Analytical and Bioanalytical Chemistry

, Volume 411, Issue 17, pp 3963–3977 | Cite as

(Un)targeted hair metabolomics: first considerations and systematic evaluation on the impact of sample preparation

  • Lisa Eisenbeiss
  • Andrea E. SteuerEmail author
  • Tina M. Binz
  • Markus R. Baumgartner
  • Thomas Kraemer
Research Paper


The interest in metabolomic studies has rapidly increased over the past few years. Changes of endogenous compounds are typically detected in plasma or urine. However, the use of hair allows for long-term monitoring of metabolomic changes and has recently started being applied to metabolomic studies. Within the proposed study, we aimed for a systematical investigation of different pre-analytical parameters on detected metabolites from different chemical classes in hair. For this purpose, three different parameters were varied: (1) multi-step decontamination (dichloromethane (DCM), acetone, H2O, acetone; H2O, acetone, DCM, acetone; and H2O, methanol/acetone), (2) homogenization (pulverization vs. cutting into snippets), and (3) extraction (acetonitrile (ACN)/buffer pH 4 vs. ACN/H2O vs. ACN/buffer pH 8.5). To include as many metabolites as possible, samples were analyzed by high-resolution time of flight mass spectrometry coupled to liquid chromatography (HPLC-HRMS) and additionally by gas chromatography high-resolution mass spectrometry (GC-HRMS) followed by untargeted-like data processing, respectively. The application of different decontamination procedures yielded similar results, although pointing to a trend towards increased washing-out effects if protic solvents were used as a first washing step. Pulverization of hair samples was favorable in terms of detected and tentatively identified metabolites. Evaluation of extraction solvents showed differences in extraction yield for the majority of investigated metabolites, yet, a prediction of metabolite extraction according to their pKa values was not possible. Overall, successive decontamination with DCM, acetone, H2O, and acetone; homogenization by pulverization; and extraction with ACN/H2O produced reliable results.

Graphical abstract


Hair analysis Metabolomics Decontamination Extraction LC-MS/MS GC-MS 



The authors would like to express their gratitude to Emma Louise Kessler, MD, for her generous legacy; she donated to the Institute of Forensic Medicine at the University of Zurich, Switzerland, for research purposes.

Compliance with ethical standards

Hair samples were collected from two healthy volunteers who provided written informed consent. According to Swissethics (Humanforschungsgesetz), no further ethical approval from the cantonal ethic commission is necessary if the research is not aiming to investigate diseases or functions of the human body as is the case in the current study.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2019_1873_MOESM1_ESM.pdf (231 kb)
ESM 1 (PDF 231 kb)


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Copyright information

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

Authors and Affiliations

  1. 1.Department of Forensic Pharmacology and ToxicologyZurich Institute of Forensic MedicineZurichSwitzerland
  2. 2.Center for Forensic Hair AnalyticsZurich Institute of Forensic MedicineZurichSwitzerland

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