The EN 455-3:2023 standard is a cornerstone in the EN 455 series for single-use medical gloves. It defines the requirements and test methods for biological evaluation, ensuring that gloves are biocompatible, chemically safe, and suitable for medical use.
The 2023 revision modernizes the standard by introducing explicit obligations for chemical disclosure, testing of residual chemicals, and alignment with EN ISO 10993-1:2020, the globally recognized framework for biological evaluation within a risk-management process.
Overview
EN 455-3:2023 supersedes EN 455-3:2015 and introduces key structural and conceptual updates.
The revised edition now formally references EN ISO 10993-1:2020 and incorporates requirements consistent with the EU Medical Device Regulation (EU 2017/745).
Its purpose is to ensure that every glove placed on the market is assessed not only for physical integrity (as per EN 455-1 and -2) but also for chemical and biological safety throughout its lifecycle, from raw materials to sterilization and packaging.
Biological Evaluation and Chemical Safety
The foundation of EN 455-3:2023 is its reference to EN ISO 10993-1:2020, which mandates biological evaluation as part of an integrated risk-management process.
Manufacturers must identify and assess all potential biological risks that may arise from the materials used in the glove or from processing chemicals that remain as residues.
The standard requires the manufacturer to provide a comprehensive list of all chemical substances that are:
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intentionally added during manufacturing, or
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known to remain in the final product.
These include accelerators, antioxidants, biocides, and stabilizers that may pose health hazards such as Type IV allergic reactions or reproductive toxicity.
A prominent example is Wingstay L, a phenolic antioxidant that appears on the EU’s REACH Joint Submission list of reproductive toxicants.
Such substances must be declared and assessed as part of the glove’s biological safety evaluation, and manufacturers must demonstrate that exposure levels are either eliminated or controlled below acceptable limits.
Testing of Residual Chemicals
EN 455-3:2023 expands the scope of biological evaluation by linking it directly to quantitative testing of residual chemicals.
While the standard itself focuses on disclosure and general requirements, it refers to the EN ISO 10993 series for biological testing, especially:
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ISO 10993-5 (cytotoxicity),
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ISO 10993-10 (irritation and sensitization), and
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ISO 10993-17 and -18 (chemical characterization and allowable limits).
The 2023 revision is designed to be complemented by EN 455-5:2025, a newly introduced standard that defines specific analytical methods for measuring extractable residues under simulated-use conditions.
EN 455-5 uses high-performance liquid chromatography (HPLC) and other validated methods to quantify potentially allergenic or toxic substances such as thiurams, carbamates, thiazoles, and phenolic antioxidants.
Testing for residual chemicals provides objective evidence that medical gloves do not release harmful substances during use.
The results of these tests contribute directly to the biological safety assessment required under ISO 10993-1 and must be documented in the glove’s technical file.
Key Testing and Reporting Requirements
EN 455-3:2023 retains core test categories from the previous edition but clarifies thresholds, procedures, and documentation duties.
Endotoxins
Sterile gloves labeled as “low endotoxin” must comply with the European Pharmacopoeia 2.6.14 method and show an endotoxin content ≤ 20 EU per glove pair.
This is crucial for applications such as neurosurgery or implant procedures, where pyrogenic reactions pose serious clinical risks.
Powder Residues
Powder-free gloves must not exceed 2 mg of removable surface powder per glove, as measured according to EN ISO 21171:2006.
The standard reiterates that powdered gloves are discouraged due to well-documented risks of respiratory and wound complications.
Several countries, including the USA and Japan, have banned their medical use.
Leachable Proteins
For gloves containing natural rubber latex, manufacturers must monitor water-soluble extractable proteins using the Modified Lowry Method (Annex A of EN 455-3).
Results must remain below validated process limits, and supporting data must be available for inspection.
Annex B and C of the standard also describe immunological and chromatographic methods for further protein and amino-acid analysis.
Labelling Requirements
Glove labelling must conform to EN ISO 15223-1:2021 and clearly indicate:
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that the glove is for single use,
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if it contains natural rubber latex, and
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any known allergens or sensitizing chemicals used in production.
Where accelerator-free or chemical-reduced variants are marketed, the manufacturer must substantiate such claims through testing and risk assessment data.
Reporting and Documentation
All testing results must be compiled in a test report that identifies the applied methods, process limits, and test outcomes.
Any declaration that a chemical is “absent” must be supported by evidence that it has not been used at any stage of production and that no precursors capable of forming it are present.
Connection to EN 455-5:2025
EN 455-5:2025, “Extractable Chemical Residues,” is the natural extension of EN 455-3.
It introduces standardized methods for quantitative chemical analysis of gloves and defines procedures for:
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extraction under simulated-use conditions,
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chemical separation, and
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quantification of residual substances via HPLC or comparable analytical techniques.
Its annexes provide detailed guidance on measuring Type IV allergenic accelerators, antioxidants, and other extractables.
By adopting EN 455-5 testing as part of their ongoing risk management, manufacturers can substantiate claims of low residual content and ensure compliance with both ISO 10993 and MDR requirements for chemical safety.
Normative Framework
The biological evaluation of medical gloves now operates within a cohesive network of European and international standards:
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EN 455-1:2020 – Freedom from holes.
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EN 455-2:2020 – Physical properties.
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EN 455-3:2023 – Biological evaluation, requirements, and testing.
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EN 455-4:2009 – Shelf-life determination.
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EN 455-5:2025 – Extractable chemical residues.
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EN ISO 10993-1:2020 – Biological evaluation of medical devices within a risk management process.
Together, these standards ensure that mechanical integrity, chemical purity, and biological compatibility are assessed systematically across the glove’s design, manufacture, and shelf life.
Conclusion
The 2023 revision of EN 455-3 marks a major advance in the European regulatory landscape for medical gloves.
By aligning with EN ISO 10993-1 and integrating future testing from EN 455-5, the standard shifts the focus from simple compliance to comprehensive biological risk management.
Manufacturers are expected to:
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Identify and disclose all relevant chemical constituents,
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Evaluate their toxicological profiles,
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Quantify extractable residues, and
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Maintain traceable, validated evidence of conformity.
Ultimately, EN 455-3:2023 reinforces the industry’s commitment to safety, transparency, and scientific rigor, ensuring that every glove not only meets its functional purpose but also protects the health of those who wear and rely on it.