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

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

Food Testing — 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.

Food testing shows that a product is safe for consumption and contains what its label states. The family works along three axes: microbiological safety, chemical safety and nutritional composition. All three require different laboratory infrastructure, a different body of methods and a different turnaround time.

The most distinctive feature of the field is TIME PRESSURE. A steel specimen can wait on a shelf; a raw milk or fresh meat sample cannot. The time the sample was taken, the transport temperature and how long it takes to reach the laboratory directly determine the validity of the result. In food microbiology the cold chain record is therefore an inseparable part of the test report.

On the microbiological side there are two different questions and they must not be confused. DETECTION of a pathogen is a qualitative question: is Salmonella present in the sample or not? ENUMERATION of indicator organisms is a quantitative question and an indicator of production hygiene. The absence of a pathogen in a product does not show that hygiene is good; if the total mesophilic aerobic count is high, there is a problem in the process.

On the chemical side, pesticide residues, mycotoxins, heavy metals, additives, veterinary drug residues and process contaminants (acrylamide, furan, PAH) are measured. Multi-residue methods in this area (QuEChERS-based LC-MS/MS and GC-MS/MS) can screen hundreds of active substances in a single analysis; limits are defined separately for each product–substance pair.

On the composition side, protein, fat, moisture, ash, fibre, sugar profile, fatty acid composition, vitamin and mineral content are determined. These values are the basis of the nutrition declaration on the label and the tolerance ranges are defined in legislation — the difference between the declared value and the measured value must stay within that tolerance.

There is NO CE marking on food, and this shapes how the field works. Conformity is demonstrated through operating a HACCP-based food safety system, complying with labelling rules and not exceeding contaminant, residue and microbiological limits. A test result produces evidence not for a certificate but for a functioning system.

Allergen management carries particular weight. The presence of an undeclared allergen in a product is one of the most frequent causes of food recalls. Analysis here runs together with cleaning validation: it is shown that the line has been cleaned adequately after a production run containing an allergen and that the risk of cross-contamination is under control.

Voluntary schemes have become effectively mandatory in this field. Retail chains require their suppliers to hold a certificate under a GFSI-recognised scheme (BRCGS, IFS, FSSC 22000); without it, getting onto the shelf is practically impossible.

Method groups in this field

Pathogen detection
Detection of Salmonella, Listeria monocytogenes, Campylobacter, Cronobacter and Shiga toxin-producing E. coli; culture-based and molecular methods.
Indicator counts
Total mesophilic aerobic count, Enterobacteriaceae, coliforms and E. coli, coagulase-positive staphylococci, yeasts and moulds, lactic acid bacteria, sulphite-reducing Clostridia.
Pesticide and veterinary residues
Multi-residue screening by LC-MS/MS and GC-MS/MS with QuEChERS extraction.
Mycotoxins and contaminants
Aflatoxins, ochratoxin A, deoxynivalenol, patulin; heavy metals (ICP-MS), acrylamide and PAH compounds.
Nutritional composition
Kjeldahl protein, fat and fatty acid profile, moisture, ash, dietary fibre, sugar profile, vitamin and mineral analyses.
Allergens and authenticity
Gluten, milk, egg, nut and soy allergen analyses; molecular methods for species identification and adulteration detection.

Commonly confused in this field

Sampling is half the result. A sample transported at the wrong temperature or delayed gives an invalid result even if it is analysed correctly in the laboratory.
The absence of a pathogen does not show that hygiene is good; indicator counts give separate information and the two are read together.
A result from a sample taken from a lot can be generalised to the whole lot only if the sampling plan was set up statistically.
Limits are defined in separate regulations by product group and are updated regularly; a conformity decision based on an outdated limit table may be wrong.
For exports, the limits of the destination country may differ from EU limits; the same product may be compliant in one market and non-compliant in another.

Which decisions these results support

Verification of the HACCP plan and process monitoring
The basis for the nutrition declaration and allergen information on the label
Health certificates before export and compliance with destination market limits
Supplier and lot-based acceptance control
Shelf-life studies and determining storage conditions
Producing evidence for audits under GFSI-recognised schemes

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.