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Improved particle‐packed HPLC/MS microchips for proteomic analysis
Author(s) -
Trusch Maria,
Ehlert Steffen,
Bertsch Andreas,
Kohlbacher Oliver,
Hildebrand Diana,
Schlüter Hartmut,
Tallarek Ulrich
Publication year - 2010
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.201000474
Subject(s) - chromatography , chemistry , reproducibility , high performance liquid chromatography , sample preparation , particle size , analytical chemistry (journal) , fraction (chemistry) , particle (ecology) , proteomics , mass spectrometry , biochemistry , oceanography , gene , geology
Abstract The influence of packing process parameters (packing pressure, application of ultrasound) and the stationary phase particle size (3.5 and 5 μm) on the chromatographic performance of HPLC/MS chips was systematically investigated for proteomic samples. First, reproducibility and detection limits of the separation were evaluated with a low‐complexity sample of tryptic BSA peptides. The influence of adsorbent packing quality on protein identification was then tested with a typical proteomics sample of high complexity, a human plasma protein fraction (Cohn fraction IV‐4). All HPLC/MS chips provided highly reproducible separations of these proteomic samples, but improved packing conditions and smaller particle sizes resulted in chromatograms with narrower peaks and correspondingly higher signal intensities. Improved separation performance increased the peak capacity, the number of identified peptides, and thus the sequence coverage in the proteomic samples, particularly for low sample amounts.