Screening of food supplements for toxic pyrrolizidine alkaloids

Pyrrolizidine alkaloids (PA) and PA-N-oxides (PANO) are a group of more than 660 secondary plant metabolites with hepatotoxic, carcinogenic and mutagenic effects in animals and humans. The phytotoxins can enter the food chain by transfer of PA/PANO between plants via the soil, unintended co-harvesting of PA/PANO-producing weeds, as well as by honeybees collecting pollen and nectar of these plants. Thus, bee- and plant-based products, e.g. (herbal) teas, spices and culinary herbs were identified to be a main source of consumers’ exposure to PA/PANO. Consequently, food supplements based on those ingredients may as well be contaminated with PA/PANO, but so far there are only very few studies available on this topic. Therefore, the current study investigated 50 herbal and bee product-based food supplements available in German retail pharmacies, drugstores, and online on the occurrence of 44 PA/PANO. In total, 19 samples contained PA/PANO with sum contents ranging from 0.1 to 105.1 ng/g in solid samples and from 0.03 to 2.20 ng/mL in liquid preparations. Considering the recommended daily consumption, the sum contents were of no or little concern for the health risk of adults, whereas in case of children the contents of singular samples could significantly contribute to the overall PA/PANO exposure

Distribution of T-2 toxin and HT-2 toxin during experimental feeding of yellow mealworm (Tenebrio molitor)

Within the European Union (EU), edible insects need to be approved as “Novel Food” according to Regulation (EU) 2015/2283 and must comply with the requirements of European food law with regard to microbiological and chemical food safety. Substrates used for feeding insects are susceptible to the growth of Fusarium spp. and consequently to contamination with trichothecene mycotoxins. Therefore, the current study aimed to investigate the influence of T-2 and HT-2 toxins on the larval life cycle of yellow mealworm (Tenebrio molitor (L.)) and to study the transfer of T-2, HT-2, T-2 triol and T-2 tetraol in the larvae. In a 4-week feeding study, T. molitor larvae were kept either on naturally (oat flakes moulded with Fusarium sporotrichioides) or artificially contaminated oat flakes, each at two levels (approximately 100 and 250 μg/kg total T-2 and HT-2). Weight gain and survival rates were monitored, and mycotoxins in the feeding substrates, larvae and residues were determined using LC-MS/MS. Larval development varied between the diets and was 44% higher for larvae fed artificially contaminated diets. However, the artificially contaminated diets had a 16% lower survival rate. No trichothecenes were detected in the surviving larvae after harvest, but T-2 and HT-2 were found both in the dead larvae and in the residues of naturally and artificially contaminated diets

A sensitive LC-MS/MS method for isomer separation and quantitative determination of 51 pyrrolizidine alkaloids and two tropane alkaloids in cow's milk

1,2-Unsaturated pyrrolizidine alkaloids (PA), their corresponding N-oxides (PANO), and tropane alkaloids (TA) are toxic secondary plant metabolites. Their possible transfer into the milk of dairy cows has been studied in feeding trials;however, only few data on the occurrence of these toxins in milk are available. In this study, the development of a sensitive analytical approach for the simultaneous detection and quantification of a broad range of 54 PA/PANO as well as of the TA atropine and scopolamine in milk of dairy cows is presented. The method optimisation focused on sensitivity and separation of PA/PANO isomers. Milk samples were extracted using liquid-liquid extraction with aqueous formic acid and n-hexane, followed by a cation-exchange solid-phase extraction for purification. Reversed phase liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis was performed using alkaline solvent conditions. Validation proved low limits of detection and quantification of 0.005 to 0.054 mu g/L and of 0.009 to 0.123 mu g/L, respectively. For 51 of the 54 tested PA/PANO and both TA, the recovery rates ranged from 64 to 127% with repeatability (RSDr) values below 15% at concentration levels of 0.05 and 0.50 mu g/L and below 8% at a concentration level of 3.00 mu g/L. Only three PANO did not match the validation criteria and were therefore regarded as semiquantitative. The final method was applied to 15 milk samples obtained from milk vending stations at farms and from local marketers in Bavaria, Germany. In three of the milk samples, traces of PA were detected

Untersuchungen zu Verhalten, Vorkommen und Risikobewertung toxischer Pyrrolizidinalkaloide in verschiedenen Lebens- und Futtermittelmatrices

Pyrrolizidine alkaloids (PA) and PA N-oxides (PANO) are toxic secondary plant metabolites contaminating food and feed. Analysis of honeys by different analytical approaches highlighted uncertainties of results. Additionally, the work demonstrated a decrease of PANO in fresh honey presumably due to enzymes. A new LC-MS/MS method for sensitive determination of 44 PA/PANO in important tea matrices was developed. Application of the method on a large set of spices and herbs combined with a risk assessment revealed exposure being of concern, in particular in case of children.Pyrrolizidinalkaloide (PA) und PA-N-Oxide (PANO) sind toxische Kontaminanten pflanzlichen Ursprungs. Honiganalysen mittels verschiedener Methodiken zeigten gewisse Ergebnisunsicherheiten, zudem wurde die vermutlich enzymatisch bedingte PANO Abnahme in frischem Honig belegt. Eine neue LC-MS/MS Methode zum sensitiven Nachweis von 44 PA/PANO in bedeutenden Teematrices wurde entwickelt und zusätzlich auf eine große Anzahl von Gewürzen und Kräutern angewandt. Die Risikobewertung offenbarte Matrices von Belang für die Lebensmittelsicherheit, insbesondere für Kinder

Pyrrolizidine alkaloids and tropane alkaloids in milk samples from individual dairy farms of the German federal states of Bavaria and Schleswig-Holstein

1,2-Dehydro-pyrrolizidine alkaloids (PA), their corresponding N-oxides (PANO) and tropane alkaloids (TA), are toxic plant metabolites. If plant material, containing these toxins, is present in the feed of dairy cows these toxins can be transferred into milk. Here, milk was sampled directly from dairy farms in the German federal states of Bavaria and Schleswig-Holstein in 2020–2022 in order to investigate a possible contamination of milk at the production stage. In total, 228 milk samples were analysed for 54 PA/PANO and two TA by a sensitive LC–ESI-MS/MS method. In addition, a subset of milk samples (n = 85) was independently analysed for TA by a cooperating laboratory for verification. PA/PANO were found in 26 samples (11%) with a low median sum content of the contaminated samples of 0.024 µg/L. The highest level of contamination was 5.6 µg/L. Senecionine-, lycopsamine- and heliotrine-type PA/PANO were detected. In four samples (1.8%), atropine was determined up to 0.066 µg/L. The toxin levels in the milk samples hardly contributed to the total daily exposure. These data are first-time results on contamination rates and levels occurring in milk from individual dairy farms, based on a large sample number.</p