ASTM F3321-19

Standard Guide for Methods of Extraction of Test Soils for the Validation of Cleaning Methods for Reusable Medical Devices

SIGNIFICANCE AND USE
5.1 This guide may be used by medical device manufacturers as part of their design plan and implementation of the validation of the cleaning instructions of their reusable medical devices.  
5.2 This guide helps medical device manufacturers to identify the best method(s) for extracting simulated-use test soil (see Guide F3208), thereby evaluating whether the medical device can be adequately cleaned.  
5.3 Methods describing various techniques for extracting soil are given.  
5.4 Guidance is further given as to how to validate the method(s) for extraction.
SCOPE
1.1 This guide provides methods and considerations for extracting test soil(s) from reusable medical device(s) that occurs during simulated use validation, clinical use of the device(s) and after the device(s) have been through a cleaning process.  
1.2 This is a part of a series of ASTM guides for validating cleaning instructions. The scope of the first guide in the series is regarding selecting appropriate test soils (Guide F3208). The second in the series (Guide F3293) describes methods that are used to inoculate medical devices with simulated-use test soil(s). This third in the series describes methods for extracting test soils in order to measure residual soil remaining on medical devices after the performance of cleaning procedures.  
1.3 Units—The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.4 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.  
1.5 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-2019
• ICS: 11.080.99 - Other standards related to sterilization and disinfection
• Technical Committee: F04 - Medical and Surgical Materials and Devices
• Drafting Committee: F04.15 - Material Test Methods

Relations

• Refers: ASTM F3208-20 - Standard Guide for Selecting Test Soils for Validation of Cleaning Methods for Reusable Medical Devices
• Effective Date: 15-Jul-2020
• Refers: ASTM F3208-19 - Standard Guide for Selecting Test Soils for Validation of Cleaning Methods for Reusable Medical Devices
• Effective Date: 15-Nov-2019
• Refers: ASTM F3208-18 - Standard Guide for Selecting Test Soils for Validation of Cleaning Methods for Reusable Medical Devices
• Effective Date: 01-Oct-2018
• Refers: ASTM F3293-18 - Standard Guide for Application of Test Soils for the Validation of Cleaning Methods for Reusable Medical Devices
• Effective Date: 01-May-2018
• Refers: ASTM F2459-18 - Standard Test Method for Extracting Residue from Metallic Medical Components and Quantifying via Gravimetric Analysis
• Effective Date: 01-Feb-2018
• Refers: ASTM F3208-17 - Standard Guide for Selecting Test Soils for Validation of Cleaning Methods for Reusable Medical Devices
• Effective Date: 15-Jan-2017
• Refers: ASTM E1097-12 - Standard Guide for Determination of Various Elements by Direct Current Plasma Atomic Emission Spectrometry
• Effective Date: 01-Jun-2012
• Refers: ASTM F2459-12 - Standard Test Method for Extracting Residue from Metallic Medical Components and Quantifying via Gravimetric Analysis
• Effective Date: 01-Mar-2012
• Refers: ASTM F619-03(2008) - Standard Practice for Extraction of Medical Plastics
• Effective Date: 01-Aug-2008
• Refers: ASTM E2314-03(2008) - Standard Test Method for Determination of Effectiveness of Cleaning Processes for Reusable Medical Instruments Using a Microbiologic Method (Simulated Use Test)
• Effective Date: 01-May-2008
• Refers: ASTM E1097-07 - Standard Guide for Direct Current Plasma-Atomic Emission Spectrometry Analysis
• Effective Date: 01-Nov-2007
• Refers: ASTM E2520-07 - Standard Practice for Verifying Minimum Acceptable Performance of Trace Explosive Detectors
• Effective Date: 01-Feb-2007
• Refers: ASTM F2459-05 - Standard Test Method for Extracting Residue from Metallic Medical Components and Quantifying via Gravimetric Analysis
• Effective Date: 01-Aug-2005
• Refers: ASTM F619-03 - Standard Practice for Extraction of Medical Plastics
• Effective Date: 01-Nov-2003
• Refers: ASTM E1097-03 - Standard Guide for Direct Current Plasma Emission Spectrometry Analysis
• Effective Date: 01-Oct-2003

Overview

ASTM F3321-19: Standard Guide for Methods of Extraction of Test Soils for the Validation of Cleaning Methods for Reusable Medical Devices establishes best practices for extracting test soils from reusable medical devices as part of cleaning validation protocols. Medical device manufacturers and laboratories use this guide to select, perform, and validate extraction methods on devices that have gone through simulated use, actual clinical use, and post-cleaning processes. Proper extraction methods are crucial for evaluating whether residual soils, such as proteins or microbial contaminants, have been effectively removed, supporting patient safety and regulatory compliance.

Key Topics

• Extraction Methods: The standard details various validated procedures for removing test soil from medical devices, including flushing, swabbing, shaking, immersion, vortexing, sonication, Soxhlet extraction, brushing, and destructive testing. Each technique is suited for specific device designs and levels of soil adherence.
• Validation of Extraction: The guide provides criteria and steps for validating extraction efficiency, ensuring that the chosen method effectively recovers residual markers for accurate quantitative analysis.
• Controls and Replicates: Positive and negative device and sample controls are emphasized to support the robustness of validation studies. Guidance on determining the appropriate number of replicates is also included.
• Detection and Quantification: The document explains the concepts of limit of detection (LOD) and limit of quantification (LOQ), ensuring sensitivity and reliability of residual analysis post-cleaning.
• Reporting Requirements: Clear guidance is provided on documenting the extraction method, data supporting extraction efficacy, and the history of test articles used for validation.

Applications

• Reusable Medical Device Manufacturers: Use this standard during product development and risk management to validate cleaning instructions and ensure devices can be reliably cleaned between uses.
• Testing Laboratories: Apply the guide when assessing the cleanliness of reusable medical devices, supporting regulatory submissions or quality assurance programs.
• Regulatory Compliance: Following ASTM F3321-19 supports alignment with internationally recognized principles, including those set forth by the World Trade Organization Technical Barriers to Trade (TBT) Committee, and complementary FDA and ISO guidelines.
• Healthcare Facilities: Aid in evaluating cleaning processes for complex devices, ensuring effective reprocessing and minimizing risk of cross-contamination.

Related Standards

Adopting ASTM F3321-19 is often done in conjunction with other standards, such as:

• ASTM F3208: Guide for Selecting Test Soils for Validation of Cleaning Methods for Reusable Medical Devices.
• ASTM F3293: Guide for Application of Test Soils for the Validation of Cleaning Methods.
• ASTM F2459: Test Method for Extracting Residue from Metallic Medical Components and Quantifying via Gravimetric Analysis.
• ANSI/AAMI/ISO 15883-1: Washer-disinfectors – General requirements.
• AAMI TIR30: Compendium of processes, materials, test methods, and criteria for cleaning reusable medical devices.
• FDA Guidance for Industry and FDA Staff, “Processing/Reprocessing Medical Devices in Health Care Settings.”

Practical Value

Following ASTM F3321-19 ensures that cleaning validation for reusable medical devices is scientifically sound and internationally accepted. Proper implementation helps:

• Demonstrate device cleanability through validated, reproducible extraction methods.
• Document cleaning performance for regulatory submissions.
• Mitigate infection risk by ensuring residual soils and contaminants are effectively detected and quantified.
• Adapt protocols for a wide range of device types and test soils, supporting innovation and patient safety in a global marketplace.

By adhering to this standard, manufacturers and laboratories can ensure that their cleaning validation studies are thorough, reproducible, and meet current best practices for medical device cleanliness.

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: F3321 − 19
Standard Guide for
Methods of Extraction of Test Soils for the Validation of
Cleaning Methods for Reusable Medical Devices
This standard is issued under the fixed designation F3321; 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 Direct Current Plasma Atomic Emission Spectrometry
E2314 Test Method for Determination of Effectiveness of
1.1 This guide provides methods and considerations for
Cleaning Processes for Reusable Medical Instruments
extracting test soil(s) from reusable medical device(s) that
Using a Microbiologic Method (Simulated Use Test)
occurs during simulated use validation, clinical use of the
E2520 Practice for Measuring and Scoring Performance of
device(s) and after the device(s) have been through a cleaning
Trace Explosive Chemical Detectors
process.
F619 Practice for Extraction of Medical Plastics
1.2 This is a part of a series ofASTM guides for validating
F2459 Test Method for Extracting Residue from Metallic
cleaning instructions. The scope of the first guide in the series
Medical Components and Quantifying via Gravimetric
is regarding selecting appropriate test soils (Guide F3208).The
Analysis
second in the series (Guide F3293) describes methods that are
F3127 GuideforValidatingCleaningProcessesUsedDuring
used to inoculate medical devices with simulated-use test
the Manufacture of Medical Devices
soil(s).This third in the series describes methods for extracting
F3208 Guide for Selecting Test Soils for Validation of
test soils in order to measure residual soil remaining on
Cleaning Methods for Reusable Medical Devices
medical devices after the performance of cleaning procedures.
F3293 Guide forApplication of Test Soils for the Validation
of Cleaning Methods for Reusable Medical Devices
1.3 Units—The values stated in SI units are to be regarded
as standard. No other units of measurement are included in this 2.2 AAMI Documents:
AAMI TIR17 Compatibility of materials subject to steriliza-
standard.
tion
1.4 This standard does not purport to address all of the
AAMI TIR30 A compendium of processes, materials, test
safety concerns, if any, associated with its use. It is the
methods, and acceptance criteria for cleaning reusable
responsibility of the user of this standard to establish appro-
medical devices
priate safety, health, and environmental practices and deter-
ANSI/AAMI/ISO 15883-1 Washer-disinfectors – Part 1:
mine the applicability of regulatory limitations prior to use.
General requirements, terms and definitions and tests
1.5 This international standard was developed in accor-
2.3 FDA Document:
dance with internationally recognized principles on standard-
FDA Guidance for Industry and FDA Staff, Processing/
ization established in the Decision on Principles for the
Reprocessing Medical Devices in Health Care Settings:
Development of International Standards, Guides and Recom-
Validation Methods and Labeling, June 6, 2017
mendations issued by the World Trade Organization Technical
2.4 Other Documents:
Barriers to Trade (TBT) Committee.
EPA, SW-846 Test Method 3540C Soxhlet Extraction
Evotech® endoscope cleaner and reprocessor (ECR) simu-
2. Referenced Documents
lated use and clinical use evaluation of cleaning efficacy,
2.1 ASTM Standards:
BMC Infectious Diseases 201010:200, Alfa, M.J.,
E1097 Guide for Determination of Various Elements by
DeGagne, P., Olson, N, Fatima, I . 2010
3. Terminology
This guide is under the jurisdiction of ASTM Committee F04 on Medical and
3.1 Definitions:
Surgical Materials and Devices and is the direct responsibility of Subcommittee
F04.15 on Material Test Methods.
Current edition approved Oct. 1, 2019. Published October 2019. DOI: 10.1520/ Available from Association for the Advancement of Medical Instrumentation
F3321-19. (AAMI), 4301 N. Fairfax Dr., Suite 301, Arlington, VA 22203-1633, http://
For referenced ASTM standards, visit the ASTM website, www.astm.org, or www.aami.org.
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM https://www.fda.gov/downloads/medicaldevices/deviceregulationandguidance/
Standards volume information, refer to the standard’s Document Summary page on guidancedocuments/ucm253010.pdf
the ASTM website. https://www.epa.gov/hw-sw846/sw-846-test-method-3540c-soxhlet-extraction
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
F3321 − 19
3.1.1 extraction, n—procedure for sampling a medical de- exhaustive extraction is that a significant fraction of soil may
vice to recover residuals that will then be used for quantitative still remain even after exhaustive recovery ceases to yield
testing for cleaning markers such as protein, hemoglobin, etc. measurableresults.Stepsshouldbetakentodetermineifthisis
indeed the case. The inspection or testing used to verify that
3.1.2 limit of detection (LOD), n—lowest quantity of a
soil is removed by extraction should be capable of detecting
substance that can be distinguished from the absence of that
very low surface concentrations of soil remaining at levels that
substance within a stated confidence limit (Practice E2520-15).
would significantly change the extraction efficiency calcula-
LOD is also generally defined as 3 times the standard deviation
tions. This could be a step during validation of the extraction
of the blank (Guide F3127-16).
method (Section 6.3).
3.1.3 limit of quantification (LOQ), n—lowest concentration
6.1.2 Establish any potential interference effects between
at which an instrument can measure reliably with a defined
the extraction media and the test methods, to ensure that the
error and confidence level (Guide E1097-17). LOQ is also
extraction method accurately represents the measurement of
generally defined as 10 times the standard deviation of the
what is being tested. The correct use of extraction volumes
blank (Guide F3127-16).
shouldbeassessedaspartofthevalidationtoavoiddilutingthe
3.1.4 test soil, n—single substance or a mixture of sub-
sample and reporting inaccurate results.The extraction volume
stances that reflect the contaminants likely to be encountered
used to remove the test soil for the test method used to detect
during the use of the device for its intended clinical procedure
the test marker should not be compromised as this could lead
(Guide F3208).
to a false negative (FDA, Reprocessing, 2017, Section A.3.c).
6.2 For all testing, choose a justifiable number of replicate
4. Summary of Guide
samples to support the validity of any instructions based on the
4.1 This guide describes techniques for extracting residuals
tests being performed (FDA, Reprocessing, 2017 Section VIII
from soiled medical devices during validation testing of the
Part A). Consider performing a preliminary experiment to
instructions for medical device reprocessing by a healthcare
establish the number of test articles needed to support the
facility.
validation of cleaning instructions.
4.2 This guide describes method(s) for validating the ex-
6.2.1 There are two types of positive controls that are
traction method, including use of positive and negative con-
performed during a cleaning validation, one that uses a
trols.
contaminated test article to determine the baseline for the test
articles before processing, referred to as a positive device
4.3 This guide also describes methods for extracting
control. The other control is one that serves to assure that the
samples from clinically used medical devices as a means of
chemical assays used to evaluate the cleanliness of the test
determining the clinical relevance of the simulated-use testing.
articles are capable of detecting trace amounts of the test soils
5. Significance and Use
used for the validation, referred as a positive sample control.
The positive device control utilizes an unprocessed contami-
5.1 This guide may be used by medical device manufactur-
nated test article that is extracted and tested for residuals. To
ers as part of their design plan and implementation of the
conductthepositivesamplecontrol,theamountofsoilmarkers
validation of the cleaning instructions of their reusable medical
in the test soil should be determined. The positive sample
devices.
control should then be spiked with a small, known amount of
5.2 This guide helps medical device manufacturers to iden-
test soil and undergo the same extraction conditions as the test
tify the best method(s) for extracting simulated-use test soil
samples. This control assesses whether extraction conditions
(see Guide F3208), thereby evaluating whether the medical
(e.g., duration of testing, temperature, plastic ware) causes a
device can be adequately cleaned.
decrease in the amount of measured soil in the sample. If the
5.3 Methods describing various techniques for extracting
amount of soil measured in the positive sample control is
soil are given. significantly decreased relative to the amount of soil that was
added, consider revising the extraction conditions to minimize
5.4 Guidance is further given as to how to validate the
sample loss. For validation, the positive control is extracted
method(s) for extraction.
multiple times through an exhaustive extraction to establish an
extraction efficiency.
6. General Considerations
6.2.2 It is recommended to perform at least one (1) positive
6.1 Medicaldevicemanufacturers,aspartoftheirvalidation
control for a study. Multiple positive controls can be tested if
testing, need to:
testing occurs over multiple days. However, it is not necessary
6.1.1 Use a validated extraction method(s). Extraction
to perform an exhaustive extraction every day of the study.
method(s) that are effective at removing constitutes of the test
6.2.3 Test articles are contaminated, cleaned, and subjected
soil, including key markers (protein, etc.) to be measured, need
to only one extraction. Exhaustive extraction is not necessary
to be validated. Exhaustive extraction or equivalent methods
for cleaned devices as levels of residuals would be expected to
are to be used to validate the efficiency of the extraction
be below or at the LOD of the residual being evaluated.
method.Ensurethattheextractionmethod(s)usedtodetermine
the residuals after the cleaning process will allow for a 6.3 The medical device manufacturer must validate the
quantitative method to be used for measuring residual soil extraction method, (FDAReprocessing, 2017, Section VIII.A.
(with the chosen marker(s) to be us
...