MUSIC ACID TEST

Objective: To produce dicarboxylic acid from monosaccharides.

At the end of the practical, you should be able to:

1. Define mucic acid test and understand its reaction
2. Observe the crystals under the microscope
3. Understand the biochemical reaction and its application

Introduction
Monosaccharides are the simplest carbohydrates and are classified according to whether
they are aldehyde or ketone derivatives, as well as the number of atoms contained in the
molecule. Single hexoses, glucose and galactose require no digestion and can be absorbed
directly into the bloodstream. Hexoses contain six carbon atoms, and are found in foods, while
pentoses, ribose and deoxyribose contain five carbon atoms and are produced during the
metabolism of foodstuffs.
Three common sugars—glucose, galactose, and fructose, share the same molecular
formula: C6H12O6. Because of their six carbon atoms, each is a hexose. Although all three share
the same molecular formula, the arrangement of atoms differs in each case.
Galactose is not normally found in nature, but is mostly hydrolyzed from the disaccharide
lactose, which is found in milk, as part of a disaccharide made by glycosidic linkage to a glucose
molecule. The lactose disaccharide from milk is a major energy source for almost all animals,
including human. Although not very water-soluble, and less sweet than glucose, it forms part of
glycolipids and glycoproteins, which is found in many tissues. The body can change glucose to
galactose in order to enable the mammary glands to produce lactose.
Galactose, lactose and other carbohydrates can be oxidized by concentrated nitric acid to
give mucic acid which will form crystals if the solution is allowed to stand undisturbed
overnight. Mucic acid is almost insoluble in cold water. The nitric acid oxidizes galactose to an
isomer of tetrahydroxyadipic (tetrahydroxyhexanedioic) acid that crystallizes out from water
under the conditions of the test. The particular isomer resulting from galactose is called mucic
acid. Other sugars, such as glucose, also give similar acids, called by other common names, but
they are water-soluble under conditions of the test.
Lactose is a disaccharide (double sugar), galactosyl(1->4)glucose. The nitric acid first
catalyzes hydrolysis of lactose to glucose and galactose. Then the nitric acid oxidizes both of
those sugars to the tetrahydroxy acids. Mucic acid crystallizes out, which is a positive test. The
acid from glucose remains in solution. Sucrose is also a double sugar of glucose and fructose.
The nitric acid catalyzes hydrolysis to those two sugars, but neither gives mucic acid on
oxidation. The application of this test is the determination of blood group and detection of
galactose in urine.
Although the dicarboxylic acids do not occur in appreciable amounts as components of
animal or vegetal lipids, they are in general important metabolic products of fatty acids since
they originate from them by oxidation. Dicarboxylic acids are suitable substrates for preparation
of organic acids for the pharmaceutical and food industries. Furthermore, they are useful
materials for the preparation of fragrances, polyamides, adhesives, lubricants, and polyesters.

PROCEDURE

1. List down all the apparatus and materials needed for this practical
2. Construct a simplified version of the methodology in a flowchart form
3. Observe the result under the microscope

Apparatus
Test tubes
Water bath (boiling point)
Glass rod
Glass Slides
Microscope

Material
Concentrated Nitric Acid (HNO3)
Galactose
Glucose
Lactose
Unknown Sample

Method
Take about 50mg galactose, 50mg glucose, 50mg lactose and an unknown separately in test
tubes. Add 1mL distilled water and 1mL concentrated HNO3 to each tube. Heat the tubes in a
boiling water bath for about 1 hour. Add 5mL distilled water and let the tubes to stand and cool
slowly. Colourless needle like crystals will indicate the presence of galactose. Collect some of
the crystals using a glass rod and examine with a microscope.

Results & Discussion

1. Observe and describe the crystal formation.
2. Discuss the principle of the components and reaction of the test.
3. Describe the application of this test in daily life.
Conclusion
Conclude the results obtained from this experiment.

SAFETY PRECAUTIONS
Nitric Acid is corrosive, appropriate gloves, safety glasses and lab coat should be worn during
the experiment. It should be handled inside a fume hood. Avoid inhalation and any contact to the
skin.