Mass spectrometry has become the technique of choice for the assessment of a high variety of molecules in
complex food matrices. It is best suited for monitoring the evolution of digestive processes in vivo and in vitro.
However, considering the variety of equipment available in different laboratories and the diversity of sample
preparation methods, instrumental settings for data acquisition, statistical evaluations, and interpretations of
results, it is difficult to predict a priori the ideal parameters for optimal results. The present work addressed this
uncertainty by executing an inter-laboratory study with samples collected during in vitro digestion and presenting
an overview of the state-of-the-art mass spectrometry applications and analytical capabilities available for
studying food digestion. Three representative high-protein foods – skim milk powder (SMP), cooked chicken
breast and tofu – were digested according to the static INFOGEST protocol with sample collection at five different
time points during gastric and intestinal digestion. Ten laboratories analysed all digesta with their in-house
equipment and applying theirconventional workflow.
The compiled results demonstrate in general, that soy proteins had a slower gastric digestion and the presence
of longer peptide sequences in the intestinal phase compared to SMP or chicken proteins, suggesting a higher
resistance to the digestion of soy proteins. Differences in results among the various laboratories were attributed
more to the peptide selection criteria than to the individual analytical platforms. Overall, the combination of
mass spectrometry techniques with suitable methodological and statistical approaches is adequate for contributing
to the characterisation of the recently defined digestome.