Coaxial Electrospray Ionization for the Study of Rapid In-source Chemistry

  • Brynn N. Sundberg
  • Anthony F. LagalanteEmail author
Research Article


Coaxial electrospray has been used effectively for several dual-emitter applications, but has not been utilized for the study of rapid in-source chemistry. In this paper, we report the fabrication of a coaxial, micro-volume dual-emitter through the modification of a manufacturer’s standard electrospray probe. This modification creates rapid mixing inside the Taylor cone and the ability to manipulate fast reactions using a variety of solvents and analytes. We demonstrate its potential as a low-cost, dual-emitter assembly for diverse applications through three examples: relative ionization in a biphasic electrospray, hydrogen-deuterium exchange, and protein supercharging.

Graphical Abstract


Coaxial electrospray Hydrogen-deuterium exchange Protein folding Biphasic electrospray ionization Supercharging 


  1. 1.
    Jaworek, A.: Electrostatic micro- and nanoencapsulation and electroemulsification: a brief review. J. Microencapsul. 25, 443–468 (2008)CrossRefPubMedGoogle Scholar
  2. 2.
    Reneker, D.H., Chun, I.: Nanometre diameter fibres of polymer, produced by electrospinning. Nanotechnology. 7, 216–223 (1996)CrossRefGoogle Scholar
  3. 3.
    Tetler, L.W., Cooper, P.A., Powell, B.: Influence of capillary dimensions on the performance of a coaxial capillary electrophoresis electrospray mass-spectrometry Interface. J. Chromatogr. A. 700, 21–26 (1995)CrossRefGoogle Scholar
  4. 4.
    Hashimoto, M., Ishihama, Y., Tomita, M., Soga, T.: Microelectrospray interface with coaxial sheath flow for high-resolution capillary electrophoresis/mass spectrometry separation. Rapid Commun. Mass Spectrom. 21, 3579–3584 (2007)CrossRefPubMedGoogle Scholar
  5. 5.
    Edmonds, C.G., Loo, J.A., Barinaga, C.J., Udseth, H.R., Smith, R.D.: Capillary electrophoresis electrospray ionization-mass spectrometry. J. Chromatogr. 474, 21–37 (1989)CrossRefGoogle Scholar
  6. 6.
    Jiang, L.F., Moini, M.: Development of multi-ESI-sprayer, multi-atmospheric-pressure-inlet mass spectrometry and its application to accurate mass measurement using time-of-flight mass spectrometry. Anal. Chem. 72, 20–24 (2000)CrossRefPubMedGoogle Scholar
  7. 7.
    Li, Z., Zhang, S.H., Cai, Y., Wu, Q.H., Chen, H.: Hollow fiber-based solid-liquid phase microextraction combined with theta capillary electrospray ionization mass spectrometry for sensitive and accurate analysis of methamphetamine. Anal. Methods. 8, 7800–7807 (2016)CrossRefGoogle Scholar
  8. 8.
    Li, Y.F., Zhang, N., Zhou, Y.M., Wang, J.N., Zhang, Y.M., Wang, J.Y., Xiong, C.Q., Chen, S.M., Nie, Z.X.: Induced dual-nanospray: a novel internal calibration method for convenient and accurate mass measurement. J. Am. Soc. Mass Spectrom. 24, 1446–1449 (2013)CrossRefPubMedGoogle Scholar
  9. 9.
    Liu, C.C., Alary, J.F., Vollmerhaus, P., Kadkhodayan, M.: Design, optimisation, and evaluation of a sheath flow interface for automated capillary electrophoresis-electrospray-mass spectrometry. Electrophoresis. 26, 1366–1375 (2005)CrossRefPubMedGoogle Scholar
  10. 10.
    Zhang, J.T., Wang, H.Y., Zhu, W., Cai, T.T., Guo, Y.L.: Solvent-assisted electrospray ionization for direct analysis of various compounds (complex) from low/nonpolar solvents and eluents. Anal. Chem. 86, 8937–8942 (2014)CrossRefPubMedGoogle Scholar
  11. 11.
    Chen, H.W., Venter, A., Cooks, R.G.: Extractive electrospray ionization for direct analysis of undiluted urine, milk and other complex mixtures without sample preparation. Chem. Commun. 2042–2044 (2006)Google Scholar
  12. 12.
    Chang, D.Y., Lee, C.C., Shiea, J.: Detecting large biomolecules from high-salt solutions by fused-droplet electrospray ionization mass spectrometry. Anal. Chem. 74, 2465–2469 (2002)CrossRefPubMedGoogle Scholar
  13. 13.
    Wu, C., Siems, W.F., Hill, H.H.: Secondary electrospray ionization ion mobility spectrometry/mass spectrometry of illicit drugs. Anal. Chem. 72, 396–403 (2000)CrossRefPubMedGoogle Scholar
  14. 14.
    Girod, M., Moyano, E., Campbell, D.I., Cooks, R.G.: Accelerated bimolecular reactions in microdroplets studied by desorption electrospray ionization mass spectrometry. Chem. Sci. 2, 501–510 (2011)CrossRefGoogle Scholar
  15. 15.
    Lu, Y., Liu, F., Lion, N., Girault, H.H.: Dual-channel electrospray microchip. J. Am. Soc. Mass Spectrom. 24, 454–457 (2013)CrossRefPubMedGoogle Scholar
  16. 16.
    Mark, L.P., Gill, M.C., Mahut, M., Derrick, P.J.: Dual nano-electrospray for probing solution interactions and fast reactions of complex biomolecules. Eur J Mass Spectrom. 18, 439–446 (2012)CrossRefGoogle Scholar
  17. 17.
    Rob, T., Wilson, D.J.: Time-resolved mass spectrometry for monitoring millisecond time-scale solution-phase processes. Eur J Mass Spectrom. 18, 205–214 (2012)CrossRefGoogle Scholar
  18. 18.
    Mortensen, D.N., Williams, E.R.: Investigating protein folding and unfolding in electrospray nanodrops upon rapid mixing using theta-glass emitters. Anal. Chem. 87, 1281–1287 (2015)CrossRefPubMedGoogle Scholar
  19. 19.
    Fisher, C.M., Kharlamova, A., McLuckey, S.A.: Affecting protein charge state distributions in nano-electrospray ionization via in-spray solution mixing using theta capillaries. Anal. Chem. 86, 4581–4588 (2014)CrossRefPubMedGoogle Scholar
  20. 20.
    Prudent, M., Mendez, M.A., Girault, H.H.: Biphasic electrospray ionization for the study of interfacial complexes. Anal. Sci. 24, 1399–1404 (2008)CrossRefPubMedGoogle Scholar
  21. 21.
    Ling, X., Ding, C.F.: Online electrochemistry/electrospray mass spectrometry with a coaxial probe for investigation of electrochemical derivatization of anthracene with dodecylamine. Acta Phys. -Chim. Sin. 28, 2616–2624 (2012)Google Scholar
  22. 22.
    Rashid, S., Overton, S., Mazigh, B., Mayer, P.M.: Dual-spray hydrogen/deuterium exchange (HDX) reactions: a new method of probing protein structure. Rapid Commun. Mass Spectrom. 30, 1505–1512 (2016)CrossRefPubMedGoogle Scholar
  23. 23.
    Cotham, V.C., Shaw, J.B., Brodbelt, J.S.: High-throughput bioconjugation for enhanced 193 nm photodissociation via droplet phase initiated ion/ion chemistry using a front-end dual spray reactor. Anal. Chem. 87, 9396–9402 (2015)CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Ingram, A.J., Boeser, C.L., Zare, R.N.: Going beyond electrospray: mass spectrometric studies of chemical reactions in and on liquids. Chem. Sci. 7, 39–55 (2016)CrossRefPubMedGoogle Scholar
  25. 25.
    Ramjit, H.G., Newton, R., Guare, J.P.: A novel coaxial electrospray ionization method for characterizing hexacosanoylceramides by Fourier transform ion cyclotron resonance mass spectrometry. Rapid Commun. Mass Spectrom. 19, 1257–1262 (2005)CrossRefPubMedGoogle Scholar
  26. 26.
    Palmer, M.E., Tetler, L.W., Wilson, I.D.: Hydrogen/deuterium exchange using a coaxial sheath-flow interface for capillary electrophoresis/mass spectrometry. Rapid Commun. Mass Spectrom. 14, 808–817 (2000)CrossRefPubMedGoogle Scholar
  27. 27.
    Hemling, M.E., Conboy, J.J., Bean, M.F., Mentzer, M., Carr, S.A.: Gas-phase hydrogen-deuterium exchange in electrospray-ionization mass-spectrometry as a practical tool for structure elucidation. J. Am. Soc. Mass Spectrom. 5, 434–442 (1994)CrossRefPubMedGoogle Scholar
  28. 28.
    Kostyukevich, Y., Kononikhin, A., Popov, I., Nikolaev, E.: Conformational changes of ubiquitin during electrospray ionization as determined by in-ESI source H/D exchange combined with high-resolution MS and ECD fragmentation. J. Mass Spectrom. 49, 989–994 (2014)CrossRefPubMedGoogle Scholar
  29. 29.
    Tolonen, A., Turpeinen, M., Uusitalo, J., Pelkonen, O.: A simple method for differentiation of monoisotopic drug metabolites with hydrogen-deuterium exchange liquid chromatography/electrospray mass spectrometry. Eur. J. Pharm. Sci. 25, 155–162 (2005)CrossRefPubMedGoogle Scholar
  30. 30.
    Wolff, J.C., Laures, A.M.F.: ‘On-the-Fly’ hydrogen/deuterium exchange liquid chromatography/mass spectrometry using a dual-sprayer atmospheric pressure ionisation source. Rapid Commun. Mass Spectrom. 20, 3769–3779 (2006)CrossRefPubMedGoogle Scholar
  31. 31.
    Lam, W., Ramanathan, R.: In electrospray ionization source hydrogen/deuterium exchange LC-MS and LC-MS/MS for characterization of metabolites. J. Am. Soc. Mass Spectrom. 13, 345–353 (2002)CrossRefPubMedGoogle Scholar
  32. 32.
    Jeong, E.S., Cha, E., Cha, S., Kim, S., Bin Oh, H., Kwon, O.S., Lee, J.: Online simultaneous hydrogen/deuterium exchange of multitarget gas-phase molecules by electrospray ionization mass spectrometry coupled with gas chromatography. Anal. Chem. 89, 12284–12292 (2017)Google Scholar
  33. 33.
    Jansson, E.T., Lai, Y.H., Santiago, J.G., Zare, R.N.: Rapid hydrogen-deuterium exchange in liquid droplets. J. Am. Chem. Soc. 139, 6851–6854 (2017)CrossRefPubMedGoogle Scholar
  34. 34.
    Deng, Y.Z., Zhang, Z.Q., Smith, D.L.: Comparison of continuous and pulsed labeling amide hydrogen exchange/mass spectrometry for studies of protein dynamics. J. Am. Soc. Mass Spectrom. 10, 675–684 (1999)CrossRefPubMedGoogle Scholar
  35. 35.
    Iavarone, A.T., Jurchen, J.C., Williams, E.R.: Supercharged protein and peptide ions formed by electrospray ionization. Anal. Chem. 73, 1455–1460 (2001)CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Li, X.Y., Li, Z.X., Xie, B., Sharp, J.S.: Supercharging by m-NBA improves ETD-based quantification of hydroxyl radical protein footprinting. J. Am. Soc. Mass Spectrom. 26, 1424–1427 (2015)CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Iavarone, A.T., Paech, K., Williams, E.R.: Effects of charge state and cationizing agent on the electron capture dissociation of a peptide. Anal. Chem. 76, 2231–2238 (2004)CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Lomeli, S.H., Yin, S., Loo, R.R.O., Loo, J.A.: Increasing charge while preserving noncovalent protein complexes for ESI-MS. J. Am. Soc. Mass Spectrom. 20, 593–596 (2009)CrossRefPubMedGoogle Scholar
  39. 39.
    Nshanian, M., Lakshmanan, R., Chen, H., Loo, R.R.O., Loo, J.A.: Enhancing sensitivity of liquid chromatography–mass spectrometry of peptides and proteins using supercharging agents. Int. J. Mass Spectrom. 427, 157–164 (2018)CrossRefPubMedGoogle Scholar
  40. 40.
    Miladinovic, S.M., Fornelli, L., Lu, Y., Piech, K.M., Girault, H.H., Tsybin, Y.O.: In-spray supercharging of peptides and proteins in electrospray ionization mass spectrometry. Anal. Chem. 84, 4647–4651 (2012)CrossRefPubMedGoogle Scholar
  41. 41.
    Mendes, M.A., Souza, B.M., Marques, M.R., Palma, M.S.: The effect of glycerol on signal supression during electrospray ionization analysis of proteins. Spectrosc Int J. 18, 339–345 (2004)CrossRefGoogle Scholar

Copyright information

© American Society for Mass Spectrometry 2018

Authors and Affiliations

  1. 1.Department of ChemistryVillanova UniversityVillanovaUSA

Personalised recommendations