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Decay Mechanisms of Protonated 4-Quinolone Antibiotics After Electrospray Ionization and Ion Activation

  • Research Article
  • Published:
Journal of The American Society for Mass Spectrometry

Abstract

This study presents a detailed experimental investigation of charge isomers of protonated 4-quinolone antibiotics molecules formed during electrospray ionization (ESI) with proposed dissociation mechanisms after collisional activation. Piperazinyl quinolones have been previously shown to exhibit erratic behavior during tandem MS analyses of biological samples, which originated from varying ratios of two isomeric variants formed during ESI. Here, a combination of ESI-collision-induced dissociation (CID), differential ion mobility spectrometry (DMS), high resolution MS, and density functional theory (DFT) was used to investigate the underlying mechanisms of isomer formation and their individual dissociation behaviors. The study focused on ciprofloxacin; major findings were confirmed using structurally related 4-quinolones. DFT calculations showed a reversal of basicity for piperazinyl quinolones between liquid and gas phase. We provide an experimental comparison and theoretical treatment of factors influencing the formation ratio of the charge isomers during ESI, including solvent pH, protic/aprotic nature of solvent, and structural effects such as pK a and proton affinity. The actual dissociation mechanisms of the isomers of the protonated molecules were studied by separating the individual isomers via DMS-MS, which allowed type-specific CID spectra to be recorded. Both primary CID reactions of the two charge isomers originated from the same carboxyl group by charge-remote (CO2 loss) and charge-mediated (H2O loss) fragmentation of the piperazinyl quinolones, depending on whether the proton resides on the more basic keto or the piperazinyl group, followed by a number of secondary dissociation reactions. The proposed mechanisms were supported by calculated energies of precursors, transition states, and products for competing pathways.

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Acknowledgments

D.A.V. and B.K. acknowledge joint funding from a bilateral research program of the German Academic Exchange Service (DAAD) and the Croatian Ministry of Science, Education, and Sports (MZOŠ). B.K. thanks the Computing Center of the University of Zagreb (SRCE) for computational time on the ISABELLA cluster. D.A.V. acknowledges general research support by the Alfried Krupp von Bohlen und Halbach Stiftung.

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Authors and Affiliations

  1. Quantum Chemistry Group, Ruđer Bošković Institute, Zagreb, Croatia

    Borislav Kovačević

  2. Institute of Bioanalytical Chemistry, Department of Chemistry, Saarland University, Saarbrücken, Germany

    Pascal Schorr, Yulin Qi & Dietrich A. Volmer

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  1. Borislav Kovačević
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  2. Pascal Schorr
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  3. Yulin Qi
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  4. Dietrich A. Volmer
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Correspondence to Dietrich A. Volmer.

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Kovačević, B., Schorr, P., Qi, Y. et al. Decay Mechanisms of Protonated 4-Quinolone Antibiotics After Electrospray Ionization and Ion Activation. J. Am. Soc. Mass Spectrom. 25, 1974–1986 (2014). https://doi.org/10.1007/s13361-014-0972-2

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