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ASTM E3351-22

(Test Method)

Standard Test Method for Detection of Nitric Oxide Production In Vitro

Standard Test Method for Detection of Nitric Oxide Production <emph type="bdit">In Vitro</emph >

SIGNIFICANCE AND USE
5.1 This test method is designed to evaluate nanomaterial capacity to induce nitric oxide production by macrophages.  
5.2 Activated macrophages generate large quantities of NO. NO generated from activated macrophages is a cytostatic/cytotoxic agent (3-6).  
5.3 The production of NO in excessive amounts leads to the generation of peroxynitrite by its spontaneous reaction with superoxide. Peroxynitrite causes tissue injury through its capability to damage lipids, proteins, and DNA (2).  
5.4 NO is a proinflammatory mediator and it is an important marker for activation of inflammation (5, 6).  
5.5 Testing the capacity of a nanomaterial to induce NO production in vitro helps in predicting the nanomaterial’s biocompatibility through anticipating and understanding the potential problems that might be encountered during its in vivo administration.
SCOPE
1.1 This test method delivers a protocol for a quantitative measure of nitrite (NO2–), a stable end-product of nitric oxide (NO), in cell culture medium due to exposure to nanomaterial(s).  
1.2 NO has a critical role in several pathological conditions in addition to its role in many physiological processes.  
1.3 This test method uses murine macrophage cell line RAW 264.7 as an in vitro model.  
1.4 The nitrite is measured in the cell culture medium by a colorimetric analysis using Griess reagent as shown in Fig. 1.
FIG. 1 Summary of Nitric Oxide Production Assay  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

General Information

Status
Published
Publication Date
30-Jun-2022
ICS
11.100.10 - In vitro diagnostic test systems
Technical Committee
E56 - Nanotechnology
Drafting Committee
E56.08 - Nano-Enabled Medical Products
Current Stage
Ref Project

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ASTM E3351-22 - Standard Test Method for Detection of Nitric Oxide Production <emph type="bdit">In Vitro</emph >ASTM E3351-22 - Standard Test Method for Detection of Nitric Oxide Production <emph type="bdit">In Vitro</emph >
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ASTM E3351-22 - Standard Test Method for Detection of Nitric Oxide Production <emph type="bdit">In Vitro</emph >
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This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: E3351 − 22
Standard Test Method for
1
Detection of Nitric Oxide Production In Vitro
This standard is issued under the fixed designation E3351; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision.Anumber in parentheses indicates the year of last reapproval.A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 3. Terminology
1.1 This test method delivers a protocol for a quantitative 3.1 Definitions:
–
measure of nitrite (NO ), a stable end-product of nitric oxide 3.1.1 Cal—calibration standards
2
(NO), in cell culture medium due to exposure to nanomateri-
3.1.2 C —maximum serum concentration
max
al(s).
3.1.3 CV—coefficient of variation
1.2 NO has a critical role in several pathological conditions
3.1.4 DEA NONOate—diethylamine NONOate/AM
in addition to its role in many physiological processes.
3.1.5 DMSO—dimethyl sulfoxide
1.3 This test method uses murine macrophage cell line
3.1.6 FBS—fetal bovine serum
RAW 264.7 as an in vitro model.
3.1.7 Int.—intermediate
1.4 The nitrite is measured in the cell culture medium by a
colorimetric analysis using Griess reagent as shown in Fig. 1. 3.1.8 LPS—lipopolysaccharide
1.5 This standard does not purport to address all of the
3.1.9 PBS—phosphate buffered saline
safety concerns, if any, associated with its use. It is the
3.1.10 PDFT—percent difference from theoretical
responsibility of the user of this standard to establish appro-
3.1.11 RPMI—Roswell Park Memorial Institute
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use. 3.1.12 QC—quality control
1.6 This international standard was developed in accor-
3.1.13 SD—standard deviation
dance with internationally recognized principles on standard-
3.1.14 w/v—weight to volume ratio
ization established in the Decision on Principles for the
Development of International Standards, Guides and Recom-
4. Summary of Test Method
mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee. 4.1 This test method is used to assess the capability of
nanomaterials to induce nitric oxide production by macro-
2. Referenced Documents
phages in vitro(see Fig. 1).
2
2.1 ASTM Standards:
4.2 TheNOmoleculehasashorthalf-lifeandreactsquickly
E2490Guide for Measurement of Particle Size Distribution
with free oxygen, oxygen radicals, redox metals and even with
of Nanomaterials in Suspension by Photon Correlation
oxygenated hemoglobin to generate other reactive nitrogen
Spectroscopy (PCS) –
intermediates which decomposes to form nitrite (NO ) and
2
– 3
E2834Guide for Measurement of Particle Size Distribution
nitrate (NO ) (1). NO molecule can react with oxygenated
3
of Nanomaterials in Suspension by NanoparticleTracking –
hemoglobin to produce nitrate (NO ) (1, 2).
3
Analysis (NTA)
4.3 This test method describes a protocol for assessing and
F1877Practice for Characterization of Particles
measuring nitrite as a replacement marker and quantitative
F1903Practice for Testing for Cellular Responses to Par-
indicator of NO production.
ticles in vitro
4.4 In this test method, nitrite is measured in cell culture
medium using the Griess reagent.
1
This test method is under the jurisdiction of ASTM Committee E56 on
Nanotechnology and is the direct responsibility of Subcommittee E56.08 on
4.5 The upper limit of nitrite quantification is 250µM and
Nano-Enabled Medical Products.
the lower limit of quantification is 1.95µM.
Current edition approved July 1, 2022. Published July 2022. DOI: 10.1520/
E3351-22.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
3
Standards volume information, refer to the standard’s Document Summary page on Theboldfacenumbersinparenthesesrefertothelistofreferencesattheendof
the ASTM website. the standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
E3351 − 22
FIG. 1 Summary of Nitric Oxide Production Assay
5. Significance and Use 6. Materials
5.1 This test method is designed to evaluate nanomaterial
6.1 Pipettes covering the range of 0.05mL to 10mL.
capacity to induce nitric oxide production by macrophages.
6.2 Flat bottom 96-well plates.
5.2 Activated macrophages generate large quantities of NO.
6.3 24-well plates.
NO generated from activated macrophages is a cytostatic/
6.4 Polypropylene tubes, 5mL, and 15mL.
cytotoxic agent (3-6).
5.3 Theprod
...

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5.3 Information shall be evaluated in a flexible manner consistent with the needs of the authorizing program. This requires proper characterization of the current problem. For example, compounds may be ranked relative to the environmental criteria of the prospective alternatives, the replacement compound, and within bo...
SCOPE
1.1 This guide is intended to determine the relative environmental influence of new substances, consistent with the research and development (R&D) level of effort and is intended to be applied in a logical, tiered manner that parallels both the available funding and the stage of research, development, testing, and evaluation. Specifically, conservative assumptions, relationships, and models are recommended early in the research stage, and as the technology is matured, empirical data will be developed and used. Munition constituents are included and may include propellants, oxidizers, explosives, binders, stabilizers, metals, dyes, and other compounds used in the formulation to produce a desired effect. Munition systems range from projectiles, grenades, rockets/missiles, training simulators, to smokes and obscurants. Given the complexity of issues involved in the assessment of environmental fate and effects and the diversity of the systems used, this guide is broad in scope and not intended to address every factor that may be important in an environmental context. Rather, it is intended to reduce uncertainty at minimal cost by considering the most important factors related to human health and environmental impacts of energetic materials. This guide provides an outline for collecting data useful in a relative ranking procedure to provide the systems scientist with a sound basis for prospectively determining a selection of candidates based on environmental and human health criteria. The general principles in this guide are applicable to substances other than energetics if intended to be used in a similar manner with similar exposure profiles.  
1.2 The scope of this guide includes:  
1.2.1 Energetic and other new/novel materials and compositions in all stages of research, development, test and evaluation.  
1.2.2 Environmental assessment, including:
1.2.2.1 Human and ecological effects of the unexploded energetics and ...

  • Guide
    9 pages
    English language
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  • Guide
    9 pages
    English language
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ASTM
ASTM D8042-18(2023)(Test Method)
Test Method for Shrinkage Temperature of Wet Blue and Wet White

SIGNIFICANCE AND USE
5.1 This test method is designed to determine the temperature at which the Wet Blue or Wet White specimen experiences shrinkage. In this test method, shrinkage occurs as a result of hydrothermal denaturation of the collagen protein molecules which make up the fiber structure of the Wet Blue or Wet White. The shrinkage temperature of Wet Blue or Wet White is influenced by many different factors, most of which appear to affect the number and nature of crosslinking interactions between adjacent polypeptide chains of the collagen protein molecules. The value of the shrinkage temperature of Wet Blue or Wet White is commonly used as an indicator of the type of tannage or the degree of tannage, or both, of that particular Wet Blue or Wet White (especially for the more hydrothermally stable tannages such as chrome tannage).
SCOPE
1.1 This test method covers the determination of the shrinkage temperature of all types of Wet Blue and Wet White. The heating medium is water when the shrinkage temperature is at or below 98 °C. The heating medium is a glycerine-water solution when the shrinkage temperature is above 98 °C.  
1.2 The values stated in SI units are to be regarded as the standard. The values in parentheses are for information only.  
1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.4 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

  • Standard
    3 pages
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