ASTM E2524-08(2013)

Standard Test Method for Analysis of Hemolytic Properties of Nanoparticles (Withdrawn 2022)

SIGNIFICANCE AND USE
5.1 This test method is one of a series of tests listed in Practice F748 and ISO 10993-4 to assess the biocompatibility of materials contacting blood in medical applications.  
5.2 This test method is similar to Practice F756 but modified to accommodate nanoparticulate materials.
SCOPE
1.1 This test method covers assessing the effect of nanoparticulate materials on the integrity of red blood cells.  
1.2 This test method uses diluted whole blood incubated with nanoparticulate material and the hemoglobin released from damaged red blood cells is determined.  
1.3 This test method is similar to Practice F756 with the volumes reduced to accommodate nanoparticulate material.  
1.4 This test method is part of the in-vitro preclinical characterization and is important for nanoparticulate material that will contact the blood in medical applications.  
1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.6 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 and health practices and determine the applicability of regulatory limitations prior to use.

General Information

• Status: Historical
• Publication Date: 31-Aug-2013
• ICS: 11.040.20 - Transfusion, infusion and injection equipment
• ICS: 71.100.01 - Products of the chemical industry in general
• Technical Committee: E56 - Nanotechnology
• Drafting Committee: E56.03 - Environment, Health, and Safety

Standards Content (Sample)

NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation: E2524 − 08 (Reapproved 2013)
Standard Test Method for
1
Analysis of Hemolytic Properties of Nanoparticles
This standard is issued under the fixed designation E2524; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number 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 covers assessing the effect of nanopar-
3.1 Acronyms:
ticulate materials on the integrity of red blood cells. 3.1.1 Cal—calibration standard
3.1.2 CMH—cyanmethemoglobin
1.2 This test method uses diluted whole blood incubated
with nanoparticulate material and the hemoglobin released
3.1.3 DPBS—Dulbecco’s phosphate-buffered saline
from damaged red blood cells is determined.
3.1.4 PEG—polyethylene glycol
1.3 This test method is similar to Practice F756 with the
3.1.5 PFH—plasma-free hemoglobin
volumes reduced to accommodate nanoparticulate material.
3.1.6 QC—quality controls
1.4 This test method is part of the in-vitro preclinical
3.1.7 TBH—total blood hemoglobin
characterization and is important for nanoparticulate material
3.1.8 TBHd—blood sample diluted to 10 mg 6 1 mg/mL
that will contact the blood in medical applications.
1.5 The values stated in SI units are to be regarded as
4. Summary of Test Method
standard. No other units of measurement are included in this
4.1 Thistestmethoddescribesaprotocolforassessingacute
standard.
in-vitro damage to red blood cells (that is, hemolysis) caused
1.6 This standard does not purport to address all of the
by exposure to nanoparticles.
safety concerns, if any, associated with its use. It is the
4.2 This test method is based on the quantitative determi-
responsibility of the user of this standard to establish appro-
nation of hemoglobin released into PFH as a percentage of the
priate safety and health practices and determine the applica-
TBH concentration when blood is exposed to nanoparticulate
bility of regulatory limitations prior to use.
materials.
2. Referenced Documents 4
4.3 Using an established colorimetric assay, hemoglobin
2
2.1 ASTM Standards:
and its derivatives, such as sulfhemoglobin, are oxidized to
F748 PracticeforSelectingGenericBiologicalTestMethods
methemoglobin by ferricyanide in the presence of alkali. A
for Materials and Devices
stable CMH concentration is measured using a plate reader
F756 Practice for Assessment of Hemolytic Properties of
spectrophotometer set at 540 nm.
Materials
4.4 Hemoglobin standards are used to create a standard
F1877 Practice for Characterization of Particles
curve covering the range from 0.025 to 0.8 mg/mLand prepare
F1903 Practice for Testing For Biological Responses to
quality control samples at low (0.0625-mg/mL), mid (0.125-
Particles In Vitro
mg/mL), and high (0.625-mg/mL) concentrations to monitor
3
2.2 ISO Standard:
assay performance. The required sample volume is 100 µLper
ISO 10993-4 Biological Evaluation of Medical Devices Part
test replicate.
4: Selection of Tests for Interactions with Blood
4.5 The results are expressed as percent hemolysis to
1 evaluate the acute in-vitro hemolytic properties of nanopar-
This test method is under the jurisdiction of ASTM Committee E56 on
Nanotechnology and is the direct responsibility of Subcommittee E56.03 on ticles.
Environment, Health, and Safety.
Current edition approved Sept. 1, 2013. Published September 2013. Originally
5. Significance and Use
approved in 2008. Last previous edition approved in 2008 as E2524 – 08. DOI:
5.1 This test method is one of a series of tests listed in
10.1520/E2524-08R13.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or Practice F748 and ISO 10993-4 to assess the biocompatibility
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
of materials contacting blood in medical applications.
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website.
3 4
Available fromAmerican National Standards Institute (ANSI), 25 W. 43rd St., International Committee for Standardization in Haemotology, Journal of
4th Floor, New York, NY 10036, http://www.ansi.org. Clinical Pathology, Vol 31, 1978, pp. 139–143.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
E2524 − 08 (2013)
TABLE 2 Quality Controls
5.2 ThistestmethodissimilartoPracticeF756butmodified
to accommodate nanoparticulate materials. Nominal
Level Conc., Preparation Procedure
mg/mL
6. Apparatus
QC 1 0.625 1.5 mL of stock solution + 0.42 mL of CMH reagent
6.1 Pipettes covering range from 0.05 to 10 mL.
QC 2 0.125 200 µL of QC1
...