CE-MS method development for peptides analysis, especially hepcidin, an iron metabolism marker.
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.
Martin
Gaëlle B
GB
Department of Analytical Pharmaceutical Chemistry, Institute of Pharmacy, University of Liège, Liège, Belgium.
Mansion
François
F
Servais
Anne-Catherine
AC
Debrus
Benjamin
B
Rozet
Eric
E
Hubert
Philippe
P
Crommen
Jacques
J
Fillet
Marianne
M
D000085:Acetates / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000032:analysis*; D002417:Cattle; D019075:Electrophoresis, Capillary / Q000379:methods*; D064451:Hepcidins; D006736:Horses; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015999:Multivariate Analysis; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000032:analysis*; D011506:Proteins / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013696:Temperature
D000085:Acetates; D000097:Acetonitriles; D023181:Antimicrobial Cationic Peptides; C578702:HAMP protein, human; D064451:Hepcidins; D010446:Peptide Fragments; D011506:Proteins; C018824:ammonium acetate; C032159:acetonitrile