Report Reducing Sugar
Report Reducing Sugar
Report Reducing Sugar
APPARATUS
Material :
a) 200mL beaker
b) Pipette
c) Volumetric flask.
d) cuvvete
e) spectrophotometer.
Chemicals :
a) Low-alkalinity copper reagent sodium potassium tartrate (40g) and
anhydrous (28g) were dissolved in about 700 mL of water. A solution
of 10% cupric acid sulphate pentahydrate in water (80mL) was
added in small amounts with stirring. 180 g anhydrous was added in
small amounts and was dilute to 1 litre. After 1 day of standing, the
clear supernatant solution was used.
b) Arsenomolybdate reagent- 25g of ammonium molybdate in 450 mL
water was added 21mL of 95% sulphuric acid, followed by 3.0g of
disodium hydrogen arsenate heptahydrate dissolved in 25mL water.
The mixed solution was incubated 25 hours at 37OC and stored in a
glass- stoppered brown bottle.
c) Standard glucose solution stock solution was diluted containing
1mg/mL glucose in sturated benzoic acid solution to give a standard
solution of 100 mg/mL and 300 mg/mL
Sample : carbonated drinks.
PROCEDURES
A : sample preparation
RESULT
Glucose
solution
(100g/ml)
Glucose
solution
(300g/ml)
Distilled
water (ml)
Absorbance
reading
Tube 1
0.0
Tube 2
0.5
Tube 3
1.0
Tube 4
1.5
Tube 5
2.0
Tube 6
-
Tube 7
-
1.0
1.5
2.0
1.5
1.0
0.5
0.0
1.0
0.5
0.055
0.297
0.507
0.779
1.029
1.641
1.962
f(x) = 0x + 0.04
R = 0.98
1.5
Absorbance 520nm
1
0.5
0
0
Amount (ml)
Absorbance
A1
A2
B1
B2
0.5
0.5
1.0
1.0
Sample A2
X
Sample B1
X
Sample B2
X
0.244
0.246
0.428
0.406
= 50 x 10-6g x 50 x 10 x 4
= 0.100 g/ml
A2
= 50.5 x 10-6g x 50 x 10 x 4
= 0.101 g/ml
Average
= (0.100 g/ml + 0.101g/ml) / 2
= 0.1005g/ml
= 96 x 10-6g x 50 x 10 x 2
= 0.096 g/ml
B2
= 90.5 x 10-6g x 50 x 10 x 2
= 0.0905 g/ml
Average
= (0.096 g/ml + 0.0905 g/ml) / 2
= 0.09325 g/ml
Final concentration of carbonated drinks reducing sugar is ;= 0.1005 g/ml + 0.09325 g/ml) / 2
= 0.097 g/ml
DISCUSSION
For this experiment, the Somogyi Nelson method have been used in
determining the concentration of reducing sugar in the sample which is
carbonated drink. This method is commonly used in determining the reducing
sugar rather than the DNS method because of its effectiveness for all types of
sample. That is because, it does not matter about the color of the sample, it can
be easily applied in determination of reducing sugar.
For this experiment, the sample that we used is Coke drink which is a kind
of carbonated drink. There several steps in doing this method and it started with
removing the gas of that carbonated drink. Then, the sample solution is diluted
with the distilled water for neutralization. After the several dilution of the sample,
the sample is added with copper reagent and arsenomolybdate reagent. After
mixing and incubated at room temperature for several minutes, the absorbance
of the sample can be read by spectrophotometer at 520nm wavelength.
In principle, during the first step, glucose (or a reducing sugar) is oxidised
using a solution of Cu(II) ion which in the process is reduced to Cu(I). In the
second step the Cu(I) ions are then oxidised back to Cu(II) using a colourless
hetero-polymolybdate complex, which is, in the process, reduced to give the
characteristic blue colour. Finally the absorption of the hetero-poly molybdenum
blue is measured using a colorimeter and compared to standards prepared from
reacting sugar solutions of known concentration, to determine the amount of
reducing-sugar present.
When the reading of absorbance is got, the concentrations of reducing
sugar can be determined by refer to the reducing sugar standard curve that has
been produced before. In order to achieve the exact concentration of the
reducing sugar, we need to multiply the result with the dilution factor. And after
all, the concentration of reducing sugar of coke drink that we got is only 0.097
g/ml. From the result we got, the concentration of reducing sugar that we
determined is not really high.
CONCLUSION
The concentration of reducing sugar of the carbonated drink is not really
high which is only about 0.097 g/ml. However, if compared to the other type of
drinks, the amount of reducing sugar of carbonated drink is much higher.
REFFERENCES
1) http://en.wikipedia.org/wiki/Reducing_sugar
2) http://www.eplantscience.com/index_files/plant
%20protocols/Carbohydrates/reducing_sugars_by_NelsonSomogyi_method.php