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Development of a sensitive analytical method for determining 44 pyrrolizidine alkaloids in teas and herbal teas via LC-ESI-MS/MS

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Abstract

Pyrrolizidine alkaloids (PA) and PA-N-oxides (PANO) are a large group of secondary plant metabolites comprising more than 660 compounds. Exhibiting geno- and hepatotoxic properties, they are responsible for multiple cases of food and feed poisoning over the last 100 years. For food and feed safety reasons, relevant PA/PANO should be monitored extensively in the main sources of PA/PANO intake. In this study, a sensitive analytical method was developed for detecting a broad range of 44 commercially available PA/PANO compounds, and in-house validation procedures were performed for several (herbal) teas. Various extraction solvents and procedures, as well as solid phase extraction materials for sample clean-up and analyte concentration, were tested to establish the methods’ efficiency and effectiveness. Chromatographic conditions were optimised to obtain the best possible separation of isomers for the 44 PA/PANO analytes. The final method was proven very sensitive and accurate, with detection limits ranging from 0.1 to 7.0 μg/kg and precisions between 0.7 and 16.1%. For 40 of the analytes, the recovery rates ranged from 60.7 to 128.8%. The applicability and trueness of the method were examined by analysing tea samples from a local supermarket and comparing them to a reference material. At least one PA/PANO analyte was detected in 17 of the 18 samples under investigation, and the sum contents of the samples ranged from 0.1 to 47.9 μg/kg. Knowledge of the PA/PANO composition in a sample can be used to indicate the botanical origin of the impurity and, thus, the geographical region of cultivation.

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Abbreviations

AcIm:

Acetylintermedine

AcImN:

Acetylintermedine-N-oxide

AcLy:

Acetylycopsamine

AcLyN:

Acetylycopsamine-N-oxide

BfR:

German Federal Institute for Risk Assessment

BMDL10 :

Benchmark dose lower confidence limit 10%

Ec:

Erucifoline

EcN:

Erucifoline-N-oxide

Em:

Echimidine

EmN:

Echimidine-N-oxide

Eu:

Europine

EuN:

Europine-N-oxide

EFSA:

European Food Safety Authority

ESI+:

Positive electrospray ionisation

HILIC:

Hydrophilic interaction liquid chromatography

HPLC:

High-performance liquid chromatography

Ht:

Heliotrine

HtN:

Heliotrine-N-oxide

Ic:

Indicine

IcN:

Indicine-N-oxide

Ig:

Integerrimine

IgN:

Integerrimine-N-oxide

Im:

Intermedine

ImN:

Intermedine-N-oxide

Jb:

Jacobine

Jb:

Jacobine-N-oxide

Jl:

Jacoline

JlN:

Jacoline-N-oxide

Lc:

Lasiocarpine

LcN:

Lasiocarpine-N-oxide

LC-MS/MS:

Liquid chromatography tandem mass spectrometry

LOD:

Limit of detection

LOQ:

Limit of quantification

Ly:

Lycopsamine

LyN:

Lycopsamine-N-oxide

Mc:

Monocrotaline

McN:

Monocrotaline-N-oxide

Mk:

Merenskine

MkN:

Merenskine-N-oxide

MOE:

Margin of exposure

MRM:

Multiple reaction monitoring

Mx:

Merepoxine

MxN:

Merepoxine-N-oxide

MS:

Mass spectrometry

MS3:

Multiple tandem mass spectrometry

PA:

Pyrrolizidine alkaloid

PANO:

Pyrrolizidine alkaloid-N-oxide

RP:

Reversed phase

Rs:

Retrorsine

RsN:

Retrorsine-N-oxide

RSD:

Relative standard deviation

S/N:

Signal-to-noise ratio

Sc:

Senecionine

ScN:

Senecionine-N-oxide

SCX:

Strong cation exchange

Sl:

Sceleratine

SlN:

Sceleratine-N-oxide

Sp:

Seneciphylline

SPE:

Solid phase extraction

SpN:

Seneciphylline-N-oxide

Sk:

Senkirkine

Sv:

Senecivernine

SvN:

Senecivernine-N-oxide

Td:

Trichodesmine

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Acknowledgements

The authors gratefully acknowledge the skilful technical assistance of Carmen Piller, Michaela Freitag and Helmut Ziemann during the course of the study.

Funding

This IGF Project No. 19010 of the FEI was supported via AiF within the programme for promoting the Industrial Collective Research (IGF) of the German Ministry of Economic Affairs and Energy (BMWi), based on a resolution of the German Parliament. This project was additionally supported by the Adalbert-Raps Foundation (Kulmbach, Germany) and the Brigitte and Wolfram Gedek-Foundation (Ismaning, Germany).

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Authors and Affiliations

  1. Chair of Food Safety, Faculty of Veterinary Medicine, Ludwig-Maximilians-University of Munich, Schoenleutnerstr. 8, 85764, Oberschleissheim, Germany

    Florian Kaltner, Manfred Gareis & Christoph Gottschalk

  2. Chair of Analytical Food Chemistry, TUM School of Life Science Weihenstephan, Technical University of Munich, Maximus-von-Imhof-Forum 2, 85354, Freising, Germany

    Florian Kaltner, Benedikt Stiglbauer & Michael Rychlik

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  1. Florian Kaltner
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  2. Benedikt Stiglbauer
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  3. Michael Rychlik
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  4. Manfred Gareis
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  5. Christoph Gottschalk
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Correspondence to Florian Kaltner.

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Kaltner, F., Stiglbauer, B., Rychlik, M. et al. Development of a sensitive analytical method for determining 44 pyrrolizidine alkaloids in teas and herbal teas via LC-ESI-MS/MS. Anal Bioanal Chem 411, 7233–7249 (2019). https://doi.org/10.1007/s00216-019-02117-1

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  • DOI: https://doi.org/10.1007/s00216-019-02117-1

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