Reference : Specific detection of blood derived products in animal feed using UPLC-MS/MS
Scientific congresses and symposiums : Paper published in a book
Life sciences : Veterinary medicine & animal health
Life sciences : Agriculture & agronomy
Specific detection of blood derived products in animal feed using UPLC-MS/MS
Lecrenier, Marie-Caroline mailto [Université de Liège - ULiège > > > Doct. sc. vété. (Bologne)]
Dieu, Marc []
Veys, Pascal []
Planque, Mélanie []
Baeten, Vincent []
Abstracts of lectures and posters of the 11th Conference RME 2016
RME 2016 - 11th Rapid Methods Europe Conference
7-9 Novembre 2016
The Nederlands
[en] Blood derived products are valuable animal products used in feed for their nutritional value and their positive effects on growth and health. Nevertheless, since the BSE crisis, their use is strictly regulated. Blood meal and blood products (hemoglobin powder and plasma powder) of bovine origin are totally prohibited. Blood meal and blood products of porcine origin are authorised in aquafeed, whereas only blood products are allowed to be used in feed intended for other non-ruminant. The detection of the type of protein and the species of origin is therefore crucial to ensure feed safety.
With the current official methods for the detection of PAPs, light microscopy and PCR, it is not always possible to specifically identify this type of protein source. The objective of our work was to set-up a routine mass spectrometry method for a qualitative detection of blood meal and hemoglobin powder of bovine and porcine origin in feed. The method was based on the detection of species-specific peptides biomarkers identified in a previous study by a non-targeted approach. All biomarkers were peptides of α and β hemoglobin subunits.
Proteins were extracted using a TCA/acetone protein precipitation protocol followed by purification with a 2-D Clean-Up Kit (GE Healthcare, USA). After an in-solution trypsin digestion step, analyses were performed by liquid chromatography (Acquity system, Waters, UK) coupled with a triple quadrupole mass spectrometer (Xevo TQS, Waters, UK) with electrospray ionisation. The acquisition and processing of data was carried out by MassLynx software (v. 4.1, Waters) and multiple reaction monitoring (MRM) design was made using the open-source software Skyline (
Reference hemoglobin powder was used to select MRM transitions for each peptide biomarkers and to optimise their collision energy. Commercial feed material and compound feed containing or free from blood derived products were then analysed. Selected transitions were present in materials or feed known to contained blood derived product of the same origin and absent in the others. Artificially contaminated feed with various contamination levels were also analysed in order to evaluate the influence of matrix composition and to experimentally determine the limit of detection. A first estimation of the LOD was around 0.05 % w/w which was below the LOD imposed by the EC for animal proteins detection method.
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