sampling and analysis of grain cleaning byproducts for truly rapid
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Sampling and Analysis of Grain Cleaning Byproducts for Truly Rapid Screening of the Mycotoxin Deoxynivalenol in WheatSheryl A. Tittlemier,1 D Sobering2, K Bowler2, T Zirdum2, D Gaba1, JM Chan1, M Roscoe1, R Blagden1, L Campbell31Grain Research Laboratory, Canadian Grain Commission, Winnipeg, MB, Canada2Industry Services, Canadian Grain Commission, Winnipeg, MB, Canada3National Monitoring Unit, Canadian Grain Commission, Winnipeg, MB, Canada
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Canadian Grain Commission certifies the quality, safety
and weight of Canadian grain that is delivered to domestic and export markets quality and quantity
assurance programs scientific research to
understand all aspects of grain quality and safety
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Fusarium head blight
“scab”, “FHB” disease caused by infection
with Fusarium fungal species bleached heads and shriveled
chalky/pinkish kernels (FDK) decreased yield and quality a variety of mycotoxins can be
produced
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Deoxynivalenol (DON)
main mycotoxin produced by F. graminearum
“vomitoxin” maximum limits in wheat: 2 mg/kg Codex (raw grain, proposed) 1 -2 mg/kg Canada (under review) 1.25 – 1.75 mg/kg EU 1.1 mg/kg Japan (provisional standard) 1 mg/kg US (finished wheat products)
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Analysis – chemical testing
primary sampling
laboratorysample
test portion
officialsample
secondary sampling
chemical analysis
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Analysis – chemical testing
primary sampling
laboratorysample
test portion
secondary sampling
chemical analysis
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Piggyback on existing analyses dockage
dockage analyzed during grain inspection dockage = wheat with long rootlets, unthreshed wheat
heads or spikelets, chaff, dust, loose hulls, fragments of stem or rachis, soft earth pellets, broken kernels, bunt balls, stones, wild oats, etc.
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Piggyback on existing analyses dockage
dockage analyzed during grain inspection dockage must be removed from grain by use of
approved cleaning equipment so grain can be assigned a grade (Canada Grain Act)
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Mycotoxins are present in dockage
Cleaning byproduct N samples DON
(mg/kg)OTA
(mg/kg)ZEA
(mg/kg)
chaff and other light material 1 0.75 0.019 0.045
broken grain and weed seeds 3 1.42 – 1.86 0.004 – 0.007 <0.010 – 0.019
waste 9 0.21 – 2.94 <0.001 – 0.014 <0.010
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Piggyback on existing analyses dockage
dockage analyzed during grain inspection dockage must be removed from grain by use of
approved cleaning equipment so grain can be assigned a grade (Canada Grain Act)
dockage is a readily available sample matrix
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Dockage removal
Carter Day dockage tester
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Dockage removal
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Dockage fractions contain mycotoxins
small dockage
largedockage
light dockage
cleanedgrain
mg/kg
0.00
0.05
0.10
0.15
0.20
0.25
0.0
0.1
0.2
0.3
0.4
0
5
10
15
20
25
30
35
OTA
ZEA
DON
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Light dockage is a small proportion of wheat
average percentage of mass of un‐cleaned wheat
clean wheat
large dockage
light dockage
small dockage
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Pilot: DON in light dockage and wheat
single railcar composites collected by CGC inspectors using automatic
samplers at terminal elevator facilities samples with FDK and dockage selected because of
increased likelihood of detecting DON homogenized and divided into two portions using
Boerner divider one half (~ 1 kg) run through dockage tester to
collect light dockage other half was kept as whole, un-cleaned grain
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Sample analysis
grind 1 kg 50 g test portion
NeogenVeratox DON
ELISA
Whole un-cleaned grain:
all light dockage
NeogenVeratox DON
ELISA
Light dockage:
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Performance of ELISA: un-cleaned wheat
ELISA (mg/kg)
0 2 4 6 8 10 12
GC
-MS
(mg/
kg)
0
2
4
6
8
10
12
14
y = 0.907x + 0.0503R² = 0.881
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Performance of ELISA: light dockage
ELISA (mg/kg)0 5 10 15 20 25
GC
-MS
(mg/
kg)
0
5
10
15
20
25
y = 1.059x - 0.1557R² = 0.986
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DON in whole grain (mg/kg)0 1 2 3 4 5
DO
N in
ligh
t doc
kage
(mg/
kg)
0
10
20
30
40
y = 5.8688x + 0.4324R² = 0.6066
DON in whole grain is correlated to DON in light dockage
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Conclusions
use of light dockage for screening DON in wheat is promising benefits increased sensitivity save time by eliminating grinding and prep steps save resources by removing need for grinders uses available material does not require additional equipment
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