Allergen Analysis — ELISA / PCR / LC-MS/MS and Allergens in Cosmetics · Cosmetics Testing

What Is Allergen?

Allergen analysis covers two separate areas. The presence and quantity of allergens listed in Annex II of FOOD (EU) 1169/2011 are determined by ELISA, real-time PCR or LC-MS/MS; R5 ELISA for gluten is widely used. IN COSMETICS, perfume allergens, which are required to be labeled in (EC) 1223/2009 Annex III, are measured by GC-MS.

Purpose of Allergen

The aim is to verify the accuracy of the label declaration and the effectiveness of cross-contamination control. Undeclared allergen is one of the most common causes of food recalls.

Which Products Is Allergen Applied To?

Products declared gluten-free
Baked goods and snacks
Products declared dairy-free and hazelnut-free
Ready meals and sauces
Perfumed cosmetic products

In Which Sectors Is Allergen Used?

FoodAgricultureRetailCosmetics

Standards Related to Allergen

TS EN 15634 series (allergen detection in food — general assumptions and PCR methods)
AOAC 2012.01 (Determination of gluten by R5 ELISA)
TS EN ISO 21571 (nucleic acid extraction)
ISO/TS 20224 series (animal species determination, real-time PCR)
TS EN 16274 series (Determination of allergenic odorous substances in cosmetics by GC-MS)

Allergen Conditions

Method selection: Protein hedefliyse ELISA/LC-MS, DNA hedefliyse PCR
Processing effect: Heat treatment may denature the protein and reduce the ELISA result
Gluten: By R5 ELISA; Defined threshold for 'gluten-free' declaration
Kozmetik alerjen: Annex III listed substances are screened by GC-MS

Equipment for Allergen

ELISA plate reader and incubator
Real-time PCR system
LC-MS/MS
GC-MS (for cosmetic allergens)
Homogenizer and grinder

Sample Requirements for Allergen

Homogenized representative sample
Line swab samples may also be taken if cross-contamination is investigated.
Processing parameters must be declared
Perfume composition and INCI list must be presented in cosmetics

Duration of Allergen

ELISA results are available relatively quickly. PCR and LC-MS/MS analyzes take several working days; Cosmetic allergen screening takes similar time due to GC-MS measurement.

Allergen Procedure

1The target allergen and the appropriate method are determined.
2The sample is homogenized and extracted.
3In ELISA, antibody-based measurement is made.
4In PCR, DNA extraction and species-specific amplification are applied.
5Target peptides are measured in LC-MS/MS.
6In cosmetics, allergenic scented substances are screened with GC-MS.

Allergen Acceptance Criteria

In food (EU) 1169/2011, Annex II allergens must be declared highlighted in the ingredient list. It sets the (EU) 828/2014 defined threshold for the 'gluten-free' claim. Annex III of Cosmetics (EC) 1223/2009 requires that listed perfume allergens be separately stated on the label if they are present in above certain concentrations.

Evaluation of Results

A negative result is limited to the detection limit of the method and does not guarantee 'allergen-free'. Additionally, processing may alter the target protein, decreasing the ELISA result; In this case, PCR or LC-MS/MS is more reliable. Cross-contamination management is achieved through production separation and cleaning verification, not analysis.

Allergen Report

Sample description and target allergen list
The method applied and its rationale
Result and detection limit for each allergen
Processing parameters information
Comparing legislation threshold or statement

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 authorisations

Related Regulations

Regulation (EU) 1169/2011 on Food Information to Consumers Annex II
Gluten-free declaration Regulation (EU) 828/2014
(EC) 1223/2009 Cosmetic Products Regulation Annex III

Kozmetik Testleri — What Does This Testing Field Cover?

The section below describes the field that the Allergen 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 Allergen

Shouldn't we have an analysis instead of a 'may contain' warning?

Analysis is part of demonstrating that the risk of cross-contamination is managed, but it is not sufficient on its own. The warning statement must be based on a genuine risk assessment; The analysis supports, not replaces, this assessment.

Why is ELISA low in heat-treated products?

Heat treatment changes the structure of the protein and can disrupt the regions recognized by the antibody. In this case, the result may be lower than the truth; DNA-targeted PCR or peptide-targeted LC-MS/MS would be more appropriate.