Food Allergen Detection: ELISA versus Mass Spectrometry

The eight major foods linked to allergies are peanut and tree nut, milk, egg, soy, wheat, fish and shellfish. Such allergens can be detected using a variety of methods, however, this article compares ELISA to mass spectrometry.

Food allergens can be detected using Mass Spectrometry or ELISATassaneeT | Shutterstock

While foods containing the above eight ingredients need to be labeled in certain countries, this is only required if these are added intentionally. Therefore, this means that traces which could have come from other food items being prepared in the same factory may be present in foods which are made without these ingredients, e.g. due to shared surfaces. Consequently, there is a need for food allergen detection.

Why is it important to test food for allergens?

Manufacturers do place “may contain…” advisory notices and one study found that 43% of chocolate with an advisory label showed traces of peanuts. However, the same study also showed that 25% of chocolate without an advisory label were found to contain peanuts.

A different study found that 75% of products with an advisory label showed traces of milk, while some other products without an advisory label (or with “dairy free” label) also showed traces of milk. This shows that, while these advisory labels are accurate in some instances, there are other cases where these are unnecessary.  

Over-labelling with advisory notices has the potential to create a scenario whereby these are ignored. Hence, more accurate labeling would be preferred, which would stem from the accurate quantification of allergens.

How is the testing for food allergens done?

In Enzyme-Linked Immunosorbent Assay's (ELISAs), antibodies, an enzyme, and its substrate are used to detect proteins found in food which can be allergens to some people; i.e. proteins found in peanuts. There are variations in the ELISA methods which differ in the number of antibodies used and whether the target protein is “captured” first or not.

Once the antibodies have been added to the reaction and allowed to bind, the substrate for the enzyme linked to the last antibody is added. This results in a change which can be measured, and these measurements can be used to quantify the amount of target protein present in the food.

Mass Spectrometry is a technique which measures the different masses of molecules within a sample, including proteins. These molecules are initially vaporized and is then hit by electrons. This gives each molecule or fragments of molecules a charge; if electrons are lost from the molecule, the molecule becomes positively charged, while the addition of an electron to the molecule will result in a negative charge.

These ions are then sorted by charge and size according to how fast they travel, and also how much they are deflected by when a magnetic field is applied. Mass spectrometry can also be coupled to chromatography, such as liquid chromatography (LC-MS).

One big difference between ELISA and mass spectrometry is that ELISA detects the presence of proteins, while mass spectrometry detects the presence of peptides, which are fragments derived from proteins. ELISA is limited to the detection of one protein, while multiple peptides can be detected by mass spectrometry.

What are the advantages and disadvantages of each method?

It has been reported that ELISA is sensitive, and has the ability to detect allergens at levels of ~0.1 – 5mg kg-1. It is difficult, however, to know at what quantity the protein becomes an allergen, due to differences in individual reactions. Another advantage of ELISA is that it is relatively easy to use and there are readily available standards and validation tools.

A disadvantage of ELISA is that, if the target protein has been altered (perhaps by heating), then the antibodies used may no longer be able to recognize the target protein. This results in a false negative, which could have dangerous consequences for the consumer. Conversely, if the target protein is similar to other proteins found in food, an ELISA could report a false positive result as the allergen is not present.

Differences in preparing the food for testing may also affect the ELISA and thus the final results. Other disadvantages include the fact that different antibodies and standards can be used for the same target protein, which may lead to different results.

As mentioned before, mass spectrometry can detect multiple target peptides at once, while retaining the same level of sensitivity as ELISA. However, there can be some levels of assumption when quantifying peptides from mass spectrometry data, therefore this could be a disadvantage. Another advantage is that the structure of proteins is not important, as mass spectrometry looks at its peptides.

The main disadvantage of mass spectrometry is that if the starting material is highly complex, then the sensitivity may actually be impacted. This could potentially be overcome by first removing dominant proteins and peptides in order to look for other proteins which are not as dominant.

Further Reading

Last Updated: Mar 26, 2019

Dr. Maho Yokoyama

Written by

Dr. Maho Yokoyama

Dr. Maho Yokoyama is a researcher and science writer. She was awarded her Ph.D. from the University of Bath, UK, following a thesis in the field of Microbiology, where she applied functional genomics to Staphylococcus aureus . During her doctoral studies, Maho collaborated with other academics on several papers and even published some of her own work in peer-reviewed scientific journals. She also presented her work at academic conferences around the world.


Please use one of the following formats to cite this article in your essay, paper or report:

  • APA

    Yokoyama, Maho. (2019, March 26). Food Allergen Detection: ELISA versus Mass Spectrometry. News-Medical. Retrieved on July 15, 2024 from

  • MLA

    Yokoyama, Maho. "Food Allergen Detection: ELISA versus Mass Spectrometry". News-Medical. 15 July 2024. <>.

  • Chicago

    Yokoyama, Maho. "Food Allergen Detection: ELISA versus Mass Spectrometry". News-Medical. (accessed July 15, 2024).

  • Harvard

    Yokoyama, Maho. 2019. Food Allergen Detection: ELISA versus Mass Spectrometry. News-Medical, viewed 15 July 2024,


The opinions expressed here are the views of the writer and do not necessarily reflect the views and opinions of News Medical.
Post a new comment

While we only use edited and approved content for Azthena answers, it may on occasions provide incorrect responses. Please confirm any data provided with the related suppliers or authors. We do not provide medical advice, if you search for medical information you must always consult a medical professional before acting on any information provided.

Your questions, but not your email details will be shared with OpenAI and retained for 30 days in accordance with their privacy principles.

Please do not ask questions that use sensitive or confidential information.

Read the full Terms & Conditions.