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ISO 22196 – Antibacterial Testing of plastics and other non - porous surfaces
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Summary
ISO 22196 testing measures the antibacterial activity of treated plastics and other non-porous surfaces under controlled laboratory conditions. The method quantifies bacterial reduction by comparing treated and untreated samples, providing standardized log reduction (R) values. ISO 22196 test data is used to support antibacterial performance claims, regulatory documentation, and product development for packaging, medical,consumer product and surface applications.
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Quick understanding about test
ISO 22196: 2011 – Determination of Antibacterial Efficacy of Plastic Surfaces
- Staphylococcus aureus (ATCC 6538P)
- Escherichia coli(ATCC 8739)
- Bacterial suspensions are applied to treated and untreated samples.
- Samples are incubated at 35°C for 24 hours.
- For untreated samples, neutralizers are added immediately after incubation, followed by 10-fold serial dilutions
- For treated samples, neutralizers are added after incubation of 24 hours, followed by 10-fold serial dilutions.
- Then, both the samples ( test and control ) are incubated at 35 °C for 40 hours to 48 hours.
- After incubation, CFU is counted for plates containing 300 colonies
- Antibacterial activity is evaluated by comparing CFU reduction in treated samples against controls
- Ensures products meet international standards for market acceptance.
- It supports product development.
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What Is ISO 22196?
ISO 22196 is an internationally accepted test standard for assessing the antibacterial efficacy of treated plastic materials and other non porous surfaces. It is used when a product contains an antibacterial agent and needs quantitative evidence of bacterial reduction. The test results can be used by manufacturers to support their antibacterial performance claims and to prepare technical documentation for regulatory and compliance requirements.
ISO-22196 test is a harmonized version of the Japanese standard JIS Z 2801 which provides guidelines for testing the antibacterial efficacy of plastic material.
Applicable Products Under ISO 22196 Testing
ISO 22196 is applicable to plastics and other non-porous materials such as
- Coatings
- Steel
- acrylic fiber
- Glass
- Metals
- Ceramics
- and tiles, etc.
ISO 22196 test methods and requirements
Sample preparation
- Number of samples: 3 treated samples and 6 untreated samples.
- Size and shape : Both treated and untreated samples should be 50 ± 2 mm × 50 ± 2 mm in size, thickness ≤ 10 mm. Other sizes/shapes are acceptable if the surface area covered by the inoculum film is between 400–1600 mm².
- Contamination control : All the test specimens are kept separately in sterile petri plates to avoid cross-contamination.
Test Organisms Used in ISO 22196
Mandatory strains
- Staphylococcus aureus (ATCC 6538P)
- Escherichia coli(ATCC 8739)
Additional strains available for testing at MIS:
- Klebsiella pneumoniae (ATCC 4352)
- Pseudomonas aeruginosa (ATCC 15442)
- Enterococcus hirae (ATCC 10541)
- MRSA (ATCC 33591)
- Salmonella enterica (ATCC 10708)
ISO 22196 test procedure
- The test bacterial inoculum is inoculated onto the treated and untreated test samples and covered with a piece of inert film of size 40 mm × 40 mm to prevent the spread of inoculum from the edges.
- These petri plates are then incubated at specific temperature conditions (35°C) and humidity for 24 hours.
- For untreated samples (control):
- 10 ml of SCDLP (Soya Caesin Digest Lecithin Polysorbate) broth (which serves as a neutralizer) is added to three untreated samples immediately after inoculation.
- For the remaining three untreated samples, 10 mL of SCDLP is added following a 24-hour incubation period.
- Further, a 10-fold serial dilution is performed with the recovered neutralizing solution to enumerate the viable bacteria.
- Tryptic Soya Agar (TSA) is used as the culture media for the growth of bacteria and the plates are incubated for 40-48 hours at 35 °C.
- For treated samples (test):
- 10 ml of SCDLP broth (used as a neutralizer) is added to Petri plates having samples after the incubation of 24 hours.
- Further, 10-fold serial dilution is performed and viable bacteria are enumerated by Plate Count method.
- Plates are incubated at 35 °C for 40 hours to 48 hours. After incubation, CFU (Colony Forming Unit) is counted on plates (sample and control) containing 30 – 300 colonies.
- The bactericidal activity of the testing material is determined by comparing the reduction in viable bacterial count of treated samples with control samples.
Passing Criteria & Log Reduction
ISO 22196 expresses antibacterial performance as antibacterial activity (R), which represents the log reduction of viable bacteria on treated surfaces compared to untreated controls after the test period (typically 24 hours).
Higher the R value, stronger is the antibacterial activity.
Factors affecting the validity of ISO 22196 test method
1. Impact of contact time
2. Effect of surface topography
3. Influence of temperature and humidity
ISO 22196 vs Other Standards
| Feature | ISO 22196 | ASTM E2180 |
|---|---|---|
| Purpose | To determine the Antibacterial Activity on Plastics and Other Non-Porous Surfaces | To determine the Activity of Incorporated Antimicrobial Agent(s) in Polymeric or Hydrophobic Materials |
| Standards body | International Organization for Standardization (ISO) | ASTM International |
| Scope of application | Antibacterial activity on treated plastics and other non-porous surfaces | Antimicrobial agents incorporated into hydrophobic/polymeric materials (e.g., plastics, shower curtains, medical devices) |
| Mandatory Test organisms | Staphylococcus aureus (ATCC 6538P), Escherichia coli (ATCC 8739) | Staphylococcus aureus (ATCC 6538P) Yeast and fungal conidia may be tested |
| Inoculation method | Bacteria mixed into a warm agar slurry that forms a thin, even layer (“pseudo-biofilm”) | Bacterial suspension (liquid inoculum) is placed on the surface and covered with a thin film to ensure even contact |
| Surface types suited | Plastics and other non-porous materials such as coatings, steel, acrylic fiber, glass, metals, ceramics, and tiles, etc. | Best for hydrophobic/polymeric surfaces including textured or difficult-to-wet materials |
| Result interpretation/Passing criteria | Quantitative antibacterial activity (bacteriostatic/bactericidal) compared to control | Quantitative reduction of microbes on treated vs. untreated material |
Strengths and limitations of ISO 22196 test method
Strengths of ISO 22196 test method
- Provides measurable, reproducible results through standardized microbial concentrations, nutrient conditions, and triplicate testing, ensuring statistical reliability.
- Testing scope extends well beyond plastics to include a wide range of non-porous materials, making ISO 22196 applicable across many sectors.
- Assesses both bacteriostatic and bactericidal effects, offering a complete picture of a surface’s antimicrobial performance under controlled conditions.
- Allows method adjustments (e.g., inoculum volume) for diverse surfaces and uses defined pass/fail criteria, ensuring consistent, comparable, and objective assessment.
Limitations of ISO 22196 test method
- The ISO 22196 test doesn’t reflect real-world conditions, as it keeps a wet microbial suspension on the surface for 24 hours,while in reality, microbes dry quickly, reducing true contact time with the antimicrobial surface.
- Excludes several material types such as building materials, treated textiles, and photocatalytic surfaces, restricting its use to specific non-porous substrates.
- Results can be influenced by test modifications (e.g., inoculum load, contact time), and the high-humidity, controlled environment may overestimate actual antimicrobial performance
Customized ISO 22196 Testing Services by MIS
Testing on a wide range of non-porous materials (plastics, coatings, metals).
Tailored bacterial strain selection beyond ISO 22196 specifications.
Simulated real-world conditions for increased reliability.
Regulatory support for compliance and market approval.
Why Choose Our Lab for ISO 22196 Testing
At Microbe Investigations Switzerland (MIS), we provide ISO 22196 (and ASTM equivalent) testing services to evaluate the antibacterial activity of plastics and other non-porous surfaces.
Our expert team ensures precise, reproducible, and standard-compliant results, helping you verify surface efficacy claims and demonstrate product performance.
Request ISO 22196 Testing
Looking to validate antibacterial activity of your product in accordance with ISO 22196? Contact us today to discuss your testing requirements.
Frequently Asked Questions
DR. Martinoz Scholtz
ISO 22196 reports antibacterial activity (R) as the log reduction of bacteria on treated surfaces versus untreated controls after incubation (typically 24 hours). Higher R indicates stronger antibacterial performance.
Yes, ISO 22196 is widely accepted to support antibacterial performance claims for plastics and non-porous surfaces.
Yes, ISO 22196 is the internationally adopted version of JIS Z 2801 and is commonly used to support SIAA (Kohkin) antibacterial certification.
ISO 22196 uses optimized laboratory conditions, including high humidity and constant moisture, to maximize bacterial survival and contact with the surface.
Yes, ISO 22196 test report data can be used to support antibacterial performance claims for medical or food-contact plastics.
Alternatives include ASTM E2149 (dynamic, real-use simulation), ASTM E2180 (polymeric materials), ISO 21702 (antiviral surfaces), and ISO 846 for evaluation of the action of microorganisms.
Contact time, surface topography and environmental conditions like temperature and humidity can impact the effectiveness of the antibacterial treatment.
ISO 22196 certification gives a confirmation of the antibacterial efficacy of plastic surfaces, and once products pass for this standard, they satisfy international hygiene requirements, which boosts customer confidence and aids in regulatory compliance.
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