Use of a Continuous Stirred Tank Reactor for the Determination of Electrospray Response Factors and Its Application to Underivatized Sugars Under Various Solvent Parameters

  • Jonathan B. Thacker
  • Kevin A. Schug
Research Article


The relationship between the electrospray ionization (ESI)-mass spectrometric (MS) response of an analyte and its concentration has been well studied for permanently charged and basic analytes in the positive ionization mode, but there has been a lack of research effort for other analytes, and for the negative ionization mode, in general. In this study, this relationship was investigated for various adducts and deprotonated species of glucose, sucrose, and raffinose using a continuous stirred tank reactor (CSTR) coupled with ESI-tandem MS to obtain a continuum of response factors across a wide concentration range in both the positive and negative ionization modes with a single injection under 18 different combinations of solvents and additives. Profiles of response factors vs. concentrations varied widely and were dependent upon the analyte and solvent parameters. The use of ammonium trifluoroacetate resulted in the highest response factors for methanol-based and acetonitrile-based solvents in the positive and negative ionization modes, respectively. Ammonium acetate, ammonium formate, and sodium chloride in 80:20 acetonitrile:water in the negative ionization mode resulted in good linearities, useful for quantitative analysis. In the positive ionization mode, response factors tended to increase with an increase in the molecular weight of the analyte, and acetonitrile was generally found to decrease response factors. We have also demonstrated the ability of CSTR-ESI-MS to visualize ionization suppression in the presence of co-analytes. These data should be useful for liquid chromatography-ESI-MS method development for sugar analysis, to help guide the choice of mobile phase that will result in high sensitivity and linearity.

Graphical Abstract


Electrospray ionization Continuous stirred tank reactor Sugars Carbohydrates Saccharides Solvent effects 



The authors gratefully acknowledge Restek Corporation (Bellefonte, PA, USA) for their financial support of this study.

Supplementary material

13361_2018_2112_MOESM1_ESM.pdf (1.1 mb)
ESM 1 (PDF 1146 kb)


  1. 1.
    Enke, C.G.: A predictive model for matrix and analyte effects in electrospray ionization of singly-charged ionic analytes. Anal. Chem. 69, 4885–4893 (1997)CrossRefGoogle Scholar
  2. 2.
    Tang, L., Kebarle, P.: Dependence of ion intensity in electrospray mass spectrometry on the concentration of the analytes in the electrosprayed solution. Anal. Chem. 65, 3654–3668 (1993)CrossRefGoogle Scholar
  3. 3.
    Mallet, C.R., Lu, Z., Mazzeo, J.R.: A study of ion suppression effects in electrospray ionization from mobile phase additives and solid-phase extracts. Rapid Commun. Mass Spectrom. 18, 49–58 (2004)CrossRefGoogle Scholar
  4. 4.
    Cole, R.B., Harrata, A.K.: Solvent effect on analyte charge state, signal intensity, and stability in negative ion electrospray mass spectrometry; implications for the mechanism of negative ion formation. J. Am. Soc. Mass Spectrom. 4, 546–556 (1993)CrossRefGoogle Scholar
  5. 5.
    Cech, N.B., Enke, C.G.: Relating electrospray ionization response to nonpolar character of small peptides. Anal. Chem. 72, 2717–2723 (2000)CrossRefGoogle Scholar
  6. 6.
    Jemal, M., Hawthorne, D.J.: Effect of high performance liquid chromatography mobile phase (methanol versus acetonitrile) on the positive and negative ion electrospray response of a compound that contains both an unsaturated lactone and a methyl sulfone group. Rapid Commun. Mass Spectrom. 13, 61–66 (1999)CrossRefGoogle Scholar
  7. 7.
    Cech, N.B., Enke, C.G.: Practical implications of some recent studies in electrospray ionization fundamentals. Mass Spectrom. Rev. 20, 362–387 (2001)CrossRefGoogle Scholar
  8. 8.
    Cole, R.B.: Some tenets pertaining to electrospray ionization mass spectrometry. J. Mass Spectrom. 35, 763–772 (2000)CrossRefGoogle Scholar
  9. 9.
    Schug, K., McNair, H.M.: Adduct formation in electrospray ionization. Part 1: common acidic pharmaceuticals. J. Sep. Sci. 25, 759–766 (2002)CrossRefGoogle Scholar
  10. 10.
    Schug, K., McNair, H.M.: Adduct formation in electrospray ionization mass spectrometry. II. Benzoic acid derivatives. J. Chromatogr. A. 985, 531–539 (2003)CrossRefGoogle Scholar
  11. 11.
    Yoshino, K., Takao, T., Murata, H., Shimonishi, Y.: Use of the derivatizing agent 4-aminobenzoic acid 2-(diethylamino)ethyl ester for high-sensitivity detection of oligosaccharides by electrospray ionization mass spectrometry. Anal. Chem. 67, 4028–4031 (1995)CrossRefGoogle Scholar
  12. 12.
    Ahn, Y.H., Yoo, J.S.: Malononitrile as a new derivatizing reagent for high-sensitivity analysis of oligosaccharides by electrospray ionization mass spectrometry. Rapid Commun. Mass Spectrom. 12, 2011–2015 (1998)CrossRefGoogle Scholar
  13. 13.
    Ikonomou, M.G., Blades, A.T., Kebarle, P.: Investigations of the electrospray interface for liquid chromatography/mass spectrometry. Anal. Chem. 62, 957–967 (1990)CrossRefGoogle Scholar
  14. 14.
    Raffaelli, A., Bruins, A.P.: Factors affecting the ionization efficiency of quaternary ammonium compounds in electrospray/ionspray mass spectrometry. Rapid Commun. Mass Spectrom. 5, 269–275 (1991)CrossRefGoogle Scholar
  15. 15.
    Kohler, M., Leary, J.A.: LC/MS/MS of carbohydrates with postcolumn addition of metal chlorides using a triaxial electrospray probe. Anal. Chem. 67, 3501–3508 (1995)CrossRefGoogle Scholar
  16. 16.
    Hulthe, G., Stenhagen, G., Fogelqvist, E.: Comparison of dynamic fast atom bombardment/liquid secondary ion mass spectrometry and electrospray mass spectrometry coupled to reversed-phase liquid chromatography for the determination of oligosaccharides in seawater. J. Chromatogr. A. 777, 141–153 (1997)CrossRefGoogle Scholar
  17. 17.
    Rogatsky, E., Jayatillake, H., Goswami, G., Tomuta, V., Stein, D.: Sensitive LC MS quantitative analysis of carbohydrates by Cs+ attachment. J. Am. Soc. Mass Spectrom. 16, 1805–1811 (2005)CrossRefGoogle Scholar
  18. 18.
    Liu, Y., Urgaonkar, S., Verkade, J.G., Armstrong, D.W.: Separation and characterization of underivatized oligosaccharides using liquid chromatography and liquid chromatography–electrospray ionization mass spectrometry. J. Chromatogr. A. 1079, 146–152 (2005)CrossRefGoogle Scholar
  19. 19.
    Wan, E.C.H., Yu, J.Z.: Determination of sugar compounds in atmospheric aerosols by liquid chromatography combined with positive electrospray ionization mass spectrometry. J. Chromatogr. A. 1107, 175–181 (2006)CrossRefGoogle Scholar
  20. 20.
    Jiang, Y., Cole, R.B.: Oligosaccharide analysis using anion attachment in negative mode electrospray mass spectrometry. J. Am. Soc. Mass Spectrom. 16, 60–70 (2005)CrossRefGoogle Scholar
  21. 21.
    Thacker, J.B., Schug, K.A.: Effects of solvent parameters on the electrospray ionization tandem mass spectrometry response of glucose. Rapid Commun. Mass Spectrom. 32, 1191–1198 (2018)CrossRefGoogle Scholar
  22. 22.
    Santos, I.C., Waybright, V.B., Fan, H., Ramirez, S., Mesquita, R.B.R., Rangel, A.O.S.S., Fryčák, P., Schug, K.A.: Determination of noncovalent binding using a continuous stirred tank reactor as a flow injection device coupled to electrospray ionization mass spectrometry. J. Am. Soc. Mass Spectrom. 26, 1204–1212 (2015)CrossRefGoogle Scholar
  23. 23.
    Zhu, J., Cole, R.B.: Ranking of gas-phase acidities and chloride affinities of monosaccharides and linkage specificity in collision-induced decompositions of negative ion electrospray-generated chloride adducts of oligosaccharides. J. Am. Soc. Mass Spectrom. 12, 1193–1204 (2001)CrossRefGoogle Scholar
  24. 24.
    Kruve, A., Kaupmees, K., Liigand, J., Oss, M., Leito, I.: Sodium adduct formation efficiency in ESI source. J. Mass Spectrom. 48, 695–702 (2013)CrossRefGoogle Scholar
  25. 25.
    Angyal, S.J.: Complex formation between sugars and metal ions. Pure Appl. Chem. 35, 131–146 (1973)CrossRefGoogle Scholar
  26. 26.
    Verardo, G., Duse, I., Callea, A.: Analysis of underivatized oligosaccharides by liquid chromatography/electrospray ionization tandem mass spectrometry with post-column addition of formic acid. Rapid Commun. Mass Spectrom. 23, 1607–1618 (2009)CrossRefGoogle Scholar
  27. 27.
    McIntosh, T.S., Davis, H.M., Matthews, D.E.: A liquid chromatography–mass spectrometry method to measure stable isotopic tracer enrichments of glycerol and glucose in human serum. Anal. Biochem. 300, 163–169 (2002)CrossRefGoogle Scholar
  28. 28.
    Kruve, A., Kaupmees, K.: Adduct formation in ESI/MS by mobile phase additives. J. Am. Soc. Mass Spectrom. 28, 887–894 (2017)CrossRefGoogle Scholar

Copyright information

© American Society for Mass Spectrometry 2018

Authors and Affiliations

  1. 1.Department of Chemistry and BiochemistryThe University of Texas at ArlingtonArlingtonUSA

Personalised recommendations