Hand hygiene is a key pillar in preventing cross-contamination, both in healthcare and in the food, cosmetics, and pharmaceutical industries. In this context, the EN 1499 and EN 1500 standards define the requirements and methodologies to evaluate the effectiveness of hand hygiene products. This article offers a comprehensive guide to both standards, highlighting their key differences, the importance of dilute soft soap as a reference product, and their application in proficiency testing programs.
EN 1499 and EN 1500: What Are They and What Are They For?
The EN 1499 and EN 1500 standards are harmonized European documents that establish the methods for evaluating the antimicrobial activity of hand hygiene products. While both aim to assess disinfection efficacy, they apply in different contexts:
- EN 1499: Evaluates the effectiveness of liquid or solid soaps intended for hygienic handwashing with water. It applies to products that remove microorganisms through mechanical action.
- EN 1500: Measures the effectiveness of alcohol-based hand rubs used for hygienic hand disinfection without water. It is the reference standard for alcohol-based products.
Both standards define a comparative test between a product under test and a reference product (dilute soft soap), using human volunteers and a non-pathogenic bacterial strain.
The Role of Escherichia coli K12 in the Tests
Both EN 1499 and EN 1500 use the strain Escherichia coli K12 as the test organism, a safe and representative bacterium for contamination simulations.
This strain is harmless to volunteers but allows realistic contamination conditions on hands to be simulated, enabling evaluation of the logarithmic reduction after washing or rubbing.
Proficiency or interlaboratory tests, such as those organized by SHAPYPRO, use E. coli K12 to evaluate the technical competence of laboratories applying EN 1499 and EN 1500.
Dilute Soft Soap: The Reference Product
Both standards state that the reference product for the tests is dilute soft soap (Potassium Linseed oil).
According to section 5.2.2.6 of both standards:
“The soft soap shall be prepared by diluting the reference soft soap (potassium type) to 20% (w/w) just before use.”
This product must meet the specifications outlined in Annex D of each standard, detailing the physicochemical characteristics, pH requirements, viscosity, and purity of the soap.
At SHAPYPRO, we offer a standardized dilute soft soap ready for use in tests according to EN 1499 and EN 1500, manufactured under strict quality controls.
Key Differences Between EN 1499 and EN 1500
Although both standards share the use of volunteers, E. coli K12, and dilute soft soap as a reference, their most notable differences include:
| Characteristic | EN 1500 | EN 1499 |
| Objective | Evaluate bactericidal efficacy of hand hygiene products | |
| Test microorganism | Escherichia coli K12 | |
| Neutralizer validation | Requires validation of an appropriate neutralizer | |
| Experimental design | Includes a specific number of volunteers to ensure statistically significant results | |
| Hand movement sequence | Standardized movements to ensure complete product coverage | |
| Type of product evaluated | Hand rub products (alcohol-based gels) | Hygienic hand wash products (liquid disinfectant soaps) |
| Test procedure | Application and rubbing of the product without rinsing | Washing with water and soap followed by rinsing |
| Contact time | 30-60 seconds without rinsing | 60 seconds of washing followed by 10 seconds of rinsing |
| Expected results | Logarithmic reduction compared to 60% propan-2-ol | Logarithmic reduction compared to a standard soap wash |
Proficiency Testing According to EN 1499 and EN 1500
SHAPYPRO organizes interlaboratory proficiency tests based on EN 1500 and EN 1499, allowing laboratories to demonstrate technical competence and meet ISO/IEC 17025 requirements.
In the 2025 test, participants will evaluate:
- The test product (provided by the laboratory)
- The reference product (SHAPYPRO dilute soft soap)
- Reduction of E. coli K12 after application according to protocol
Results are analyzed using z-score, according to ISO 13528, and each laboratory receives a detailed evaluation report.
Requirements for Preparing Dilute Soft Soap
According to Annex D, the soft soap must:
- Be potassium-based
- Be free of contaminants
- Have a pH between 9.0 and 10.0
- Be diluted just before use
The quality of this product is critical to ensure reproducibility and comparability of the results obtained during testing.
Summary Table of Dilute Soft Soap Use
| Standard | Test purpose | Reference product | Comparison type |
| EN 1499 | Hygienic handwash with water | Dilute soft soap | Superiority |
| EN 1500 | Hygienic hand rub without water | 60% Propan-2-ol | Non-inferiority |
Regulatory Importance of EN 1499 and EN 1500
The application of these standards is essential to:
- Validate the effectiveness of hand hygiene products
- Meet regulatory requirements for biocides (EU Regulation 528/2012)
- Ensure quality in hospital and industrial processes
- Participate in public tenders that require regulatory compliance
SHAPYPRO as a Comprehensive Provider
At SHAPYPRO, we offer:
- Standardized dilute soft soap
- Test design according to EN 1499 and EN 1500
- Organization of accredited interlaboratory comparisons
- Personalized evaluation reports
You can contact us or read other related articles like our guide on ISO 13528 and z-score.
Conclusion
EN 1499 and EN 1500 are essential reference standards for ensuring the efficacy of hand hygiene products. Thanks to the standardized use of dilute soft soap and strains like E. coli K12, they allow for objective and reproducible product comparisons.
With support from laboratories like SHAPYPRO, manufacturers and institutions can comply with regulations, validate their products, and demonstrate competence through proficiency testing programs.