💊 Microbiological purity of cosmetics
In short
Microbiological limits for finished cosmetic products according to PN-EN ISO 17516. The test is performed according to PN-EN ISO 17516:2014-11; Total aerobic mesophilic microorganisms — children under 3, eye area, mucous membranes: ≤ 1 × 10² CFU per 1 g or 1 ml (out of limit: > 200 CFU). The procedure comprises 5 steps; it is used for: Assessment of the microbiological quality of finished cosmetic products against the limits of Table 1, Products specifically intended for children under three, the eye area and mucous membranes — limit 1 × 10² CFU per 1 g or 1 ml, Other cosmetic products — limit 1 × 10³ CFU per 1 g or 1 ml.
At a glance
- Standard: PN-EN ISO 17516:2014-11
- Category: Pharma and Cosmetics
- Procedure steps: 5
- Total aerobic mesophilic microorganisms — children under 3, eye area, mucous membranes: ≤ 1 × 10² CFU per 1 g or 1 ml (out of limit: > 200 CFU)
- Total aerobic mesophilic microorganisms — other products: ≤ 1 × 10³ CFU per 1 g or 1 ml (out of limit: > 2000 CFU)
- Specified microorganisms: Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans — absent in 1 g or 1 ml
Overview
STATUS (as of 02.11.2026). PN-EN ISO 17516:2014-11 — Polish version (published 28.11.2017), implements EN ISO 17516:2014 and ISO 17516:2014 [IDT]; the catalogue card carries no withdrawal notice. ISO 17516:2014 (first edition, 30.09.2014) — in the ISO catalogue at stage 90.92 (“to be revised”); work on a new edition is under way (draft prEN ISO 17516, in the Slovenian catalogue oSIST prEN ISO 17516:2026), whose content we do not take into account here. Committee ISO/TC 217 “Cosmetics”.
SCOPE AND PRINCIPLE (Clauses 1 and 3). The standard applies to all cosmetics and assists in the assessment of the microbiological quality of finished products. Cosmetics, their raw materials and manufacturing conditions are not required to be sterile, but the microorganisms in a product must not harm the consumer or the quality of the product during intended or foreseeable use — therefore quantitative and qualitative limits are established. Products with a low microbiological risk (according to ISO 29621) need not be tested. The manufacturer should follow the good manufacturing practices of ISO 22716.
LIMITS (Table 1). Total aerobic mesophilic microorganisms — bacteria plus yeasts and moulds growing under the conditions of ISO 21149 and ISO 16212: not more than 1 × 10² CFU per 1 g or 1 ml in products specifically intended for children under three years of age, for the eye area or for mucous membranes, and not more than 1 × 10³ CFU per 1 g or 1 ml in other products. Because of the variability of the plate count method (according to USP Chapter 61 or Ph. Eur. 2.6.12), a result is considered out of limit only above 200 CFU (first group) and above 2000 CFU (other products) per 1 g or 1 ml. Specified microorganisms — Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans: absent in 1 g or 1 ml of product, in every group.
INTERPRETATION (Clause 4 and Annex A). The number of non-specified microorganisms should remain stable or decrease over the product life. Their presence is not considered objectionable if they are unable to grow in the product — as shown by a risk assessment including a preservation efficacy test (e.g. ISO 11930) or by demonstrating that the product does not support microbial growth (ISO 29621). Results are assessed according to the normative flowchart in Annex A.
TEST METHODS. When testing is needed, it is performed according to the method standards: ISO 21148 (general instructions for microbiological examination of cosmetics), ISO 21149 (count of aerobic mesophilic bacteria), ISO 16212 (count of yeasts and moulds), ISO 18415 (detection of specified and non-specified microorganisms), ISO 21150 (Escherichia coli), ISO 22717 (Pseudomonas aeruginosa), ISO 22718 (Staphylococcus aureus) and ISO 18416 (Candida albicans). The standard notes that when bacterial colonies grow on Sabouraud dextrose agar, this medium with antibiotics may be used.
LEGISLATION. Regulation (EC) No 1223/2009 requires the cosmetic product safety report to contain the microbiological specifications of the substances, mixtures and product, with particular attention to cosmetics used around the eyes, on mucous membranes, on damaged skin, on children under three, on elderly persons and persons with compromised immune responses (Annex I Part A point 3). The Commission guidelines 2013/674/EU distinguish products with a low microbiological risk (microbiological quality tests of the finished product are not necessary — with a scientific justification), single-use and non-openable products (only microbiological quality tests) and all other products.
WHAT WE DO NOT GIVE HERE. Culture media, diluents and neutralizers, incubation conditions, enrichment and confirmation tests — they are specified in the method standards listed above, whose texts we did not have. The content of the draft new edition of ISO 17516.
Method principle
The product (except products with a low microbiological risk according to ISO 29621) is tested by the methods of ISO 21149 and ISO 16212 (counts) and ISO 18415, ISO 21150, ISO 22717, ISO 22718 and ISO 18416 (specified microorganisms). The results are compared with the limits of Table 1 — 1 × 10² or 1 × 10³ CFU per 1 g or 1 ml depending on the product group and the absence of the four microorganisms in 1 g or 1 ml — according to the flowchart in Annex A.
Applications
- Assessment of the microbiological quality of finished cosmetic products against the limits of Table 1
- Products specifically intended for children under three, the eye area and mucous membranes — limit 1 × 10² CFU per 1 g or 1 ml
- Other cosmetic products — limit 1 × 10³ CFU per 1 g or 1 ml
- Microbiological specifications of the product in the safety report (Regulation (EC) No 1223/2009, Annex I Part A point 3)
- Assessment of out-of-limit results taking into account the variability of the plate count method (above 200 or 2000 CFU per 1 g or 1 ml)
Key parameters
| Parameter | Value |
|---|---|
| Total aerobic mesophilic microorganisms — children under 3, eye area, mucous membranes | ≤ 1 × 10² CFU per 1 g or 1 ml (out of limit: > 200 CFU) |
| Total aerobic mesophilic microorganisms — other products | ≤ 1 × 10³ CFU per 1 g or 1 ml (out of limit: > 2000 CFU) |
| Specified microorganisms | Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans — absent in 1 g or 1 ml |
| The total count includes | bacteria plus yeasts and moulds growing aerobically under the conditions of ISO 21149 and ISO 16212 |
| Products with a low microbiological risk | no testing (ISO 29621) |
| Method standards | ISO 21148, ISO 21149, ISO 16212, ISO 18415, ISO 21150, ISO 22717, ISO 22718, ISO 18416 |
Standard
- Standard number
- PN-EN ISO 17516:2014-11
- Title (PL)
- Kosmetyki — Mikrobiologia — Limity mikrobiologiczne
- Title (EN)
- Cosmetics — Microbiology — Microbiological limits
Step-by-step procedure
-
Is testing needed
Check whether the product has a low microbiological risk according to ISO 29621 — if so, testing need not be performed (Clause 1).
-
Microbial count
Determine the count of aerobic mesophilic bacteria according to ISO 21149 and of yeasts and moulds according to ISO 16212; their sum is the total aerobic mesophilic microorganism count.
-
Specified microorganisms
Detect Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans in 1 g or 1 ml of product according to ISO 21150, ISO 22717, ISO 22718 and ISO 18416 (general rules — ISO 18415).
-
Comparison with the limits
Compare the total count with the limit for the product group (1 × 10² or 1 × 10³ CFU per 1 g or 1 ml); consider a result out of limit only above 200 or 2000 CFU because of the variability of the plate count method.
-
Interpretation
Assess the result according to the flowchart in Annex A; the presence of non-specified microorganisms does not disqualify the product if they cannot grow in it (risk assessment according to ISO 29621 or ISO 11930).
Required equipment and apparatus
| Equipment | Example | Indicative price |
|---|---|---|
| Microbiological laboratory equipment | according to the method standards ISO 21148, ISO 21149, ISO 16212, ISO 18415, ISO 21150, ISO 22717, ISO 22718 and ISO 18416 — ISO 17516 does not specify apparatus | — |
Reagents, media and consumables
| Reagent | CAS | Details |
|---|---|---|
| Culture media, diluents and neutralizers | — | according to the method standards; ISO 17516 only states that if bacteria grow on Sabouraud dextrose agar, this medium with antibiotics may be used |
Health and safety (OHS)
- ISO 17516 contains no safety requirements — the rules for working with microorganisms, including the specified ones, are set by the laboratory on the basis of a risk assessment and the method standards
Frequently asked questions
Which standard describes this test?
The test is performed according to PN-EN ISO 17516:2014-11 — “Cosmetics — Microbiology — Microbiological limits”.
How does this method work?
The product (except products with a low microbiological risk according to ISO 29621) is tested by the methods of ISO 21149 and ISO 16212 (counts) and ISO 18415, ISO 21150, ISO 22717, ISO 22718 and ISO 18416 (specified microorganisms). The results are compared with the limits of Table 1 — 1 × 10² or 1 × 10³ CFU per 1 g or 1 ml depending on the product group and the absence of the four microorganisms in 1 g or 1 ml — according to the flowchart in Annex A.
What is the measuring range and accuracy?
Total aerobic mesophilic microorganisms — children under 3, eye area, mucous membranes: ≤ 1 × 10² CFU per 1 g or 1 ml (out of limit: > 200 CFU); Total aerobic mesophilic microorganisms — other products: ≤ 1 × 10³ CFU per 1 g or 1 ml (out of limit: > 2000 CFU); Specified microorganisms: Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans — absent in 1 g or 1 ml; The total count includes: bacteria plus yeasts and moulds growing aerobically under the conditions of ISO 21149 and ISO 16212.
How long does the test take?
The procedure comprises 5 steps. The standard does not give a duration for every stage — the laboratory's own procedure decides.
What equipment is required?
Microbiological laboratory equipment.
Where is this test used?
Assessment of the microbiological quality of finished cosmetic products against the limits of Table 1; Products specifically intended for children under three, the eye area and mucous membranes — limit 1 × 10² CFU per 1 g or 1 ml; Other cosmetic products — limit 1 × 10³ CFU per 1 g or 1 ml; Microbiological specifications of the product in the safety report (Regulation (EC) No 1223/2009, Annex I Part A point 3); Assessment of out-of-limit results taking into account the variability of the plate count method (above 200 or 2000 CFU per 1 g or 1 ml).
What safety precautions apply?
ISO 17516 contains no safety requirements — the rules for working with microorganisms, including the specified ones, are set by the laboratory on the basis of a risk assessment and the method standards.
Which laboratory can perform this test?
The test is performed by laboratories accredited to ISO/IEC 17025. On LabCoda you can find them by the standard number PN-EN ISO 17516.