Analytical and Bioanalytical Chemistry

, Volume 411, Issue 2, pp 479–491 | Cite as

Quantitative analysis of chemoresistance-inducing fatty acid in food supplements using UHPLC–ESI-MS/MS

  • Jörg Schlotterbeck
  • Malgorzata Cebo
  • Agnes Kolb
  • Michael LämmerhoferEmail author
Research Paper


Polyunsaturated fatty acids are important signaling molecules. A recent study reported hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid, 12-oxo-5Z,8E,10E-heptadecatrienoic acid, and (12S)-hydroxy-5Z,8E,10E-heptadecatrienoic acid as chemotherapy resistance-inducing factors when tumor cells were treated with cisplatin. Marine-based food supplements like fish oil or algae extracts are rich in polyunsaturated fatty acids and can contain large amounts of hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid. Thus, it was concluded that oral uptake of hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid might induce chemoresistance as shown in a mouse model. Cancer patients tend to consume food supplements containing polyunsaturated fatty acids on a regular basis. The uptake of hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid and (12S)-hydroxy-5Z,8E,10E-heptadecatrienoic acid should be controlled, because even low concentrations of 0.5 ng mL-1 showed chemoresistance-inducing effects in animal experiments. For accurate analysis of hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid and (12S)-hydroxy-5Z,8E,10E-heptadecatrienoic acid a validated method was developed by using ultrahigh-performance liquid chromatography hyphenated to quadrupole time of flight mass spectrometry via electrospray ionization and sample preparation by solid-phase extraction (SPE) with 3-aminopropyl silica. A combined targeted/untargeted approach was utilized using MS/MS by data-independent acquisition with SWATH and applied to commercial food supplements (refined fish oil, fish oil capsules, algae oil capsules, and flaxseed capsules). Accurate quantification of hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid and (12S)-hydroxy-5Z,8E,10E-heptadecatrienoic acid on the MS/MS level with simultaneous untargeted fatty acid screening revealed additional information. The LODs for hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid and (12S)-hydroxy-5Z,8E,10E-heptadecatrienoic acid were 0.036 ng mL-1 and 0.054 ng mL-1, respectively. Since hexadeca-4Z,7Z,10Z,13Z-tetraenoic acid was present in the samples in large amounts and (12S)-hydroxy-5Z,8E,10E-heptadecatrienoic was not expected to be present in high concentrations, two calibration ranges, namely, 0.5–20 ng mL-1 and 5–200 ng mL-1, were validated. An untargeted screening identified 18–39 free fatty acids being present in the lipid extracts of the food supplement samples.

Graphical abstract


Platinum-induced fatty acids Chemotherapy resistance-inducing fatty acid Fish oil Data-independent acquisition SWATH 



We acknowledge the financial support by the “Struktur- und Innovationsfonds Baden-Württemberg (SI-BW)”, the German Science Foundation (DFG no. INST 37/821-1 FUGG).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2018_1468_MOESM1_ESM.pdf (159 kb)
ESM 1 (PDF 158 kb)


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Copyright information

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

Authors and Affiliations

  • Jörg Schlotterbeck
    • 1
  • Malgorzata Cebo
    • 1
  • Agnes Kolb
    • 1
  • Michael Lämmerhofer
    • 1
    Email author
  1. 1.Institute of Pharmaceutical Sciences, Pharmaceutical (Bio-)AnalysisUniversity of TübingenTübingenGermany

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