Liquid chromatography–tandem mass spectrometry analysis of pyrrolizidine alkaloids in polish honey

Ewelina Kowalczyk

doi:10.2478/jvetres-2026-0046

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Liquid chromatography–tandem mass spectrometry analysis of pyrrolizidine alkaloids in polish honey

Journal of Veterinary Research

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By: Ewelina Kowalczyk

Open Access

|Aug 2026

Abstract

Introduction

Pyrrolizidine alkaloids (PAs) are among the most widespread naturally occurring toxins. Since they are primarily produced by flowering plants, there is a high likelihood of their transfer into honey. Given the toxic properties of PAs, their presence in honey can significantly compromise the safety of the product. Therefore, monitoring the occurrence of PAs in honey is essential.

Material and Methods

A total of 65 honey samples collected in Poland were analysed for the presence of 42 PAs. The samples included multifloral, acacia, rapeseed and forest honeys. Pyrrolizidine alkaloids were quantified using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS).

Results

Quantifiable amounts of PAs were detected in 10 of the honey samples. At least one of the monitored PAs was present in 39 samples, although most were below the method’s limit of quantification. The highest concentration observed was 46.9 μg/kg.

Conclusion

A newly validated LC-MS/MS method was successfully applied to determine PAs in honey from Poland. Most of the analysed samples were free of the monitored PAs, confirming the general safety of Polish honey.

References

Barreiros J., Cepeda A., Franco C., Nebot C., Vázquez B.: Analysis of minerals in honey and their nutritional implications. J Food Compos Anal 2024, 136, 106733, doi: 10.1016/j.jfca.2024.106733.
Open DOI Search in Google Scholar Back to article
Boppré M.: The ecological context of pyrrolizidine alkaloids in food, feed and forage: An overview. Food Addit Contam Part A 2011, 28, 260–281, doi: 10.1080/19440049.2011.555085.
Open DOI Search in Google Scholar Back to article
Boppré M., Colegate S.M., Edgar J.A.: Pyrrolizidine alkaloids of Echium vulgare honey found in pure pollen. J Agric Food Chem 2005, 53, 594–600, doi: 10.1021/jf0484531.
Open DOI Search in Google Scholar Back to article
Chen T., Mei N., Fu P.P.: Genotoxicity of pyrrolizidine alkaloids. J Appl Toxicol 2010, 30, 183–196, doi: 10.1002/jat.1490.
Open DOI Search in Google Scholar Back to article
Directorate-General for Health and Food Safety of the European Commission: Analytical quality control and method validation procedures for pesticide residues analysis in food and feed (SANTE/11312/2021 v2). Directorate-General for Health and Food Safety, 2021.
Search in Google Scholar Back to article
Eurachem/Co-operation on International Traceability in Analytical Chemistry: Guide: CG 4 Quantifying Uncertainty in Analytical Measurement, third edition, edited by S.L.R. Ellison, A. Williams, Eurachem/CITAC, 2012.
Search in Google Scholar Back to article
European Commission: Commission Regulation (EU) 2020/2040 of 11 December 2020 amending Regulation (EC) No. 1881/2006 as regards maximum levels of pyrrolizidine alkaloids in certain foodstuffs. OJEU L 2020, 63, 420, 14/12/2020, 1–5.
Search in Google Scholar Back to article
European Commission: Commission Regulation (EU) 2023/915 of 25 April 2023 on maximum levels for certain contaminants in food and repealing Regulation (EC) No. 1881/2006. OJEU L 2023, 66, 119, 05/05/2023, 103–157.
Search in Google Scholar Back to article
European Food Safety Authority Panel on Contaminants in the Food Chain (CONTAM), Alexander J., Benford D., Boobis A., Ceccatelli S., Cottrill B., Cravedi J.-P., Di Domenico A., Doerge D., Dogliotti E., Edler L., Farmer P., Filipič M., Fink-Gremmels J., Fürst P., Guérin T., Knutsen H.K., Machala M., Mutti A., Rose M., Schlatter J., van Leeuwen R.: Scientific Opinion on pyrrolizidine alkaloids in food and feed. EFSA J 2011, 9, 1–134, doi: 10.2903/j.efsa.2011.2406.
Open DOI Search in Google Scholar Back to article
European Food Safety Authority Panel on Contaminants in the Food Chain (CONTAM), Alexander J., Barregård L., Bignami M., Ceccatelli S., Cottrill B., DiNovi M., Edler L., Grasl-Kraupp B., Hogstrand C., Hoogenboom L.R., Knutsen H.K., Nebbia C.S., Oswald I.P., Petersen A., Rogiers V.M., Rose M., Roudot A.-C., Schwerdtle T., Vleminckx C., Vollmer G., Wallace H.: Scientific Report: Dietary exposure assessment to pyrrolizidine alkaloids in the European population. EFSA J 2016, 14, 1–50, doi: 10.2903/j.efsa.2016.4572.
Open DOI Search in Google Scholar Back to article
European Food Safety Authority Panel on Contaminants in the Food Chain (CONTAM), Knutsen H.K., Alexander J., Barregård L., Bignami M., Brüschweiler B., Ceccatelli S., Cottrill B., DiNovi M., Edler L., Grasl-Kraupp B., Hogstrand C., Hoogenboom L., Nebbia C.S., Oswald I.P., Petersen A., Rose M., Roudot A.C., Schwerdtle T., Vleminckx C., Vollmer G., Wallace H., Ruiz Gomes J.A., Binaglia M.: Statement: Risks for human health related to the presence of pyrrolizidine alkaloids in honey, tea, herbal infusions and food supplements. EFSA J 2017, 15, 4908, doi: 10.2903/j.efsa.2017.4908.
Open DOI Search in Google Scholar Back to article
Fu P.P., Xia Q., Chou M.W., Lin G.: Detection, hepatotoxicity, and tumorigenicity of pyrrolizidine alkaloids in Chinese herbal plants and herbal dietary supplements. J Food Drug Anal 2007, 15, 400–415, doi: 10.38212/2224-6614.2392.
Open DOI Search in Google Scholar Back to article
Gottschalk C., Huckauf A., Dübecke A., Kaltner F., Zimmermann M., Rahaus I., Beuerle T.: Uncertainties in the determination of pyrrolizidine alkaloid levels in naturally contaminated honeys and comparison of results obtained by different analytical approaches. Food Addit Contam Part A 2018, 35, 1366–1383, doi: 10.1080/19440049.2018.1468929.
Open DOI Search in Google Scholar Back to article
Huybrechts B., Callebaut A.: Pyrrolizidine alkaloids in food and feed on the Belgian market. Food Addit Contam Part A 2015, 32, 1939–1951, doi: 10.1080/19440049.2015.1086821.
Open DOI Search in Google Scholar Back to article
Kempf M., Reinhard A., Beuerle T.: Pyrrolizidine alkaloids (PAs) in honey and pollen – legal regulation of PA levels in food and animal feed required. Mol Nutr Food Res 2010, 54. 158–168, doi: 10.1002/mnfr.200900529.
Open DOI Search in Google Scholar Back to article
Kowalczyk E., Kwiatek K.: Pyrrolizidine alkaloids in honey: determination with liquid chromatography–mass spectrometry method. J Vet Res 2018, 62, 173–181, doi: 10.2478/jvetres-2018-0027.
Open DOI Search in Google Scholar Back to article
Kowalczyk E., Kwiatek K.: Simultaneous determination of pyrrolizidine and tropane alkaloids in honey by liquid chromatography–mass spectrometry. J Vet Res 2022, 66, 235–243, doi: 10.2478/jvetres-2022-0032.
Open DOI Search in Google Scholar Back to article
Lindigkeit I.R., Biller A., Buch M., Schiebel H.M., Boppré M., Hartmann T.: The two faces of pyrrolizidine alkaloids: the role of the tertiary amine and its N-oxide in chemical defense of insects with acquired plant alkaloids. Proc Natl Acad Sci U S A 1997, 94, 6795–6800, doi: 10.1073/pnas.94.13.6795.
Open DOI Search in Google Scholar Back to article
Lucatello L., Merlanti R., Rossi A., Montesissa C., Capolongo F.: Evaluation of some pyrrolizidine alkaloids in honey samples from the Veneto region (Italy) by LC-MS/MS. Food Anal Methods 2016, 9, 1825–1836, doi: 10.1007/s12161-015-0364-7.
Open DOI Search in Google Scholar Back to article
Macel M., Vrieling K., Klinkhamer P.G.L.: Variation in pyrrolizidine alkaloid patterns of Senecio jacobaea. Phytochemistry 2004, 65, 865–873, doi: 10.1016/j.phytochem.2004.02.009.
Open DOI Search in Google Scholar Back to article
Mulder P.P.J., Sánchez P.L., These A., Preiss-Weigert A., Castellari M.: Occurrence of pyrrolizidine alkaloids in food. EFSA Support Publ 2017, 12, 859E, doi: 10.2903/sp.efsa.2015.en-859.
Open DOI Search in Google Scholar Back to article
Roncada P., Isani G., Peloso M., Dalmonte T., Bonan S., Caprai E.: Pyrrolizidine alkaloids from monofloral and multifloral Italian honey. Int J Environ Res Public Health 2023, 20, 5410, doi: 10.3390/ijerph20075410.
Open DOI Search in Google Scholar Back to article
Than K.A., Stevens V., Knill A., Gallagher P.F., Gaul K.L., Edgar J.A., Colegate S.M.: Plant-associated toxins in animal feed: screening and confirmation assay development. Anim Feed Sci Technol 2005, 121, 5–21, doi: 10.1016/j.anifeedsci.2005.02.005.
Open DOI Search in Google Scholar Back to article
Xia Q., He X., Shi Q., Lin G., Fu P.P.: Quantitation of DNA reactive pyrrolic metabolites of senecionine – a carcinogenic pyrrolizidine alkaloid by LC/MS/MS analysis. J Food Drug Anal 2020, 28, 167–174, doi: 10.1016/j.jfda.2019.12.001.
Open DOI Search in Google Scholar Back to article

Articles in this issue

DOI: https://doi.org/10.2478/jvetres-2026-0046 | Journal eISSN: 2450-8608 (formerly 2300-3235)

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Language: English

Submitted on: Mar 2, 2026

Accepted on: Aug 21, 2026

Published on: Aug 27, 2026

Published by: National Veterinary Research Institute in Pulawy

In partnership with: Paradigm Publishing Services

Keywords:

food contaminants,

honey,

LC-MS/MS,

pyrrolizidine alkaloids,

toxic plant secondary metabolites

Related subjects:

Life sciences,

Molecular biology,

Microbiology and virology,

Life sciences, other,

Medicine,

Veterinary medicine

© 2026 Ewelina Kowalczyk, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution 4.0 License.

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