspx

 Class 1 substances are distinct chemicals with known, non-variable
molecular structures

  http://www.nano.gov/NNI_EHS_research_needs.pdf

    HYPERLINK
"http://publicaa.ansi.org/sites/apdl/Documents/Forms/AllItems.aspx"
http://publicaa.ansi.org/sites/apdl/Documents/Forms/AllItems.aspx

 Class 1 substances are distinct chemicals with known, non-variable
molecular structures

  http://www.nano.gov/NNI_EHS_research_needs.pdf

 PAGE

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TABLE OF CONTENTS (Continued)

Page

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B-  PAGE  7

Compilation of Panelists’ Preliminary Comments

C-  PAGE  39


[Federal Register Volume 78, Number 15 (Wednesday, January 23, 2013)]
[Rules and Regulations]
[Pages 4790-4792]
[FR Doc No: 2013-01239]

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DEPARTMENT OF HOMELAND SECURITY

Coast Guard

33 CFR Part 165

[Docket Number USCG-2012-0938]
RIN 1625-AA87

Security Zone, Potomac and Anacostia Rivers; Washington, DC

AGENCY: Coast Guard, DHS.

ACTION: Temporary final rule.

-----------------------------------------------------------------------

SUMMARY: The Coast Guard is establishing a temporary security zone
encompassing certain waters of the Potomac River and Anacostia River.
This action is necessary to safeguard persons and property, and prevent
terrorist acts or incidents. This rule prohibits vessels and people
from entering the security zone and requires vessels and persons in the
security zone to depart the security zone, unless specifically exempt
under the provisions in this rule or granted specific permission from
the Coast Guard Captain of the Port Baltimore. This action is intended
to temporarily restrict vessel traffic in portions of the Potomac and
Anacostia Rivers during the event.

DATES: This rule is effective from January 15, 2013 until January 24,
2013.

ADDRESSES: Documents mentioned in this preamble are part of docket
[USCG-2012-0938]. To view documents mentioned in this preamble as being
available in the docket, go to http://www.regulations.gov, type the
docket number in the ``SEARCH'' box and click ``SEARCH.'' Click on Open
Docket Folder on the line associated with this rulemaking. You may also
visit the Docket Management Facility in Room W12-140 on the ground
floor of the Department of Transportation West Building, 1200 New
Jersey Avenue SE., Washington, DC 20590, between 9 a.m. and 5 p.m.,
Monday through Friday, except Federal holidays.

FOR FURTHER INFORMATION CONTACT: If you have questions on this rule,
call or email Mr. Ronald L. Houck, at Sector Baltimore Waterways
Management Division, U.S. Coast Guard; telephone<PHONE_NUMBER>, email
<EMAIL_ADDRESS>If you have questions on viewing or submitting
material to the docket, call Renee V. Wright, Program Manager, Docket
Operations, telephone<PHONE_NUMBER>.

SUPPLEMENTARY INFORMATION:

Table of Acronyms

DHS Department of Homeland Security
FR Federal Register
NPRM Notice of Proposed Rulemaking

A. Regulatory History and Information

    On October 24, 2012, we published a notice of proposed rulemaking
(NPRM) entitled ``Security Zone, Potomac and Anacostia Rivers;
Washington, DC'' in the Federal Register (77 FR 64943). After the NPRM
was published in the Federal Register, however, the Coast Guard
determined that the boundary of the proposed security zone on the south
between the Virginia shoreline and the District of Columbia shoreline
along latitude 38[deg]51'00'' N needed to be relocated farther
downstream to and along latitude 38[deg]50'00'' N. On November 28,
2012, we published a supplemental notice of proposed rulemaking (SNPRM)
entitled ``Security Zone, Potomac and Anacostia Rivers; Washington,
DC'' in the Federal Register (77 FR 70964). We received one comment on
the proposed rules. No public meeting was requested, and none was held.
    Under 5 U.S.C. 553(d)(3), the Coast Guard finds that good cause
exists for making this rule effective less than 30 days after
publication in the Federal Register. Due to the need for immediate
action, the restriction of vessel traffic is necessary to protect life,
property and the environment; therefore, a 30-day notice is
impracticable. Delaying the effective date would be contrary to the
safety zone's intended objectives of protecting persons and vessels
involved in the event and its associated activities, and enhancing
public and maritime safety.

B. Basis and Purpose

    On January 20, 2013, the U.S. Presidential Inauguration swearing-in
ceremony will take place at the U.S. Capitol in Washington, DC.
Activities associated with the Presidential Inauguration include
several Inaugural ceremonies, balls, parades and receptions in the
District of Columbia, which are scheduled to occur from January 15,
2013 through January 24, 2013. During these activities, gatherings of
high-ranking United States officials and the public-at-large are
expected to take place. These activities are located along navigable
waterways within the Captain of the Port Baltimore's Area of
Responsibility. The Coast Guard has given each Coast Guard Captain of
the Port the ability to implement comprehensive port security regimes
designed to safeguard human life, vessels, and waterfront facilities
while still sustaining the flow of commerce.
    The Captain of the Port Baltimore is establishing a security zone
to address the aforementioned security concerns and to take steps to
prevent the catastrophic impact that a terrorist attack against the
large gatherings of high-ranking United States officials, the public-
at-large, and surrounding waterfront areas and communities would have.
The security zone is necessary to safeguard life and property on the
navigable waters before, during, and after activities associated with
the Presidential Inauguration and will help the Coast Guard prevent
vessels or persons from bypassing the security measures established on
shore for the events and engaging in waterborne terrorist actions
during the highly-publicized events.

C. Discussion of Comments, Changes and the Final Rule

    The Coast Guard received one comment in response to the NPRM. No
public meeting was requested and none was held. What follows is a
review of,

and the Coast Guard's response to, the issue that was presented by the
commenter concerning the proposed regulations.
    The commenter, Mr. David A. Bell, a resident of Hamburg, NY, stated
his support for the Coast Guard's proposed temporary security zone.
    The security zone is tailored to impose a minimum adverse affect on
port operations and waterway users located within certain waters of the
Potomac River and Anacostia River at Washington, DC during the event
and its associated activities.

D. Regulatory Analyses

    We developed this rule after considering numerous statutes and
executive orders related to rulemaking. Below we summarize our analyses
based on these statutes and executive orders.

1. Regulatory Planning and Review

    This rule is not a significant regulatory action under section 3(f)
of Executive Order 12866, Regulatory Planning and Review, as
supplemented by Executive Order 13563, Improving Regulation and
Regulatory Review, and does not require an assessment of potential
costs and benefits under section 6(a)(3) of Executive Order 12866 or
under section 1 of Executive Order 13563. The Office of Management and
Budget has not reviewed it under those Orders. There is no vessel
traffic associated with recreational boating and commercial fishing
expected during the effective period, and vessels may seek permission
from the Captain of the Port Baltimore to enter and transit the zone.

2. Impact on Small Entities

    The Regulatory Flexibility Act of 1980 (RFA), 5 U.S.C. 601-612, as
amended, requires federal agencies to consider the potential impact of
regulations on small entities during rulemaking. The term ``small
entities'' comprises small businesses, not-for-profit organizations
that are independently owned and operated and are not dominant in their
fields, and governmental jurisdictions with populations of less than
50,000. The Coast Guard certifies under 5 U.S.C. 605(b) that this rule
will not have a significant economic impact on a substantial number of
small entities. This rule may affect the following entities, some of
which might be small entities: the owners or operators of vessels
intending to operate or transit through or within the security zone
during the enforcement period. Although the security zone will apply to
the entire width of the Potomac and Anacostia Rivers, traffic may be
allowed to pass through the zone with the permission of the Captain of
the Port Baltimore. Before the effective period, maritime advisories
will be widely available to the maritime community. Additionally, given
the time of year this event is scheduled, the vessel traffic is
expected to be minimal.

3. Assistance for Small Entities

    Under section 213(a) of the Small Business Regulatory Enforcement
Fairness Act of 1996 (Public Law 104-121), we want to assist small
entities in understanding this rule. If the rule would affect your
small business, organization, or governmental jurisdiction and you have
questions concerning its provisions or options for compliance, please
contact the person listed in the FOR FURTHER INFORMATION CONTACT,
above.
    Small businesses may send comments on the actions of Federal
employees who enforce, or otherwise determine compliance with, Federal
regulations to the Small Business and Agriculture Regulatory
Enforcement Ombudsman and the Regional Small Business Regulatory
Fairness Boards. The Ombudsman evaluates these actions annually and
rates each agency's responsiveness to small business. If you wish to
comment on actions by employees of the Coast Guard, call 1-888-REG-FAIR
(1-888-734-3247). The Coast Guard will not retaliate against small
entities that question or complain about this rule or any policy or
action of the Coast Guard.

4. Collection of Information

    This rule will not call for a new collection of information under
the Paperwork Reduction Act of 1995 (44 U.S.C. 3501-3520).

5. Federalism

    A rule has implications for federalism under Executive Order 13132,
Federalism, if it has a substantial direct effect on the States, on the
relationship between the national government and the States, or on the
distribution of power and responsibilities among the various levels of
government. We have analyzed this rule under that Order and determined
that this rule does not have implications for federalism.

6. Protest Activities

    The Coast Guard respects the First Amendment rights of protesters.
Protesters are asked to contact the person listed in the FOR FURTHER
INFORMATION CONTACT section to coordinate protest activities so that
your message can be received without jeopardizing the safety or
security of people, places or vessels.

7. Unfunded Mandates Reform Act

    The Unfunded Mandates Reform Act of 1995 (2 U.S.C. 1531-1538)
requires Federal agencies to assess the effects of their discretionary
regulatory actions. In particular, the Act addresses actions that may
result in the expenditure by a State, local, or tribal government, in
the aggregate, or by the private sector of $100,000,000 (adjusted for
inflation) or more in any one year. Though this rule will not result in
such an expenditure, we do discuss the effects of this rule elsewhere
in this preamble.

8. Taking of Private Property

    This rule will not cause a taking of private property or otherwise
have taking implications under Executive Order 12630, Governmental
Actions and Interference with Constitutionally Protected Property
Rights.

9. Civil Justice Reform

    This rule meets applicable standards in sections 3(a) and 3(b)(2)
of Executive Order 12988, Civil Justice Reform, to minimize litigation,
eliminate ambiguity, and reduce burden.

10. Protection of Children

    We have analyzed this rule under Executive Order 13045, Protection
of Children from Environmental Health Risks and Safety Risks. This rule
is not an economically significant rule and does not create an
environmental risk to health or risk to safety that may
disproportionately affect children.

11. Indian Tribal Governments

    This rule does not have tribal implications under Executive Order
13175, Consultation and Coordination with Indian Tribal Governments,
because it does not have a substantial direct effect on one or more
Indian tribes, on the relationship between the Federal Government and
Indian tribes, or on the distribution of power and responsibilities
between the Federal Government and Indian tribes.

12. Energy Effects

    This action is not a ``significant energy action'' under Executive
Order 13211, Actions Concerning Regulations That Significantly Affect
Energy Supply, Distribution, or Use.

13. Technical Standards

    This rule does not use technical standards. Therefore, we did not
consider the use of voluntary consensus standards.

14. Environment

    We have analyzed this rule under Department of Homeland Security
Management Directive 023-01 and Commandant Instruction M16475.lD, which
guide the Coast Guard in complying with the National Environmental
Policy Act of 1969 (NEPA) (42 U.S.C. 4321-4370f), and have determined
that this action is one of a category of actions that do not
individually or cumulatively have a significant effect on the human
environment. This rule involves establishing a temporary security zone.
This rule is categorically excluded from further review under paragraph
34(g) of Figure 2-1 of the Commandant Instruction. An environmental
analysis checklist and a categorical exclusion determination are
available in the docket where indicated under ADDRESSES. We seek any
comments or information that may lead to the discovery of a significant
environmental impact from this rule.

List of Subjects in 33 CFR Part 165

    Harbors, Marine safety, Navigation (water), Reporting and
recordkeeping requirements, Security measures, Waterways.

    For the reasons discussed in the preamble, the Coast Guard amends
33 CFR part 165 as follows:

PART 165--REGULATED NAVIGATION AREAS AND LIMITED ACCESS AREAS

0
1. The authority citation for part 165 continues to read as follows:

    Authority: 33 U.S.C. 1231; 46 U.S.C. Chapter 701, 3306, 3703; 50
U.S.C. 191, 195; 33 CFR 1.05-1, 6.04-1, 6.04-6, and 160.5; Pub. L.
107-295, 116 Stat. 2064; Department of Homeland Security Delegation
No. 0170.1.

0
2. Add Sec.  165.T05-0938 to read as follows:

Sec.  165.T05-0938  Security Zone, Potomac and Anacostia Rivers;
Washington, DC.

    (a) Location. The following area is a security zone:
    (1) All waters of the Potomac River, from shoreline to shoreline,
bounded on the north by the Francis Scott Key (U.S. Route 29) Bridge at
mile 113.0, downstream to and bounded on the south between the Virginia
shoreline and the District of Columbia shoreline along latitude
38[deg]50'00'' N, including the waters of the Georgetown Channel Tidal
Basin; and
    (2) All waters of the Anacostia River, from shoreline to shoreline,
bounded on the north by the 11th Street (I-295) Bridge at mile 2.1,
downstream to and bounded on the south by its confluence with the
Potomac River. All coordinates refer to datum NAD 1983.
    (b) Regulations. The general security zone regulations found in 33
CFR 165.33 apply to the security zone created by this temporary
section, Sec.  165.T05-0938.
    (1) All persons are required to comply with the general regulations
governing security zones found in 33 CFR 165.33.
    (2) Entry into or remaining in this zone is prohibited unless
authorized by the Coast Guard Captain of the Port Baltimore. Vessels
already at berth, mooring, or anchor at the time the security zone is
implemented do not have to depart the security zone. All vessels
underway within this security zone at the time it is implemented are to
depart the zone.
    (3) Persons desiring to transit the area of the security zone must
first obtain authorization from the Captain of the Port Baltimore or
his designated representative. Permission may be requested prior to
activation of the zone. To seek permission to transit the area, the
Captain of the Port Baltimore and his designated representatives can be
contacted at telephone number<PHONE_NUMBER> or on Marine Band Radio VHF-
FM channel 16 (156.8 MHz). The Coast Guard vessels enforcing this
section can be contacted on Marine Band Radio VHF-FM channel 16 (156.8
MHz). Upon being hailed by a U.S. Coast Guard vessel, or other Federal,
State, or local agency vessel, by siren, radio, flashing light, or
other means, the operator of a vessel shall proceed as directed. If
permission is granted, all persons and vessels must comply with the
instructions of the Captain of the Port Baltimore or his designated
representative and proceed at the minimum speed necessary to maintain a
safe course while within the zone.
    (c) Enforcement. The U.S. Coast Guard may be assisted in the patrol
and enforcement of the zone by Federal, State, and local agencies.
    (d) Definitions. As used in this section:
    Captain of the Port Baltimore means the Commander, U.S. Coast Guard
Sector Baltimore, Maryland.
    Designated representative means any Coast Guard commissioned,
warrant, or petty officer who has been authorized by the Captain of the
Port Baltimore to assist in enforcing the security zone described in
paragraph (a) of this section.
    (e) Effective period. This section will be enforced from 8 a.m. on
January 15, 2013 through 10 p.m. on January 24, 2013.

    Dated: January 10, 2013.
Kevin C. Kiefer,
Captain, U.S. Coast Guard, Captain of the Port Baltimore.
[FR Doc. 2013-01239 Filed 1-22-13; 8:45 am]
BILLING CODE 9110-04-P


[Federal Register Volume 77, Number 136 (Monday, July 16, 2012)]
[Rules and Regulations]
[Pages 41670-41671]
[FR Doc No: 2012-17302]

-----------------------------------------------------------------------

DEPARTMENT OF COMMERCE

Bureau of Industry and Security

15 CFR Part 774

The Commerce Control List

CFR Correction

    In Title 15 of the Code of Federal Regulations, Parts 300 to 799,
revised as of January 1, 2012, in supplement no. 1 to part 774, make
the following corrections:
    1. In Category 0:
    A. On page 663, in 0A981, add ``N/A'' behind ``LVS:''.
    B. On page 665, in 0A985, add the heading ``License Requirements''
above ``Reason for Control''.
    C. On page 665, in 0A986, correct the table under ``License
Requirements'' to read as set forth below.
    D. On page 671, in 0B986, add ``, North Korea,'' between ``Iraq''
and ``Rwanda'' in UN Reason for Control.
    2. In Category 1:
    A. On page 676, in 1A004, add ``(1)'' after the colon, at the
beginning of ``Related Definitions''.
    B. On page 682, in 1B001, remove ``Note: 1B001.c does not control
textile machinery not modified for the above end-uses.''
    C. On page 707, in 1C351, after ``Related Definitions;'' remove ``*
* *''

and add paragraphs (1) and (2) as set forth below.
    3. In Category 2:
    A. On page 734, in 2B009, remove the text after ``Related
Definitions'' and add ``N/A'' in its place.
    B. On page 734, in 2B009, revise the Technical Note to read
``Technical Note: For the purpose of 2B009, machines combining the
function of spin-forming and flow-forming are regarded as flow-forming
machines.''
    C. On page 757, in 2E003, in the Notes to Table on Deposition
Techniques, in note 15, add the word ``are'' after ``Dielectric
layers''.
    D. On page 759, in 2E018, in the ``Reasons for Control'', remove
``CC, RS,'', and remove ``License Requirement Notes: See Sec.  743.1 of
the EAR for reporting requirements for exports under License
Exceptions.''
    E. On page 759, in 2E101, add ``(1)'' after the colon at the
beginning of ``Related Controls''.

0
The text to be revised and added is set forth below:

Supplement No. 1 to Part 774--The Commerce Control List

* * * * *

Category 0

* * * * *
0A986 Shotgun shells, except buckshot shotgun shells, and parts.
* * * * *

------------------------------------------------------------------------
                  Control(s)                         Country chart
------------------------------------------------------------------------
AT applies to entire entry. A license is
 required for items controlled by this entry
 to North Korea for anti-terrorism reasons.
 The Commerce Country Chart is not designed
 to determine AT licensing requirements for
 this entry. See Sec.   742.19 of the EAR for
 additional information.
FC applies to entire entry...................  FC Column 1.
UN applies to entire entry...................  Iraq, North Korea, and
                                                Rwanda.
------------------------------------------------------------------------

* * * * *

Category 1

* * * * *
1C351 Human and zoonotic pathogens and ``toxins'', as follows (see
List of Items Controlled).
* * * * *
    Related Definitions: (1) For the purposes of this entry
``immunotoxin'' is defined as an antibody-toxin conjugate intended
to destroy specific target cells (e.g., tumor cells) that bear
antigens homologous to the antibody. (2) For the purposes of this
entry ``subunit'' is defined as a portion of the ``toxin''.
* * * * *
[FR Doc. 2012-17302 Filed 7-13-12; 8:45 am]
BILLING CODE 1505-01-D