Skip to main content
Log in

A rapid UHPLC-MS/MS screening method for the detection of the addition of porcine blood plasma to emulsion-type pork sausages

  • Research Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

The adulteration of meat products by the undeclared addition of commercially available blood plasma powder is quite conceivable due to low costs, high protein contents (about 70%), and advantageous functional properties. This applies particularly to pork, which has the highest meat production rate in the European Union. Evidence of this type of food fraud has been rather difficult to identify due to the lack of appropriate analytical methods, especially when adding plasma to meat of the same animal species. Consequently, a rapid UHPLC-MS/MS method for the detection of porcine blood plasma in emulsion-type pork sausages was developed. After protein extraction and tryptic digestion in a quick and simple one-pot process, species-specific marker peptides for porcine blood cell proteins (four markers) and plasma proteins (12 markers) were measured by UHPLC-MS/MS. Emulsion-type pork sausages were produced from a variety of raw materials that differed in the age or sex of the slaughtered pigs. Sausages were spiked with 0.5, 1, 1.5, 2, 3, or 5% meat substitution by one of two plasma powders, or produced as corresponding blank samples, and subjected to different thermal treatments as full or semi-preserves. Four plasma peptides were identified for the overall sample that allowed detection down to 0.7% meat substitution from the sum of their peak areas, with 5% error probability for both false positives and negatives.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Hees H, Tschudi P. Blut und Knochenmark. In: Mosimann W, Kohler T, editors. Zytologie, Histologie und mikroskopische Anatomie der Haussäugetiere. Berlin, Hamburg: Verlag Paul Parey; 1990. p. 98–113.

    Google Scholar 

  2. Warriss PD. Meat science: an introductory text. Wallingford: CABI Publishers; 2010.

    Google Scholar 

  3. Gatnau R. Spray dried porcine plasma as a source of protein and immunoglobulins for weanling pigs. Retrospective theses and dissertations. 16877. 1990. https://lib.dr.iastate.edu/rtd/16877. Accessed 2 May 2019.

  4. European Commission (2019): https://ec.europa.eu/agriculture/sites/agriculture/files/market-observatory/meat/pigmeat/doc/dashboard-pig_en.pdf. Accessed 2 June 2019.

  5. Tändler K, Stiebing A. Utilisation of blood, bloodplasma and bloodserum as human food. 24th International Congress of Meat Science and Technology, Kulmbach, Germany; 1978.

  6. Scheunemann R. Das 5. Viertel – Die sinnvolle Verwertung von Schlachtnebenprodukten (SNP). RFL. 2011;11:377–8.

    Google Scholar 

  7. Bah CSF, Bekhit AEA, Carne A, McConnell MA. Slaughterhouse blood: an emerging source of bioactive compounds. Compr Rev Food Sci F. 2013;12:314–31.

    Article  CAS  Google Scholar 

  8. Tarte R. Meat-derived protein ingredients. In Tarte R, editor. Ingredients in meat products: properties, functionality and applications. Springer Science + Business Media, LLC; 2009. pp. 145–171.

  9. European Union. (2009). Regulation (EC) No. 1069/2009 of the European Parliament and of the Council of 21 October 2009 laying down rules as regards animal by-products and derived products not intended for human consumption and repealing Regulation (EC) No 1774/2002 (Animal by-products Regulation). Official Journal of the European Union. 14.11.2009. L 300/1–33.

  10. European Union. (2004). Regulation (EC) No. 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for on the hygiene of foodstuffs. Official Journal of the European Union. 30.04.2004. L 139/55–205.

  11. Ockerman HW, Hansen CL. Animal by-product processing & utilization. Boca Raton: CRC Press; 2000. p. 79–81.

    Google Scholar 

  12. Feiner G. Meat products handbook—practical science and technology. Boca Raton: CRC Press; 2006. p. 95–7.

    Book  Google Scholar 

  13. Hargin KD. Authenticity issues in meat and meat products. Meat Sci. 1996;43:277–89.

    Article  Google Scholar 

  14. Davila E, Pares D, Cuvelier G, Relkin P. Heat-induced gelation of porcine blood plasma proteins as affected by pH. Meat Sci. 2007;76:216–25.

    Article  CAS  PubMed  Google Scholar 

  15. Lu GH, Chen TC. Application of egg white and plasma powders as muscle food binding agents. J Food Eng. 1999;42:147–51.

    Article  Google Scholar 

  16. Hurtado S, Daga I, Espigule E, Pares D, Saguer E, Toldra M, et al. Use of porcine blood plasma in “phosphate-free frankfurters”. Procedia Food Sci. 2011;1:477–82.

    Article  CAS  Google Scholar 

  17. Kim S, Jin S, Choi J. Effects of the addition of blood plasma proteins on physico-chemical properties of emulsion-type pork sausage during cold storage. J Sci Food Agric. 2017;97:4501–7.

    Article  CAS  PubMed  Google Scholar 

  18. European Union. (2011). Regulation (EU) No. 1169/2011 of the European Parliament and of the Council of 25 October 2011 on the provision of food information to consumers, amending Regulations (EC) No. 1924/2006 and (EC) No. 1925/2006 of the European Parliament and of the Council, and repealing Commission Directive 87/250/EEC, Council Directive 90/496/EEC, Commission Directive 1999/10/EC, Directive 2000/13/EC of the European Parliament and of the Council, Commission Directives 2002/67/EC and 2008/5/EC and Commission Regulation (EC) No. 608/2004. Official journal of the European Union. 22.11.2011. Annex VI, Part A, 5.

  19. Leitsätze für Fleisch und Fleischerzeugnisse (2015). https://www.bmel.de/SharedDocs/Downloads/Ernaehrung/Lebensmittelbuch/LeitsaetzeFleisch.pdf?__blob=publicationFile. Accessed 2 June 2019.

  20. Maxstadt JJ, Pollmann RM. Spectrophotometric determination of sulfites, benzoates, sorbates, ascorbates, and added blood in ground beef—collaborative study. J Assoc Off Anal Chem. 1980;63:667–74.

    CAS  PubMed  Google Scholar 

  21. Bjarno OC. Multicomponent analysis of meat-products. J Assoc Off Anal Chem. 1981;64:1392–6.

    CAS  PubMed  Google Scholar 

  22. Bauer F, Stachelberger H. Detection of blood plasma in heat-treated meat products by ultrathin-layer isoelectric focusing. Z Lebensm Unters For. 1984;178:86–9.

    Article  CAS  Google Scholar 

  23. Otto W, Sinell HJ. Immunochemical detection of dried blood-plasma in heat processed mixtures with meat. Berl Munch Tierarztl. 1989;102:14–8.

    CAS  Google Scholar 

  24. Ofori JA, Hsieh YHP. Monoclonal antibodies as probes for the detection of porcine blood-derived food ingredients. J Agric Food Chem. 2016;64:3705–11.

    Article  CAS  PubMed  Google Scholar 

  25. Grundy HH, Reece P, Sykes MD, Clough JA, Audsley N, Stones R. Method to screen for the addition of porcine blood-based binding products to foods using liquid chromatography/triple quadrupole mass spectrometry. Rapid Commun Mass Sp. 2008;22:2006–8.

    Article  CAS  Google Scholar 

  26. Jira W, Schwägele F. A sensitive HPLC-MS/MS method for the simultaneous detection of microbial transglutaminase, and bovine and porcine fibrinogen/thrombin in restructured meat. Food Chem. 2017;237:841–8.

    Article  CAS  PubMed  Google Scholar 

  27. LFGB §64: Official Collection of Methods according to §64 of the German Food and Feed Code: L 06.00-3 (8/2014), L 06.00-4 (4/2007), L 06.00-6 (8/2014), and L 06.00-7 (8/2014).

  28. Jira W, Münch S. A sensitive HPLC-MS/MS screening method for the simultaneous detection of barley, maize, oats, rice, rye and wheat proteins in meat products. Food Chem. 2019;275:214–23.

    Article  CAS  PubMed  Google Scholar 

  29. Lecrenier MC, Marbaix H, Dieu M, Yeys P, Saegerman C, Raes M, et al. Identification of specific bovine blood biomarkers with a non-targeted approach using HPLC ESI tandem mass spectrometry. Food Chem. 2016;213:417–24.

    Article  CAS  PubMed  Google Scholar 

  30. Orduna AR, Husby E, Yang CT, Ghosh D, Beaudry F. Detection of meat species adulteration using high-resolution mass spectrometry and a proteogenomics strategy. Food Addit Contam A. 2017;34:1110–20.

    Article  CAS  Google Scholar 

  31. 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.

    Article  CAS  Google Scholar 

  32. Johnson PE, Baumgartner S, Aldick T, Bessant C, Giosafatto V, Heick J, et al. Current perspectives and recommendations for the development of mass spectrometry methods for the determination of allergens in food. J AOAC Int. 2011;94:1026–33.

    CAS  PubMed  Google Scholar 

  33. Johnson KD, Clark A, Marshall S. A functional comparison of ovine and porcine trypsins. Comp Biochem Physiol B. 2002;131:423–31.

    Article  Google Scholar 

  34. Elbers ARW, Counotte GHM, Tielen MJM. Haematological and clinicochemical blood profiles in slaughter pigs. Vet Q. 1992;14:57–62.

    Article  CAS  PubMed  Google Scholar 

  35. Warriss PD. Exsanguination of animals at slaughter and the residual blood content of meat. Vet Rec. 1984;115:292–5.

    Article  CAS  PubMed  Google Scholar 

  36. Halliday DA. Blood—a source of proteins. Process Biochem. 1973;8:15–7.

    Google Scholar 

  37. Hsieh YP, Ofori JA. Food-grade proteins from animal by-products: their usage and detection methods. In: Nollet LM, Toldra F, editors. Handbook of analysis of edible animal by-products. New York: CRC Press; 2011. p. 13–41.

    Chapter  Google Scholar 

  38. Stoyke M, Becker R, Brockmeyer J, Jira W, Popping B, Uhlig S, et al. German government official methods board points the way forward: launch of a new working group for protein analysis to detect food fraud and food allergens. J AOAC Int. 2019;102:1–6.

    Article  Google Scholar 

Download references

Acknowledgments

The authors would like to thank Sebastian Zimmermann for his valuable support in veterinarian questions, Yvonne Höhn and Liane Weber for their excellent technical assistance, and Manfred Behrschmidt, Josef Haida, and Marco Zäh for the professional production of the various sausages.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Wolfgang Jira.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(PDF 140 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Stader, C., Judas, M. & Jira, W. A rapid UHPLC-MS/MS screening method for the detection of the addition of porcine blood plasma to emulsion-type pork sausages. Anal Bioanal Chem 411, 6697–6709 (2019). https://doi.org/10.1007/s00216-019-02043-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-019-02043-2

Keywords

Navigation