Ergocristine

CAS# 511-08-0

Ergocristine

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Chemical structure

Ergocristine

3D structure

Chemical Properties of Ergocristine

Cas No. 511-08-0 SDF Download SDF
PubChem ID 31116 Appearance White powder
Formula C35H39N5O5 M.Wt 609.7
Type of Compound Alkaloids Storage Desiccate at -20°C
Solubility Soluble in chloroform, ethanol and pyridine
Chemical Name (6aR,9R)-N-[(1S,2S,4R,7S)-7-benzyl-2-hydroxy-5,8-dioxo-4-propan-2-yl-3-oxa-6,9-diazatricyclo[7.3.0.02,6]dodecan-4-yl]-7-methyl-6,6a,8,9-tetrahydro-4H-indolo[4,3-fg]quinoline-9-carboxamide
SMILES CC(C)C1(C(=O)N2C(C(=O)N3CCCC3C2(O1)O)CC4=CC=CC=C4)NC(=O)C5CN(C6CC7=CNC8=CC=CC(=C78)C6=C5)C
Standard InChIKey HEFIYUQVAZFDEE-MKTPKCENSA-N
Standard InChI InChI=1S/C35H39N5O5/c1-20(2)34(37-31(41)23-16-25-24-11-7-12-26-30(24)22(18-36-26)17-27(25)38(3)19-23)33(43)40-28(15-21-9-5-4-6-10-21)32(42)39-14-8-13-29(39)35(40,44)45-34/h4-7,9-12,16,18,20,23,27-29,36,44H,8,13-15,17,19H2,1-3H3,(H,37,41)/t23-,27-,28+,29+,34-,35+/m1/s1
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.
We recommend that you prepare and use the solution on the same day. However, if the test schedule requires, the stock solutions can be prepared in advance, and the stock solution must be sealed and stored below -20℃. In general, the stock solution can be kept for several months.
Before use, we recommend that you leave the vial at room temperature for at least an hour before opening it.
About Packaging 1. The packaging of the product may be reversed during transportation, cause the high purity compounds to adhere to the neck or cap of the vial.Take the vail out of its packaging and shake gently until the compounds fall to the bottom of the vial.
2. For liquid products, please centrifuge at 500xg to gather the liquid to the bottom of the vial.
3. Try to avoid loss or contamination during the experiment.
Shipping Condition Packaging according to customer requirements(5mg, 10mg, 20mg and more). Ship via FedEx, DHL, UPS, EMS or other couriers with RT, or blue ice upon request.

Source of Ergocristine

The extract of tall fescue (Lolium arundinaceum) seed infected by

Biological Activity of Ergocristine

DescriptionErgocristine, a dopamine agonist, can block prolactin release without any noticeable latent period.

Ergocristine Dilution Calculator

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Preparing Stock Solutions of Ergocristine

1 mg 5 mg 10 mg 20 mg 25 mg
1 mM 1.6402 mL 8.2008 mL 16.4015 mL 32.803 mL 41.0038 mL
5 mM 0.328 mL 1.6402 mL 3.2803 mL 6.5606 mL 8.2008 mL
10 mM 0.164 mL 0.8201 mL 1.6402 mL 3.2803 mL 4.1004 mL
50 mM 0.0328 mL 0.164 mL 0.328 mL 0.6561 mL 0.8201 mL
100 mM 0.0164 mL 0.082 mL 0.164 mL 0.328 mL 0.41 mL
* Note: If you are in the process of experiment, it's necessary to make the dilution ratios of the samples. The dilution data above is only for reference. Normally, it's can get a better solubility within lower of Concentrations.

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References on Ergocristine

Epimerization of ergot alkaloids in feed.[Pubmed:32637710]

Heliyon. 2020 Jun 30;6(6):e04336.

Chronic intake of cereals contaminated with ergot alkaloids can cause ergotism and result in the loss of toes and fingers or even death. Today, due to common risk management practices, ergotism is rare as a human disease but remains a problem in livestock husbandry. Each alkaloid coexists under two forms (R and S), though only the R-form presents toxic effects. The epimerization occurs spontaneously but the mechanisms remain globally unknown. Therefore, different processing methods were evaluated for their respective influences on the epimerization. The results suggest that ergotamine and ergosine are very stable ergot alkaloids, and neither their concentrations, nor their respective R/S ratios, are significantly influenced by heating, protic solvents or UV light. In contrast, for Ergocristine, ergokryptine, ergocornine and ergometrine, heating can decrease the concentrations of these alkaloids and heat, protic solvents and UV light influence the R/S ratio towards the S-form, though the respective influence on the epimerization of these compounds is variable. In addition, the total concentration of all ergot alkaloids is reduced through heating. However, all these effects are not strong enough to change the composition of ergot alkaloids in feed substantially and to transform toxic feed into non-toxic feed.

Ergot Alkaloids in Wheat and Rye Derived Products in Italy.[Pubmed:31052444]

Foods. 2019 May 1;8(5). pii: foods8050150.

Genus Claviceps is a plant pathogen able to produce a group of toxins, ergot alkaloids (EAs), whose effects have been known since the Middle Ages (ergotism). Claviceps purpurea is the most important representative specie, known to infect more than 400 monocotyledonous plants including economically important cereal grains (e.g., rye, wheat, triticale). EAs are not regulated as such. Maximum limits are in the pipeline of the EU Commission while at present ergot sclerotia content is set by the Regulation (EC) No. 1881/2006 in unprocessed cereals (0.05% as a maximum). This study aimed to investigate the presence of the six principal EAs (ergometrine, ergosine, ergocornine, alpha-ergocryptine, ergotamine and Ergocristine) and their relative epimers (-inine forms) in rye- and wheat-based products. Of the samples, 85% resulted positive for at least one of the EAs. Wheat bread was the product with the highest number of positivity (56%), followed by wheat flour (26%). Rye and wheat bread samples showed the highest values when the sum of the EAs was considered, and durum wheat bread was the more contaminated sample (1142.6 mug/kg). These results suggest that ongoing monitoring of EAs in food products is critical until maximum limits are set.

Liquid chromatography-mass spectrometry-based determination of ergocristine, ergocryptine, ergotamine, ergovaline, hypoglycin A, lolitrem B, methylene cyclopropyl acetic acid carnitine, N-acetylloline, N-formylloline, paxilline, and peramine in equine hair.[Pubmed:31009898]

J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Jun 1;1117:127-135.

Ingestion of hypoglycin A (HGA) in maple seeds or alkaloids produced by symbiotic fungi in pasture grasses is thought to be associated with various syndromes in grazing animals. This article describes analytical methods for monitoring long-term exposure to HGA, its metabolite MCPA-carnitine, as well as Ergocristine, ergocryptine, ergotamine, ergovaline, lolitrem B, N-acetylloline, N-formylloline, peramine, and paxilline in equine hair. After extraction of hair samples separation was achieved using two ultra high performance liquid chromatographic systems (HILIC or RP-C18, ammonium formate:acetonitrile). A benchtop orbitrap instrument was used for high resolution tandem mass spectrometric detection. All analytes were sensitively detected with limits of detection between 1pg/mg and 25pg/mg. Irreproducible extraction or ubiquitous presence in horse hair precluded quantitative validation of lolitrem B/paxilline and N-acetylloline/N-formylloline, respectively. For the other analytes validation showed no interferences in blank hair. Other validation parameters were as follows: limits of quantification (LOQ), 10 to 100pg/mg; recoveries, 18.3 to 91.0%; matrix effects, -48.2 - 24.4%; linearity, LOQ - 1000pg/mg; accuracy, -14.9 - 6.4%, precision RSDs Ergocristine, ergocryptine, ergotamine, ergovaline, HGA, MCPA-carnitine, and peramine in horse hair. Applicability was proven for N-acetylloline and N-formylloline by analyzing hair of 13 horses.

Fate of Ergot Alkaloids during Laboratory Scale Durum Processing and Pasta Production.[Pubmed:30935118]

Toxins (Basel). 2019 Mar 31;11(4). pii: toxins11040195.

The fate of ergot alkaloids during the milling of durum and subsequent production and cooking of pasta was examined. Durum samples containing varying amounts of ergot sclerotia (0.01(-)0.1% by mass) were milled, and all milling product was analyzed for 10 ergot alkaloids using liquid chromatography with tandem mass spectrometry. Spaghetti was prepared from the semolina obtained during milling. Ergocristine, ergocristinine, and ergotamine were the predominant ergot alkaloids observed in the milling fractions and spaghetti. Approximately 84% of the total ergot alkaloid mass of the whole grain durum resided in the milling product fractions associated with the outer kernel layers (bran, shorts, feeds). No consistent loss of ergot alkaloids was observed during the production or cooking of spaghetti. However, changes in the ratio of R- to S-enantiomers occurred during the milling and cooking of spaghetti. Products containing bran, shorts, and feeds, as well as cooked spaghetti, contained a higher proportion of the less biologically active S-enantiomers. The results of this study emphasize the need to monitor R- and S-enantiomers, and to consider food and feed products, as opposed to whole grain, when assessing any exposure of consumers to ergot alkaloids.

Links Between Genetic Groups, Host Specificity, and Ergot-Alkaloid Profiles within Claviceps purpurea (Fr.) Tul. on Slovenian Grasses.[Pubmed:30673578]

Plant Dis. 2018 Jul;102(7):1334-1340.

In the present study, the genetic relationships and ergot-alkaloid production of the fungus Claviceps purpurea on grasses were investigated, to determine any associations between grass host specificity, ergot-alkaloid production, and geographic origin. C. purpurea sclerotia were obtained from wild and cultivated grasses along a 300-km climatic gradient, from sub-Mediterranean to continental climates. Twenty-one infected grass samples provided 39 sclerotia for analysis of the ergot alkaloids ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, and Ergocristine, and their "-inine" epimers, using liquid chromatography-tandem mass spectrometry. C. purpurea ribosomal DNA underwent molecular classification to determine any grass host or geographic specificity of ergot-alkaloid composition for the different operational taxonomic units. Molecular analysis of sclerotia ribosomal DNA showed three genetic groups, with some associations with specific grass host taxonomic groups. The ergot-alkaloid composition data were in agreement with the data obtained by molecular methods. The most frequent ergot-alkaloid epimers were Ergocristine, and ergosine. The total ergot-alkaloid concentrations in sclerotia varied from 59 to 4,200 mg kg(-1), which corresponds to 0.059 to 4.2 mg kg(-1) in animal feed (assuming ergot alkaloids at 1,000 mg kg(-1) sclerotia). Therefore, grasses can be associated with significant levels of ergot alkaloids. In addition, the ergot-alkaloid compositions of C. purpurea sclerotia can be different for infections with different C. purpurea genetic groups, because these show different ergot-alkaloid compositions.

Comparative study on the metabolism of the ergot alkaloids ergocristine, ergocryptine, ergotamine, and ergovaline in equine and human S9 fractions and equine liver preparations.[Pubmed:30623698]

Xenobiotica. 2019 Oct;49(10):1149-1157.

1. Ergopeptine alkaloids like ergovaline and ergotamine are suspected to be associated with fescue toxicosis and ergotism in horses. Information on the metabolism of ergot alkaloids is scarce, especially in horses, but needed for toxicological analysis of these drugs in urine/feces of affected horses. The aim of this study was to investigate the metabolism of ergovaline, ergotamine, Ergocristine, and ergocryptine in horses and comparison to humans. 2. Supernatants of alkaloid incubations with equine and human liver S9 fractions were analyzed by reversed-phase liquid-chromatography coupled to high-resolution tandem mass spectrometry with full scan and MS(2) acquisition. Metabolite structures were postulated based on their MS(2) spectra in comparison to those of the parent alkaloids. All compounds were extensively metabolized yielding nor-, N-oxide, hydroxy and dihydro-diole metabolites with largely overlapping patterns in equine and human liver S9 fractions. However, some metabolic steps e.g. the formation of 8'-hydroxy metabolites were unique for human metabolism, while formation of the 13/14-hydroxy and 13,14-dihydro-diol metabolites were unique for equine metabolism. Incubations with equine whole liver preparations yielded less metabolites than the S9 fractions. 3. The acquired data can be used to develop metabolite-based screenings for these alkaloids, which will likely extend their detection windows in urine/feces from affected horses.

Development and validation of an ultrahigh performance liquid chromatography-high resolution tandem mass spectrometry assay for nine toxic alkaloids from endophyte-infected pasture grasses in horse serum.[Pubmed:29779692]

J Chromatogr A. 2018 Jul 27;1560:35-44.

Endophyte fungi (e.g. Epichloe ssp. and Neotyphodium ssp.) in symbiosis with pasture grasses (e.g. Festuca arundinacaea and Lolium perenne) can produce toxic alkaloids, which are suspected to be involved in equine diseases such as fescue toxicosis, ryegrass staggers, and equine fescue oedema. The aim of this study was, therefore, to develop and validate a quantification method for these and related alkaloids: Ergocristine, ergocryptine, ergotamine, ergovaline, lolitrem B, lysergic acid, N-acetylloline, N-formylloline, peramine, and paxilline in horse serum. Horse serum samples (1.5mL) were worked up by solid-phase extraction (OASIS HLB). The extracts were analyzed by ultra high performance liquid chromatography-high resolution tandem mass spectrometry (UHPLC-HRMS/MS). Chromatographic separation was achieved by gradient elution with ammonium formate buffer and acetonitrile on a RP18 column (100x2.1mm; 1.7mum). HRMS/MS detection was performed on a QExactive Focus instrument with heated positive electrospray ionization and operated in the parallel reaction monitoring (PRM) mode. Method validation included evaluation of selectivity, matrix effect, recovery, linearity, limit of quantification (LOQ), limit of detection (LOD), accuracy, and stability. With exception of lolitrem B solid phase extraction yielded high recoveries (73.6-104.6%) for all analytes. Chromatographic separation of all analytes was achieved with a run time of 25min. HRMS/MS allowed sensitive detection with LODs ranging from 0.05 to 0.5ng/mL and LOQs from 0.1 to 1.0ng/mL. Selectivity experiments showed no interferences from matrix or IS, but N-acetylloline and N-formylloline were found to be ubiquitous in horse serum. Newborn calf serum was therefore used as surrogate matrix for the validation study. Calibration ranges were analyte-dependent and in total covered concentrations from 0.1 to 50ng/mL. Lolitrem B and paxilline could be sensitively detected, but did not meet quantification requirements. For the other analytes, accuracy and precision were shown for 3 different concentrations (QC low, medium, high) with acceptable bias (-10, 5%-7.9%) and precision (CV 2.6%-12.5%). Matrix effects varied from 55.0% to 121% (RSD 7.8-18.5%) and were adequately compensated by IS. Matrix effects of N-acetylloline and N-formylloline could only be estimated in newborn calf serum (n=1) and ranged from 52.5-88.3%. All analytes were stable under autosampler conditions and over 3 freeze and thaw cycles. Applicability was proven by analyzing authentic horse serum samples (n=24). In conclusion, the presented method allows a sensitive detection of ergocrisitine, ergocryptine, ergotamine, ergovaline, lolitrem B, lysergic acid, N-acetylloline, N-formylloline, peramine, and paxilline in horse serum and reliable quantification of all but lolitrem B and paxilline.

Tall fescue ergot alkaloids are vasoactive in equine vasculature.[Pubmed:29293720]

J Anim Sci. 2017 Nov;95(11):5151-5160.

Mares grazing endophyte-infected () tall fescue () typically exhibit reproductive dysfunction rather than problems associated with peripheral vasoconstriction as a primary sign of the fescue toxicosis syndrome. Research using Doppler ultrasonography demonstrated that consumption of endophyte-infected tall fescue seed causes measurable vasoconstriction in the medial palmar artery. The objective of this study was to evaluate contractile responses of medial palmar artery and vein to increasing concentrations of various tall fescue alkaloids. Medial palmar arteries and veins were collected immediately following euthanasia from 23 horses of mixed breed, age, and gender from both forelimbs, and uterine arteries were collected from females ( = 12). Vessels were separated, cleaned of excess connective and adipose tissue, divided into 2- to 3-mm cross-sections, and suspended in a multimyograph chamber with continuously oxygenated Krebs-Henseleit buffer (95% O/5% CO; pH 7.4; 37 degrees C). Following a 90-min equilibration and recovery from reference compound exposure, increasing concentrations of norepinephrine, 5-hydroxytryptamine, ergotamine, and ergonovine for the palmar artery and vein and uterine artery and ergovaline, ergocryptine, Ergocristine, ergocornine, and lysergic acid for the palmar artery and vein were added to assess vasoactivity. Data were normalized as a percentage of contractile response induced by the reference compound addition and analyzed as a completely randomized design. Both norepinephrine and serotonin were vasoactive in all 3 types of blood vessels. Neither ergotamine nor ergonovine were vasoactive in the uterine artery. All alkaloids tested with the palmar artery and vein produced a contractile response, except that neither the palmar artery nor the palmar vein responded to lysergic acid ( > 0.05). Ergovaline was the most vasoactive ergot alkaloid in both the palmar artery and the palmar vein ( < 0.05) followed by ergonovine, whereas out of the 4 remaining ergopeptine alkaloids tested, Ergocristine induced the lowest contractile response. Although horses do not outwardly appear to be affected by peripheral vasoconstriction as observed in cattle, these data indicate that tall fescue alkaloids are vasoactive and suggest that potential exists for peripheral vascular effects of tall fescue alkaloids in horses. This does not appear to be the case for the uterine artery, and future research should be directed at understanding how ergot alkaloids cause equine reproductive dysfunction.

Effects of Continuously Feeding Diets Containing Cereal Ergot Alkaloids on Nutrient Digestibility, Alkaloid Recovery in Feces, and Performance Traits of Ram Lambs.[Pubmed:29257065]

Toxins (Basel). 2017 Dec 19;9(12). pii: toxins9120405.

Allowable limits for cereal ergot alkaloids in livestock feeds are being re-examined, and the objective of this study was to compare nutrient digestibility, growth performance and carcass characteristics of ram lambs fed a range of alkaloid concentrations, including the maximum currently allowed in Canada (2 to 3 ppm). Four pelleted diets were fed: control, with no added alkaloids; 930; 1402; and 2447 ppb alkaloids based on total R and S epimers. Eight ram lambs (30.0 +/- 3.1 kg) were used to examine the impacts of dietary treatments on nutrient digestibility and alkaloid recovery from feces. Concentrations of dietary alkaloids evaluated did not affect nutrient digestibility or N metabolism. Excepting ergocornine and ergocryptine, recovery of alkaloids in feces varied among periods, suggesting that individual lambs may differ in their ability to metabolize Ergocristine, ergometrine, ergosine, ergotamine and their S epimers. In a second experiment, ram lambs (n = 47, 30 +/- 8 kg) were randomly assigned to a diet and weighed weekly until they achieved a slaughter weight of >/= 45 kg (average 9 weeks; range 6 to 13 weeks). Intake of DM did not differ (p = 0.91) among diets, although lambs fed 2447 ppb alkaloids had a lower (p < 0.01) ADG than did lambs receiving other treatments. The concentration of serum prolactin linearly declined (p < 0.01) with increasing alkaloids. Feeding 2447 ppb total alkaloids negatively impacted growth, while feeding 1402 ppb did not harm growth performance, but reduced carcass dressing percentage. Due to different concentrations of alkaloids affecting growth and carcass characteristics in the present study, determining allowable limits for total dietary alkaloids will require a better understanding of impacts of alkaloid profiles and interactions among individual alkaloids.

Human and animal dietary exposure to ergot alkaloids.[Pubmed:32625563]

EFSA J. 2017 Jul 6;15(7):e04902.

The ergot alkaloids (EAs) are mycotoxins produced by several species of fungi in the genus Claviceps. In Europe, Claviceps purpurea is the most widespread species and it commonly affects cereals such as rye, wheat, triticale, barley, millets and oats. Food and feed samples used to estimate human and animal dietary exposure were analysed for the 12 main C. purpurea EAs: ergometrine, ergosine, ergocornine, ergotamine, Ergocristine, ergocryptine (alpha- and beta-isomers) and their corresponding -inine (S)-epimers. The highest levels of EAs were reported in rye and rye-containing commodities. In humans, mean chronic dietary exposure was highest in 'Toddlers' and 'Other children' with maximum UB estimates of 0.47 and 0.46 mug/kg bw per day, respectively. The 95th percentile exposure was highest in 'Toddlers' with a maximum UB estimate of 0.86 mug/kg bw per day. UB estimations were on average fourfold higher than LB estimations. Average acute exposure (MB estimations) ranged from 0.02 mug/kg bw per day in 'Infants' up to 0.32 mug/kg bw per day estimated in 'Other children'. For the 95th percentile acute exposure, the highest estimate was for a dietary survey within the age class 'Other children' (0.98 mug/kg bw per day). Dietary exposure estimates for animals, assuming a mean concentration scenario, varied between 0.31-0.46 mug/kg bw per day in beef cattle and 6.82-8.07 mug/kg bw per day (LB-UB) in piglets, while exposure estimates assuming a high concentration scenario (95th percentile) varied between 1.43-1.45 mug/kg bw per day and 16.38-16.61 mug/kg bw per day (LB-UB) in the same species. A statistically significant linear relationship between the content of sclerotia and the levels of EAs quantified was observed in different crops (barley, oats, rye, triticale and wheat grains). However, the absence of sclerotia cannot exclude the presence of EAs as samples with no sclerotia identified showed measurable levels of EAs ('false negatives').

Occurrence of ergot alkaloids in wheat from Albania.[Pubmed:28332434]

Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2017 Aug;34(8):1333-1343.

The occurrence of ergot alkaloids in wheat harvested in Albania was investigated. A total of 71 samples of winter wheat collected in 2014 and 2015 were analysed for the 12 most important ergot alkaloids using liquid chromatography-tandem mass spectrometry. In the harvesting year 2014, 48.6% of samples were contaminated with ergot alkaloids, whereas in 2015 only 19.4% of samples were contaminated. In 2014, the concentrations of total ergot alkaloids ranged from 17.3 to 975.4 mug kg(-)(1), and in 2015 they ranged from 10.3 to 390.5 mug kg(-1). The samples contained from one to nine ergot alkaloids. The most frequent were ergometrine, ergosine and Ergocristine, and the least frequent were ergocryptine, ergocryptinine and ergocorninine.

Localization of ergot alkaloids in sclerotia of Claviceps purpurea by matrix-assisted laser desorption/ionization mass spectrometry imaging.[Pubmed:27873003]

Anal Bioanal Chem. 2017 Feb;409(5):1221-1230.

The fungus Claviceps purpurea produces highly toxic ergot alkaloids and accumulates these in the hardened bodies of fungal mycelium. These so-called sclerotia, or ergot bodies, replace the crop seed of infected plants, which can include numerous important food- and feedstuff such as rye and wheat. While several studies have explored details of the infection process and development of ergot bodies, little information is available on the spatial distribution of the mycotoxins in the sclerotia. Here we used matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) at a lateral resolution of 35 mum to visualize the distribution of two representative alkaloids, Ergocristine and ergometrine, produced by Ecc93 and Gal 310 variants of C. purpurea, respectively, after infection of rye. To improve cryosectioning of this fragile biological material tissue with complex texture, we developed a practical embedding protocol based on cellulose polymers. The MALDI-MS images recorded from the so produced intact tissues sections revealed that ergometrine exhibited a relatively homogeneous distribution throughout the ergot body, whereas Ergocristine was found to be enriched in the proximal region. This finding can be correlated to the morphological development of sclerotia as ergot alkaloids are only produced in the sphacelial stage. The ability to localize toxins and other secondary metabolites in intact sections of crop-infecting fungi with high lateral resolution renders MALDI-MSI a powerful tool for investigating biosynthetic pathways and for obtaining a deeper understanding of the parasite-host interaction. Graphical abstract Workflow for identification and spatial localization of ergot alkaloids in infected rye grains.

Development and Validation of an Analytical Method for Determination of Ergot Alkaloids in Animal Feedingstuffs with High Performance Liquid Chromatography-fluorescence Detection.[Pubmed:27760024]

Pol J Vet Sci. 2016 Sep 1;19(3):559-565.

A high performance liquid chromatography combined with fluorescence detection (HPLC-FLD) method was developed for determination of five ergot alkaloids (EA): ergometrine, ergotamine, ergocornine, ergocrypine and Ergocristine in animal feedingstuffs. The method was based on the application of QuEChERS salts for extraction and modified QuEChERS dispersive SPE for the cleanup step. Alkaloids separation was performed on a C18, 250 mm x 4.6 mm, 5 mum column with the mobile phase containing ammonium carbonate and acetonitrile. The excitation and emission wavelengths were 330 and 420 nm respectively. The method was validated according to the Commission Decision 2002/657/EC and all parameters are in agreement with the requirements of the Decision. Linearity was determined for the concentration range of 25-400 mug/kg. The coefficient of determination (R2) for all curves was from 0.985 to 0.996. The limit of detection (LOD) was in the range 3.23 to 6.53 mug/kg and the limit of quantification (LOQ) from 11.78 to 13.06 mug/kg. The decision limit (CCalpha) ranged from 29.56 to 43.08 mug/kg and detection capability (CCbeta) from 40.65 to 51.01 mug/kg. The highest coefficient of variation (CV) for repeatability was 14.3% and for reproducibility 15.4%.

Contamination with ergot bodies (Claviceps purpurea sensu lato) of two horse pastures in Northern Germany.[Pubmed:27495979]

Mycotoxin Res. 2016 Nov;32(4):207-219.

Because the occurrence of Claviceps in European pastures may have been overlooked to cause serious health problem for grazing animals, we documented the degree of Claviceps contamination in two horse pastures and estimated whether the horses could have ingested a critical quantity of alkaloids. We counted the Claviceps sclerotia and determined alkaloid levels using high performance liquid chromatography with fluorescence detection. Depending on the location, the number of sclerotia varied from 0.09 to 0.19 per square meter (central area) and from 0.23 to 55.8 per square meter (border strips). Alkaloid levels in individual sclerotia also varied in different genera of grasses, ranging from 0.98 +/- 0.17 mug/kg in Agrostis sp. to 25.82 +/- 9.73 mug/kg in Dactylis sp., equivalent to 0.98 mug/kg and 7.26 mg/kg. Sclerotia from Dactylis contained high levels of ergosine (0.209 % +/- 0.100 %) and Ergocristine (0.374 % +/- 0.070 %). Depending on the localization in pastures, alkaloid levels in forage (dry matter, DM) ranged from 16.1 to 45.4 mug/kg in central areas and from 23.9 to 722 mug/kg in border strips. The amount of alkaloids that a horse could have ingested depended on its daily DM uptake, which was higher in the central areas (5.85 kg/day) than in the border strips (2.73 or 0.78 kg/day). In the central areas, this amount of alkaloids ranged from 94.2 to 265.9 mug/day; and in the border strips, from 65.3 (in 2.73 kg DM/day) to as much as 563.8 mug/day (in 0.78 kg DM/day). All these amounts are higher than the European averages for alkaloids ingested by horses via feedstuffs.

Rapid Screening of Ergot Alkaloids in Sclerotia by MALDI-TOF Mass Spectrometry.[Pubmed:27455930]

J AOAC Int. 2016 Jul;99(4):895-898.

Ergot is a common disease of wheat and other cereal grains that is predominantly caused by Claviceps purpurea in the field, often affecting crop yield in addition to the environment. Infected grain can be contaminated with dark sclerotia, which contain fungal metabolites such as ergot alkaloids. The occurrence of ergot alkaloids in cereal grain is a major health concern for humans and livestock. Effective and rapid screening of these mycotoxins is crucial for producers, processors, and consumers of cereal-based food and feed grain. Established methods of ergot alkaloid screening based on LC-MS or GC-MS require laborious processes. A novel method using matrix-assisted laser desorption ionization (MALDI)-time-of-flight (TOF) MS was developed to identify four ergot alkaloids. Using dihydroxybenzoic acid as the matrix, ergosine, ergocornine, ergocryptine, and Ergocristine were readily detected in individual sclerotia of C. purpurea. The accuracy of the identified ergot alkaloids was further confirmed by tandem MS analysis. MALDI-TOF MS is suitable for high-throughput screening of ergot alkaloids because it permits rapid and accurate identification, simple sample preparation, and no derivatization or chromatographic separation.

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