Determination of Pyrrolizidine Alkaloids in Honey with Sensitive Gas Chromatography-Mass Spectrometry Method
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Determination of Pyrrolizidine Alkaloids in Honey with Sensitive Gas Chromatography-Mass Spectrometry Method Ewelina Kowalczyk 1
&
Zbigniew Sieradzki 1 & Krzysztof Kwiatek 1
Received: 1 May 2017 / Accepted: 23 November 2017 # The Author(s) 2017. This article is an open access publication
Abstract Pyrrolizidine alkaloids (PAs) are secondary metabolites produced by plants as a chemical defense against herbivores. Plants containing PAs are widely distributed in almost all geographical regions posing a risk of honey contamination. To provide safety of honey and decrease the potential risk for the consumers, a sensitive method based on gas chromatography-mass spectrometry enabling determination of a content of 1,2-unsaturated PAs in honey was developed. Honey samples were purified on MCX cartridges, and PAs were eluted with a solvent mixture consisting of ethyl acetate, methanol, ammonia, and triethylamine. Subsequently, 1,2-unsaturated alkaloids were reduced to their common backbone structures and derivatized with heptafluorobutyric anhydride. The method was validated according to SANTE 2015. All received parameters are in consistence with the document requirements as recovery ranged from 73.1 to 93.6%. The repeatability and reproducibility were calculated as relative standard deviation and ranged from 3.9 to 8.6% and from 10.6 to 17.8%, respectively. The limit of quantification was determined as 1 μg kg−1. Good linearity of the method was obtained with the coefficient of determination R2 > 0.99. The method was applied to 40 Polish and 14 Asian honey sample analyses. Keywords Pyrrolizidine alkaloids . Honey . Food . GC-MS
Introduction Honey is used and consumed worldwide for its sweet taste, nutritive, and pro-health properties. Honey, however, is produced in a natural environment, which is why it can be contaminated with many natural substances (Dubreil-Chéneau et al. 2013). Certain types of flower nectar and pollen have been reported to result in honey that is psychoactive or that can lead to toxicity (Islam et al. 2014). The natural toxic substances that can be transferred to honey are, among others, pyrrolizidine alkaloids (PAs). PAs are considered to be the most widely spread toxins of natural origin. They are produced by many plants, estimated as 3% of all flowering plants; however, the main sources of PAs are the families Boraginaceae, Asteraceae, Fabaceae, and Apocynaceae (Kempf et al. 2008). Toxicity of the compounds
* Ewelina Kowalczyk [email protected] 1
Department of Hygiene of Animal Feedingstuffs, National Veterinary Research Institute, Partyzantów Avenue 57, 24-100 Puławy, Poland
involves unsaturation of the 1,2 position and esterification of at least one of the hydroxyl groups with an acid and increases with the degree of branching and formation of long cyclic diesters (Mudge et al. 2015). However, PAs are pro-toxins as they require metabolic activation to exert toxic effects. Bioactivation takes place in the liver and is induced by the cytochrome P450 (CYP) monooxygenases. Forme
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