What Is Nitrosamine?
Nitrosamine analysis determines the presence and amount of N-nitroso compounds in a product. Nitrosamines are not added directly as a raw material; they form during manufacture or storage from the reaction of secondary amines with nitrosating agents. The analysis can therefore cover both nitrosamines and the potential to form nitrosatable substances.
Purpose of Nitrosamine
The purpose is to show the product is free of nitrosamines banned or restricted due to their carcinogenic potential. In cosmetics, Annex II of (EC) 1223/2009 lists nitrosamines among banned substances.
Which Products Is Nitrosamine Applied To?
- Cosmetic formulations (particularly with amine-containing emulsifiers)
- Pacifiers, feeding bottle teats and teethers
- Rubber balloons and toys
- Rubber seals and gloves
- Rubber products with mouth contact
In Which Sectors Is Nitrosamine Used?
Standards Related to Nitrosamine
- TS EN 71-12 (N-nitrosamines and N-nitrosatable substances in toys)
- TS EN 12868 (child use and care articles — nitrosamines in elastomer soothers)
- TS EN 13130 series (relevant substances in food-contact materials)
Nitrosamine Conditions
- Extraction
- With artificial saliva or the solvent defined in the standard
- Measurement
- LC-MS/MS or GC with a thermal energy analyser
- Two parameters
- Nitrosamines and nitrosatable substances are reported separately
- Unit
- mg/kg
Equipment for Nitrosamine
- LC-MS/MS
- GC-TEA or GC-MS/MS
- Water bath and shaker
- Solid-phase extraction system
Sample Requirements for Nitrosamine
- Specimen as offered for sale
- For rubber products, the mouth-contact area must be sampled separately
- Formulation and raw material information must be provided
- The specimen must be sent under suitable conditions without delay
Duration of Nitrosamine
Because of the extraction and chromatographic measurement, analysis takes several working days. If both nitrosamines and nitrosatable substances are required, two separate preparations are made, extending the duration.
Nitrosamine Procedure
- 1The specimen is prepared and weighed.
- 2It is extracted for nitrosamines with the solution defined in the standard.
- 3A nitrosation procedure is applied for nitrosatable substances.
- 4The extracts are cleaned up and concentrated.
- 5Measurement is performed by LC-MS/MS or GC.
- 6Results are reported separately for the two parameters.
Nitrosamine Acceptance Criteria
In cosmetics, Annex II of (EC) 1223/2009 bans nitrosamines; for technically unavoidable trace amounts, the regulation's general provisions apply and are addressed in the safety assessment. For products such as toys and soothers, EN 71-12 and EN 12868 define limit values. The applicable limit is determined by the product's scope.
Evaluation of Results
Because nitrosamines can form over time, a single analysis does not demonstrate compliance for the shelf life; re-assessment after storage is meaningful, particularly for amine-containing formulations. Because the nitrosatable-substance result shows the potential for formation, it is more informative from a preventive standpoint.
Nitrosamine Report
- Specimen identification and component analysed
- Standard and extraction method applied
- Nitrosamine results and limits of quantification
- Nitrosatable substance results
- Regulatory limit compared against
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
- (EC) 1223/2009 Cosmetic Products Regulation Annex II
- 2009/48/EC Toy Safety Directive
- (EC) 1935/2004 Regulation on Materials Intended to Come into Contact with Food
Kozmetik Testleri — What Does This Testing Field Cover?
The section below describes the field that the Nitrosamine 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 Nitrosamine
We don't add nitrosamines to our formulation — how could they appear?
Nitrosamines are not added — they form. It is enough for raw materials containing secondary amines (some emulsifiers, alkanolamines) to be present together with nitrite or nitrosating contaminants. Raw material selection and purity are therefore decisive.
Why is there a separate standard for rubber soothers?
A soother stays in the mouth for a long time and is in continuous contact with saliva. EN 12868 defines the artificial saliva extraction representing this use and the relevant limits; a general rubber analysis does not represent this condition.