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

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%.

Automated summary

A UHPLC-MS/MS method was developed for the simultaneous detection of various legume proteins in meat products. Marker peptides for alfalfa, broad bean, chickpea, and lentil were identified and used for quantification. Emulsion-type sausages were produced for matrix calibration, confirming the reliability and accuracy of the method in quantifying the proteins.