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

In: World Mycotoxin Journal
Authors:
T. Grusie Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive Saskatoon Saskatchewan Canada.

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V. Cowan Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive Saskatoon Saskatchewan Canada.
Toxicology Centre, University of Saskatchewan, 44 Campus Drive Saskatoon Saskatchewan Canada.

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J. Singh Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive Saskatoon Saskatchewan Canada.

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J. McKinnon Department of Animal and Poultry Science, University of Saskatchewan, 51 Campus Drive Saskatoon Saskatchewan Canada.

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B. Blakley Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive Saskatoon Saskatchewan Canada.
Toxicology Centre, University of Saskatchewan, 44 Campus Drive Saskatoon Saskatchewan Canada.

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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.

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