Last Update: January 24, 2007 Part I

M 21
ESTIMATION OF PERCENTAGE OF GLUCOSE (Benedicts Method)

PRINCIPLE :
Glucose is estimated by the quantitative reduction of Benedicts quantitative reagent. Glucose, due
to its reducing properties, reduces copper sulphate in hot alkaline medium. Because of the presence of
potassium thiocyanate in the Benedicts quantitative reagent a white precipitate of cuprous thiocyanate
instead of the usual red precipitate of cuprous oxide, is formed on reduction of Cu ++ by glucose. The little
amount of potassium ferrocyanide present in the reagent keeps cuprous oxide in solution. Sodium citrate
present in the reagent keeps CuCO2 (formed by the inteaction of CuSO4 with Na2CO3) in solution. The
alkali used is Na2CO3 which is likely to cause less destruction of sugar.

REQUIREMENTS :
1. Glucose solution (supplies)
2. Distilled Water
3. Benedicts Quantitative Reagent
Weigh 200g of sodium citrate, 200g of sodium carbonate and 125g of potassium thiocyanate and
place them in a litre beaker. Add about 500ml of distilled water and boil vigorously for one hour with
occasional stirring. Cool and filter. Collect the filtrate in litre Hask.
Weigh 18g of pure copper sulphate and dissolve it in about 100-150 ml of water. Transfer this
copper sulphate solution to the litre flask containing the litre flask containing the above solution. Add 5ml
of 5% potassium ferrocyanide solution and mix thoroughly. Make the volume of the solution upto the litre
mark with distilled water.
4. Anydrous Na2CO3.

PROCEDURE :
Take 10 ml of supplied glucose solution in a 100 ml volumetric flask and make the volume upto the
mark with distilled water. Mix thoroughly and transfer the diluted solution to a 25 ml burette. Note the
initial burette reading.
Pipette 10 ml of Benedicts quantitative reagent and transfer carefully to a 100 ml conical flask.
Add about 1 g of anhydrous Na2CO3 and place the conical flask over a tripod stand. Boil the reagent gently
on a low flame, and while boiling add the diluted glucose solution drop by drop from the burette.
The disappearance of blue colour and appearance of pale white precipitate indicates the end point of
titration. Note the final burette reading.

RESULT :
No. of Burette reading in ml Volume of dilute Mean volume of dilute
observations Initial Final glucose required glucose required (ml)
(ml)
1.
2.
3.

Calculation :
10 ml Benedicts Quantitative Reagent =0.02g of glucose.
If the mean titration reading is X ml., the percentage of glucose in the given solution
.02 100 10
g
x