Correlation and variability between weighing, counting and analytical methods to determine ergot (Claviceps purpurea) contamination of grain

2017 ◽  
Vol 10 (3) ◽  
pp. 209-218 ◽  
Author(s):  
T. Grusie ◽  
V. Cowan ◽  
J. Singh ◽  
J. McKinnon ◽  
B. Blakley
Keyword(s):  
Particle Size ◽  
Ergot Alkaloid ◽  
Total Alkaloid ◽  
High Performance ◽  
Ergot Alkaloids ◽  
Sample Volume ◽  
Small Sample ◽  
Claviceps Purpurea ◽  
Liquid Chromatography Tandem Mass ◽  
The Impact

Ergot alkaloid mycotoxins produced by the fungus Claviceps purpurea, are contaminants of cereal crops and grasses. The objectives of this study were to determine the correlation between number of ergot sclerotia and weight compared to the total ergot alkaloid concentration, to evaluate the effect of grinding process (i.e. particle size (PS)) on ergot alkaloid analysis using high performance liquid chromatography – tandem mass spectrometry, and to determine the impact of sample volume on analytical variability. This study demonstrated that correlations exist between both ergot sclerotia count (R2=0.7242, P<0.001) and ergot sclerotia weight (R2=0.9618, P<0.001) compared to the total alkaloid concentration of 6 ergot alkaloids. However, at alkaloid ergot concentrations below 350 µg/kg grain, ergot sclerotia count (R2=0.0002, P=0.956) and ergot sclerotia weight (R2=0.0064, P=0.769) were not correlated to the total alkaloid concentration. A lower variability (P=0.041), defined by coefficient of variation (CV), was observed using a commercial UDY cyclone sample mill (PS=192 µm, CV=9 µg/kg) as compared to a household coffee grinder (PS=516 µm, CV=66 µg/kg). Total amount and concentration of individual ergot alkaloids varied (P<0.05) among sclerotia of similar weight. For the analytical method, CV was numerically reduced as sample volume increased (97% CV for 75 ml to 64% CV for 1000 ml; mean of all concentrations) but increased as sample concentration declined (17% CV for 81,678 µg/kg to 284% for 35 µg/kg; mean of all sample volumes). This implies that analysis of small sample volumes at low ergot alkaloid concentrations may result in highly variable and potentially misleading results. In conclusion, number of ergot sclerotia and weight are unreliable indicators of alkaloid content at ergot concentrations below 350 µg/kg and particle size influences the variability. An analytical approach with fine grinding (mean PS<200 µm, 85% particles <400 µm) of a large sample should be used to assess low-level ergot contamination.

scholarly journals Assessment of ergot (Claviceps purpurea) exposure in pregnant and postpartum beef cows

2018 ◽  
Vol 98 (4) ◽  
pp. 688-700 ◽  
Author(s):  
T. Grusie ◽  
V. Cowan ◽  
J. Singh ◽  
J. McKinnon ◽  
B. Blakley
Keyword(s):  
Ergot Alkaloid ◽  
Post Partum ◽  
Ergot Alkaloids ◽  
Maximum Size ◽  
Beef Cows ◽  
Plasma Prolactin ◽  
Claviceps Purpurea ◽  
Ambient Temperatures ◽  
Beef Cow ◽  
The Impact

Cows were fed ration for 9 wk containing 5, 48, 201, and 822 μg kg−1 ergot alkaloids. The objective was to evaluate the impact of ergot consumption in beef cow–calf operations. Ergot alkaloids up to 822 μg kg−1 did not alter the weight of peripartum and postpartum beef cows (P = 0.93) or nursing calves (P = 0.08), rectal temperature (P = 0.16), or plasma prolactin concentrations (P = 0.30) at moderate ambient temperatures. Ergot did not influence the time (>1 ng mL−1; P = 0.79) or the progesterone concentration (P = 0.38) at the time of first postpartum rise or the size of the first (14 ± 0.6 mm; P = 0.40) and second (13 ± 0.5 mm; P = 0.41) follicles to ovulate. The maximum size of the first postpartum corpus luteum (CL) was 4 mm larger in the 822 μg kg−1 ergot group compared with the control (P = 0.03) for the first ovulation post partum, but not for the second (P = 0.11). There was no effect of ergot exposure on the number of days until the appearance of the first (43 ± 4 d; P = 0.95) or second (52 ± 4 d; P = 0.98) CL post partum. Ergot alkaloid concentrations up to 822 μg kg−1 did not affect pregnancy rates (X2 = 0.36). In conclusion, ergot alkaloid exposure for 9 wk to concentrations as high as 822 μg kg−1 did not alter performance in pregnant and postpartum beef cattle at moderate ambient temperatures.

The effects of selected factors on measured ergot alkaloid content in Claviceps purpurea-infected hexaploid and durum wheat

2016 ◽  
Vol 9 (4) ◽  
pp. 555-564 ◽  
Author(s):  
S.A. Tittlemier ◽  
D. Drul ◽  
M. Roscoe ◽  
J.G. Menzies
Keyword(s):  
Durum Wheat ◽  
Host Plant ◽  
Spring Wheat ◽  
Ergot Alkaloid ◽  
Ergot Alkaloids ◽  
Alkaloid Content ◽  
Claviceps Purpurea ◽  
Wheat Line ◽  
Hard Red Spring Wheat ◽  
Plant Line

Four wheat genotypes, including the ergot-susceptible durum ‘AC Avonlea’ and hard red spring wheat ‘AC Cadillac’, as well as the resistant durum wheat line 9260B-173A and the hard red spring wheat line ‘Kenya Farmer’ wereinoculated with different Claviceps purpurea isolates. Honeydew and sclerotia were collected and analysed for 10 ergot alkaloids. Total concentrations of the 10 ergot alkaloids ranged from 16 µg/kg in honeydew to 1,798 mg/kg insclerotia. Ergonovine and ergosine were the predominant alkaloids in honeydew obtained from plants inoculated with various isolates, whereas ergocristine and ergocryptine were the main alkaloids observed in sclerotia. Bothhost plant and C. purpurea isolate were significant factors affecting total ergot alkaloid concentrations in sclerotia. Irrespective of host plant line, all mean total ergot alkaloid concentrations were higher in sclerotia produced from the EI-2 isolate (695-1,010 mg/kg), as compared to EI-4 (255-594 mg/kg). The mass of total ergot alkaloids was alsopositively correlated with the mass of individual sclerotia produced from these two C. purpurea isolates, with the slope of the regression higher for the EI-2 isolate. The total ergot alkaloid concentrations in sclerotia from various plants inoculated with the same C. purpurea isolate differed; however, the resistance of host plant line did notappear to be consistent with ergot alkaloid content in sclerotia. Concentrations of total ergot alkaloids were highestand lowest in sclerotia from the two lines that are both classified as ‘resistant’, suggesting that the mechanism ofresistance for these lines is not restriction on the production of ergot alkaloids in sclerotia.

scholarly journals Genetic Reprogramming of the Ergot Alkaloid Pathway of Metarhizium brunneum

2020 ◽  
Vol 86 (19) ◽  
Author(s):  
Kyle A. Davis ◽  
Jessi K. Sampson ◽  
Daniel G. Panaccione
Keyword(s):  
Ergot Alkaloid ◽  
High Performance ◽  
Neurological Diseases ◽  
Ergot Alkaloids ◽  
Lysergic Acid ◽  
Efficient Production ◽  
Natural Sources ◽  
Content Type ◽  
Metarhizium Brunneum ◽  
Alkaloid Synthesis

ABSTRACT Ergot alkaloids are important specialized fungal metabolites that are used to make potent pharmaceuticals for neurological diseases and disorders. Lysergic acid (LA) and dihydrolysergic acid (DHLA) are desirable lead compounds for pharmaceutical semisynthesis but are typically transient intermediates in the ergot alkaloid and dihydroergot alkaloid pathways. Previous work with Neosartorya fumigata demonstrated strategies to produce these compounds as pathway end products, but their percent yield (percentage of molecules in product state as opposed to precursor state) was low. Moreover, ergot alkaloids in N. fumigata are typically retained in the fungus as opposed to being secreted. We used clustered regularly interspaced short palindromic repeat (CRISPR)–CRISPR-associated protein 9 (Cas9) and heterologous expression approaches to engineer these compounds in Metarhizium brunneum, representing an alternate expression host from a different lineage of fungi. The relative percent yields of LA (86.9%) and DHLA (72.8%) were much higher than those calculated here for previously engineered strains of N. fumigata (2.6% and 2.0%, respectively). Secretion of these alkaloids also was measured, with averages of 98.4% of LA and 87.5% of DHLA being secreted into the growth medium; both values were significantly higher than those measured for the N. fumigata derivatives (both of which were less than 5.6% secreted). We used a similar approach to engineer a novel dihydroergot alkaloid in M. brunneum and, through high-performance liquid chromatography-mass spectrometry (LC-MS) analyses, provisionally identified it as the dihydrogenated form of lysergic acid α-hydroxyethylamide (dihydro-LAH). The engineering of these strains provides a strategy for producing novel and pharmaceutically important chemicals in a fungus more suitable for their production. IMPORTANCE Ergot alkaloids derived from LA or DHLA are the bases for numerous pharmaceuticals with applications in the treatment of dementia, migraines, hyperprolactinemia, and other conditions. However, extraction of ergot alkaloids from natural sources is inefficient, and their chemical synthesis is expensive. The ability to control and redirect ergot alkaloid synthesis in fungi may allow more efficient production of these important chemicals and facilitate research on novel derivatives. Our results show that Metarhizium brunneum can be engineered to efficiently produce and secrete LA and DHLA and, also, to produce a novel derivative of DHLA not previously found in nature. The engineering of dihydroergot alkaloids, including a novel species, is important because very few natural sources of these compounds are known. Our approach establishes a platform with which to use M. brunneum to study the production of other ergot alkaloids, specifically those classified as lysergic acid amides and dihydroergot alkaloids.

scholarly journals Assessment of the performance of the inter-arrival time algorithm to identify ice shattering artifacts in cloud particle probe measurements

2015 ◽  
Vol 8 (2) ◽  
pp. 761-777 ◽  
Author(s):  
A. Korolev ◽  
P. R. Field
Keyword(s):  
Particle Size ◽  
Arrival Time ◽  
Time Algorithm ◽  
Sample Volume ◽  
Ice Crystals ◽  
Cloud Particle ◽  
Measured Concentration ◽  
Arrival Times ◽  
Microphysical Properties ◽  
The Impact

Abstract. Shattering presents a serious obstacle to current airborne in situ methods of characterizing the microphysical properties of ice clouds. Small shattered fragments result from the impact of natural ice crystals with the forward parts of aircraft-mounted measurement probes. The presence of these shattered fragments may result in a significant overestimation of the measured concentration of small ice crystals, contaminating the measurement of the ice particle size distribution (PSD). One method of identifying shattered particles is to use an inter-arrival time algorithm. This method is based on the assumption that shattered fragments form spatial clusters that have short inter-arrival times between particles, relative to natural particles, when they pass through the sample volume of the probe. The inter-arrival time algorithm is a successful technique for the classification of shattering artifacts and natural particles. This study assesses the limitations and efficiency of the inter-arrival time algorithm. The analysis has been performed using simultaneous measurements of two-dimensional (2-D) optical array probes with the standard and antishattering "K-tips" collected during the Airborne Icing Instrumentation Experiment (AIIE). It is shown that the efficiency of the algorithm depends on ice particle size, concentration and habit. Additional numerical simulations indicate that the effectiveness of the inter-arrival time algorithm to eliminate shattering artifacts can be significantly restricted in some cases. Improvements to the inter-arrival time algorithm are discussed. It is demonstrated that blind application of the inter-arrival time algorithm cannot filter out all shattered aggregates. To mitigate against the effects of shattering, the inter-arrival time algorithm should be used together with other means, such as antishattering tips and specially designed algorithms for segregation of shattered artifacts and natural particles.

Proportions of predominant Ergot alkaloids (Claviceps purpurea) detected in Western Canadian grains from 2014 to 2016

2018 ◽  
Vol 11 (2) ◽  
pp. 259-264 ◽  
Author(s):  
T. Grusie ◽  
V. Cowan ◽  
J. Singh ◽  
J. McKinnon ◽  
B. Blakley
Keyword(s):  
Ergot Alkaloid ◽  
Ergot Alkaloids ◽  
Cereal Crops ◽  
Western Canada ◽  
Claviceps Purpurea ◽  
Total Alkaloids ◽  
Relative Composition ◽  
Equivalence Factor ◽  
Total Alkaloid Content ◽  
Toxicological Significance

Ergot alkaloids, produced by the fungus Claviceps purpurea, are contaminants of cereal crops. Depending on various factors, the relative composition of individual ergot alkaloids can differ among samples. The objective was to determine if the percentage of individual ergot alkaloids were similar across different cereal grains (barley n=39, rye n=7, triticale n=9, wheat n=94) collected in Western Canada over different years. Ergocristine was the predominant alkaloid accounting for half of the total alkaloids in all grain types. This study documented that barley, rye, triticale and wheat collected across Western Canada had similar percentages of ergocornine (6±1%, P=0.201), ergocristine (48±2%, P=0.939), ergocryptine (17±2%, P=0.302) and ergosine (5±0.5%, P=0.239). There were differences between grain types for ergometrine (P=0.027) and ergotamine (P=0.011), which ranged between 6 to 13% and 11 to 24%, respectively, of the total alkaloid content in different cereals. Both barley and wheat alkaloid percentages were similar between 2015 and 2016; ergocornine (7±1%, P=0.969), ergocristine (47±2%, P=0.680), ergocryptine (18±2%, P=0.572), ergometrine (8±1%, P=0.080), ergosine (15±1%, P=0.119) and ergotamine (P=0.189). The ergocornine percentage was higher in wheat (P=0.017) as compared to barley for 2015/2016 samples. Ergometrine was higher in barley (P=0.002) as compared to wheat for 2015/2016 samples. While two of the alkaloid proportions varied statistically, overall proportions of the six ergot alkaloids were comparable among the four grain types collected across Western Canada. If proportions of ergot alkaloids are similar across a region, then it may be deemed acceptable to recommend a maximum total ergot alkaloid concentration for that region. However, areas that exhibit variation among the ergot alkaloid proportions, individual ergot alkaloid guidelines based on a toxic equivalence factor, may be more appropriate. In contrast, since major differences were not seen between years or grain type, from a producer perspective there may be limited biological/toxicological significance for individual alkaloid guidelines.

scholarly journals Phytochemical and High-Performance Liquid Chromatography Analysis of Extract of Vernonia cinerea

2019 ◽  
Vol 9 (1) ◽  
pp. 229-232
Author(s):  
Ruchi Acharya ◽  
Bhawna Sharma ◽  
Ruchi Singh ◽  
Prabhat Jain
Keyword(s):  
High Performance ◽  
Sample Volume ◽  
Small Sample ◽  
Phytochemical Analysis ◽  
Flavonoid Content ◽  
Hydroalcoholic Extract ◽  
Phytochemical Content ◽  
Chromatography Analysis ◽  
Hplc Profiles ◽  
Vernonia Cinerea

The present study aims to screen and quantify hydroalcoholic extract for phytochemical content and HPLC profiles for standardization. HPLC was carried out using a RP-C18 analytical column with a mobile phase composed of acetonitrile: methanol (50:50 v/v) and was isocratically eluted at a flow rate of 1 mL min-1. A small sample volume of 20 μL was used for each sample run, being injected into the HPLC system. The chromatogram was monitored with UV detection at a wavelength of 256 nm. Phytochemical screening of the extract showed the presence of flavonoids, amino acids, carbohydrates and protiens in hydroalcoholic extract. Quantification of total flavonoids showed that hydroalcoholic extract of Vernonia cinerea had flavonoid content 0.547 mg/100mg equivalent to quercetin. The data presented here could be used for the standardization of hydroalcoholic extract of Vernonia cinerea, either for future studies or in herbal drug formulations. Keywords: Vernonia cinerea, HPLC profiling, Phytochemical analysis, Hydroalcoholic extract.

scholarly journals Simultaneous Determination of Ergot Alkaloids in Swine and Dairy Feeds Using Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry

Toxins ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 724
Author(s):  
Saranya Poapolathep ◽  
Narumol Klangkaew ◽  
Zhaowei Zhang ◽  
Mario Giorgi ◽  
Antonio Francesco Logrieco ◽  
...  
Keyword(s):  
High Performance ◽  
Ergot Alkaloids ◽  
The European Union ◽  
Chromatography Tandem Mass Spectrometry ◽  
Occurrence Data ◽  
Performance Recovery ◽  
Liquid Chromatography Tandem Mass ◽  
Swine Feed ◽  
Feed Samples

Ergot alkaloids (EAs) are mycotoxins mainly produced by the fungus Claviceps purpurea. EAs are known to affect the nervous system and to be vasoconstrictors in humans and animals. This work presents recent advances in swine and dairy feeds regarding 11 major EAs, namely ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, ergocristine, ergosinine, ergotaminine, ergocorninine, ergocryptinine, and ergocristinine. A reliable, sensitive, and accurate multiple mycotoxin method, based on extraction with a Mycosep 150 multifunctional column prior to analysis using UHPLC-MS/MS, was validated using samples of swine feed (100) and dairy feed (100) for the 11 targeted EAs. Based on the obtained validation results, this method showed good performance recovery and inter-day and intra-day precision that are in accordance with standard criteria to ensure reliable occurrence data on EA contaminants. More than 49% of the swine feed samples were contaminated with EAs, especially ergocryptine(-ine) (40%) and ergosine (-ine) and ergotamine (-ine) (37%). However, many of the 11 EAs were not detectable in any swine feed samples. In addition, there were contaminated (positive) dairy feed samples, especially for ergocryptine (-ine) (50%), ergosine (-ine) (48%), ergotamine (-ine), and ergocristine (-ine) (49%). The mycotoxin levels in the feed samples in this study almost complied with the European Union regulations.

scholarly journals An Old Yellow Enzyme Gene Controls the Branch Point between Aspergillus fumigatus and Claviceps purpurea Ergot Alkaloid Pathways

2010 ◽  
Vol 76 (12) ◽  
pp. 3898-3903 ◽  
Author(s):  
Christine M. Coyle ◽  
Johnathan Z. Cheng ◽  
Sarah E. O'Connor ◽  
Daniel G. Panaccione
Keyword(s):  
Aspergillus Fumigatus ◽  
Ergot Alkaloid ◽  
Ergot Alkaloids ◽  
Aldehyde Group ◽  
Ring Structure ◽  
Ring Closure ◽  
Claviceps Purpurea ◽  
Wild Type ◽  
Old Yellow Enzyme ◽  
Clavicipitaceous Fungi

ABSTRACT Ergot fungi in the genus Claviceps and several related fungal groups in the family Clavicipitaceae produce toxic ergot alkaloids. These fungi produce a variety of ergot alkaloids, including clavines as well as lysergic acid derivatives. Ergot alkaloids are also produced by the distantly related, opportunistic human pathogen Aspergillus fumigatus. However, this fungus produces festuclavine and fumigaclavines A, B, and C, which collectively differ from clavines of clavicipitaceous fungi in saturation of the last assembled of four rings in the ergoline ring structure. The two lineages are hypothesized to share early steps of the ergot alkaloid pathway before diverging at some point after the synthesis of the tricyclic intermediate chanoclavine-I. Disruption of easA, a gene predicted to encode a flavin-dependent oxidoreductase of the old yellow enzyme class, in A. fumigatus led to accumulation of chanoclavine-I and chanoclavine-I-aldehyde. Complementation of the A. fumigatus easA mutant with a wild-type allele from the same fungus restored the wild-type profile of ergot alkaloids. These data demonstrate that the product of A. fumigatus easA is required for incorporation of chanoclavine-I-aldehyde into more-complex ergot alkaloids, presumably by reducing the double bond conjugated to the aldehyde group, thus facilitating ring closure. Augmentation of the A. fumigatus easA mutant with a homologue of easA from Claviceps purpurea resulted in accumulation of ergot alkaloids typical of clavicipitaceous fungi (agroclavine, setoclavine, and its diastereoisomer isosetoclavine). These data indicate that functional differences in the easA-encoded old yellow enzymes of A. fumigatus and C. purpurea result in divergence of their respective ergot alkaloid pathways.

scholarly journals High-Performance Liquid Chromatography/Tandem Mass Spectrometry Method for the Determination of Arbidol in Human Plasma

2011 ◽  
Vol 94 (4) ◽  
pp. 1100-1105 ◽  
Author(s):  
Xin Xiong ◽  
Suodi Zhai
Keyword(s):  
Human Plasma ◽  
High Performance ◽  
Internal Standard ◽  
Sample Volume ◽  
Tandem Mass ◽  
Spectrometry Method ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Butyl Methyl Ether

Abstract An HPLC/MS/MS method for the determination of arbidol in human plasma was developed. Arbidol and internal standard (loratadine) were extracted from alkaline plasma with tert-butyl methyl ether and analyzed on a Zorbax SB C18 column (30 × 2.1 mm id, 3.5 µm particle size). The detection was by monitoring arbidol at m/z 479.1 → 434.1 and the internal standard at m/z 383.2 → 337.2. The method was validated according to U.S. Food and Drug Administration guidelines. The calibration curve was linear over the range of 0.5–500 ng/mL using a 100 µL sample volume. The intraday and interday precisions were less than 6.5%, and acceptable values were obtained for accuracy, recovery, and sensitivity. The developed method was selective, simple, sensitive, and easily applicable.

SOME ASPECTS OF PHOSPHATE METABOLISM OF CLAVICEPS PURPUREA (FR.) TUL.: II. RELATIONSHIP BETWEEN NUCLEIC ACIDS SYNTHESIS AND ERGOT ALKALOID PRODUCTION

1961 ◽  
Vol 7 (6) ◽  
pp. 883-888 ◽  
Author(s):  
C. de Waart
Keyword(s):  
Nucleic Acid ◽  
Ergot Alkaloid ◽  
Ribonucleic Acid ◽  
Ergot Alkaloids ◽  
Acid Synthesis ◽  
Claviceps Purpurea ◽  
Phosphate Metabolism ◽  
Alkaloid Production ◽  
Phosphorus Containing ◽  
Insoluble Phosphate

A study was made of the distribution of the phosphorus-containing compounds in the phosphate pool of stationary cultures of Claviceps purpurea (Fr.) Tul. Exogenous KH2PO4 was mainly converted to ribonucleic acid. Conditions favorable to increased nucleic acid synthesis increased the yield of ergot alkaloids. The proportion of nucleic acid and acid-insoluble phosphate fraction appeared to be an important factor influencing the ergot alkaloid production.

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