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Background: Omics technologies have been widely applied in different fields, among which, proteomics is gaining increasing interest for its application to the authenticity of food products. MS, typically coupled with LC, represents a key technique for proteomics-related studies dedicated to fish and other seafood products by using a bottom-up approach. Objective and Methods: In this paper, the optimization of an untargeted proteomics-based method using LC separation and MS detection relying on a quadrupole time-of-flight mass spectrometer is described and applied to the analysis of Canadian farmed and wild-type salmon, followed by statistical analysis based on principal component (PC) analysis. Results and Conclusions: This untargeted approach, using a data-independent acquisition MS scheme, demonstrated the ability to effectively discriminate salmon belonging to the two classes. Furthermore, selected peptides showing high loadings on PC1 could represent potential candidate peptide markers able to discriminate farmed from wild-type salmon samples in the future.

The detection of food fraud and undeclared food allergens is one of the major challenges for competent authorities. Because adulterations are continuously adapted to the methods used to uncover them, the accomplishment of this task has become increasingly difficult over time. In recent years, various new promising methods for the detection of multiple food adulterants and multiple food allergens have been developed. Some of them utilize LC-MS to identify specific marker peptides. However, these methods have yet to be validated and standardized. For this reason, the German officials have established a working group with the objective of validating methods through multilaboratory validation studies. The experts of the working group also aim for the first time to standardize validated methods and to develop general validation criteria. This manuscript will highlight the current work of the group. For this purpose, an overview is given on the principles and applications of the new mass spectrometric methods. Moreover, requirements and the present work of other institutions regarding method validation are described.

Allergic reactions to food are among the major food safety concerns in industrialized countries, and it is estimated that approximately 5% of the population suffers from immunoglobulin-E-mediated food allergy. High-resolution mass spectrometry has become one of the most important techniques for the molecular characterization of allergens, including structural modification, degradation in the gastrointestinal environment, or identification of suitable marker peptides for the development of novel analytical approaches, in the past decade. This perspective aims to briefly summarize the current situation and discuss future developments.

The prevalence of allergic reactions to food is believed to be increasing in industrialized countries worldwide. One of the major tasks in risk management is, therefore, the analytical surveillance of allergen contamination in food and targeted proteomics using MS, which is of hugely growing interest due to its specificity and sensitivity and the possibility to analyze multiple allergens in parallel. Though approximately 200 different foods have been described as having the potential to elicit allergic reactions, current regional labeling requirements are focused on the 5-14 priority allergens that elicit the vast majority of severe reactions or that pose a risk as hidden allergens in food production. MS-based detection methods have been published for the majority of priority allergens, and this review provides an overview of the different methodological approaches, namely multiple-reaction monitoring-, high-resolution MS-, and triple-stage MS-based methods. In addition, requirements for the identification and validation of specific marker peptides and the influence of thermal processing and structural heterogeneity of allergens are discussed.

Crustacean shellfish allergy ranks among the most frequent and severe food allergies for adults, demanding rugged and sensitive analytical routine methods. The objective of this study was therefore to develop a mass spectrometric approach for the detection of contamination with shrimp and lobster, two economically important types of crustaceans, in complex food matrices. Following a biomarker approach, we identified proteotypic peptides and developed a multiple reaction monitoring (MRM) method allowing for the identification and differentiation of shrimp and lobster in the food matrix at concentrations down to 0.1%. To further enhance sensitivity, we employed the MRM-cubed (MRM(3)) mode, which allowed us to detect crustaceans down to concentrations of 25 μg/g (crustacean/food, 0.0025%). We hereby present the first mass spectrometric method for the detection of shrimp and lobster in food matrices.

Food allergies have emerged as a global problem over the last few decades; therefore, reliable and sensitive analytical methods to ensure food safety for allergic consumers are required. The application of mass spectrometry is of growing interest in this field and several procedures based on low resolution tandem mass spectrometry using single tryptic peptides as analytical targets have recently been described. However, a comprehensive survey of marker peptides for the development of multi-methods is still missing, as is a consensus guide to marker identification. In this study, we therefore report a consistent approach to the development of liquid chromatography-mass spectrometry (LC-MS) multi-screening methods for the detection of allergens in food matrices. Proteotypic peptides were identified by a shotgun proteomics approach and verified through a thorough investigation of specificity and sensitivity. On the basis of this procedure, we identified 44 suitable tryptic marker peptides from six allergenic nut species and developed the first analytical LC-MS method for the detection of trace nut contaminations in processed foods using high resolution mass spectrometry (HRMS). The analysis of spiked matrix samples gave limits of detection (LODs) below 10 μg/g for several nuts; these LODs are comparable with routinely used methods such as ELISA and PCR. Notably, the HRMS approach can be used in an untargeted fashion to identify multiple allergens also retrospectively. In conclusion, we present here the so far largest consensus set of analytical markers from nut allergens and to the best of our knowledge the first multi-allergen method based on LC-HRMS.

The application of mass spectrometry-based proteomics to address different aspects of Food Quality and Food Safety is of hugely growing interest. This article reviews recent developments and applications in the field of food proteomics with special attention on food authentication, process-induced modification, nutriproteomics and different aspects of food safety such as allergen detection and celiac disease. [ABSTRACT FROM AUTHOR]

Abstract Food allergies are a growing worldwide concern and the contamination of products with food allergens represents a significant health risk to allergic consumers. With the introduction of reference doses, quantitative methods are needed for the monitoring of allergen levels, and the potential of LC-MS/MS is of hugely growing interest. In this study, we demonstrate that relevant food matrices (bakery products and chocolates) and thermal food processing substantially influence the quantification of 18 marker peptides from various nut and peanut allergens via targeted proteomics. In addition, we characterize the individual release kinetics of marker peptides and provide examples for metastable marker peptide candidates. Matrix recovery rates overall ranged between 15 and 250% with the observed variation being linked to the individual peptide structure as well as to specific matrix interferences. In contrast, thermal processing considerably influences the detectability of allergens on the protein level as different marker peptides from the identical parent allergen are similarly affected, leading to a loss in signal of up to 83% in extreme cases after a 45-min simulated baking. Provided data are finally used for evaluation of different calibrators as well as the overall potential and challenges of LC-MS for the absolute quantification of food allergens. Significance With the scientific discussion moving towards a risk-based management of food allergens, including the establishment of threshold doses, robust methods for the absolute quantification of allergens in food samples are urgently needed. Because the currently used antibody- and DNA-based technologies show severe limitations in terms of specificity and reproducibility, LC-MS has emerged as a promising alternative. Its application to absolute quantification, however, first requires an understanding of the various impacts that affect quantification results, including different food matrices, sample preparation, and thermal processing of foodstuffs. Knowledge of these factors, which are assessed as part of a comprehensive survey in this study, is also an important prerequisite to evaluate means of calibration for an LC-MS-based quantification of food allergens. Graphical abstract Unlabelled Image Highlights • Comparative analysis of release kinetics of marker peptides from parent allergens. • Characterization of individual matrix interference with allergen markers. • Assessment of thermal processing on allergen recovery. • Pitfalls and potential solutions for absolute allergen quantification. [ABSTRACT FROM AUTHOR]

[Display omitted] • A range of raw and processed pork/beef (P/B) products were prepared and analysed. • HPLC-MS/MS (MRM mode) was used for relative quantification of meat composition. • Species-specific and global peptide markers were used for validating P/B contents. • Combined analysis of the peptides can be used for meat composition authentication. We used global and species-specific peptide markers for a relative quantitative determination of pork and beef in raw and processed meat products made of the two meat species. Four groups of products were prepared (i.e., minced raw meats, sausages, raw and fried burgers) in order to represent products with different extents of food processing. In each group, the products varied in the pork/beef proportions. All products were analysed by multiple reaction monitoring mass spectrometry (MRM–MS) for the presence/concentration of pork- and beef-specific peptide markers, as well as global markers – peptides widely distributed in muscle tissue. The combined MRM-MS analysis of pork-specific peptide HPGDFGADAQGAMSK, beef-specific peptide VLGFHG and global marker LFDLR offered the most reliable validation of declared pork/beef compositions across the whole range of meat products. Our work suggests that a simultaneous analysis of global and species-specific peptide markers can be used for composition authentication in commercial pork/beef products. [ABSTRACT FROM AUTHOR]