What Is Microbial Load?
Microbiological load analysis in cosmetics consists of two parts: total viable aerobic mesophilic microorganism count, and detection of specified microorganisms the regulation prohibits (P. aeruginosa, S. aureus, C. albicans, E. coli). ISO 17516 defines the framework for microbiological limits, while other ISO standards define the methods.
Purpose of Microbial Load
The purpose is to show that the product was not contaminated during manufacturing and filling, and remains microbiologically safe over its shelf life. Results are among the bases of the cosmetic product safety report (CPSR).
Which Products Is Microbial Load Applied To?
- Creams, lotions and emulsions
- Shampoos and shower gels
- Make-up products
- Baby care products
- Eye-area products
In Which Sectors Is Microbial Load Used?
Standards Related to Microbial Load
- ISO 17516 (cosmetics — microbiological limits)
- ISO 21149 (enumeration and detection of aerobic mesophilic bacteria)
- ISO 16212 (enumeration of yeast and mould)
- ISO 22717 / ISO 22718 / ISO 18416 / ISO 21150 (detection of specified microorganisms)
Microbial Load Conditions
- Scope
- Total aerobic mesophilic count, yeast-mold count and search for specific microorganisms
- neutralization
- A neutralizer is used to eliminate the effects of preservatives in the product.
- Method suitability
- It has been previously shown that the product does not suppress fertility
- Product category
- Separate, stricter expectations for under-three and eye area products
Equipment for Microbial Load
- Class II biosafety cabinet
- Incubators
- Membrane filtration setup
- Homogeniser and dilution equipment
- Colony counting setup
Sample Requirements for Microbial Load
- The product must be sent in its original, unopened packaging.
- Batch number, manufacturing and expiry dates must be stated.
- The ingredients list (INCI) and formulation information, if any, must be shared.
- The required quantity varies by number of analyses; confirmation must be obtained before shipment.
- Light- and temperature-sensitive products must be transported under suitable conditions.
Duration of Microbial Load
The analysis is roughly a week's work, including incubation and confirmation steps.
Microbial Load Procedure
- 1A method suitability study is performed and the neutraliser selected.
- 2The sample is weighed and homogenised with a diluent containing the neutraliser.
- 3Inoculation onto suitable media is done for the total count.
- 4A separate medium is inoculated for yeast and mould.
- 5Enrichment and selective inoculation are applied for the specified microorganisms.
- 6Plates are incubated at the defined temperature and duration.
- 7Colonies are counted, suspect ones confirmed, and the result reported.
Microbial Load Acceptance Criteria
The acceptance criterion for cosmetic products does not come from a single test but from the product's safety assessment report and the regulation. EU 1223/2009 and the Cosmetics Regulation define prohibited and restricted substances, microbiological expectations, and labelling requirements; this page therefore does not give a fixed limit.
Evaluation of Results
The result is assessed together with the limits in ISO 17516 and the product's category. Expectations are stricter for products aimed at children under three and eye-area products. Absence of the specified microorganisms is essential.
Microbial Load Report
- Product name, batch number, and packaging information
- Sample receipt date and storage conditions
- Method applied and reference standard
- Test conditions and conditioning, if any
- Measurement results and units
- Measurement uncertainty (if requested)
- A statement that the result applies only to the specimen submitted for testing
Accreditation
EUROLAB holds internationally recognised accreditation obtained from accreditation bodies party to the IAF and ILAC multilateral recognition arrangements, under ISO/IEC 17025 for laboratory testing, ISO/IEC 17020 for inspection and surveillance, and ISO/IEC 17021-1 and ISO/IEC 17065 for system and product certification. This chain ensures the reports and certificates issued are accepted worldwide.
Whether this test can be offered within accredited scope depends on your product and the standard applied; since accreditation is method-based, the scope is confirmed in writing at the quotation stage.
Scope and authorisationsRelated Regulations
- EU 1223/2009 Cosmetic Products Regulation
- Cosmetics Regulation (Türkiye)
- Cosmetic Product Safety Report (CPSR) requirements
- ISO 22716 (good manufacturing practice in cosmetics)
Kozmetik Testleri — What Does This Testing Field Cover?
The section below describes the field that the Microbial Load test belongs to; the conditions and acceptance criteria of this particular test are given in the sections above. The scope specific to your product is confirmed in writing at the quotation stage.
Cosmetics testing shows that a product stays microbiologically safe throughout its use, delivers the performance it claims and contains no restricted substances. There is NO PRODUCT APPROVAL in this family: no authority approves a cosmetic product; responsibility rests entirely with the manufacturer and with the responsible person established in the EU.
This responsibility model shapes how the field works. Before the product is placed on the market a product information file (PIF) is prepared, a safety assessment (CPSR) is carried out and notification is completed. Inspection comes afterwards and asks for the file; if the file is incomplete the product is withdrawn from the market. Test reports form the evidence base of that file.
On the microbiological side there are two different questions and each is answered by a separate test. The CURRENT microbiological quality of the product is measured by enumeration and testing for specified microorganisms. The EFFECTIVENESS of the preservative system is examined by deliberately inoculating the product with microorganisms (challenge test): can the product resist repeated contamination throughout its use? A jar of cream is touched by a finger at every use; the preservative system must cope with that.
On the performance side, claim substantiation studies are carried out. For sunscreens, determination of SPF and UVA protection and measurement of water resistance are mandatory and are the basis of the value declared on the label. Evidence is also produced by in vivo or in vitro methods for claims such as moisturisation, appearance of wrinkles, hair care and colour retention.
On the chemical side, banned and restricted substances are screened: heavy metals, prohibited colorants, preservative concentrations, UV filter levels and contaminants such as nitrosamines. Products containing nanomaterials carry a separate notification obligation.
The BAN ON ANIMAL TESTING is the most defining constraint of the field. Animal testing cannot be carried out on finished products or ingredients, and a product tested on animals cannot be placed on the market. Alternative in vitro methods (reconstructed human skin models, cell culture based tests) are therefore at the centre of the field, and toxicological assessment rests largely on existing data, read-across approaches and in vitro results.
Stability and packaging compatibility studies determine shelf life and the period after opening (PAO). The interaction of the product with its packaging is also assessed: an active substance may migrate into the packaging, and substances that should not be present may pass from the packaging into the product.
On the process side, good manufacturing practice is demonstrated through ISO 22716; this is separate from product testing but a mandatory part of the file.
Method groups in this field
Microbiological quality
Enumeration of aerobic mesophilic bacteria, yeasts and moulds; testing for Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans and E. coli.
Preservative efficacy
Resistance of the preservative system to repeated contamination by challenge test (ISO 11930).
Sun protection
In vivo SPF determination, in vitro UVA protection determination, water resistance and critical wavelength measurements.
Chemical analysis
Heavy metals, banned and restricted substances, quantification of preservatives and UV filters, contaminant screening.
Safety assessment
In vitro irritation and sensitisation methods, toxicological risk assessment, exposure calculations.
Stability and compatibility
Real-time and accelerated stability, packaging compatibility, PAO determination, sensory and physicochemical monitoring.
Commonly confused in this field
- There is NO CE marking on cosmetics; conformity is demonstrated through the PIF, the safety assessment and notification.
- The responsible person must be established in the EU; unless the exporter arranges this, the product cannot be placed on the market.
- Microbiological quality and preservative efficacy are different tests; a product that comes out clean may still have a weak preservative system.
- The animal testing ban covers ingredients too; animal test data generated for another market creates a problem of use.
- An SPF claim must be supported by measurement; a formulation change requires a new measurement.
- A change of packaging may require the stability and compatibility studies to be repeated.
Which decisions these results support
- The product information file and the safety assessment
- Verification of the preservative system design
- Substantiation of SPF and efficacy claims
- Determining shelf life and the period after opening
- Compliance with destination market restrictions for export
Related Tests
Frequently Asked Questions About Microbial Load
Why is a neutraliser necessary?
When preservatives in the product carry over into the growth medium, they inhibit microbial growth and give a falsely clean result. The neutraliser eliminates this effect; its effectiveness is demonstrated by the method suitability study.
Is it needed for water-free products too?
Microbiological growth risk is low in water-free products. ISO 17516 and risk assessment accept that analysis frequency can be reduced for these products; the decision belongs to the safety assessor.