Glucose molecule

Glucose Homeostasis
Glucose

Carbon
Oxygen
Hydrogen
Cuantitative analysis of glucose by chemical method
Cuantitative analysis of glucose by enzymatic method

Glucose oxidase catalyse the oxidation of Beta D- glucose present in the plasma to D-gluconic acid with the formation
of hydrogen peroxide. In the second reaction, the hydrogen peroxide produced is then broken down to oxygen and
water by a peroxidase enzyme. Oxygen then react with an oxygen acceptor such as a mix of phenol and 4-
aminoantipyrine which itself converted to a coloured compound (quinonimine), the amount of which can be measured
colorimetrically at 505 nm. The second reaction is called Trinder Reaction. The pink intensity color is proportional to
the glucose concentration present in the samples.
Protocol: Glucose Oxidase Method
 Protocol: Glucose Oxidase Method

To elaborate the standard curve, each group will prepare the next glucose solutions in 6 labeled
test tubes (A-F). For that, the student will make dilutions of the 0.2% (p/v) commercial glucose in
distilled water.
 Protocol: Glucose Oxidase Method
Follow the instructions given in the below table. The final volume of each tube is 2.5 mL. Do not forget
that the zero and the samples do not have standard glucose:

We will use only one plastic cuvette for all the measurements. Firstly, we will calibrate with the zero
absorbance (0 test tube). For the rest of measurements, we will measure from the lower (light colour)
to the higher concentration (strong colour) at 505 nm. Use the table below to annotate the
absorbance values.
 Protocol: Glucose Oxidase Method

https://www.youtube.com/watch?v=DyGIvs9zrVA
Laboratory work
Laboratory work