Simultaneous Determination of Animal Products from Ruminant, Pig, Poultry, and Fish in Feedingstuff by Targeted High-Resolution LC–MS/MS

Abstract

The prohibition of processed animal proteins (PAPs) has been relaxed gradually since 2007. The official control method for PAPs in feedingstuff, a combination of light microscopy (LM) followed by PCR, is no longer sufficient. Thus, a targeted LC−MS/MS method was developed, which enables a tissue-specific distinction between egg and dairy products, gelatine, and PAPs derived from blood or muscle tissue of the species ruminants, pigs, poultry, and fish. Tissue-specific proteins were analyzed after tryptic digestion to peptides with high-resolution ESI-QTOF-MS. A targeted method was developed based on untargeted proteomics approaches and the selection of specific peptides (45 unique peptides in total). Proficiency testing of blank and spiked samples revealed excellent results for trueness and selectivity. Furthermore, sensitivity was achieved at a level of 0.1% (w/w) for assessed peptides. Summing up, the developed method seems to be suitable for routine analysis after verification by ring trials.