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Bioavailability and biotransformation of sulforaphane and erucin metabolites in different biological matrices determined by LC–MS–MS

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Abstract

The food-related isothiocyanate sulforaphane (SFN), a hydrolysis product of the secondary plant metabolite glucoraphanin, has been revealed to have cancer-preventive activity in experimental animals. However, these studies have often provided inconsistent results with regard to bioavailability, bioaccessibility, and outcome. This might be because the endogenous biotransformation of SFN metabolites to the structurally related erucin (ERN) metabolites has often not been taken into account. In this work, a fully validated liquid chromatography tandem mass spectrometry (LC–MS–MS) method was developed for the simultaneous determination of SFN and ERN metabolites in a variety of biological matrices. To reveal the importance of the biotransformation pathway, matrices including plasma, urine, liver, and kidney samples from mice and cell lysates derived from colon-cancer cell lines were included in this study. The LC–MS–MS method provides limits of detection from 1 nmol L−1 to 25 nmol L−1 and a mean recovery of 99 %. The intra and interday imprecision values are in the range 1–10 % and 2–13 %, respectively. Using LC–MS–MS, SFN and ERN metabolites were quantified in different matrices. The assay was successfully used to determine the biotransformation in all biological samples mentioned above. For a comprehensive analysis and evaluation of the potential health effects of SFN, it is necessary to consider all metabolites, including those formed by biotransformation of SFN to ERN and vice versa. Therefore, a sensitive and robust LC–MS–MS method was validated for the simultaneous quantification of mercapturic-acid-pathway metabolites of SFN and ERN.

Biotransformation of sulforaphane and erucin metabolites in mice and cell culture

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Abbreviations

ACN:

Acetonitrile

DMEM:

Dulbecco’s modified Eagle’s medium

DMSO:

Dimethyl sulfoxide

ERN:

Erucin

ERN–Cys:

ERN–cysteine

ERN–GSH:

ERN–glutathione

ERN–NAC:

ERN–N-acetylcysteine

ESI:

Electrospray ionization

FA:

Formic acid

GSH:

Glutathione

HPLC:

High-performance liquid chromatography

LC–MS–MS:

Liquid chromatography–tandem mass spectrometry

LOQ:

Limit of quantification

MeOH:

Methanol

NMR:

Nuclear magnetic resonance

R 2 :

Coefficient of determination

SFN:

Sulforaphane

SFN–Cys:

SFN–cysteine

SFN–CysGly:

SFN–cysteinylglycine

SFN–NAC:

SFN–N-acetylcysteine

SPE:

Solid-phase extraction

TFA:

Trifluoroacetic acid

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Authors and Affiliations

  1. Hamburg School of Food Science, Institute of Food Chemistry, University of Hamburg, 20146, Hamburg, Germany

    Stefanie Platz & Sascha Rohn

  2. Department of Food Chemistry and Toxicology, Institute of Applied Bioscience, Karlsruhe Institute of Technology, 76131, Karlsruhe, Germany

    Ann Liza Piberger & Andrea Hartwig

  3. Department of Gastrointestinal Microbiology, German Institute of Human Nutrition Potsdam-Rehbrücke, 14558, Nuthetal, Germany

    Julia Budnowski, Michael Blaut & Laura Hanske

  4. Institute for Environmental Health Sciences and Hospital Infection Control, University Medical Center, 79106, Freiburg, Germany

    Corinna Herz & Evelyn Lamy

  5. Department of Quality, Leibniz-Institute of Vegetable and Ornamental Crops, Großbeeren/Erfurt e.V., 14979, Großbeeren, Germany

    Monika Schreiner

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  1. Stefanie Platz
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  2. Ann Liza Piberger
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  3. Julia Budnowski
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  4. Corinna Herz
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  8. Evelyn Lamy
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Correspondence to Sascha Rohn.

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Platz, S., Piberger, A.L., Budnowski, J. et al. Bioavailability and biotransformation of sulforaphane and erucin metabolites in different biological matrices determined by LC–MS–MS. Anal Bioanal Chem 407, 1819–1829 (2015). https://doi.org/10.1007/s00216-015-8482-z

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  • DOI: https://doi.org/10.1007/s00216-015-8482-z

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