Transfer and toxicokinetic modeling of non-dioxin-like polychlorinated biphenyls (ndl-PCBs) into accidentally exposed dairy cattle and their calves - A case report.

Polychlorinated biphenyls (PCBs) are persistent environmental pollutants that accumulate in tissues of exposed animals and humans. This case report refers ton=3 dairy cows accidentally exposed to non-dioxin-like PCBs (ndl-PCBs) of unknown origin on a German farm. At study start they had a cumulative total of 122-643 ng/g fat in milk and 105-591 ng/g fat in blood, consisting mainly of PCBs 138, 153, and 180. Two cows calved during the study and their calves were raised on their mothers' milk, resulting in cumulative exposure until slaughter. A physiologically based toxicokinetic model was developed to describe the fate of ndl-PCBs in the animals. The toxicokinetic behavior of ndl-PCBs was simulated in individual animals, including transfer of contaminants into calves via milk and placenta. Both the simulations and experimental data indicate that contamination via both routes is significant. In addition, the model was used to estimate kinetic parameters for risk assessment.

Cause and Effect Analysis between Influencing Factors Related to Environmental Conditions, Hunting and Handling Practices and the Initial Microbial Load of Game Carcasses.

Environmental, hunting and handling factors affect the microbial load of hunted game and the resulting meat products. The aim of this study was to systematically investigate the influence of several factors on the initial microbial load (IML) of game carcasses during the early hunting chain. Eviscerated roe deer body cavities (n = 24) were investigated in terms of total viable count and the levels of Pseudomonas spp., Lactobacillus spp., Enterobacteriaceae and Escherichia coli (E. coli). Furthermore, a risk analysis based on the obtained original IML data, literature search and a Failure Mode and Effects Analysis (FMEA) was performed. The IML could be explained in a regression model by factors including the higher body weight (BW), damaged gastrointestinal tract by the shot, ambient temperature or rain. The levels of Lactobacillus spp. (p = 0.0472), Enterobacteriaceae (p = 0.0070) and E. coli (p = 0.0015) were lower on the belly flap surface when gloves were used during evisceration. The literature search revealed that studies examining influencing factors (IF) on the IML of game carcasses found contradictory effects of the comparable IF on IML. Potential handling failures may lead to a higher IML of game carcasses during the early hunting chain ranked by FMEA. Several handling practices for game carcasses are recommended, such as ensuring efficient cooling of heavier BW carcasses to limit bacterial growth or eviscerating heavier carcasses before lighter ones.

Standardised Sampling Approach for Investigating Pathogens or Environmental Chemicals in Wild Game at Community Hunts.

Wildlife may host pathogens and chemicals of veterinary and public health relevance, as well as pathogens with significant economic relevance for domestic livestock. In conducting research on the occurrence and distribution of these agents in wildlife, a major challenge is the acquisition of a sufficient number of samples coupled with efficient use of manpower and time. The aim of this article is to present the methodology and output of a sampling approach for game animals, which was implemented from 2017/18 to 2020/21 at drive hunts in Brandenburg, Germany. The central element was a framework agreement with the BImA, whereby federal forest officials and other hunters collected most of the samples during field dressing. Further samples of game carcasses were obtained by scientists during subsequent gathering at a collection point. Altogether, 3185 samples from 938 wild ungulates of four species were obtained for various studies analysing-in this case-food-borne agents in game animals. Sampling was representative and reflected the proportional distribution of ungulate species hunted in Brandenburg. Hunting district and hunting season strongly influenced hunting bag and hence sampling success. This sampling approach was demonstrated to be a suitable basis for monitoring programs, that can be adapted to other regions.

Unraveling Congener-Specific Transfer of Non-Dioxin-Like Polychlorinated Biphenyls (ndl-PCBs) from Feed into Chicken Meat.

Non-dioxin-like polychlorinated biphenyls (ndl-PCBs) are a subclass of persistent bioaccumulative pollutants able to enter the food chain. We investigated the transfer of ndl-PCBs from contaminated feed into meat and liver of fattening chickens. A total of 48 chicks were divided into five treatment and one control groups. Treated animals were fed with contaminated diets (11.7 ± 0.4 µg/kg sum of indicator ndl-PCBs; 88% dry matter (DM)) before slaughter for different subperiods of time: 16, 23, 28, 32, and 36 days for groups 1-5, respectively. One day after the end of each subperiod, three animals per group were slaughtered to determine the congener-specific ndl-PCB content. All remaining animals were fed the control feed until slaughter on day 37 to probe depuration. We used these data to generate congener-specific physiologically based toxicokinetic (PBTK) models for indicator ndl-PCBs. The models show that PCBs 28, 138, 153, and 180 form a more slowly eliminated cluster (with an observed transfer rate into meat over 74% and observed half-lives over 8.7 days) than PCBs 52 and 101 (with a transfer rate under 13% and half-lives under 2.6 days). Our simulations show that ndl-PCB levels in feed lower than 3.9 (long 56-day) or 4.4 µg/kg (short 37-day fattening period) would be necessary to ensure the current maximum level in muscle meat (fat basis), according to EU Regulations 1881/2006 and 1259/2011. The PBTK models are made available in the Python and Food Safety Knowledge Exchange formats.

Introducing the Rapid Alert Supply Network Extractor (RASNEX) tool to mine supply chain information from food and feed contamination notifications in Europe.

BACKGROUND: During food or feed contamination events, it is of utmost importance to ensure their rapid resolution to minimize impact on human health, animal health and finances. The existing Rapid Alert System for Food and Feed (RASFF) is used by the European Commission, national competent authorities of member countries and the European Food Safety Authority to report information on any direct or indirect human health risk arising from food or feed, or serious risks to animal health or the environment in relation to feed. Nevertheless, no methods exist to to collectively evaluate this vast source of supply chain information. METHODS: To aid in the extraction, evaluation and visualization of the data in RASFF notifications, we present the Rapid Alert Supply Network Extractor (RASNEX) open-source tool available from https://doi.org/10.5281/zenodo.4322555 freely. Among RASNEX's functions is the graphical mapping of food and feed supply chain operators implicated in contamination events. RASNEX can be used during ongoing events as a support tool for risk analysis using RASFF notifications as input. RESULTS: In a first use case, we showcase the functionality of RASNEX with the RASFF notification on a 2017/2018 contamination event in eggs caused by the illegal use of fipronil. The information in this RASFF notification is used to visualize nine different flows of main and related food products. In a second use case, we combine RASFF notifications from different types of food safety hazards (Salmonella spp., fipronil and others) to obtain wider coverage of the visualized egg supply network compared to the first use case. Actors in the egg supply chain were identified mainly for Italy, Poland and Benelux. Other countries (although involved in the egg supply chain) were underrepresented. CONCLUSIONS: We hypothesize that biases may be caused by inconsistent RASFF reporting behaviors by its members. These inconsistencies may be counteracted by implementing standardized decision-making tools to harmonize decisions whether to launch a RASFF notification, in turn resulting in a more uniform future coverage across European food and feed supply chains with RASNEX.

Marination increases the bioavailability of lead in game meat shot with lead ammunition.

As a consequence of the toxicological lead characteristics, a reduction of its exposure should consider all sources. Game meat might contain elevated levels of lead due to the use of lead ammunition. The aim of the present study was to investigate the effects of acidic marination on the bioavailability of ammunition-derived lead in game meat (Roe deer), using the growing pig as an animal model. Furthermore, the study should provide evidence that the large-area scattering of lead particles leads to noticeable differences in the individual lead intake per game meat portion. Pigs of group A (n 7) received lead-shot game meat, which was cooked in water. Pigs of group B (n 7) received lead-shot game meat, which was first marinated (wine and vinegar) and then cooked. The lead content of both game meat preparations was equal with 0⋅77-0⋅79 mg Pb/portion. Pigs of group C (n 4) received lead-free game meat, which was also marinated and cooked. Additionally, lead acetate was administered intravenously to group D pigs (n 4). Blood samples were taken on elevated time points before and after game meat intake/i.v.-application. The acidic marination increased the bioavailability of orally ingested lead, resulting in significantly higher blood lead concentrations. The bioavailability of lead was 2⋅7 % when game meat was just cooked and 15 % when the meat was marinated before. The considerable variation of the individual blood lead concentrations suggests that an inhomogeneous distribution of ammunition-derived lead particles (in terms of size and number) causes individually non-comparable lead intakes from the consumption of game meat.

Transfer of Per- and Polyfluoroalkyl Substances (PFAS) from Feed into the Eggs of Laying Hens. Part 2: Toxicokinetic Results Including the Role of Precursors.

A feeding study was performed to examine the bioaccumulation of per- and polyfluoroalkyl substances (PFAS) in laying hens' tissues and plasma and feed-to-egg transfer rates and half-lives. A 25 day exposure was followed by a 42 day depuration period. A target analysis revealed substantial amounts of the precursors N-methyl and N-ethyl perfluorooctane sulfonamidoacetic acid (Me- and EtFOSAA), perfluorooctane sulfonamidoacetic acid (FOSAA), and perfluorooctane sulfonamide (FOSA). In tissues and eggs, the highest bioaccumulation was found for PFHxS, PFHpS, PFOS, and PFOA. Low levels of PFHxS (all samples), PFOS, and FOSAA (in yolk) were measurable even after the depuration period. The egg elimination half-lives of PFOS and aforementioned precursors were estimated to be 4.3 days, while the transfer rates of PFOS and all precursors taken together were 0.99. The transfer rate of PFOA was around 0.49. PFHxS and PFHpS showed apparent transfer rates of >100%, which is hypothesized to indicate the presence of precursors.

Transfer of Per- and Polyfluoroalkyl Substances (PFAS) from Feed into the Eggs of Laying Hens. Part 1: Analytical Results Including a Modified Total Oxidizable Precursor Assay.

The group of per- and polyfluoroalkyl substances (PFAS) comprises thousands of chemicals, which are used in various industrial applications and consumer products. In this study, a feeding experiment with laying hens and feed grown on a contamination site was conducted, and PFAS were analyzed in the feed and eggs to assess the transfer of PFAS into eggs. A targeted analysis of perfluoroalkyl acids (PFAAs) and different sulfonamides was performed. Additionally, the total oxidizable precursor (TOP) assay was modified by fully oxidizing small amounts of the samples instead of oxidizing their extracts in order to overcome potential losses during extraction. Targeted analysis showed the presence of known PFAAs and four sulfonamides in the feed and egg yolk samples. In the plant-based feed, short-chain PFAAs, methyl and ethyl perfluorooctane sulfonamidoacetic acid (Me- and EtFOSAA), and perfluorooctane sulfonic acid (PFOS) were the most abundant PFAS. In the eggs, PFOS, FOSAA, and its alkylated homologues showed the highest concentrations. The TOP assay revealed the presence of substantial amounts of precursors with different chain lengths from C4 to C8. The highest relative increase of PFOA after oxidation was observed in egg yolk from the end of the exposure period (828%). The results of this study demonstrate the transfer of PFAAs and their precursors into hens' eggs and emphasize the contribution of (known and unidentified) precursors to the overall PFAS burden in edible products. The modified TOP assay approach was shown to be a powerful tool to better assess the total burden of samples with PFAS.

In vitro metabolism of pyrrolizidine alkaloids - Metabolic degradation and GSH conjugate formation of different structure types.

Pyrrolizidine alkaloid (PA) forming plants are found worldwide and may contaminate food products at levels being of concern for human health. Due to the high biodiversity of PA producing plants many different types of PA structures are formed. PAs themselves are not toxic but require metabolic activation to exert toxicity. To investigate if the structure of the PAs affects their in vitro metabolism, we incubated a set of 22 PAs and compared the degradation rates and the amount of formed glutathione (GSH) conjugates. With human liver microsomes, no metabolic degradation of monoesters was found. Degradation rates of diester PAs tended to correlate with their hydrophilicity, whereby the more polar and branched-chained PAs exhibited lower degradation. There was a trend towards higher degradation rates in the presence of rat liver microsomes, but the GSH conjugate levels were similar. Although an effective degradation seems to be related with high GSH conjugate levels, no clear correlation between both parameters could be deduced. For both species no GSH conjugates, or only trace amounts, were formed from monoesters. However, for both open-chained as well as cyclic diesters GSH conjugates were detected and determined levels were comparable for both ester types without major structure-dependent differences.

Physiologically based toxicokinetic models and in silico predicted partition coefficients to estimate tetrachlorodibenzo-p-dioxin transfer from feed into growing pigs.

Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous, toxic, persistent and bioaccumulative organic pollutant. TCDD can potentially enter the food chain through contaminated food of animal origin as a consequence of feed contamination. Prediction of the TCDD transfer from feed into animal products is thus important for human health risk assessment. Here, we develop several physiologically based toxicokinetic (PBTK) models of TCDD transfer from contaminated feed into growing pigs (Sus scrofa) exposed to doses ranging from 24.52 to 3269.25 ng of TCDD. We test the consequences of explicit dose-dependent absorption (DDA) versus the indirect effects of a self-induced liver metabolism (SIM). The DDA and SIM models showed similar fit to experimental data, although currently it is not possible to unequivocally make statement on a mechanistic preference. The performance of both toxicokinetic models was successfully evaluated using the 1999 Belgian case of contaminated fats for feeding. In combination with toxicokinetic models of other dioxin congeners, they can be used to formulate maximum allowance levels of dioxins in feedstuffs for pigs. Additionally, the implementation of in silico-predicted partition coefficients was explored as a useful alternative to predict TCDD tissue distribution in low-dose scenarios without recurring to animal experiments.

Comparison of lead levels in edible parts of red deer hunted with lead or non-lead ammunition.

Game meat may contain elevated concentrations of lead especially if lead-containing ammunition is used for hunting. Then a health risk is possible for consumer groups with high game meat intake. The lead concentrations in three edible parts (marketable meat from the area close to the wound channel, saddle and haunch) of meat from red deer (Cervus elaphus) between animals hunted either with lead or non­lead ammunition were compared. Furthermore, lead levels in game meat of lead-shot red deer were compared with those of lead-shot roe deer and lead-shot wild boar. Ninety red deer were shot and killed in the context of this study (64 with lead and 26 with non­lead ammunition). Since the lead concentration for a number of the samples was below the limit of detection or the limit of quantification, statistical methods for left-censored data were applied. The median concentrations of lead in game meat did not differ significantly between lead shot and non­lead shot animals. However, when we analyzed the more elevated lead concentrations, they were significantly higher in edible parts of animals shot with lead ammunition than non-lead ammunition. The highest concentrations were found in samples from edible meat from the area close to the wound channel (max 3442 mg Pb/kg), followed by the saddle (max 1.14 mg Pb/kg) and with the lowest levels in the haunch (max 0.09 mg Pb/kg). A comparison of game species revealed that the lead concentration in haunch and saddle of lead shot red deer was higher than in the corresponding samples of lead shot roe deer. Our results have shown that by the use of non-lead ammunition, a significant reduction of the lead concentration especially in edible parts near the wound channel is possible.

Studies on the influence of sampling on the levels of dioxins and PCB in fish.

Beaked redfish (Sebastes mentella) and Greenland halibut (Reinhardtius hippoglossoides) were collected in waters around Svalbard (Barents Sea) to study the influence of different muscle separation/filleting techniques at small and medium fishes on the dioxin/PCB content. Sampling of both species included preparation techniques such as fillets with or without belly flaps, commercially trimmed fillets and cutting into anterior and posterior cutlets. In case of Greenland halibut also the whole edible muscle part and middle cutlets were studied. All samples analysed were far below the maximum level of 6.5 pg/g wet weight (ww) WHO-PCDD/F-PCB-TEQ and 75 ng/g ww ndl-PCB. Trimmed fillets of beaked redfish had the lowest fat content and the lowest level of dioxins and PCB (1.70%, WHO-PCDD/F-PCB-TEQ = 0.320 pg/g ww). The respective posterior cutlets showed the highest fat content and highest levels of dioxins and PCB (2.66%, WHO-PCDD/F-PCB-TEQ = 0.729 pg/g ww). Levels of dioxins and PCB in Greenland halibut samples were generally higher and ranged between WHO-PCDD/F-PCB-TEQ = 0.784 pg/g ww (fillets without bells flaps, fat content = 8.83%) and WHO-PCDD/F-PCB-TEQ = 2.022 pg/g ww (edible part whole muscle, fat content = 8.62%). The results show a considerable influence of the different sampling methods on the dioxin and PCB levels of the species analysed.

Lead content in wild game shot with lead or non-lead ammunition - Does "state of the art consumer health protection" require non-lead ammunition?

The toxicity of lead has been known for a long time, and no safe uptake level can be derived for humans. Consumers' intake via food should therefore be kept as low as possible. Game meat can contain elevated levels of lead due to the use of lead ammunition for hunting. A risk assessment conducted in 2010 by the German Federal Institute for Risk Assessment including various consumption scenarios revealed a possible health risk for extreme consumers of game meat hunted with lead ammunition (i.e. hunters and members of hunters' households). Babies, infants, children and women of childbearing age were identified as vulnerable group with regards to the developmental neurotoxicity of lead. It was noted, that a sound data base was required in order to refine the assessment. Therefore, the research project "Safety of game meat obtained through hunting" (LEMISI) has been conducted in Germany, with the aims of determining the concentrations of lead (as well as of copper and zinc) brought into the edible parts of game meat (roe deer (Capreolus capreolus) and wild boar (Sus scrofa)) due to using either lead or non-lead hunting ammunition, whilst concurrently taking geogenic (i.e. "background") levels of lead into account. Compared to non-lead ammunition, lead ammunition significantly increased lead concentrations in the game meat. The use of both lead and non-lead ammunition deposited copper and zinc in the edible parts of game meat, and the concentrations were in the range of those detected regularly in meat of farm animals. For the average consumer of game meat in Germany the additional uptake of lead only makes a minor contribution to the average alimentary lead exposure. However, for consumers from hunters' households the resulting uptake of lead-due to lead ammunition-can be several times higher than the average alimentary lead exposure. Non-lead bullets in combination with suitable game meat hygienic measures are therefore recommended in order to ensure "state of the art consumer health protection".

In vitro biotransformation of pyrrolizidine alkaloids in different species. Part I: Microsomal degradation.

Pyrrolizidine alkaloids (PA) are secondary metabolites of certain flowering plants. The ingestion of PAs may result in acute and chronic effects in man and livestock with hepatotoxicity, mutagenicity, and carcinogenicity being identified as predominant effects. Several hundred PAs sharing the diol pyrrolizidine as a core structure are formed by plants. Although many congeners may cause adverse effects, differences in the toxic potency have been detected in animal tests. It is generally accepted that PAs themselves are biologically and toxicologically inactive and require metabolic activation. Consequently, a strong relationship between activating metabolism and toxicity can be expected. Concerning PA susceptibility, marked differences between species were reported with a comparatively high susceptibility in horses, while goat and sheep seem to be almost resistant. Therefore, we investigated the in vitro degradation rate of four frequently occurring PAs by liver enzymes present in S9 fractions from human, pig, cow, horse, rat, rabbit, goat, and sheep liver. Unexpectedly, almost no metabolic degradation of any PA was observed for susceptible species such as human, pig, horse, or cow. If the formation of toxic metabolites represents a crucial bioactivation step, the found inverse conversion rates of PAs compared to the known susceptibility require further investigation.

Copper and zinc content in wild game shot with lead or non-lead ammunition - implications for consumer health protection.

The aim of this study was to examine the contamination of game meat with copper and zinc and establish whether the use of alternative (non-lead) ammunition can lead to higher or unsafe levels of copper and zinc in the meat of roe deer, wild boar and red deer. The research project "Safety of game meat obtained through hunting" (LEMISI) was conducted in Germany with the purpose of examining the entry of lead as well as copper and zinc into the meat of hunted game when using either lead or non-lead ammunition. The outcome of this study shows that the usage of both lead-based ammunition and alternative non-lead ammunition results in the entry of copper and zinc into the edible parts of the game. Using non-lead ammunition does not entail dangerously elevated levels of copper and zinc, so replacing lead ammunition with alternative ammunition does not introduce a further health problem with regard to these metals. The levels of copper and zinc in game meat found in this study are in the range found in previous studies of game. The content of copper and zinc in game meat is also comparable to those regularly detected in meat and its products from livestock (pig, cattle, sheep) for which the mean human consumption rate is much higher. From the viewpoint of consumer health protection, the use of non-lead ammunition does not pose an additional hazard through copper and zinc contamination. A health risk due to the presence of copper and zinc in game meat at typical levels of consumer exposure is unlikely for both types of ammunition.

Hunting of roe deer and wild boar in Germany: Is non-lead ammunition suitable for hunting?

BACKGROUND: Non-lead hunting ammunition is an alternative to bullets that contain lead. The use of lead ammunition can result in severe contamination of game meat, thus posing a health risk to consumers. With any kind of ammunition for hunting, the terminal effectiveness of bullets is an animal welfare issue. Doubts about the effectiveness of non-lead bullets for a humane kill of game animals in hunting have been discussed. The length of the escape distance after the shot has been used previously as an indicator for bullet performance. OBJECTIVE: The object of this study was to determine how the bullet material (lead or non-lead) influences the observed escape distances. METHODS: 1,234 records of the shooting of roe deer (Capreolus capreolus) and 825 records of the shooting of wild boar (Sus scrofa) were evaluated. As the bullet material cannot be regarded as the sole cause of variability of escape distances, interactions of other potential influencing variables like shot placement, shooting distance, were analyzed using conditional regression trees and two-part hurdle models. RESULTS: The length of the escape distance is not influenced by the use of lead or non-lead ammunition with either roe deer or wild boar. With roe deer, the length of the escape distance is influenced significantly by the shot placement and the type of hunting. Increasing shooting distances increased the length of the escape distance. With wild boar, shot placement and the age of the animals were found to be a significant influencing factor on the length of the escape distance. CONCLUSIONS: The length of the escape distance can be used as an indicator for adequate bullet effectiveness for humane killings of game animals in hunting.Non-lead bullets already exist which have an equally reliable killing effect as lead bullets.

[Persistent organic contaminants in food : Exposure, hazard potential, and health assessment]. / Persistente organische Kontaminanten in Lebensmitteln : Exposition, Gefährdungspotenzial und gesundheitliche Bewertung.

Environmental emissions of organic contaminants are caused by man-made and natural combustion processes, industrial production facilities, and the release from products. Food represents the main source of human exposure for some of these compounds. This is the case for three groups of persistent organic contaminants: (1) per- and polyfluoroalkyl substances (PFAS), (2) polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), and for (3) polycyclic aromatic hydrocarbons (PAHs). The issues regarding PCDD/F emissions were already recognized as a problem in the 1970s and have since then been effectively regulated; the impact of PFAS as global anthropogenic environmental contaminants was identified much later.A system of toxicity equivalency factors (TEF system) was established for the assessment of the toxic potency of a mixed exposure to PCDD/F and certain PCBs. For the health assessment and regulation of PAHs and PFAS, no such system has been implemented so far. For PFAS, a re-evaluation of the present tolerable daily intake values (TDI values) is currently being discussed, as new insights into toxicology and epidemiology have been gained.The persistence in the environment of the compound groups discussed here leads to entry into the food chain over long periods of time, even if access into the environment is minimized. This requires a long-term continuation of the monitoring of food stuffs and forward-looking risk assessment approaches and regulatory measures.

Occurrence of fatty acid esters of 3-MCPD, 2-MCPD and glycidol in infant formula.

The discovery of fatty acid esters of monochloropropanediol (MCPD) and glycidol generated during the refinement process in vegetable fats and oils caused concerns about possible adverse health effects. As these fats are components of infant formula, the current investigation of the MCPD and glycidyl ester contents in infant formula was necessary to update the data for risk assessment purposes. For the analysis of 3-MCPD, 2-MCPD and glycidyl esters in infant formula, an existing method for fats and oils had to be modified and validated. The fat fraction containing MCPD and glycidyl esters was extracted from infant formula by accelerated solvent extraction (ASE). The extracted fat was then analysed according to an established method for fats and oils. Glycidyl esters are converted to monobrompropanediol (3-MBPD) esters, MCPD and 3-MBPD esters hydrolysed subsequently and after derivatisation detected by GC-MS. Seven different products of infant formula, covering two types and five lots each, altogether 70 samples, were bought in retail markets and analysed. In all samples, 3-MCPD and glycidyl esters could be detected. Both 3-MCPD and glycidyl esters' concentration levels were found to be lower in comparison with earlier investigations described in the literature. The occurrence of 2-MCPD esters in infant formula was investigated for the first time and revealed concentrations about half of 3-MCPD ester concentrations.

A case of human poisoning by grayanotoxins following honey ingestion: elucidation of the toxin profile by mass spectrometry.

High-resolution mass spectrometry (HRMS) was applied for the detection of grayanotoxins (GrTx) in a contaminated honey sample. This sample was provided by a hospital due to a suspicion of intoxication after a patient had shown the typical symptoms of GrTx poisoning. Subsequent analysis proved the contamination with high amounts of GrTx and other toxins belonging to grayanane-type diterpenoids. This group of natural toxins is synthesised by the plant family Ericaceae and comprises more than 60 individual toxins, but only one compound is available as a reference standard. We applied a screening approach that easily confirms the presence or absence of GrTx without access to standards. By searching for predictable mass spectrometric fragment ions, including typical in-source fragments arising from collision-induced dissociation during electrospray ionisation, the complete toxin profile was screened and allowed the mass spectrometric identification of 15 individual GrTx. The potential of this approach is especially demonstrated by the fact that at least two of these toxins have not been previously described in the literature. A semi-quantitative estimation indicated a total toxin concentration of 358 mg kg(-1). An investigation of 49 honeys from the German retail market did not reveal the presence of GrTx.