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CE‐MS method development for peptides analysis, especially hepcidin, an iron metabolism marker
Author(s) -
Martin Gaëlle B.,
Mansion François,
Servais AnneCatherine,
Debrus Benjamin,
Rozet Eric,
Hubert Philippe,
Crommen Jacques,
Fillet Marianne
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200800794
Subject(s) - chemistry , formic acid , chromatography , hepcidin , selectivity , ammonium acetate , resolution (logic) , peptide , analytical chemistry (journal) , high performance liquid chromatography , biochemistry , medicine , artificial intelligence , computer science , inflammation , catalysis
A method for the resolution of a peptides mixture including hepcidin‐25, an iron metabolism marker, was developed by CE‐ESI‐MS. Several strategies were tested to optimize peptide separation, such as the addition of cyclodextrins or organic solvents in the BGE or the use of coated capillaries. Best results in terms of resolution, symmetry and efficiency were obtained with a BGE made of 500 mM ammonium acetate pH 4.5/ACN 70:30 v/v. Using the methodology of experimental design, BGE concentration, sheath liquid composition and MS‐coupling parameters were then optimized in order to obtain the best signal intensity for hepcidin. Finally, a 225 mM BGE and a sheath liquid composed of isopropanol/water 80:20 v/v containing 0.5% v/v formic acid were selected as it constitutes the best compromise for selectivity, peak shape and sensitivity.