ASTM F756-17

Standard Practice for Assessment of Hemolytic Properties of Materials

SIGNIFICANCE AND USE
5.1 The presence of hemolytic material in contact with the blood may cause loss of, or damage to, red blood cells and may produce increased levels of free plasma hemoglobin capable of inducing toxic effects or other effects which may stress the kidneys or other organs.  
5.2 This practice may not be predictive of events occurring during all types of implant applications. The user is cautioned to consider the appropriateness of the method in view of the materials being tested, their potential applications, and the recommendations contained in Practice F748.
SCOPE
1.1 This practice provides a protocol for the assessment of hemolytic properties of materials used in the fabrication of medical devices that will contact blood.  
1.2 This practice is intended to evaluate the acute in vitro hemolytic properties of materials intended for use in contact with blood.  
1.3 This practice consists of a protocol for a hemolysis test under static conditions with either an extract of the material or direct contact of the material with blood. It is recommended that both tests (extract and direct contact) be performed unless the material application or contact time justifies the exclusion of one of the tests.  
1.4 This practice is one of several developed for the assessment of the biocompatibility of materials. Practice F748 may provide guidance for the selection of appropriate methods for testing materials for a specific application. Test Method E2524 provides a protocol using reduced test volumes to assess the hemolytic properties of blood-contacting nanoparticulate materials; this may include nanoparticles that become unbound from material surfaces.  
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.  
1.7 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: 28-Feb-2017
• ICS: 11.100 - Laboratory medicine
• Technical Committee: F04 - Medical and Surgical Materials and Devices
• Drafting Committee: F04.16 - Biocompatibility Test Methods

Relations

• Refers: ASTM F748-16 - Standard Practice for Selecting Generic Biological Test Methods for Materials and Devices
• Effective Date: 01-Apr-2016
• Referred By: ASTM F2901-19 - Standard Guide for Selecting Tests to Evaluate Potential Neurotoxicity of Medical Devices
• Effective Date: 01-Mar-2017
• Referred By: ASTM F2347-15 - Standard Guide for Characterization and Testing of Hyaluronan as Starting Materials Intended for Use in Biomedical and Tissue Engineered Medical Product Applications
• Effective Date: 01-Mar-2017
• Referred By: ASTM E2524-22 - Standard Test Method for Analysis of Hemolytic Properties of Nanoparticles
• Effective Date: 01-Mar-2017
• Referred By: ASTM F2064-17 - Standard Guide for Characterization and Testing of Alginates as Starting Materials Intended for Use in Biomedical and Tissue Engineered Medical Product Applications
• Effective Date: 01-Mar-2017
• Referred By: ASTM E3219-20 - Standard Guide for Derivation of Health-Based Exposure Limits (HBELs)
• Effective Date: 01-Mar-2017
• Referred By: ASTM F2224-09(2020) - Standard Specification for High Purity Calcium Sulfate Hemihydrate or Dihydrate for Surgical Implants
• Effective Date: 01-Mar-2017
• Referred By: ASTM F3089-23 - Standard Guide for Characterization and Standardization of Polymerizable Collagen-Based Products and Associated Collagen-Cell Interactions
• Effective Date: 01-Mar-2017
• Referred By: ASTM F2848-21 - Standard Specification for Medical-Grade Ultra-High-Molecular-Weight Polyethylene Yarns
• Effective Date: 01-Mar-2017
• Referred By: ASTM F2212-20 - Standard Guide for Characterization of Type I Collagen as Starting Material for Surgical Implants and Substrates for Tissue Engineered Medical Products (TEMPs)
• Effective Date: 01-Mar-2017
• Referred By: ASTM F451-21 - Standard Specification for Acrylic Bone Cement
• Effective Date: 01-Mar-2017
• Referred By: ASTM F648-21 - Standard Specification for Ultra-High-Molecular-Weight Polyethylene Powder and Fabricated Form for Surgical Implants
• Effective Date: 01-Mar-2017
• Referred By: ASTM F2103-18 - Standard Guide for Characterization and Testing of Chitosan Salts as Starting Materials Intended for Use in Biomedical and Tissue-Engineered Medical Product Applications
• Effective Date: 01-Mar-2017
• Referred By: ASTM F2695-12(2020) - Standard Specification for Ultra-High Molecular Weight Polyethylene Powder Blended With Alpha-Tocopherol (Vitamin E) and Fabricated Forms for Surgical Implant Applications
• Effective Date: 01-Mar-2017
• Referred By: ASTM F2759-19 - Standard Guide for Assessment of the Ultra-High Molecular Weight Polyethylene (UHMWPE) Used in Orthopedic and Spinal Devices
• Effective Date: 01-Mar-2017
• Referred By: ASTM F2565-21 - Standard Guide for Extensively Irradiation-Crosslinked Ultra-High Molecular Weight Polyethylene Fabricated Forms for Surgical Implant Applications
• Effective Date: 01-Mar-2017

Standards Content (Sample)

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: F756 − 17
Standard Practice for
1
Assessment of Hemolytic Properties of Materials
ThisstandardisissuedunderthefixeddesignationF756;thenumberimmediatelyfollowingthedesignationindicatestheyearoforiginal
adoptionor,inthecaseofrevision,theyearoflastrevision.Anumberinparenthesesindicatestheyearoflastreapproval.Asuperscript
epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 2. Referenced Documents
2
1.1 This practice provides a protocol for the assessment of 2.1 ASTM Standards:
hemolytic properties of materials used in the fabrication of E691Practice for Conducting an Interlaboratory Study to
medical devices that will contact blood. Determine the Precision of a Test Method
E2524Test Method forAnalysis of Hemolytic Properties of
1.2 This practice is intended to evaluate the acute in vitro
Nanoparticles
hemolytic properties of materials intended for use in contact
F619Practice for Extraction of Medical Plastics
with blood.
F748PracticeforSelectingGenericBiologicalTestMethods
1.3 This practice consists of a protocol for a hemolysis test
for Materials and Devices
under static conditions with either an extract of the material or
direct contact of the material with blood. It is recommended
3. Terminology
that both tests (extract and direct contact) be performed unless
3.1 Definitions of Terms Specific to This Standard:
the material application or contact time justifies the exclusion
3.1.1 plasma hemoglobin—amount of hemoglobin in the
of one of the tests.
plasma.
1.4 This practice is one of several developed for the
3.1.2 % hemolysis—free plasma hemoglobin concentration
assessment of the biocompatibility of materials. Practice F748
(mg/mL) divided by the total hemoglobin concentration (mg/
may provide guidance for the selection of appropriate methods
mL) present multiplied by 100. This is synonymous with
for testing materials for a specific application. Test Method
hemolytic index.
E2524providesaprotocolusingreducedtestvolumestoassess
3.1.3 comparative hemolysis—comparison of the hemolytic
the hemolytic properties of blood-contacting nanoparticulate
index produced by a test material with that produced by a
materials;thismayincludenanoparticlesthatbecomeunbound
standard reference material such as polyethylene under the
from material surfaces.
same test conditions.
1.5 The values stated in SI units are to be regarded as
3.1.4 direct contact test—testforhemolysisperformedwith
standard. No other units of measurement are included in this
the test material in direct contact with the blood.
standard.
3.1.5 extract test—test for hemolysis performed with an
1.6 This standard does not purport to address all of the
isotonic extract of the test material in contact with blood, as
safety concerns, if any, associated with its use. It is the
described in Practice F619.
responsibility of the user of this standard to establish appro-
3.1.6 hemolysis—destructionoferythrocytesresultinginthe
priate safety and health practices and determine the applica-
liberation of hemoglobin into the plasma or suspension me-
bility of regulatory limitations prior to use.
dium.
1.7 This international standard was developed in accor-
dance with internationally recognized principles on standard- 3.1.7 negative control—material, such as polyethylene, that
ization established in the Decision on Principles for the produces little or no hemolysis (<2 % after subtraction of the
Development of International Standards, Guides and Recom-
blank) in the test procedure. It is desirable that the control
mendations issued by the World Trade Organization Technical specimens have the same configuration as the test samples.
Barriers to Trade (TBT) Committee.
3.1.8 positive control—materials capable of consistently
producing a hemolytic index (above the negative control) of at
least 5% (see 10.3). Although positive control materials have
1
ThispracticeisunderthejurisdictionofASTMCommitteeF04onMedicaland
Surgical Materials and Devicesand is the direct responsibility of Subcommittee
2
F04.16 on Biocompatibility Test Methods. For referenced ASTM standards, visit the ASTM website, www.astm.org, or
Current edition approved March 1, 2017. Published April 2017. Originally contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
approved in 1982. Last previous edition approved in 2013 as F756–13. DOI: Standards volume information, refer to the standard’s Document Summary page on
10.1520/F0756-17. the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

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F756 − 17
not been
...

This document is not an ASTM standard and is intended only to provide the user of an ASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation: F756 − 13 F756 − 17
Standard Practice for
1
Assessment of Hemolytic Properties of Materials
This standard is issued under the fixed designation F756; 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
1.1 This practice provides a protocol for the assessment of hemolytic properties of materials used in the fabrication of medical
devices that will contact blood.
1.2 This practice is intended to evaluate the acute in vitro hemolytic properties of materials intended for use in contact with
blood.
1.3 This practice consists of a protocol for a hemolysis test under static conditions with either an extract of the material or direct
contact of the material with blood. It is recommended that both tests (extract and direct contact) be performed unless the material
application or contact time justifies the exclusion of one of the tests.
1.4 This practice is one of several developed for the assessment of the biocompatibility of materials. Practice F748 may provide
guidance for the selection of appropriate methods for testing materials for a specific application. Test Method E2524 provides a
protocol using reduced test volumes to assess the hemolytic properties of blood-contacting nanoparticulate materials; this may
include nanoparticles that become unbound from material surfaces.
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.
1.7 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.
2. Referenced Documents
2
2.1 ASTM Standards:
E691 Practice for Conducting an Interlaboratory Study to Determine the Precision of a Test Method
E2524 Test Method for Analysis of Hemolytic Properties of Nanoparticles
F619 Practice for Extraction of Medical Plastics
F748 Practice for Selecting Generic Biological Test Methods for Materials and Devices
3. Terminology
3.1 Definitions:Definitions of Terms Specific to This Standard:
3.1.1 plasma hemoglobin—amount of hemoglobin in the plasma.
3.1.2 % hemolysis—free plasma hemoglobin concentration (mg/mL) divided by the total hemoglobin concentration (mg/mL)
present multiplied by 100. This is synonymous with hemolytic index.
3.1.3 comparative hemolysis—comparison of the hemolytic index produced by a test material with that produced by a standard
reference material such as polyethylene under the same test conditions.
3.1.4 direct contact test—test for hemolysis performed with the test material in direct contact with the blood.
1
This practice is under the jurisdiction of ASTM Committee F04 on Medical and Surgical Materials and Devicesand is the direct responsibility of Subcommittee F04.16
on Biocompatibility Test Methods.
Current edition approved Dec. 1, 2013March 1, 2017. Published January 2014April 2017. Originally approved in 1982. Last previous edition approved in 20082013 as
F756 – 08.F756 – 13. DOI: 10.1520/F0756-13.10.1520/F0756-17.
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1

---------------------- Page: 1 ----------------------
F756 − 17
3.1.5 extract test—test for hemolysis performed with an isotonic extract of the test material, material in contact with blood, as
described in Practice F619, in contact with the blood.
3.1.6 hemolysis—destruction of erythrocytes resulting in the liberation of hemoglobin into the plasma or suspension medium.
3.1.7 negative control—material, such as polyethylene, that produces little or no hemolysis (<2 % after subtraction of t
...