Analytical and Bioanalytical Chemistry

, Volume 410, Issue 22, pp 5653–5662 | Cite as

In house validation of a high resolution mass spectrometry Orbitrap-based method for multiple allergen detection in a processed model food

  • Rosa Pilolli
  • Elisabetta De Angelis
  • Linda MonaciEmail author
Research Paper
Part of the following topical collections:
  1. Food Safety Analysis


In recent years, mass spectrometry (MS) has been establishing its role in the development of analytical methods for multiple allergen detection, but most analyses are being carried out on low-resolution mass spectrometers such as triple quadrupole or ion traps. In this investigation, performance provided by a high resolution (HR) hybrid quadrupole-Orbitrap™ MS platform for the multiple allergens detection in processed food matrix is presented. In particular, three different acquisition modes were compared: full-MS, targeted-selected ion monitoring with data-dependent fragmentation (t-SIM/dd2), and parallel reaction monitoring. In order to challenge the HR-MS platform, the sample preparation was kept as simple as possible, limited to a 30-min ultrasound-aided protein extraction followed by clean-up with disposable size exclusion cartridges. Selected peptide markers tracing for five allergenic ingredients namely skim milk, whole egg, soy flour, ground hazelnut, and ground peanut were monitored in home-made cookies chosen as model processed matrix. Timed t-SIM/dd2 was found the best choice as a good compromise between sensitivity and accuracy, accomplishing the detection of 17 peptides originating from the five allergens in the same run. The optimized method was validated in-house through the evaluation of matrix and processing effects, recoveries, and precision. The selected quantitative markers for each allergenic ingredient provided quantification of 60–100 μgingred/g allergenic ingredient/matrix in incurred cookies.


High resolution mass spectrometry Multi-allergen detection Processed matrix Incurred samples Peptide marker In house validation 



Roberto Schena is kindly acknowledged for his technical aid in performing MS measurements. Besana group S.p.A. is also acknowledged for kindly providing hazelnuts and peanuts.


The work was funded by the project Safe & Smart—Nuove tecnologie abilitanti per la food safety e l’integrità delle filiere agro-alimentari in uno scenario globale—National CL.AN-Cluster agroalimentare nazionale programma area 2. The equipment used in this work was supported by the “Biodiversità per la valorizzazione e sicurezza delle produzioni alimentari tipiche pugliesi, BioNet-PTP” project (Cod. 73) funded by Programma Operativo Regionale Puglia FESR 2000-2006 - Risorse liberate - Obiettivo Convergenza.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Not applicable.

Supplementary material

216_2018_927_MOESM1_ESM.pdf (220 kb)
ESM 1 (PDF 219 kb)


  1. 1.
    Monaci L, Visconti A. Mass spectrometry-based proteomics methods for analysis of food allergens. Trends Anal Chem. 2009;28:581–91.CrossRefGoogle Scholar
  2. 2.
    Monaci L, Pilolli R, De Angelis E, Mamone G. Mass spectrometry in food allergen research. In: Picó Y, editor. Comprehensive analytical chemistry. Elsevier Science and Technology; 2015. pp. 359–393.Google Scholar
  3. 3.
    Heick J, Fischer M, Pöpping B. First screening method for the simultaneous detection of seven allergens by liquid chromatography mass spectrometry. J Chromatogr A. 2011;1218:938–43.CrossRefPubMedGoogle Scholar
  4. 4.
    Pilolli R, De Angelis E, Godula M, Visconti A, Monaci L. Orbitrap™ monostage MS versus hybrid linear ion trap MS: application to multi-allergen screening in wine. J Mass Spectrom 2014;49:1254–1263.Google Scholar
  5. 5.
    Parker CH, Khuda SE, Pereira M, Ross MM, Fu TJ, Fan X, et al. Multi-allergen quantitation and the impact of thermal treatment in industry-processed baked goods by ELISA and liquid chromatography-tandem mass spectrometry. J Agric Food Chem. 2015;63:10669–80.CrossRefPubMedGoogle Scholar
  6. 6.
    Korte R, Monneuse JM, Gemrot E, Metton I, Humpf H-U, Brockmeyer J. New high-performance liquid chromatography coupled mass spectrometry method for the detection of lobster and shrimp allergens in food samples via multiple reaction monitoring and multiple reaction monitoring cubed. J Agric Food Chem. 2016;64:6219–27.CrossRefPubMedGoogle Scholar
  7. 7.
    Planque M, Arnould T, Dieu M, Delahaut P, Renard P, Gillard N. Advances in ultra-high performance liquid chromatography coupled to tandem mass spectrometry for sensitive detection of several food allergens in complex and processed foodstuff. J Chromatogr A. 2016;1464:115–23.CrossRefPubMedGoogle Scholar
  8. 8.
    Korte R, Brockmeyer J. MRM3-based LC-MS multi-method for the detection and quantification of nut allergens. Anal Bioanal Chem. 2016;408:7845–55.CrossRefPubMedGoogle Scholar
  9. 9.
    De Angelis E, Pilolli R, Monaci L. Coupling SPE on-line pre-enrichment with HPLC and MS/MS for the sensitive detection of multiple allergens in wine. Food Control. 2017;73:814–20.CrossRefGoogle Scholar
  10. 10.
    Hoffmann B, Münch S, Schwägele F, Neusüß C, Jira W. A sensitive HPLC-MS/MS screening method for the simultaneous detection of lupine, pea, and soy proteins in meat products. Food Control. 2017;71:200–9.CrossRefGoogle Scholar
  11. 11.
    Huschek G, Bӧnick J, Lӧwenstein Y, Sievers S, Rawel H. Quantification of allergenic plant traces in baked products by targeted proteomics using isotope marked peptides. LWT-Food Sci Technol. 2016;74:286–93.CrossRefGoogle Scholar
  12. 12.
    Chen Q, Zhang J, Ke X, Lai S, Tao B, Yang J, et al. Quantification of bovine β-casein allergen in baked foodstuffs based on ultra-performance liquid chromatography with tandem mass spectrometry. Food Addit Contam Part A. 2015;32:25–34.CrossRefGoogle Scholar
  13. 13.
    Monaci L, Pilolli R, De Angelis E, Godula M, Visconti A. Multi allergen detection in food by micro high performance liquid chromatography coupled to a dual cell linear ion trap mass spectrometry. J Chromatogr A. 2014;1358:136–44.CrossRefPubMedGoogle Scholar
  14. 14.
    Korte R, Lepski S, Brockmeyer J. Comprehensive peptide marker identification for the detection of multiple nut allergens using a non-targeted LC–HRMS multi-method. Anal Bioanal Chem. 2016;408:3059–69.CrossRefPubMedGoogle Scholar
  15. 15.
    Monaci L, De Angelis E, Bavaro SL, Pilolli R. High-resolution Orbitrap™-based mass spectrometry for rapid detection of peanuts in nuts. Food Addit Contam: Part A. 2015;32:1607–16.CrossRefGoogle Scholar
  16. 16.
    European Commission Commission Directive 2007/68/EC. Off J Eur Union. 2007;L310:11–4.Google Scholar
  17. 17.
    Monaci L, Pilolli R, De Angelis E, Carone R, Pascale M. LC-tandem mass spectrometry as a screening tool for multiple detection of allergenic ingredients in complex foods. ACTA IMEKO. 2016;5:5–9.CrossRefGoogle Scholar
  18. 18.
    Pilolli R, De Angelis E, Monaci L. Streamlining the analytical workflow for multiplex MS/MS allergen detection in processed foods. Food Chem. 2017;221:1747–53.CrossRefPubMedGoogle Scholar
  19. 19.
    United States Department of Agriculture Agricultural Research Service. In: USDA Food Composition Databases. USA National Agricultural Library. 2017. Accessed 10 July 2017.

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Rosa Pilolli
    • 1
  • Elisabetta De Angelis
    • 1
  • Linda Monaci
    • 1
    Email author
  1. 1.Institute of Sciences of Food ProductionNational Research Council (ISPA-CNR)BariItaly

Personalised recommendations