Additional Notes: Carbohydrates (Laboratory)

Molisch’s Test

- Sensitive chemical test for all carbohydrates (even a dilutions as low as 0.001% still gives definite
positive result)

- The test depends on the formation of furfural, or one of its derivatives, when carbohydrates are
treated with mineral acid. The furfurals further condense with alpha-naphthol forming
compounds with reddish violet or purple color.

I2 reaction

- An indicator for the presence of starch. Iodine solution (iodine dissolved in an aqueous solution
of potassium iodide) reacts with starch producing a blue-black color. This is because iodine will
slip in into the coiled (helix) chain of amylose which is found in starch, producing color change.

- Glycogen does not have a helical structure like amylose, and thus, will retain the reddish-brown
color of the iodine solution.

Tests for Reducing Sugars

- Requires high pH and temperature in order to increase susceptibility to oxidation reactions [that
is why the qualitative test for carbohydrates requires heat (direct (flame) or indirect (boiling)]

Barfoed’s reagent

- Cupric acetate in acetic acid

- Is slightly acidic and is balanced so that is can only be reduced by monosaccharides, but not by
less powerful reducing sugars. Disaccharides may also react with this reagent, but the reaction is
much slower when compared to monosaccharides.
- Positive: glucose, maltose
- Negative: sucrose

Seliwanoff’s Test

- This test is based on the principle that, when heated, ketoses are more rapidly dehydrated than
aldoses.
- Positive: fructose, sucrose
- Negative: glucose, maltose

Bial’s Test

- Positive: ribose
- Negative: glucose.