Title of article :
Automated multi-dimensional liquid chromatography: sample preparation and identification of peptides from human blood filtrate
Author/Authors :
Machtejevas، نويسنده , , Egidijus and John، نويسنده , , Harald and Wagner، نويسنده , , Knut and Stنndker، نويسنده , , Ludger and Marko-Varga، نويسنده , , Gyِrgy and Forssmann، نويسنده , , Wolf-Georg and Bischoff، نويسنده , , Rainer and Unger، نويسنده , , Klaus K.، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2004
Pages :
10
From page :
121
To page :
130
Abstract :
A comprehensive on-line sample clean-up with an integrated two-dimensional HPLC system was developed for the analysis of natural peptides. Samples comprised of endogenous peptides with molecular weights up to 20 kDa were generated from human hemofiltrate (HF) obtained from patients with chronic renal failure. The (poly-)peptides were separated using novel silica-based restricted access materials with strong cation-exchange functionalities (SCX-RAM). The size-selective sample fractionation step is followed by cation-exchange chromatography as the first dimension. The subsequent second dimension of separation is based on hydrophobic interaction using four parallel short reversed-phase (RP) columns implemented via a fully automated column switching technique. More than 1000 peaks were resolved within the total analysis time of 96 min. Substances of selected peaks were sampled to analyse their molecular weights by off-line MALDI-TOF mass spectrometry and to determine their amino acid sequence by Edman degradation. The potential for comprehensive peptide mapping and identification is demonstrated.
Keywords :
Sample preparation , Identification , Human blood filtrate , Peptides
Journal title :
Journal of Chromatography B
Serial Year :
2004
Journal title :
Journal of Chromatography B
Record number :
1456621
Link To Document :
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