Analysis of peptides, proteins, protein digests, and whole human blood by capillary electrophoresis/electrospray ionization-mass spectrometry using an in-capillary electrode sheathless interface.
An in-capillary electrode sheathless interface was applied to the capillary electrophoresis/electrospray ionization-mass spectrometry (CE/ESI-MS) analysis of mixtures of small peptides, proteins, and tryptic digests of proteins. The effects of different experimental parameters on the performance of this CE/ESI-MS interface were studied. The distance of the in-capillary electrode from the CE outlet and the length of the electrode inside the capillary had no significant effects on the CE separation and ESI behavior under the experimental conditions used. However, significant enhancement of the sensitivity resulted from the use of narrower CE capillaries. Using a quadrupole mass spectrometer, an aminopropylsilane-coated capillary, and a wide scan mass-to-charge ratio range of 500-1400, detection limits of approximately 4, 1, and 0.6 fmol for cytochrome c and myoglobin were achieved for 75-, 50-, and 30-micron inner diameter capillaries, respectively. Approximately one order of magnitude lower detection limits were achieved under the multiple-ion monitoring mode. The application of the in-capillary electrode sheathless interface to real-world samples was demonstrated by CE/ESI-MS analysis of a human blood sample.
Department of Chemistry and Biochemistry, University of Texas at Austin 78712, USA.
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Cao
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Moini
D000595:Amino Acid Sequence; D001798:Blood Proteins / Q000032:analysis*; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D004852:Epoxy Compounds / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis*; D011492:Protein Hydrolysates / Q000032:analysis*; D015203:Reproducibility of Results
D001798:Blood Proteins; D004852:Epoxy Compounds; D009842:Oligopeptides; D011492:Protein Hydrolysates