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Calorimetric Determination of Glucose by The 3,5-Dinitrosalicylic Acid Method
Calorimetric Determination of Glucose by The 3,5-Dinitrosalicylic Acid Method
Calorimetric Determination of Glucose by The 3,5-Dinitrosalicylic Acid Method
:Principle
Several reagents have been employed which assay sugars by using their
reducing properties. This method tests for the presence of free carbonyl
group (C=O), the so-called reducing sugars. This involves the oxidation
of the aldehyde functional group present in, for example, glucose and the
ketone functional group in fructose. Simultaneously, 3,5-dinitrosalicylic
acid (DNS) is reduced to 3-amino-5-nitrosalicylic acid under alkaline
:conditions, as illustrated in the equation below
:Materials
:Procedure
SECTION A SECTION B
.Tube No ml. Stand. ml. H2O ml. Dinitrosalicylic bbbbbB
ml. H2O BB
.Glucose reagent
Cool the tubes thoroughly and then add 7.0 ml of distilled water to .3
each tube as indicated in section B of the previous table, Read the
extinction (Optical density) of the colored solutions at 540 nm using the
.solution in tube 1 as a blank (control)
Note: All the tubes must be cooled to room temperature before reading
.since the extinction is sensitive to temperature change
Record the readings in section B, and plot the relationship between the .4
optical density and the concentration of glucose solution. See whether
there is a linear relationship between the concentrations of glucose
.solutions and their corresponding optical densities
Use the already prepared standard curve for the determination of the .5
unknown concentration of the glucose solution provided and tissue
.extract form exp.6 or any other unknown reducing sugar sample
Name: No.
Experiment 1:
Results Sheet
The concentration of standard glucose solution : mg/ml
- Plot the standard curve of the absorbance (y- axis) against the
concentration ( x-axis )
Experiment 1:
Results Sheet