Automated multi-dimensional liquid chromatography: sample preparation and identification of peptides from human blood filtrate
(2004) In Journal of Chromatography. B 803(1). p.121-130- 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... (More)
- 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. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/284592
- author
- Machtejevas, E ; John, H ; Wagner, K ; Standker, L ; Marko-Varga, György LU ; Forssmann, WG ; Bischoff, R and Unger, KK
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- sample preparation, identification, human blood filtrate, peptides
- in
- Journal of Chromatography. B
- volume
- 803
- issue
- 1
- pages
- 121 - 130
- publisher
- Elsevier
- external identifiers
-
- wos:000220343500013
- pmid:15026005
- scopus:1542719143
- ISSN
- 1873-376X
- DOI
- 10.1016/j.jchromb.2003.07.015
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004)
- id
- 58bb7464-4b65-4461-983c-8540a1e4aa6a (old id 284592)
- date added to LUP
- 2016-04-01 12:13:17
- date last changed
- 2022-04-13 07:54:46
@article{58bb7464-4b65-4461-983c-8540a1e4aa6a, 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.}}, author = {{Machtejevas, E and John, H and Wagner, K and Standker, L and Marko-Varga, György and Forssmann, WG and Bischoff, R and Unger, KK}}, issn = {{1873-376X}}, keywords = {{sample preparation; identification; human blood filtrate; peptides}}, language = {{eng}}, number = {{1}}, pages = {{121--130}}, publisher = {{Elsevier}}, series = {{Journal of Chromatography. B}}, title = {{Automated multi-dimensional liquid chromatography: sample preparation and identification of peptides from human blood filtrate}}, url = {{http://dx.doi.org/10.1016/j.jchromb.2003.07.015}}, doi = {{10.1016/j.jchromb.2003.07.015}}, volume = {{803}}, year = {{2004}}, }