Protocol Online logo
Top : New Forum Archives (2009-): : General Lab Techniques

Detecting glucose in reactions with OD - Detecting glucose without converting (Mar/16/2009 )

Please if one can help by giving me method for detecting glucose By OD without converting glucose to any products
for examples: without using glusoe oxidase and then peroxidase
and if possible to get out the glucose concentration
please help me

-longer-

Accuracy and precision is better with enzymatic methods but you could try 0-toluidine. In hot acetic acid this reacts with glucose to form a green derivative. Set your spectrometer to 630nm. I'm not sure what the limit of detection is with this system but I think it has a similar range to the enzymatic methods.

There is (or was) also a REDOX method. The neocuproine method uses ferricyanide (Fe3+) which is coloured. In the presence of glucose this forms the ferrocyanide (Fe2+) which is clear. Read at 420nm. I don't think this method has a very high specificity. If you are analysing serum, for instance, then high creatinine and urea concentrations will give falsley elevated glucose readings.

Last thought, what about a home glucose monitoring unit. Depending on your sample it may not be validated and it may have huge CVs at the concentration of glucose you want to measure. (These are linear between fairly narow ranges, often between 2mmol/L and 20mmol/L).

Hope that helps.

-Paraboxa-

thankx Paraboxa, I am interesting about the first method,but I have small question can I get the glucose concentrations by this

thankx once again

-longer-

Yes, the reaction follows Beers law: colour intensity is proportional to concentration of glucose. You'd need to check the specificity against other sugars and the linear range.

-Paraboxa-

Anthrone reaction.

-hanming86-