The EN 14476:2025 represents one of the most significant updates in the field of chemical disinfectants and antiseptics. This European standard defines the quantitative suspension test method for assessing the virucidal activity of disinfectant products used in the medical and healthcare sectors.
The 2025 revision introduces several major modifications that directly impact testing laboratories and biocidal product manufacturers, improving the accuracy, reproducibility, and alignment of the method with other standards under the CEN/TC 216 framework.
In this article, we review the main updates, the technical requirements introduced, and their practical implications for laboratories and manufacturers. We also highlight the role of SHAPYPRO in supporting laboratories through proficiency testing (PT) and technical consultancy for the implementation of EN 14476:2025.
Context And Scope Of EN 14476
The EN 14476 standard applies to all chemical disinfectants and antiseptics that claim virucidal activity within the medical field.
It describes a laboratory-based method to evaluate whether a product is capable of effectively inactivating viruses under defined conditions that simulate practical use.
Areas of application:
- Hand and skin disinfection (alcohol-based hand rubs, surgical scrubs, antiseptic solutions).
- Surface and medical equipment disinfection (non-invasive instruments, hospital furniture).
- Clinical and diagnostic laboratory environments.
- Pharmaceutical, veterinary, and biotechnology production areas, where viral control is critical.
The goal is to ensure that disinfectant products achieve a measurable and reproducible reduction of infectious viral particles, safeguarding healthcare professionals, patients, and laboratory personnel.
EN 14476: Test Principle and Methodology
The EN 14476 method is based on a quantitative suspension test, in which a defined viral suspension is exposed to a specific concentration of the disinfectant for a set contact time under controlled conditions.
After exposure, the disinfectant’s activity is neutralized chemically, and the remaining viral infectivity is quantified by cell culture or viral titration methods.
The virucidal efficacy is expressed as a logarithmic reduction (log₁₀) in the viral titer compared to the untreated control, representing the product’s level of effectiveness.
Efficacy criteria:
A disinfectant is considered virucidal if it achieves a minimum reduction of 4 log₁₀ (corresponding to 99.99% viral inactivation) against the target virus under the defined conditions.
Reference Microorganisms And Test Viruses
The updated standard specifies a panel of mandatory and optional test viruses, representing both enveloped and non-enveloped viruses with varying resistance levels.
Mandatory test viruses:
- Poliovirus type 1 (Sabin strain) – highly resistant, non-enveloped virus.
- Adenovirus type 5 – moderately resistant, non-enveloped virus.
- Murine Norovirus (MNV) or Feline Calicivirus (FCV) – surrogate for human Norovirus.
Optional test viruses (depending on claimed activity):
- Vaccinia virus (Ankara strain) – model for enveloped viruses.
- Bovine viral diarrhea virus (BVDV) or Hepatitis C virus surrogate.
- Human Coronavirus 229E or Murine Hepatitis Virus (MHV) – representative of relevant enveloped viruses.
The choice of viruses depends on the claim made by the manufacturer:
- “Limited virucidal activity” – active against enveloped viruses.
- “Full virucidal activity” – active against both enveloped and non-enveloped viruses.
Major Updates Introduced In EN 14476:2025
The 2025 revision of EN 14476 introduces substantial changes to improve standardization, precision, and harmonization with related CEN standards.
- Alignment With Other CEN/TC 216 Standards
The revised text aligns EN 14476 with EN 14885:2023, the overarching framework defining relationships between disinfectant efficacy standards.
This harmonization ensures consistency between virucidal (EN 14476), bactericidal (EN 13727), and fungicidal (EN 13624) methods.
- Updated Test Conditions
The standard now includes refined test conditions that better reflect practical healthcare and laboratory environments:
- Contact time: 30 seconds to 60 minutes depending on use.
- Temperature: 20 °C to 40 °C.
- Interfering substances: updated to include bovine serum, albumin, and mucin to simulate clean and dirty conditions.
- Inclusion Of New Viral Strains
The Human Coronavirus 229E has been added as an optional surrogate virus for enveloped viruses, replacing MHV where appropriate, to enhance the clinical relevance of results.
- Improved Neutralization Validation
One of the most important updates is the explicit requirement to validate the neutralization process for each disinfectant tested, ensuring that the inactivation stops precisely at the defined time point.
- Standardized Reporting Format
The reporting section has been expanded and standardized, requiring detailed documentation of pH, contact time, concentration, neutralization control, and test virus used.
These updates enhance reproducibility and make EN 14476:2025 more robust for both regulatory and scientific use.
Relevance Of EN 14476 After The COVID-19 Pandemic
The COVID-19 pandemic significantly increased public and institutional awareness of virucidal efficacy. As a result, EN 14476 has become the gold standard for demonstrating antiviral performance of hand sanitizers, surface disinfectants, and antiseptic solutions.
Compliance with EN 14476 is now widely required by European accreditation bodies and health authorities, and it serves as a reference under the Biocidal Products Regulation (EU) No 528/2012 for product types PT1, PT2, and PT4 (human hygiene, surface disinfection, and food areas).
By harmonizing scientific rigor with practical use, EN 14476 ensures that disinfectants are effective against clinically relevant viruses and maintain high safety standards across the healthcare industry.
Implications For Laboratories And Manufacturers
For testing laboratories:
- Revalidate internal methods according to the updated time, temperature, and interference conditions.
- Verify neutralization procedures and ensure proper validation documentation.
- Maintain biosafety compliance for handling viral cultures.
For manufacturers:
- Review and update efficacy documentation, ensuring studies meet EN 14476:2025 requirements.
- Revise product labeling to specify “limited” or “full” virucidal activity claims.
- Collaborate with accredited laboratories applying the updated version of the standard.
Strict compliance with EN 14476:2025 not only strengthens regulatory submissions but also provides a credible market advantage in terms of verified efficacy.
Analytical Control And Reproducibility
The new edition emphasizes interlaboratory reproducibility and analytical validation as core quality criteria.
Laboratories must implement standardized controls, including:
- Verification of virus stability during testing.
- Positive and negative control runs.
- Validation of neutralizer performance for each product type.
At SHAPYPRO, we offer international proficiency testing programs (PT) for evaluating virucidal activity according to ISO/IEC 17043 principles.
Our PT programs allow laboratories to:
- Benchmark their results against other European centers.
- Detect deviations or inconsistencies in quantitative outcomes.
- Demonstrate technical competence and regulatory readiness.
Participation in these programs helps ensure that laboratories meet both EN 14476:2025 requirements and ISO accreditation standards.
Instrumental Analysis And Chemical Validation
In addition to microbiological verification, the chemical consistency of disinfectants plays a vital role in efficacy testing.
The updated standard encourages more rigorous chemical analysis of active ingredients such as alcohols, quaternary ammonium compounds, and peroxides, to confirm formulation stability and active concentration.
Advanced analytical tools, like the S2 PUMA X-ray fluorescence (XRF) system used at SHAPYPRO, enable quantitative analysis of elemental composition and trace contaminants, ensuring that the chemical integrity of disinfectants supports the microbiological results obtained under EN 14476.
This combination of instrumental precision and microbiological validation reinforces product reliability and compliance.
Conclusion: EN 14476 and Regulatory Alignment
The EN 14476:2025 standard significantly strengthens the scientific foundation for assessing virucidal activity in disinfectants, aligning the methodology with modern healthcare and biocidal requirements.
The updates improve method reproducibility, test realism, and harmonization with other antimicrobial standards, ensuring consistent and reliable evaluation across Europe.
For laboratories, this means reviewing and validating testing procedures in line with the revised protocol.
For manufacturers, it requires verifying product performance and updating claims to comply with the new regulatory framework.
At SHAPYPRO, we assist laboratories and producers in implementing the new EN 14476:2025 through technical consultancy, proficiency testing, and specialized training, helping ensure that results are traceable, comparable, and compliant with European standards.