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External calibration was performed in MS/MS mode using fragment ions of Glu-fibrinopeptide as references.
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Samples were analyzed in single ion monitoring mode using characteristic fragment ions of analytes and corresponding internal standards.
Spectra were calibrated in static mode using MS/MS fragment ions of standard supplied by the manufacturer.
Spectra were calibrated in static nanospray mode using MS/MS fragment-ions of a renin peptide standard (His immonium-ion with m/z at 110.0713, and b8-ion with m/z at 1028.5312) providing a mass accuracy of ≤50 ppm.
The binding mode of these weak ligands was characterized by a combination of NMR data and docking calculations and was confirmed using fragment homologues, highlighting the role of hydroxyl functions.
The system was externally calibrated in MS/MS mode using the parent ion and selected fragments of adrenocorticotropic hormone (ACTH) human fragment 18 39 (m/z = 2465.1983; Sigma Aldrich).
A chromatography analysis was performed in SIM mode using an ion with m/z = 143 as a marker fragment.
Fragments captured in hybridization were indexed, amplified and sequenced in a paired end 100-bp mode using an Illumina HiSeq2000 deep sequencing instrument (v3 sequencing chemistry; Illumina, San Diego, CA).
After identification of metabolites in the scan mode using the NIST data bank, quantification of labeling enrichment was done in SIM (single ion monitoring) mode in two technical replicates using the following unique fragments (m/z) containing the complete carbon skeleton of metabolites: pyruvate 174, lactate 261, alanine 260, glycine 246, serine 390, aspartate 418, glutamate 432, glutamine 431.
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