Anthrone’s Test

The Anthrone method is an example of a colorimetric method of determining the concentration of the total sugars in a
sample. Sugars react with the anthrone reagent under acidic conditions to yield a deep green colored-solution. The
sample is mixed with sulfuric acid and the anthrone reagent and then boiled until the reaction is completed. The
solution is then allowed to cool and its absorbance is measured at 620 nm. There is a linear relationship between the
absorbance and the amount of sugar that was present in the original sample. This method determines both reducing
and non-reducing sugars because of the presence of the strongly oxidizing sulfuric acid.

ANALYSIS

In our experiment, the solutions made were not boiled but instead we just let the solution to stand for sometime.
Since, this test would made to react to all of the solutions therefore all the sugars must yield a positive result. In our
experiment, only the glucose and the glycogen showed a positive result. Anthrone reagent is prepared in
concentrated sulfuric acid. Adding reagents to acid causes immediate dissociation of the reagent and
releases heat. The negative results of the rest of the sugars may be due to poor reaction of the acid to
hydrolyzed the complex sugars.

NYLANDER’S TEST

In this said experiment, we used Nylander’s reagent. This reagent is composed of a solution of Rochelle salt
(potassium sodium tartrate), potassium or sodium hydroxide, and bismuth subnitrate in water; used to test for
sugar in urine. Nylander's test is a test for reducing sugars ( principally glucose ) in usually, urine samples.
It relies on the reducing properties of glucose and related reducing sugars. The reagent is a basic tartrate
complex of a bismuth salt. When heated in the presense of a reducing sugar the
bismuth is REDUCED to the finely divided black element which precipitates. There should be no
appreciable glucose in the urine of normal individuals while diabetics often have high urine glucose which
would give a strong positive ( black precipitate ) test with Nylander's reagent.

ANALYSIS

Barfoed’s Test

Barfoed's Test is a chemical test used for detecting the presence of monosaccharides. It is based on the
reduction of copper(II) acetate to copper(I) oxide (Cu2O), which forms a brick-red precipitate. Barfoed's
reagent consists of a 0.33 molar solution of neutral copper acetate in 1% acetic acid solution. The reagent
does not keep well and it is therefore advisable to make it up when it is actually required.
(Disaccharides may also react, but the reaction is much slower.) The aldehyde group of the
monosaccharide which normally forms a cyclic hemiacetal is oxidized to the carboxylate. A number of
other substances, including sodium chloride,[3] may interfere. The test is similar to the reaction of Fehling's
solution to aldehydes.
ANALYSIS

IODINE TEST

This test distinguishes polysaccharides and other carbohydrates. Polysaccharides will react
with iodine to form a blue, red, violet, or purple product which is a positive indicator of
the presence of a polysaccharide.
Iodine test can easily determine a starch. Starches form deeply colored blue-black complex with iodine. Starches
contain α-amylose, a helix saccharide polymer, and amylopectin. The iodine molecule slips inside of the amylose coil.
Iodine forms a large complex polysaccharide with the α-amylose helix, producing the blue-black color. Simpler
oligosaccharides and monosaccharides do not form this complex with iodine. Thus, the I2/KI test can be used to
distinguish starches from other carbohydrates.

ANALYSIS