Analytical and Bioanalytical Chemistry

, Volume 408, Issue 12, pp 3319–3329 | Cite as

A fully automated and fast method using direct sample injection combined with fused-core column on-line SPE–HPLC for determination of ochratoxin A and citrinin in lager beers

  • Ivona Lhotská
  • Dalibor Šatínský
  • Lucie Havlíková
  • Petr Solich
Research Paper

Abstract

A new fast and sensitive method based on on-line solid-phase extraction on a fused-core precolumn coupled to liquid chromatography with fluorescence detection has been developed for ochratoxin A (OTA) and citrinin (CIT) determination in lager beer samples. Direct injection of 100 μL filtered beer samples into an on-line SPE–HPLC system enabled fast and effective sample extraction including separation in less than 6 min. Preconcentration of OTA and CIT from beer samples was performed on an Ascentis Express RP C18 guard column (5 × 4.6 mm), particle size 2.7 μm, with a mobile phase of methanol/0.5 % aqueous acetic acid pH 2.8 (30:70, v/v) at a flow rate of 2.0 mL min−1. The flow switch from extraction column to analytical column in back-flush mode was set at 2.0 min and the separation was performed on the fused-core column Ascentis Express Phenyl-Hexyl (100 × 4.6 mm), particle size 2.7 μm, with a mobile phase acetonitrile/0.5 % aqueous acetic acid pH 2.8 in a gradient elution at a flow rate of 1.0 mL min−1 and temperature of 50 °C. Fluorescence excitation/emission detection wavelengths were set at 335/497 nm. The accuracy of the method, defined as the mean recoveries of OTA and CIT from light and dark beer samples, was in the range 98.3–102.1 %. The method showed high sensitivity owing to on-line preconcentration; LOQ values were found to be 10 and 20 ng L−1 for OTA and CIT, respectively. The found values of OTA and CIT in all tested light, dark and wheat beer samples were significantly below the maximum tolerable limits (3.0 μg kg−1 for OTA and 2000 μg kg−1 for CIT) set by the European Union.

Graphical Abstract

The beer contamination - potential sources of mycotoxins

Keywords

On-line SPE–HPLC Ochratoxin A Citrinin Mycotoxins Beer Fused-core column 

Notes

Acknowledgements

The authors are grateful to the Charles University Grant Agency for grant GAUK no. 1316213. I. Lhotská would like to acknowledge financial support of the project of specific research, no. SVV 260 292. The work was co-financed by the project GAČR no. 15-10781S.

Compliance with ethical standards

Conflict of interest

The authors report no conflicts of interest in this work.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Ivona Lhotská
    • 1
  • Dalibor Šatínský
    • 1
  • Lucie Havlíková
    • 1
  • Petr Solich
    • 1
  1. 1.Department of Analytical Chemistry, Faculty of PharmacyCharles UniversityHradec KrálovéCzech Republic

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