ASTM F2382-18

Standard Test Method for Assessment of Circulating Blood-Contacting Medical Device Materials on Partial Thromboplastin Time (PTT)

SIGNIFICANCE AND USE
4.1 The purpose of this test method is to determine the time citrated plasma exposed to medical materials takes to form a clot when exposed to a suspension of phospholipid particles and calcium chloride. In this test method, the test article is the activator. The PTT assay is a general screening test for a medical material’s ability to activate the intrinsic coagulation pathway. Material samples that show a shortened PTT are activators of the intrinsic coagulation pathway.  
4.2 The test article, reference materials, and controls are exposed to human plasma. The plasma is tested on a coagulation device. Each sample tube is assayed in duplicate. The results are reported as a percentage of the negative control.
SCOPE
1.1 This test method covers the screening of circulating blood-contacting device materials for their ability to induce blood coagulation via the intrinsic coagulation pathway. This assay should be part of the hemocompatibility evaluation for devices and materials contacting human blood, as per ANSI/AAMI/ISO 10993-4.  
1.2 All safety policies and practices shall be observed during the performance of this test method.  
1.3 All plasma and any materials that had contact with plasma will be bagged in a biohazard bag, properly labelled with the contents, and disposed of by appropriate means. The plasma should be handled at the Biosafety Level 2 as recommended in the Centers for Disease Control/National Institutes of Health Manual Biosafety in Microbiological Laboratories.  
1.4 The normal pooled human plasma must have tested negative for Hepatitis B (HBV) or Human Immunodeficiency (HIV) viruses. The plasmas should be treated like any patient plasma using standard precautions. The plasma should be handled at the Biosafety Level 2 as recommended in the Centers for Disease Control/National Institutes of Health Manual Biosafety in Microbiological Laboratories.  
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, health, and environmental 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: 30-Sep-2018
• ICS: 11.100 - Laboratory medicine
• ICS: 11.100.30 - Analysis of blood and urine
• Technical Committee: F04 - Medical and Surgical Materials and Devices
• Drafting Committee: F04.16 - Biocompatibility Test Methods

Relations

• Replaces: ASTM F2382-17e1 - Standard Test Method for Assessment of Circulating Blood-Contacting Medical Device Materials on Partial Thromboplastin Time (PTT)
• Effective Date: 01-Oct-2018
• Referred By: ASTM E3219-20 - Standard Guide for Derivation of Health-Based Exposure Limits (HBELs)
• Effective Date: 01-Oct-2018
• Referred By: ASTM F748-16 - Standard Practice for Selecting Generic Biological Test Methods for Materials and Devices
• Effective Date: 01-Oct-2018
• Referred By: ASTM F3225-17(2022) - Standard Guide for Characterization and Assessment of Vascular Graft Tissue Engineered Medical Products (TEMPs)
• Effective Date: 01-Oct-2018
• Referred By: ASTM F2888-19 - Standard Practice for Platelet Leukocyte Count&#x2014;An <emph type="bdit">In-Vitro</emph > Measure for Hemocompatibility Assessment of Cardiovascular Materials
• Effective Date: 01-Oct-2018

Overview

ASTM F2382-18 is the Standard Test Method for the Assessment of Circulating Blood-Contacting Medical Device Materials on Partial Thromboplastin Time (PTT). Developed by ASTM International, this test method provides a framework for evaluating the blood compatibility of medical device materials, specifically by measuring their potential to trigger blood coagulation via the intrinsic pathway using the PTT assay. This assessment is critical for ensuring that materials intended for blood contact in medical devices do not induce adverse hematological reactions, such as thrombosis, making this standard integral to hemocompatibility evaluations for medical devices.

Key Topics

• Partial Thromboplastin Time (PTT) Assay: The PTT assay is used as a general screening test to determine if a material activates the intrinsic coagulation pathway. Materials that shorten the PTT are identified as activators of coagulation, indicating a potential risk for blood clot formation.

• Screening of Blood-Contacting Materials: The test is applied to medical device materials that come into direct contact with circulating blood, supporting risk assessment in line with biological evaluation requirements, such as those outlined in ANSI/AAMI/ISO 10993-4.

• Test Protocol and Result Interpretation:

  • Human plasma is exposed to the test material, then analyzed for clotting time using a coagulation analyzer.
  • Samples are tested in duplicate to ensure reliability and accuracy.
  • Results are reported as a percentage relative to a negative control, supporting comparison between test samples and reference materials.

• Safety and Biosafety Practices: Given the handling of human plasma, strict adherence to safety protocols, including the use of Biohazard Level 2 containment as per CDC/NIH guidelines, is mandated.

• Quality Control Measures: The method includes the use of positive and negative control materials to validate assay performance, replicate testing to ensure precision, and requirements for the proper handling and disposal of biological materials.

Applications

ASTM F2382-18 is widely applicable in the medical device industry, particularly in:

• Hemocompatibility Testing: It supports manufacturers and regulatory agencies in evaluating the thrombogenic potential of medical devices such as catheters, stents, vascular grafts, and extracorporeal circuits.
• Product Development and Regulatory Submissions: Results from this standard help demonstrate compliance with international regulations and risk management guidelines for blood-contacting devices, contributing data essential for product approval.
• Material Selection: The standard aids in the selection and validation of polymeric, metallic, and composite materials intended for use in devices that interface with blood.
• Benchmarking and Research: Researchers use this method to compare new materials or surface treatments for blood-contacting applications in a controlled, reproducible manner.

Related Standards

When performing hemocompatibility and blood coagulation assessments, the following standards and documents are relevant:

• ANSI/AAMI/ISO 10993-4 – Biological Evaluation of Medical Devices - Selection of Tests for Interactions with Blood
• Biosafety in Microbiological and Biomedical Laboratories (BMBL), 5th Edition, US DHHS – Guidance for safety practices in handling biological specimens

These standards, alongside ASTM F2382-18, provide comprehensive guidelines for ensuring the safety and effectiveness of medical device materials intended for use in blood contact. By adhering to these protocols, manufacturers support product safety, regulatory compliance, and the advancement of innovative blood-contacting medical devices.

Keywords: blood compatibility, partial thromboplastin time, PTT assay, blood-contacting medical device, hemocompatibility, biomedical device testing, ASTM F2382-18, intrinsic coagulation pathway, medical material testing.

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: F2382 − 18
Standard Test Method for
Assessment of Circulating Blood-Contacting Medical Device
Materials on Partial Thromboplastin Time (PTT)
This standard is issued under the fixed designation F2382; 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 mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
1.1 This test method covers the screening of circulating
blood-contacting device materials for their ability to induce
2. Referenced Documents
blood coagulation via the intrinsic coagulation pathway. This
assay should be part of the hemocompatibility evaluation for
2.1 ANSI/AAMI Standard:
devices and materials contacting human blood, as per ANSI/
ANSI/AAMI/ISO 10993-4 Biological Evaluation of Medical
AAMI/ISO 10993-4.
Devices—Part 4: Selection of Tests for Interactions with
Blood
1.2 All safety policies and practices shall be observed
during the performance of this test method.
2.2 Other Document:
U.S. Department of Health and Human Services Biosafety in
1.3 All plasma and any materials that had contact with
Microbiological and Biomedical Laboratories (BMBL),
plasma will be bagged in a biohazard bag, properly labelled
5th ed., 1999
with the contents, and disposed of by appropriate means. The
plasma should be handled at the Biosafety Level 2 as recom-
3. Terminology
mended in the Centers for Disease Control/National Institutes
of Health Manual Biosafety in Microbiological Laboratories.
3.1 Definitions of Terms Specific to This Standard:
3.1.1 activator—a medical material which demonstrates a
1.4 The normal pooled human plasma must have tested
shortened clotting time; an initiator of the intrinsic coagulation
negative for Hepatitis B (HBV) or Human Immunodeficiency
pathway.
(HIV) viruses. The plasmas should be treated like any patient
3.1.2 partial thromboplastin time (PTT) assay—a modifica-
plasma using standard precautions. The plasma should be
handled at the Biosafety Level 2 as recommended in the tion of the Activated Partial Thromboplastin Time (APTT)
Centers for Disease Control/National Institutes of Health assay; unlike the APTT test, the PTT assay uses a reagent
Manual Biosafety in Microbiological Laboratories. (rabbit brain cephalin) without activating substances such as
silica, kaolin, elagic acid. The material being tested acts as the
1.5 The values stated in SI units are to be regarded as
activator.
standard. No other units of measurement are included in this
3.1.3 read time—the time during which data is collected to
standard.
detect a clot.
1.6 This standard does not purport to address all of the
3.1.4 blank time—a period at the beginning of an assay
safety concerns, if any, associated with its use. It is the
when no data is taken. This is done to eliminate interference
responsibility of the user of this standard to establish appro-
from premixing reagents, bubbles, and so forth.
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
3.1.5 equilibration time—the time allowed for the plasma
1.7 This international standard was developed in accor-
samples to warm to 37°C. The coagulation analyzer can be set
dance with internationally recognized principles on standard-
to zero if samples are pre-warmed to this temperature.
ization established in the Decision on Principles for the
3.1.6 duplicate flag—the agreement between the results of
Development of International Standards, Guides and Recom-
duplicate samples in percent. For example, if set to “15,” the
difference between the two channels must be less than or equal
This test method is under the jurisdiction of ASTM Committee F04 on Medical
and Surgical Materials and Devices and is the direct responsibility of Subcommittee
F04.16 on Biocompatibility Test Methods. Available from American National Standards Institute (ANSI), 25 W. 43rd St.,
Current edition approved Oct. 1, 2018. Published October 2018. Originally 4th Floor, New York, NY 10036, http://www.ansi.org.
ɛ1 3
approved in 2004. Last previous edition approved in 2017 as F2382 – 17 . DOI: The BMBL 5th Edition (December 2009) is available from the Government
10.1520/F2382-18. Printing Office or https://www.cdc.gov/biosafety/publications/bmbl5/bmbl.pdf
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
F2382 − 18
to 15 %. If the variance in clot times exceeds this percentage, 8. Preparation of Apparatus
an asterisk “*” will be printed by the average results on the
8.1 Prepare each test article, the negative reference
report.
materials, marketed comparator device (if used) and controls in
triplicate. If a positive reference control material is used, a
4. Significance and Use
single replicate is acceptable. All samples are prepared based
4.1 The purpose of this test method is to determine the time on a ratio of either 4 or, preferably 6 cm of material to 1 mL
plasma and placed into polypropylene tubes. For device
citrated plasma exposed to medical materials takes to form a
clot when exposed to a suspension of phospholipid particles testing, if test sample quantity allows, use three separate
and calcium chloride. In this test method, the test article is the devices; otherwise, take three representative samples from one
device.
activator. The PTT assay is a general screening test for a
medical material’s ability to activate the intrinsic coagulation
8.2 Label duplicate polypropylene tubes and place in the ice
pathway. Material samples that show a shortened PTT are
bath.
activators of the intrinsic coagulation pathway.
8.3 Turn on the coagulation analyzer and allow it to warm
4.2 The test article, reference materials, and controls are
up to 37 6 2°C and equilibrate for at least 10 min.
exposed to human plasma. The plasma is tested on a coagula-
8.4 Program the analyzer to test under the APTT function
tion device. Each sample tube is assayed in duplicate. The
with an equilibration time of 60 s, activation time of 120 s, a
results are reported as a percentage of the negative control.
| blank time of 14 s, and a read time of 286 s.
5. Apparatus
8.5 Pre-warm analysis cuvettes (or cups, depending on
analyzer selected) at 37 6 2°C.
5.1 Polypropylene Test Tubes with Caps, 12 by 75 mm.
8.6 Pre-warm calcium chloride at 37 6 2°C.
5.2 Automatic Pipets and Tips, 100 and 1000 µL.
8.7 Rabbit Brain Cephalin (RBC) Preparation:
5.3 Ice Bath.
8.7.1 Allow the RBC to come to room temperature.
5.4 Coagulation Analyzer qualified, see A1.1.
8.7.2 Reconstitute the RBC as indicated by the RBC reagent
5.5 Agitating Water Bath, 37 6 2°C, capable of 60 rpm. manufacturer.
8.7.3 Place in an agitating water bath set at 37 6 2°C and 60
5.6 Coagulation Analyzer Cuvettes, or equivalent for spe-
rpm for 15 min to ensure complete rehydration of contents.
cific analyzer.
8.7.4 Vortex 15 s after rehydration is complete.
8.7.5 Place at 37 6 2°C.
6. Reagents and Materials
8.8 If using frozen blood plasma, quick thaw the plasma at
6.1 Calcium Chloride, 25 mM.
37 6 2°C and place on ice immediately.
6.2 Citrated Human Blood Plasma, fresh (less than 4 h from
draw) or freshly-frozen, maintained at minus 80°C, pooled.
9. Procedure
6.3 Lyophilized Rabbit Brain Cephalin (RBC).
9.1 The test material(s), reference material(s), marketed
comparator device (i
...

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.
´1
Designation: F2382 − 17 F2382 − 18
Standard Test Method for
Assessment of Circulating Blood-Contacting Medical Device
Materials on Partial Thromboplastin Time (PTT)
This standard is issued under the fixed designation F2382; 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.
ε NOTE—Corrected editorially in January 2018.
1. Scope
1.1 This test method covers the screening of circulating blood-contacting device materials for their ability to induce blood
coagulation via the intrinsic coagulation pathway. This assay should be part of the hemocompatibility evaluation for devices and
materials contacting human blood, as per ANSI/AAMI/ISO 10993-4.
1.2 All safety policies and practices shall be observed during the performance of this test method.
1.3 All plasma and any materials that had contact with plasma will be bagged in a biohazard bag, properly labelled with the
contents, and disposed of by appropriate means. The plasma should be handled at the Biosafety Level 2 as recommended in the
Centers for Disease Control/National Institutes of Health Manual Biosafety in Microbiological Laboratories.
1.4 The normal pooled human plasma must have tested negative for Hepatitis B (HBV) or Human Immunodeficiency (HIV)
viruses. The plasmas should be treated like any patient plasma using standard precautions. The plasma should be handled at the
Biosafety Level 2 as recommended in the Centers for Disease Control/National Institutes of Health Manual Biosafety in
Microbiological Laboratories.
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, health, and environmental 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
2.1 ANSI/AAMI Standard:
ANSI/AAMI/ISO 10993-4 Biological Evaluation of Medical Devices—Part 4: Selection of Tests for Interactions with Blood
2.2 Other Document:
U.S. Department of Health and Human Services Biosafety in Microbiological and Biomedical Laboratories (BMBL), 5th
ed., 1999
3. Terminology
3.1 Definitions of Terms Specific to This Standard:
3.1.1 activator—a medical material which demonstrates a shortened clotting time; an initiator of the intrinsic coagulation
pathway.
This test method is under the jurisdiction of ASTM Committee F04 on Medical and Surgical Materials and Devices and is the direct responsibility of Subcommittee
F04.16 on Biocompatibility Test Methods.
Current edition approved Sept. 1, 2017Oct. 1, 2018. Published September 2017October 2018. Originally approved in 2004. Last previous edition approved in 20102017
ɛ1
as F2382 – 04F2382 – 17 (2010). DOI: 10.1520/F2382-17E01.10.1520/F2382-18.
Available from American National Standards Institute (ANSI), 25 W. 43rd St., 4th Floor, New York, NY 10036, http://www.ansi.org.
The BMBL 5th Edition (December 2009) is available from the Government Printing Office or https://www.cdc.gov/biosafety/publications/bmbl5/bmbl.pdf
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
F2382 − 18
3.1.2 partial thromboplastin time (PTT) assay—a modification of the Activated Partial Thromboplastin Time (APTT) assay;
unlike the APTT test, the PTT assay uses a reagent (rabbit brain cephalin) without activating substances such as silica, kaolin,
elagic acid. The material being tested acts as the activator.
3.1.3 read time—the time during which data is collected to detect a clot.
3.1.4 blank time—a period at the beginning of an assay when no data is taken. This is done to eliminate interference from
premixing reagents, bubbles, and so forth.
3.1.5 equilibration time—the time allowed for the plasma samples to warm to 37°C. The coagulation analyzer can be set to zero
if samples are pre-warmed to this temperature.
3.1.6 duplicate flag—the agreement between the results of duplicate samples in percent. For example, if set to “15,” the
difference between the two channels must be less than or equal to 15 %. If the variance in clot times exceeds this percentage, an
asterisk “*” will be printed by the average results on the report.
4. Significance and Use
4.1 The purpose of this test method is to determine the time citrated plasma exposed to medical materials takes to form a clot
when exposed to a suspension of phospholipid particles and calcium chloride. In this test method, the test article is the activator.
The PTT assay is a general screening test for a medical material’s ability to activate the intrinsic coagulation pathway. Material
samples that show a shortened PTT are activators of the intrinsic coagulation pathway.
4.2 The test article, reference materials, and controls are exposed to human plasma. The plasma is tested on a coagulation
device. Each sample tube is assayed in duplicate. The results are reported as a percentage of the negative control.
5. Apparatus
5.1 Polypropylene Test Tubes with Caps, 12 by 75 mm.
5.2 Automatic Pipets and Tips, 100 and 1000 μL.
5.3 Ice Bath.
5.4 Coagulation Analyzer (Siemens BFTqualified, see A1.1II analyzer or other).
5.5 Agitating Water Bath, 37 6 2°C, capable of 60 rpm.
5.6 Coagulation Analyzer Cuvettes, or equivalent for specific analyzer.
6. Reagents and Materials
6.1 Calcium Chloride, 25 mM.
6.2 Citrated Human Blood Plasma, fresh (less than 4 h from draw) or freshly-frozen, maintained at minus 80°C, pooled.
6.3 Lyophilized Rabbit Brain Cephalin (RBC).
6.4 Positive Reference Material (Optional), see Appendix X1A1.2.
6.5 Positive Control, glass (Pasteur pipette tips or glass beads). Other qualified positive control materials such as
Buna-N-Rubber may be selected once they have demonstrated a consistent thrombogenic response.
6.6 Negative Reference Material (e.g. High Density Polyethylene, HDPE).
6.7 Marketed Comparator Device (Optional). A legally marketed, clinically acceptable device that has similar blood contact
nature and clinical use as the material/device being investigated.
NOTE 1—It may be helpful to use a positive reference control material (n=1) per assay to assure continuity between runs.
7. Hazards
7.1 The human blood plasma should be treated like any patient plasma using standard precautions. The plasma should be
handled at the Biosafety Level 2 as recommended in the US Department of Health and Human Services Biosafety in
Microbiological and Biomedical Laboratories.
8. Preparation of Apparatus
8.1 Prepare each test article, the negative reference materials, marketed comparator device (if used) and controls in triplicate.
If a positive reference control material is used, a single replicate is acceptable. All samples are prepared based on a ratio of either
4 or, preferably 6 cm of material to 1 mL plasma and placed into polypropylene tubes. For device testing, if test sample quantity
allows, use three separate devices; otherwise, take three representative samples from one device.
8.2 Label duplicate polypropylene tubes and place in the ice bath.
8.3 Turn on the coagulation analyzer and allow it to warm up to 37 6 2°C and equilibrate for at least 10 min.
F2382 − 18
8.4 Pr
...