Anthrone carbohydrate rxn questions

[Peter]

Hello,

 

I am a biological/process engineer. I am trying to scale down the anthrone/carbohydrate assay to reaction volumes in the 25-50 uL range.

 

I have a few questions.

 

Does the latent time between when one adds anthrone reagent (anthrone + 70-75% H2SO4) to the carb. source or unknown until the reactions are brought to boiling effect the assay?

 

How long is the color of the completed assay stable?

 

Does anthrone bind to cellular material thereby skewing the results?

 

Can an anthrone concentration lower than 175 mg/mL be used in this reaction?

 

Has anyone successfully scaled this assay down to be conducted in microplates?

 

Thanks,

 

Peter