Qualitative Tests For Carbohydrates

PANTOJA, ARIEL S.
1-Y2-12
BS NURSING
QUALITATIVE TESTS FOR CARBOHYDRATES

The tests of the carbohydrates are based on their reducing properties (due to the
presence of reducing aldehyde or ketone groups). Fehling’s test, Benedict's test are examples
of this. The unspecific Molisch’s test for carbohydrates is one of the examples of some tests
which are based on the formation of furfural or furfural derivatives in presence of concentrated
acids. Specific complex formation is sometimes used as a specific test for carbohydrates.
Formation of phenylhydrazone is one such example. For testing polysaccharides, iodine is
found to be very useful.
MATERIALS MAIN REAGENTS

- Test Tubes - Fehling’s A and B
- Droppers - Benedict’s
- Graduated cylinders - Seliwanoff’s
- Test Tube Rack - Molisch’s
- Marker and Paper tape - Barfoed’s
- Iodine soin
CHEMICALS
- Glucose
- Fructose
- Lactose
- Sucrose
- Starch
METHODS
Molisch's test
- A general test for carbohydrate concentrated sulfuric acid will also be used in this test.
Alcoholic alpha naphthol forms furfural and furfural derivatives, such as
hydroxymethylfurfural, by the concentrated sulphuric acid acting on the sugar. This
compound forms a reddish-violet coloured ring at the junction of the two liquids.
Molisch’s reagent is a 5% solution of alpha naphthol in alcohol.
Benedict’s Test:
- Add 2mL of Benedict's qualitative reagent to the sugar solution, and place the test tube
boiling water bath for 3 seconds. In case of reducing sugars there will be an appearance
of blue precipitate.
Fehling’s Test
- Carbohydrates with free aldehyde or ketone groups reduce copper sulphate to cuprous
oxide forming a yellow or brownish red coloured precipitate. Fehling’s reagent is
prepared freshly by mixing equal volumes of two stock solutions A and B. Solution A is
6.93 grams of CuSO4.5H2O per 3 ml of water and Solution B is 20 grams of KOH and
34.6 grams of sodium potassium tartrate (Rochelle salt) per 1 ml solution.
Barfoed’s Test
- Mix 2 ml of Barfoed’s reagent with 1 ml of carbohydrate solution in a test tube and heat
in a boiling water bath for 10 min. Appearance of a red precipitate of cupric oxide (Cu20)
indicates the presence of reducing sugar.
Seliwanoff’s Test
- To the sugar (2 ml) add 2 ml of seliwanoff's reagent. A blank without sugar should also
be prepared to judge the colour change. Place the tubes in boiling water for exactly 5
min. Note the colour change, if any, and then continue the heating for 5 minutes and
periodically observe the colour change.
Iodine Test
- This may be an adsorption complex of starch or glycogen with iodine rather than a
definite compound. Iodine reagent is 0.5 ml of iodine diluted to 5 ml with distilled water.
Potassium iodide is added to the reagent solution in order to make the iodine more
soluble in water.
DATA
Based on the reactivity with Benedict’s or Fehling’s reagent, carbohydrates are classified
Reducing sugars
Carbohydrates that can reduce Benedict’s or Fehling’s reagents are called reducing sugars
(sugar with free aldehyde or ketone group). All monosaccharides and most of the disaccharides
are reducing sugars. Some examples are Maltose and Lactose.
Non-reducing sugars
Carbohydrates that cannot reduce Benedict’s or Fehling’s reagents are called non-reducing
sugars. Sucrose is a non-reducing sugar.
ANALYSIS
Molisch’s reagent is a 10% alcoholic solution of α-naphthol. This is a common chemical
test to detect the presence of carbohydrates. Fehling’s solution A is copper sulphate solution
and Fehling’s solution B is potassium sodium tartrate. Benedict's reagent contains blue copper
(II) ions (Cu2+, cupric ions) that are reduced to copper (I) ions (Cu+, cuprous ions) by
carbohydrates. These ions form precipitate as red coloured cuprous (copper (I) oxide. Iodine
test is used to detect the presence of starch.
CONCLUSION
It gave a positive result under the Molisch test indicating that it is a carbohydrate and it
also gave a positive result under the Benedict's test indicating that it is a reducing sugar. In
addition to this it tested positive under the iodine test indicating the presence of starch. It gave
negative results under the Seliwanoff's test which indicates it is not Fructose. It also gave a
negative result under the Bial's test which indicates that it is a hexose sugar and not a pentose
sugar. As previously mentioned the starch gave a blue result under Benedict's test which is not
what was expected from a reducing sugar, as it would have been expected to produce an
orange precipitate. This may have been down to contamination but with what remains unknown.
REFERENCES
https://www.youtube.com/watch?v=rKng5-ij6kQ&feature=youtu.be

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PANTOJA, ARIEL S.

QUALITATIVE TESTS FOR CARBOHYDRATES

MATERIALS MAIN REAGENTS

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