Hello together,
I am conducting the established assay for carbohydrate detection with anthrone reagent in an aquatic organism. Normally, the color changes like expected from yellow to green, or stays yellow when no sugar is in my sample.
But in my last assays with (differently) treated organisms, the color changed from yellow to brown instead. I already renewed the reagent and all chemicals I use in the assay, but the color always changed to brown. During my last try, I examined treated and, as control for the color change, freshly caught organisms. Suprisingly, the color changed for the freshly caught to green and for the others to brown. So the assay must have worked.
Does anyone know, how this color change to brown could happen? Were you able to observe such a color change in your assays? Is it possible that my treated organisms are so much weakened that somethings else was produced consuming the sugars?
Many thanks for your ideas.
A tan color is possible with some rare deoxy sugars: Article Limitations of the Anthrone Test for Carbohydrates
That seems an unlikely explanation, but your organisms could be producing an interfering metabolite that reacts with the anthrone reagent. Some sort of solvent extraction or filtration step (e.g. a C-18 cartridge) might be necessary to remove the interfering material.
Check https://europepmc.org/articles/pmc1269144/pdf/biochemj01094-0139.pdf
Also check https://www.researchgate.net/publication/230015476_Reaction_of_Sugar_Alcohols_with_the_Anthrone_Reagent
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