Analytical and Bioanalytical Chemistry

, Volume 411, Issue 29, pp 7771–7781 | Cite as

Non-targeted mercapturic acid screening in urine using LC-MS/MS with matrix effect compensation by postcolumn infusion of internal standard (PCI-IS)

  • Robert Bloch
  • Susann-Elisabeth Schütze
  • Erik Müller
  • Stefan Röder
  • Irina Lehmann
  • Werner Brack
  • Martin KraussEmail author
Research Paper


While the targeted analysis of mercapturic acid (MA) metabolites in human urine is used to assess exposure to selected chemicals, this compound class has only rarely been addressed in non-target screening utilizing diagnostic neutral loss liquid chromatography tandem mass spectrometry (LC-MS/MS). Additionally, this type of analysis is severely affected by matrix effects (MEs) causing poor comparability of samples and distortion of signal intensities. However, MEs have been neglected in urinary MA non-target screening so far. Therefore, we developed a non-target screening method relying on neutral loss scanning for MAs using post column infusion of an isotope-labelled standard. For signal correction, we synthesized a structural analogue to MAs, N-acetyl-S-methyl-homocysteine-D3, lacking the characteristic neutral loss of the MAs. For method development, 16 structurally different model MA compounds and 20 spiked urine samples were used. Twelve out of the 16 model compounds could be analysed by the developed method. We found severe matrix effects (largely signal suppression) for the spiked model compounds, with only 34% of all peaks’ intensities changing by less than a factor of two. This could be compensated by the post column internal standard infusion with now 68% of all peaks’ intensities changing by less than a factor of two. For three compounds, an over-compensation was observed resulting in an increase of signal of up to a factor of 16. In the 20 urine samples, altogether 558 native MAs (between 74 and 175 per sample) could be detected after ME compensation. These results indicate that a large number of so far uncharacterized MAs are present in urine, which yield a potential for biomarker discovery and pattern characterisation.

Graphical Abstract


LC-MS/MS Human urine Non-target screening Postcolumn infusion 



Collision energy


Electrospray ionisation




Internal standard


Liquid chromatography tandem mass spectrometry


Lowest detectable concentration


Lifestyle and Environmental Factors and their Influence on Newborns Allergy risk


Mercapturic acid


Matrix effect


Multiple reaction monitoring


Neutral loss


Postcolumn infusion



The authors thank Margit Petre for technical assistance and Steffen Neumann for his input regarding the R script.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Participation in the study was voluntary and informed written consent was provided by the participants. The study was conducted according to international ethical guidelines for epidemiological studies. Study participants were recruited at the municipal hospital “St. Georg” in Leipzig, Germany, an academic hospital of Leipzig University. The study was approved by the ethics committee of Leipzig University (file ref.# 046-2006).

Supplementary material

216_2019_2166_MOESM1_ESM.pdf (355 kb)
ESM 1 (PDF 355 kb).


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Robert Bloch
    • 1
    • 2
  • Susann-Elisabeth Schütze
    • 1
  • Erik Müller
    • 1
    • 2
  • Stefan Röder
    • 3
  • Irina Lehmann
    • 3
    • 4
  • Werner Brack
    • 1
    • 2
  • Martin Krauss
    • 1
    Email author
  1. 1.Department of Effect-Directed AnalysisHelmholtz Centre for Environmental Research – UFZLeipzigGermany
  2. 2.Institute for Environmental Research (Biology V), Department of Ecosystem Analysis (ESA)RWTH Aachen UniversityAachenGermany
  3. 3.Department of Environmental ImmunologyHelmholtz Centre for Environmental Research – UFZLeipzigGermany
  4. 4.Berlin Institute of Health, Molecular Epidemiology UnitCharité – Universitätsmedizin BerlinBerlinGermany

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