A simplified multidimensional approach for analysis of complex biological samples: on-line LC-CE-MS.
Information on protein expression, disease biomarkers or surrogate markers and genetic disorders can nowadays be achieved from analysis of complex biological samples by liquid separation coupled to mass spectrometric (MS) detection. This paper describes fast multidimensional separation by on-line liquid chromatography (LC) and capillary electrophoresis (CE), followed by electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FTICR) MS detection. This detector provides ultrahigh resolution of the detected ions, mass accuracy at the ppm-level and high sensitivity. Most of the challenge of this system lies in the development of a new interface for the on-line coupling of LC to CE. The interface developed in poly(dimethylsiloxane) provides a RSD for injection repeatability of <3.5% and surface control for unspecific binding by deactivation with a cationic polymer, PolyE-323. We have evaluated the interface, as well as the overall system, with respect to robustness and deconvolution ability. Sequence coverage for bovine serum albumin (BSA) of 93% showed a high recovery of sample in the different transfer steps through the system. The detection limit for identification is 277 ng mL(-1) (or 280 nM) on average for peptides. In the future, we expect LC-CE-MS to be a novel strategy for elucidating the chemistry of biological matrices.
Bergström
Sara K
SK
Analytical Chemistry, Department of Physical and Analytical Chemistry, Biomedical Centre, Uppsala University, Box 599, SE-751 24 Uppsala, Sweden.
Dahlin
Andreas P
AP
Ramström
Margareta
M
Andersson
Marit
M
Markides
Karin E
KE
Bergquist
Jonas
J
D000818:Animals; D015415:Biomarkers / Q000032:analysis*; D002417:Cattle; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods; D019075:Electrophoresis, Capillary / Q000379:methods; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D012710:Serum Albumin, Bovine / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
D015415:Biomarkers; D010455:Peptides; D011506:Proteins; D012710:Serum Albumin, Bovine