Nelson-Somogyi Methods

Sugars with reducing property (arising out of the presence of a potential aldehyde or keto
groups) are called reducing sugars. Some of the reducing sugars are glucose, galactose,
lactose and maltose. The Nelson-Somogyi method is one of the classical and widely used
methods for the quantitative determination of reducing sugars.

Principle

The reducing sugars when heated with alkaline copper tartrate reduce copper from the cupric
to cuprous state and thus cuprous oxide is formed. When cuprous oxide is treated with
arsenomolybdic acid, the reduction of molybdic acid to molybdenum blue takes place. The
blue color developed then compared with a asset of standards in a colorimetri at 620 nm.
Materials
Spectrophotometer UV-Vis Genesys™ 10S UV-Vis (Thermofisher, Waltham, Massachusetts,
United States)

Procedure
1. Weigh 100 mg of the samples and extract the sugars with 80% ethanol twice (5 ml each
time)
2. Collect the supernatant and evaporate it by keeping it on a waterbath at 80°C
3. Add 10 ml aquadest and dissolve the sugars
4. Pipette out aliquots of 0,1 ml to separate test tubes
6. Pipette out 0,2;0,4;0,6;0,8 and 1 ml of the working standard solution into a series of test
tubes
7. Pipette out 2 ml distilled water in distilled water in a separate tube to set a blank
8. Add 1 ml of alkaline copper tartrate reagent to each tube
9. Place the tubes in boiling water for 10 minutes
10. Cool the tubes and add 1 ml of arsenomolybdic acid reagent to all the tubes
11. Make up the volume in each tube to 10 ml with water
12. Read the absorbance of blue colour at 620 nm after 10 min
Result

Sample Sugar Analysis Series 1 Series 2

Reducing Sugar (%) 0,06 0,06
EHA powder Total Sugar (%) 49,44 49,60
Sucrose (%) 46,91 47,07

Reducing Sugar (%) 0,01 0,01

EHA Pellet sintered
Total Sugar (%) Not detected Not detected
at 1200°C
Sucrose (%) Not detected Not detected