Analytical and Bioanalytical Chemistry

, Volume 411, Issue 2, pp 339–351 | Cite as

Comprehensive MS-based screening and identification of pharmaceutical transformation products formed during enzymatic conversion

  • Lara F. Stadlmair
  • Sylvia Grosse
  • Thomas LetzelEmail author
  • Jörg E. Drewes
  • Johanna Grassmann
Research Paper


In this study, transformation products (TPs) of diclofenac, mefenamic acid, and sotalol derived from peroxidase- and laccase-catalyzed transformations were studied with different mass spectrometry (MS)-based workflows. A straightforward pre-screening of enzymatic degradation rate was performed using a robotic nano-ESI source coupled to single quadrupole MS. Accurate mass data and information on molecular hydrophobicity were obtained from a serial coupling of reversed phase liquid chromatography (RPLC) with hydrophilic interaction liquid chromatography (HILIC) to a time-of-flight-mass spectrometer (ToF-MS). These parameters were combined with fragmentation information from product ion scan operated in enhanced mode (EPI) with precursor selection in Q3 and data from multiple reaction monitoring (MRM) modes using a hybrid triple quadrupole-linear ion trap-mass spectrometer (QqQ/LIT-MS). “Suspect” MRM modes did not provide a significant sensitivity improvement compared to EPI experiments. The complementarity of the data from different MS-based workflows allowed for an increase of identification confidence. Overall, this study demonstrated that dimerization, hydroxylation, and dehydration reactions were the predominant mechanisms found for diclofenac and mefenamic acid during enzyme-catalyzed transformation, whereas a degradation product was observed for the peroxidase-catalyzed conversion of sotalol. Results can contribute to understand enzymatic mechanisms and provide a basis for assessing risks and benefits of enzyme-based remediation.

Graphical abstract


Mass spectrometer HPLC Pharmaceuticals Oxidative enzymes Transformation products 


Funding information

This study was partially supported by the Bavarian State Ministry of the Environment and Consumer Protection.

Compliance with ethical standards


All authors are aware of and accept responsibility for this manuscript.

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

216_2018_1442_MOESM1_ESM.pdf (510 kb)
ESM 1 (PDF 510 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Lara F. Stadlmair
    • 1
  • Sylvia Grosse
    • 1
  • Thomas Letzel
    • 1
    Email author
  • Jörg E. Drewes
    • 1
  • Johanna Grassmann
    • 1
  1. 1.Chair of Urban Water Systems Engineering, Department of Civil, Geo and Environmental EngineeringTechnical University of MunichGarchingGermany

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