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Effects of sample properties and mass spectroscopic parameters on electrospray ionization mass spectra of size-fractions from a soil humic acid

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Abstract

Size-fractions from a soil humic acid were separated by preparative size-exclusion chromatography (SEC), desalted, and concentrated by ultrafiltration and vacuum centrifugation without being subjected to any freeze-drying process. After having assessed the lack of formation of any multiple-charged ions by high-resolution Fourier transform ion cyclotron resonance electrospray ionization (ESI) mass spectrometry (MS), the size-fractions were used by direct infusion to compare the molecular ion distribution by both atmospheric pressure chemical ionization (APCI)- and ESI-MS in negative mode. The weight- (Mw) and number-averaged (Mn) molecular weight obtained by ESI-MS were invariably larger than by APCI-MS for all size-fractions, thereby indicating that ESI is more efficient than APCI to evaluate the molecular mass distribution of humic samples. No substantial difference was observed when concentration and pH of unfreeze-dried humic size-fractions were varied. The negative mode was applied to assess the effect of cone voltage from −20 to −60 V on ESI of the humic size-fractions further separated through an on-line SEC column. The resulting mass spectra and Mw and Mn values suggested that the variation of cone voltage in ESI-MS affects the ionization potential of associated humic molecules more in solution rather than their fragmentation. These findings agree with previous observations which indicated a limitation of ESI in providing consistent mass detection for a complex mixture of heterogeneous humic molecules, especially when they are aggregated by a freeze-drying process.

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Acknowledgements

The authors are grateful to Dr. Mattias Witt of Bruker Daltonics, Bremen, for FT-ICR-MS analyses. The collaboration of Mrs. S. Richter in ESI- and APCI-MS analyses is gratefully appreciated.

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Authors and Affiliations

  1. Dipartimento di Scienze del Suolo, della Pianta, dell’Ambiente, delle Produzioni Animali, Università di Napoli Federico II, Via Università 100, 80055, Portici, Italy

    A. Piccolo & A. Nebbioso

  2. Institute of Environmental Research (INFU), University of Dortmund, Otto-Hahn-Strasse 6, 44227, Dortmund, Germany

    M. Spiteller

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  1. A. Piccolo
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  2. M. Spiteller
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  3. A. Nebbioso
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Correspondence to A. Piccolo.

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Piccolo, A., Spiteller, M. & Nebbioso, A. Effects of sample properties and mass spectroscopic parameters on electrospray ionization mass spectra of size-fractions from a soil humic acid. Anal Bioanal Chem 397, 3071–3078 (2010). https://doi.org/10.1007/s00216-010-3884-4

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  • DOI: https://doi.org/10.1007/s00216-010-3884-4

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