ISO 10993-5
ISO 10993-5 is an international standard for laboratory testing of whether a material kills or damages mammalian cells grown in a dish. It was developed for medical devices and does not test chemical purity or chemical identity.
Also called: Biological evaluation of medical devices - Part 5: Tests for in vitro cytotoxicity, ISO 10993-5:2009, EN ISO 10993-5:2009+A11:2025, ANSI/AAMI/ISO 10993-5:2009/(R)2022
ISO 10993-5 is part of the ISO 10993 series. It is an in vitro cytotoxicity test, meaning a lab test outside the body that checks whether a material, or substances released from it, harms cultured cells. It is used as an early biological safety check before clinical use.
The ISO 10993-5 test procedure for cell viability
The ISO 10993-5 test usually starts with an extract from the test material. That extract is placed on mammalian cells, commonly L929 mouse fibroblasts, for 24 hours. The test checks whether anything leaching from the material damages living cell structures.
Under the standard, a material is considered non-cytotoxic if cell viability stays above 70%. If viability falls below that threshold, the material is classified as cytotoxic, which signals a possible biological risk.
Limitations and misinterpretations of ISO 10993-5 for peptides
- An 'ISO 10993-5 tested' claim on a peptide Certificate of Analysis (CoA) can be misleading because the standard was developed for medical devices, not research chemicals or peptides.
- ISO 10993-5 does not prove peptide identity, sequence, or chemical purity; it only shows whether the tested sample kills cells.
- A peptide can pass even if it is the wrong molecule or contains impurities, as long as those impurities are not acutely toxic to the cells.
- Peptide identity and purity require analytical methods such as Mass Spectrometry (MS) for sequence and HPLC for purity level.
- ISO 10993-5 is a biological safety standard, not laboratory accreditation such as ISO/IEC 17025; the FDA does not accept ISO/IEC 17025 as a replacement for Good Laboratory Practice (GLP).
- A 2023 interlaboratory study found that vague ISO 10993-5 specifications can lead to nearly a 50% chance of different labs reaching conflicting cytotoxicity conclusions for the same material.
Sources
- Toxic or not toxic? The specifications of the standard ISO 10993-5 are not explicit enough to yield comparable results in the cytotoxicity assessment of an identical medical device
- Use of International Standard ISO 10993-1, "Biological ...
- Recommendations for the generation, quantification, storage and handling of peptides used for mass spectrometry-based assays
- Liquid Chromatography-High Resolution Mass Spectrometry for Peptide Drug Quality Control - PMC
- ISO 10993-5 - شرکت توسعه کیفیت
- ISO 10993-5:2009 - Biological evaluation of medical devices
- ISO 10993-5: Cytotoxicity Testing of Medical Devices | Measurlabs
- ISO 10993-5:2009(en), Biological evaluation of medical devices — Part 5: Tests for in vitro cytotoxicity
- Cytotoxicity Testing Guide for Medical Device Manufacturers
- Biocompatibility assessment of MD (ISO/TR 10993-55:2023) - efor-group
- ANSI/AAMI/ISO 10993-5:2009/(R)2022 (PDF) | AAMI
- EVS-EN ISO 10993-5:2009+A11:2025 - EVS standard evs.ee | en
- Toxic or not toxic? Interlaboratory comparison reveals almost fifty-fifty chance in the cytotoxicity assessment (ISO 10993-5) of an identical medical device
- How To Read a Certificate of Analysis (CoA) for Peptide Research
- Peptide Certificates of Analysis: How to Read and Verify ...
- MTT Test - Eurofins Medical Device Testing
- Cytotoxicity testing - Dermatest
- Decoding Lab Reports: Peptide Verification Report Metrics