Mass spectrometry detection of protein allergenic additives in emulsion-type pork sausages

Montowska, Magdalena; Fornal, Emilia; Piątek, Michał; Krzywdzińska-Bartkowiak, Mirosława

doi:10.1016/j.foodcont.2019.04.022

Cited by 17 publications

(12 citation statements)

References 22 publications

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“…Three peptides (8–18 amino acids) for each legume species were finally chosen as peptide markers from the group of the candidate marker peptides due to their highest intensities in spiked sausages (test sausage T4, see Table 1 ) applying the optimized MRM method (see Section 3.2 .). The final selected peptides QQEQQLEGELEK [ 25 ], NTLEATFNTR [ 25 , 31 ], and ISSVNSLTLPILR [ 25 ] for lupine blue, NPYHFSSQR [ 31 ] for lupine white, ELTFPGSVQEINR, LSSGDVFVIPAGHPVAVK, and LTPGDVFVIPAGHPVAVR for pea [ 25 ], GTGNLELVAVR [ 32 , 33 , 34 , 35 , 36 ], FNLAGNHEQEFLR [ 32 , 33 , 37 , 38 ], and WLGLSAEYGNLYR [ 32 , 34 ] for peanut, and SQSDNFEYVSFK [ 23 , 34 , 36 ], EAFGVNMQIVR [ 25 , 34 , 38 , 39 ], and FYLAGNQEQEFLK [ 22 , 23 , 25 , 36 ] for soy have been reported previously in the scientific literature. The identities of each of the 14 marker peptides for alfalfa, broad bean, chickpea, and lentil, as well as lupine white 2 and 3, which have not been reported in the literature until now, were confirmed by spiking in a tryptic digest of an emulsion-type sausage (test sausage T4).…”

Section: Resultsmentioning

confidence: 99%

“…In contrast to ELISA and PCR, mass spectrometric methods have the advantage of being high-throughput screening tools for the multiplex detection of target proteins in foods [ 19 , 21 ] and are less vulnerable to interferences in complex food matrices [ 19 ]. Only a few analytical methods exist for the HPLC-MS/MS-detection of legumes in meat products, which are limited to soy [ 22 , 23 , 24 ] or lupine, pea, and soy [ 25 ]. However, other legume species are also potential foreign protein sources in meat products, as mentioned before, for which no detection methods to uncover this type of meat adulteration have been available until now.…”

Section: Introductionmentioning

confidence: 99%

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A UHPLC-MS/MS Method for the Detection of Meat Substitution by Nine Legume Species in Emulsion-Type Sausages

Spörl

Speer²,

Jira

2021

Foods

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Meat substitution by legume proteins in various types of meat products is a common practice. A reliable detection and quantification of these additives is required to control food specifications, especially regarding food fraud. Consequently, a UHPLC-MS/MS method for the simultaneous detection of alfalfa (Medicago sativa), broad bean (Vicia faba), chickpea (Cicer arietinum), lentil (Lens culinaris), lupine (Lupinus albus and Lupinus angustifolius), pea (Pisum sativum), peanut (Arachis hypogaea), and soy (Glycine max) proteins in meat products was developed. After protein extraction and tryptic digestion, three marker peptides for each legume species were measured by multiple reaction monitoring (MRM) using an optimized extraction protocol. To the best of our knowledge, the marker peptides for alfalfa, broad bean, chickpea, and lentil have not been reported previously. Emulsion-type sausages with 0.1, 0.4, 0.7, 1.0, 1.3, 1.6, 1.9, 2.2, and 2.5% meat substitution by each legume species, representing the concentration range between inadvertently transferred cross-contaminations and the conscious use for meat substitution, were produced for matrix calibration. No false-positive results were recorded in blank samples. In the quantification of alfalfa, broad bean, chickpea, lentil, pea, peanut, and soy, 673 of 756 measuring data of the recovery rate in unknown sausages were in the accepted range of 80–120%.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A UHPLC-MS/MS Method for the Detection of Meat Substitution by Nine Legume Species in Emulsion-Type Sausages

Spörl

Speer²,

Jira

2021

Foods

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“…Sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) was performed to compare the protein profiles of oilseed cakes. A sample of 5 mg was solubilized with lysis buffer and SDS‐PAGE analysis was performed according to the procedure described previously 16 . The protein concentration was determined using a 2‐D Quant kit (GE Healthcare Bio‐Sciences, Fairfield, CT, USA).…”

Section: Methodsmentioning

confidence: 99%

Proteomic analysis of oilseed cake: a comparative study of species‐specific proteins and peptides extracted from ten seed species

et al. 2020

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Background In recent years there has been a visible trend among consumers to move away from consuming meat in favor of plant products. Meat producers have therefore been trying to meet the expectations of consumers by introducing new products to the food market with a greater proportion of plant ingredients. Meat products are enriched not only by the addition of vegetable oils but also by ground or whole oilseeds or their preparation. In this study, we present in‐solution tryptic digestion and an ultra‐performance liquid chromatography coupled to quadrupole time‐of‐flight mass spectrometry (UHPLC‐Q‐TOF‐MS/MS)‐based proteomics approach to investigate specific proteins and peptides of ten oilseed cakes, by‐products of cold pressing oil from coconut, evening primrose, hemp, flax, milk thistle, nigella, pumpkin, rapeseed, sesame, and sunflower seeds, for authentication purposes. Results We identified a total of 229 unique oilseed proteins. The number of specific proteins varied depending on the sample, from 4 to 48 in evening primrose and sesame. Moreover, we identified approximately 440 oilseed unique peptides in the cakes of all the analyzed oilseeds; the largest amounts were found in sesame (107 peptides), sunflower (100), pumpkin, hemp (42), rapeseed (36), and flax cake (35 peptides). Conclusions We provide novel information on unique / species‐specific peptide markers that will extend the scope of testing the authenticity of a wide range of foods. The results of this peptide discovery experiment may further contribute to the development of targeted methods for the detection and quantification of oilseed proteins in processed foods, and thus to the improvement of food quality. © 2020 Society of Chemical Industry

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“…Protein digestion was carried out as previously described [ 45 ]. Dried samples (5 mg) were rehydrated in 100 μL of 50 mM ammonium bicarbonate.…”

Section: Methodsmentioning

confidence: 99%

Multispecies Identification of Oilseed- and Meat-Specific Proteins and Heat-Stable Peptide Markers in Food Products

et al. 2021

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Consumer demand for both plant products and meat products enriched with plant raw materials is constantly increasing. Therefore, new versatile and reliable methods are needed to find and combat fraudulent practices in processed foods. The objective of this study was to identify oilseed species-specific peptide markers and meat-specific markers that were resistant to processing, for multispecies authentication of different meat and vegan food products using the proteomic LC-MS/MS method. To assess the limit of detection (LOD) for hemp proteins, cooked meatballs consisting of three meat species and hemp cake at a final concentration of up to 7.4% were examined. Hemp addition at a low concentration of below 1% was detected. The LOD for edestin subunits and albumin was 0.9% (w/w), whereas for 7S vicilin-like protein it was 4.2% (w/w). Specific heat-stable peptides unique to hemp seeds, flaxseed, nigella, pumpkin, sesame, and sunflower seeds, as well as guinea fowl, rabbit, pork, and chicken meat, were detected in different meat and vegan foods. Most of the oilseed-specific peptides were identified as processing-resistant markers belonging to 11S globulin subunits, namely conlinin, edestin, helianthinin, pumpkin vicilin-like or late embryogenesis proteins, and sesame legumin-like as well as 2S albumins and oleosin isoforms or selected enzymic proteins.

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Mass spectrometry detection of protein allergenic additives in emulsion-type pork sausages

Cited by 17 publications

References 22 publications

A UHPLC-MS/MS Method for the Detection of Meat Substitution by Nine Legume Species in Emulsion-Type Sausages

A UHPLC-MS/MS Method for the Detection of Meat Substitution by Nine Legume Species in Emulsion-Type Sausages

Proteomic analysis of oilseed cake: a comparative study of species‐specific proteins and peptides extracted from ten seed species

Multispecies Identification of Oilseed- and Meat-Specific Proteins and Heat-Stable Peptide Markers in Food Products

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