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Evidence of ochratoxin A conjugates in urine samples from infants and adults

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Abstract

Ochratoxin A (OTA), a mycotoxin with nephrotoxic and carcinogenic properties, is an important contaminant of food and feed. Analysis of OTA in human biological fluids (blood, urine, or breast milk) has documented frequent exposure to this mycotoxin, yet at quite variable levels in different population groups across the world. Urine is the preferred matrix in biomonitoring since sample collection is non-invasive and better accepted by study participants. As only a small fraction of the ingested OTA is excreted in urine, determination of urinary OTA requires sensitive analytical techniques, and phase-II-metabolites should be also considered as biomarkers of exposure. Yet, data published so far on the presence of OTA-glucuronide/sulfate in human urine have been contradictory. In this study, urines (n = 38) from two groups of breastfed infants (German and Turkish) and from German adults were now analysed for the presence of OTA glucuronides or sulfates by an indirect method, i.e. by comparing the levels of OTA (aglycone) in urines without and after enzymatic hydrolysis with ß-glucuronidase/arylsulfatase. Additionally, ochratoxin A-8-β-glucuronide and open lactone ochratoxin A-8-β-glucuronide were synthesized to serve as reference materials for metabolite analysis. Attempts for definitive confirmation of glucuronides of OTA via direct identification in LC–MS/MS analysis were hampered by the lower ionizability of the conjugates compared to the parent compound. Considerable increases in OTA levels were found after enzymatic hydrolysis in several (not all) urine samples and provide clear evidence for the excretion of OTA-conjugates. The latter observation is of importance, since OTA phase-II-metabolites may escape detection when direct methods are applied for urinary biomarker analysis. In conclusion, enzymatic hydrolysis of urine samples is highly advisable in order to avoid an underestimation of the OTA-exposure.

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Acknowledgment

The authors want to thank Iris Glaeser (IfADo) for their technical help and Dr. Meinolf Blaszkewicz for his comments on an earlier version of this manuscript. Finally, we want to thank DAAD and CONICYT for their support.

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Authors and Affiliations

  1. Institute of Environmental Sciences, University Koblenz-Landau, Fortstrasse 7, 76829, Landau in der Pfalz, Germany

    K. Muñoz

  2. Institute of Food Chemistry, Westfälische Wilhelms-Universität Münster, Corrensstrasse 45, 48149, Münster, Germany

    B. Cramer, J. Dopstadt & H.-U. Humpf

  3. Leibniz Research Centre for Working Environment and Human Factors (IfADo), Ardeystrasse 67, 44139, Dortmund, Germany

    G. H. Degen

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  1. K. Muñoz
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  2. B. Cramer
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Correspondence to K. Muñoz.

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This study was funded by Leibniz Research Centre for Working Environment and Human Factors (IfADo).

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Chromatograms of the MRM-transitions of the reference solutions for open lactone ochratoxin A-8-O-β-glucuronide and ochratoxin A-8-O-β-glucuronide as well as product ion spectra of both compounds are provided.

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Muñoz, K., Cramer, B., Dopstadt, J. et al. Evidence of ochratoxin A conjugates in urine samples from infants and adults. Mycotoxin Res 33, 39–47 (2017). https://doi.org/10.1007/s12550-016-0261-y

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