Internal Energy Deposition in Infrared Matrix-Assisted Laser Desorption Electrospray Ionization With and Without the Use of Ice as a Matrix
The internal energy deposited into analytes during the ionization process largely influences the extent of fragmentation, thus the appearance of the resulting mass spectrum. The internal energy distributions of a series of para-substituted benzyl pyridinium cations in liquid and solid-state generated by infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) were measured using the survival yield method, of which results were subsequently compared with conventional electrospray ionization (ESI). The comparable mean internal energy values (e.g., 1.8–1.9 eV at a collision energy of 15 eV) and peak widths obtained with IR-MALDESI and ESI support that IR-MALDESI are essentially a soft ionization technique where analytes do not gain considerable internal energy during the laser-induced desorption process and/or lose energy during uptake into charged electrospray droplets. An unusual fragment ion, protonated pyridine, was only found for solid IR-MALDESI at relatively high collision energies, which is presumably resulted from direct ionization of the pre-charged analytes in form of salts. Analysis of tissue with an ice layer consistently yielded ion populations with higher internal energy than its counterpart without an ice layer, likely due to a substantially enhanced number of IR absorbers with ice. Further measurements with holo-myoglobin show that IR-MALDESI-MS retains the noncovalently bound heme-protein complexes under both native-like and denaturing conditions, while complete loss of the heme group occurred in denaturing ESI-MS, showing that the softness of IR-MALDESI is equivalent or superior to ESI for biomolecules.
KeywordsInternal energy deposition IR-MALDESI Mass spectrometry imaging Survival yield method Thermometer ions Ambient ionization
All mass spectrometry measurements were carried out in the Molecular Education, Technology, and Research Innovation Center (METRIC) at North Carolina State University. The authors gratefully acknowledge the financial support received from the National Institutes of Health (R01GM087964) and North Carolina State University.
Compliance with Ethical Standards
The animal was managed in accordance with the Institute for Laboratory Animal Research Guide, and all husbandry practices were approved by North Carolina State University Institutional Animal Care and Use Committee (IACUC).
Conflict of Interest
The authors declare that they have no competing interests.
- 5.Sampson, J.S., Hawkridge, A.M., Muddiman, D.C.: Generation and detection of multiply-charged peptides and proteins by matrix-assisted laser desorption electrospray ionization (MALDESI) Fourier transform ion cyclotron resonance mass spectrometry. J. Am. Soc. Mass Spectrom. 17, 1712–1716 (2006)CrossRefGoogle Scholar
- 7.Bokhart, M.T., Rosen, E., Thompson, C., Sykes, C., Kashuba, A.D.M., Muddiman, D.C.: Quantitative mass spectrometry imaging of emtricitabine in cervical tissue model using infrared matrix-assisted laser desorption electrospray ionization. Anal. Bioanal. Chem. 407, 2073–2084 (2015)CrossRefGoogle Scholar
- 28.Hampton, C.Y., Silvestri, C.J., Forbes, T.P., Varady, M.J., Meacham, J.M., Fedorov, A.G., Degertekin, F.L., Fernández, F.M.: Comparison of the internal energy deposition of Venturi-assisted electrospray ionization and a Venturi-assisted array of micromachined ultrasonic electrosprays (AMUSE). J. Am. Soc. Mass Spectrom. 19, 1320–1329 (2008)CrossRefGoogle Scholar
- 32.Omary, M.A., Patterson, H.H.: Luminescence, Theory. Encycl. Spectrosc. Spectrom (Third Edition), 636–653 (2017)Google Scholar
- 33.Tu, A., Muddiman, D.C.: Systematic evaluation of repeatability of IR-MALDESI-MS and normalization strategies for correcting the analytical variation and improving image quality. Anal. Bioanal. Chem. 411, 5729–5743 (2019)Google Scholar
- 36.Nazari, M., Bokhart, M.T., Muddiman, D.C.: Whole-body mass spectrometry imaging by infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI). J. Vis. Exp. 109, e53942 (2016)Google Scholar
- 39.Goodwin, R.J.A., Iverson, S.L., Andren, P.E.: The significance of ambient-temperature on pharmaceutical and endogenous compound abundance and distribution in tissues sections when analyzed by matrix-assisted laser desorption/ionization mass spectrometry imaging. Rapid Commun. Mass Spectrom. 26, 494–498 (2012)CrossRefGoogle Scholar
- 42.Zhang, W., Niu, S., Chait, B.T.: Exploring infrared wavelength matrix-assisted laser desorption/ionization of proteins with delayed-extraction time-offlight mass spectrometry. J. Am. Soc. Mass Spectrom. 9, 879–884 (1998)Google Scholar
- 52.Chen, Z., Vertes, A.: Early plume expansion in atmospheric pressure midinfrared laser ablation of water-rich targets. Phys. Rev. E. 77, 036316 (2008)Google Scholar
- 56.Li, G., Cao, Q., Liu, Y., DeLaney, K., Tian, Z., Moskovets, E., Li, L.: Characterizing and alleviating ion suppression effects in atmospheric pressure matrix-assisted laser desorption/ionization. Rapid Commun. Mass Spectrom. 33, 327–335 (2019)Google Scholar