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Optimization of micro‐HPLC peak focusing for the detection and quantification of low hepcidin concentrations
Author(s) -
Mansion François,
Chiap Patrice,
Houbart Virginie,
Crommen Jacques,
Servais AnneCatherine,
Fillet Marianne
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100244
Subject(s) - chromatography , chemistry , electrospray ionization , mass spectrometry , high performance liquid chromatography , elution , electrospray , analytical chemistry (journal) , gradient elution , tandem mass spectrometry , volumetric flow rate , physics , quantum mechanics
Micro‐high‐performance liquid chromatography is a miniaturized, economic and ecological chromatographic system allowing the use of reduced size chromatographic columns. Coupled with electrospray ionization tandem mass spectrometry, this technique can be used to detect and quantify low concentrations of peptides. In this study, hepcidin was used as the model compound and analysed using octadecylsilica stationary phase by means of a gradient elution mode at a flow rate of 4 μL/min. Several parameters were studied to optimize peak focusing. Using the methodology of experimental design, the mobile‐phase gradient conditions and the sample composition were optimized in order to maximize the sensitivity and minimize retention time. Stability of the target peptide in solution was also demonstrated.