title: Applications of monolithic silica capillary columns in proteomics.
abstract: The use and applicability of silica based capillary monolithic reversed-phase columns in proteomic analysis has been evaluated by liquid chromatography-mass spectrometry (LC-MS). Chromatographic performance of the monolithic capillaries was evaluated with a tryptic digest of cytochrome C showing very good resolution and reproducibility in addition to the known advantages of a low pressure drop over a time period of 6 months. Monoliths were subsequently tested for their suitability to separate proteins and peptides from samples typically encountered in proteomic research such as in-gel digested tryptic peptide mixtures or fractions of proteolytically digested human serum. The monolithic capillaries also proved useful in the analysis of phospholipid species in bronchoalveolar lavage fluid. Compared to particle-filled conventional capillary columns, rapid and highly efficient separation of peptides and proteins was achieved using these bimodal pore size distribution columns, and good quality collision induced dissociation (CID) mass spectra were obtained on an ion trap mass spectrometer. These novel monolithic separation media are thus a promising addition to the methodological toolbox of proteomics research.
authors.affiliation: University of Groningen, Department of Bioanalysis & Toxicology, University Centre for Pharmacy, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands. b.barroso@farm.rug.nl
authors.forename: Begona
authors.initials: B
authors.lastname: Barroso
authors.forename: Dieter
authors.initials: D
authors.lastname: Lubda
authors.forename: Rainer
authors.initials: R
authors.lastname: Bischoff
mesh_terms: D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism; D004549:Elastin / Q000737:chemistry / Q000378:metabolism; D005260:Female; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D010196:Pancreatic Elastase / Q000737:chemistry / Q000378:metabolism; D010455:Peptides / Q000032:analysis*; D016062:Porosity; D040901:Proteomics / Q000379:methods; D012822:Silicon Dioxide; D002583:Uterine Cervical Neoplasms / Q000097:blood
chemical_list: D010455:Peptides; D012822:Silicon Dioxide; D045304:Cytochromes c; D004549:Elastin; D010196:Pancreatic Elastase