Summary
In recent years, the hyphenation of capillary electrophoresis (CE) and electrospray ionization mass spectrometry (ESI-MS) has been widely used for the analysis of biological molecules with high efficiency and high accuracy. However, the signal intensity of CE-ESI-MS is restricted by various parameters. This paper reports the effects on the CE-ESI-MS signal of the pH and the electrolyte concentration, the formic acid concentration of the sheath liquid, and the sheath liquid composition, using several proteins as samples. The study was performed systematically by experimental and theoretical analyses. The maximum signal intensity of three proteins (cytochrome c, insulin, bovine serum albumin) was attained with a pH 4.40 buffer containing 75 mM formic acid and 75 mM ammonium acetate. Investigation of the influence of the formic acid concentration in the sheath liquid (over the range of 0%–1%) on the ESI-MS signal of brovine serum albumin showed that the feasible amount of the formic acid in sheath liquid could improve the signal intensity of the sample ions. However, considerable band broadening was observed that should be attributed mainly to column overloading, band spreading at the interface, and scanning data acquisition.
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References
Karger, B.L.J. Chem. Ed. 1997,74, 45–48.
Li, S.F.Y.Capillary electrophoresis: principles, practice and applications, J. Chromatogr. Library, Elsevier, The Netherlands,1992.
Katz, E.D.High performance liquid chromatography: principles and methods in biotechnology, In: Scott, R.P.W.; Simpson, C.F.; Katz, E.D. (Eds).Separation Science Series, Wiley, New York, NY,1996.
Gilges, M.; Kleemiss, M.H.; Schomburg, G.Anal. Chem. 1994,66, 2038–2046.
Kjeindienst, G.; Huber, C.G.; Gjerde, D.; Yengoyan, L.; Bonn, G.K.Electrophoresis 1998,19, 262–269.
Andren, P.E.; Emmett, M.R.; Caprioli, R.M.J. Am. Soc. Mass Spectrom. 1994,5, 867–869.
Whitehouse, C.M.; Dreyer, R.N.; Yamashita, M.; Fenn, J.B.Anal. Chem. 1985,57, 675–679.
Tomer, K.B.; Deterding, L.J.; Parker, C.E.Adv. Chromatogr. 1995,35, 53–99.
Cai, J.; Henion, J.D.J. Chromatogr. A 1995,703, 667–692.
Niessen, W.M.A.; Tjarden, U.R.; Van der Greef, J.J. Chromatogr. 1993,636, 3–19.
Smith, R.D.; Udseth, H.R.; In: Lunte, S.M.; Radzik, D.M. (Eds.)Pharmaceutical and Biomedical Applications of Capillary Electrophoresis, Elsevier, Oxford,1996, p. 229.
Smith, R.D.; Udseth, H.R.; Wahl, J.H.; Goodlett, D.R.; Hofstadler, S.A.Methods Enzymol. 1996,271, 448–486.
Smith, R.D.; Wahl, J.H.; Goodlett, D.R.; Hofstadler, S.A.Anal. Chem. 1993,65, 574A-584A.
Banks, J.F.Electrophoresis 1997,18, 2255–2266.
Hugener, M.; Tinke, A.P.; Niessen, M.A.; Tjaden, U.R.; Van der Greef, J.J. Chromatogr. 1993,647, 375–385.
Lane, S.J.; Boughtflower, R.; Paterson, C.; Underwood, T.Rapid Commun. Mass Spectrom. 1995,9, 1283–1287.
Lane, S.J.; Boughtflower, R.; Paterson, C.; Morris, M.Rapid Commun. Mass Spectrom. 1996,10, 733–736.
Fore, F.; Thompson, T.J.; Karger, B.L.; Gebàuer, P.; Bocek, P.Anal. Chem. 1994,66, 4450–4458.
Moseley, M.A.; Jorgenson, J.W.; Shabanowitz, J.; Hunt, D.F.; Tomer, K.B.J. Am. Soc. Mass Spectrom. 1992,3, 289–300.
Wahl, J.H.; Smith, R.D.;J. Cap. Electrophoresis 1994,1, 62–71.
Wang, G.; Cole, R.B.Org. Mass Spectrom. 1994,29, 419–427.
Gale, D.C.; Smith, R.D.Rapid Commun. Mass Spectrom. 1993,7, 1017–1021.
Thompson, T.J.; Foret, F.; Kirby, D.P.; Vouros, P.; Karger, B.L.presented at the 41 st ASMS Conference on Mass Spectrometry and Allied Topics, San Francisco, CA, 31 May 4 June1993.
Thompson, T.J.; Foret, F.; Kirby, D.P.; Karger, B.L.; Vouros, P.presented at the 42nd ASMS Conference on Mass Spectrometry and Allied Topics, Chicago, IL, 29 May 3 June1994.
Kebarle, P.; Ho, Y. In Cole, R.B. (Editor)Electrosprary ionization Mass Spectrometry wiely, New York,1997.
Tang, L.; Kebarle, P.Anal. Chem. 1993,65, 3654–3668.
Van Berkel, G.J. In Cole, R.B. (Editor)Electrosprary ionization Mass Spectrometry, wiely, New York,1997.
Xiong, W.; Chen, L.M.; Chao, J.J. Biol. Chem. 1990,265, 2822–2827.
William, C.W.; Hideaki, Y., Jun, A.Kidney International 2000,58, 730–739.
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Liang, Z., Yang, Q., Zhang, W. et al. Effects of experimental parameters on the signal intensity of capillary electrophoresis electrospray ionization mass spectrometry in protein analysis. Chromatographia 57, 617–621 (2003). https://doi.org/10.1007/BF02491738
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DOI: https://doi.org/10.1007/BF02491738