Learn all about the JIS Z 2801 and ISO 22196 standards

We never tire of saying that TNS additives comply with the strictest national and international quality standards, such as JIS Z 2801 / ISO 22196

But do you actually know what that means?

In this infographic, we explain the main tests used to evaluate our additives on non-porous surfaces: JIS Z 2801 and ISO 22196.

What are JIS Z 2801 and ISO 22196?

JIS Z 2801 and ISO 22196 are widely recognized international technical standards for evaluating antibacterial activity on non-porous surfaces (such as plastics and laminates).

Origin of JIS Z 2801:

Initially, JIS Z 2801 was originally established by Japan through the Japanese Industrial Standards Committee (JISC), as a national standard for testing antibacterial activity on plastics and other non-porous surfaces.

The standard emerged in the early 2000s, just as Japan was leading the world in the development of antimicrobial technologies applied to the plastics, healthcare, and consumer goods industries.

The country was facing serious concerns about contamination in public facilities, food, and hospitals, which necessitated the standardization of reliable testing methods to verify antimicrobial efficacy.

Origin of ISO 22196:

Given the importance of the JIS Z 2801 standard, the International Organization for Standardization (ISO) decided to create an equivalent international version, enabling manufacturers in various countries to use a standardized testing method.

ISO 22196 was published based almost entirely on the method described in JIS Z 2801, with minor adaptations for global harmonization. ISO 22196 was published in 2007

Just to illustrate and wrap things up, in practice:

JIS Z 2801: Japanese standard

ISO 22196: validated international equivalent

Although these standards are published by different organizations, they use the same testing method and, in practice, are equivalent in the global market.

What are the JIS Z 2801 and ISO 22196 standards used for?

The JIS Z 2801 and ISO 22196 standards are used to scientifically and systematically evaluate the antimicrobial efficacy of materials.

They provide measurable evidence that a product treated with an antimicrobial additive actually inhibits bacterial growth on its surface.

In practice, they:

They provide technical assurance to the market and regulatory agencies;
They facilitate the entry of products into international markets;
They demonstrate the performance of surfaces treated with antibacterial agents;
They enhance the credibility of products made of materials such as plastics, ceramics, and other non-porous materials.

Therefore, companies that wish to market antimicrobial materials—such as those intended for hospital use—need this validation to ensure competitiveness, safety, and a technological edge.

What materials can be tested?

Among the main materials tested are, for example:

Rigid and flexible plastics: PE, PP, PVC, ABS, and many others.
Synthetic and decorative laminates;
Ceramics and glazed tiles;
Acrylics, polycarbonates, and engineering polymers;
Films and panels for high-touch surfaces;
Non-porous hospital supplies and medical devices.

What is the methodology of JIS Z 2801?

Step 1: Sample preparation

First and foremost, proper sample preparation is the first critical step. The standard specifies that:

First, the samples must have standard dimensions of 5 cm x 5 cm (flat and uniform surface);
For each microorganism tested, the laboratory must prepare:
- 3 samples treated with the antimicrobial additive;
- 6 untreated samples (control group).
The sample thickness must not exceed 1 cm

This number of replicates is required to ensure statistical significance and minimize any experimental deviations.

For example, the standard specifies that pre-sterilization cleaning must ensure that no physical contaminants or manufacturing residues remain on the surface (e.g., molding residues, dust, oils, adhesives, among others).

Some samples may require a preliminary washing with a mild detergent before using alcohol.

Step 2: Sterilization

To prevent contamination from external sources, all samples must be thoroughly sterilized before bacterial inoculation.

The most common and safest method used in the procedure is the application of 70% alcohol.
The use of alcohol preserves the integrity of the material (it does not damage polymers or laminates) and ensures the removal of residual microorganisms without altering the chemical composition of the surface.

In addition to 70% alcohol, laboratories may also use alternative methods such as:

UV light (without heating, especially for heat-sensitive plastics);

Omitting this step can directly compromise the validity of the results, since contaminants can skew the additive’s actual antimicrobial activity.

Step 3: Inoculation

In Stage 3 of the JIS Z 2801 and ISO 22196 standards, controlled contact between the biological agent and the material surface then begins.

The bacterial strains used comply with standardized international standards:
- Staphylococcus aureus ATCC 6538P (Gram-positive bacterium);
- Escherichia coli ATCC 8739 (Gram-negative bacterium);
Prepare a bacterial suspension with a standardized concentration of 2.5 × 10⁵ to 10⁵ CFU/mL, with an ideal target of 6 ×10⁵ CFU/mL. A suspension in a 1:500 dilution of nutrient broth (NB), prepared as described in the standard, should be used;
0.4 mL of the bacterial suspension is placed in the center of the 50 mm × 50 mm sample. The inoculum should cover approximately 4 cm × 4 cm of the central area of the sample;
A sterile cover film (e.g., polyester) measuring 40 mm × 40 mm is placed over the inoculum; it must cover the inoculated area exactly and prevent any leakage to the edges.

This procedure simulates actual contact between a contaminated surface and pathogenic microorganisms, establishing a standardized condition for evaluating the antimicrobial performance of the sample during incubation.

Step 4: Incubation

After inoculation, the samples are stored in a controlled environment to allow the bacteria to grow properly:

Temperature: 35 °C (±1°C)
Relative humidity: ≥ 90% RH
Duration: 24 consecutive hours
The specimens are kept in sealed Petri dishes to minimize evaporation.

This combination of heat and humidity simulates conditions conducive to bacterial growth, thereby putting the antimicrobial surface’s performance to the test.

This is where the additive begins to take effect: we will assess its effectiveness based on the number of surviving microorganisms after this incubation period.

JIS Z 2801 / ISO 22196 - Step 4: Incubation

Step 5: Neutralization and homogenization

Once the incubation cycle is complete in accordance with JIS Z 2801 and ISO 22196 standards, the recovery of bacteria for quantification begins:

The samples are neutralized with nutrient broth (the neutralizing solution is typically SCDLP), homogenized, and finally diluted and plated to determine the microbial count
It is shaken in Stomacher-type bags to ensure the uniform mechanical removal of microorganisms remaining on the surface.

This procedure ensures the efficient collection of viable organisms, enabling standardized quantitative analysis.

Step 5: Neutralization and homogenization

Step 6: Counting

Finally, the colony-forming units (CFU) obtained from the test and control samples are counted:

The final result is expressed as a logarithmic reduction®.

Group	Time (0h)	Weather (24-hour forecast)
Control (untreated)	High bacterial count	Exponential growth
Treated sample	High initial count	Almost no bacteria remaining

JIS Z 2801 and ISO 22196 - Step 6: Counting

Which microorganisms are used in the JIS Z 2801 tests?

The test uses standardized bacterial and fungal strains that represent different groups of microorganisms of clinical and environmental relevance.

The selection of these strains is intended to ensure that the performance of the tested materials is validated against pathogens that pose a genuine risk, for example, in industrial, hospital, and human-contact applications.

Below are the two most commonly used microorganisms, classified by group:

Gram-positive bacterium: Staphylococcus aureus ATCC 6538P (Gram-positive bacterium);
Gram-negative bacterium: Escherichia coli ATCC 8739 (Gram-negative bacterium).

Why is JIS Z 2801 considered the gold standard for antimicrobial surfaces?

Based on its scientific and historical validation of reproducibility, the standard is:

An international standard widely accepted by industries, laboratories, etc.;
Required by multinational brands as a technical prerequisite;
A product certified under JIS Z 2801 has much greater commercial credibility, including for export.

Testing is important to ensure that the final product functions as intended.

Would you like to learn more about testing in accordance with JIS Z 2801 / ISO 22196, or how to make your products resistant to mold and bacteria? Contact us!

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