116 / 166
y = 0.2965x + 0.2085
R² = 0.9977
y = 0.2362x + 2.4442
R² = 0.9893
y = 0.1034x + 0.8348
R² = 0.999
0.00
10.00
20.00
30.00
40.00
50.00
60.00
70.00
0
50
100
150
200
GLUTEN PPM
SPIKE PPM
GLUTEN R5 Assay Kit Relative Response by Spiked Grain Weight
Barley
Rye
Wheat
Linear (Barley)
Linear (Rye)
Linear (Wheat)
BARLEY SLOPE = 29.7%
WHEAT SLOPE = 10.3%
RYE SLOPE = 23.6%
15
ACCURACY STRATEGY
• Response of prospective method should be evaluated for 3 grains
independently.
• Experimental evaluation should be performed on grain (flour) spiked
into oat flour, and extracted as per proposed kit extraction method.
• Samples of spiked material could be made by 3
rd
party lab and
made available to researchers, along with non-ELISA “reference”
values.
• Results of these experiments have shown much lower RSDs than
we see in practice, so this experiment will not be useful for
estimating precision.
• Because of the uncertainty in everything, the acceptance criteria for
this approach will need to be very wide.
• This approach is probably best done as part of single lab validation
or single lab with independent lab verification, along with some sort
of cross-reactivity study.
16