Feb 20, 2014

Mass spectrometric characterisation of the circulating peptidome following oral glucose ingestion in control and gastrectomised patients.

BioRxiv : the Preprint Server for Biology
Jesse D BloomFiona M Gribble


Meal ingestion triggers secretion of a variety of gut and endocrine peptides, several of which are routinely measured in research studies by commercial immunoassays. We developed an LC-MS/MS based assay for parallel monitoring of multiple peptides in small volumes of human plasma, providing the benefit of analysing exact peptide sequences rather than immuno-reactivity, and potential advantages of cost and sample volumes for measuring multiple peptide hormones. The method involves acetonitrile precipitation of larger proteins, followed by solid phase extraction and nano-LC-MS/MS using an untargeted approach on an orbitrap mass spectrometer. Analysis of plasma from control subjects and patients who have undergone gastrectomy with Roux-en-Y reconstruction, revealed elevated levels of a number of peptides following glucose ingestion. These included GLP-1(7-36), GLP-1(9-36), glicentin, oxyntomodulin, GIP(1-42), GIP(3-42), PYY(1-36), PYY(3-36), neurotensin, insulin and C-peptide, as well as motilin, which decreased following glucose ingestion. Results showed good correlation with those peptides measured previously by immunoassay in the same samples. The gastrectomy group had higher, but non-glucose-dependent, circulating levels of pe...Continue Reading

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