BUTUAN DOCTORS’ COLLEGE

Butuan City

Biochemistry Laboratory
Experiment Guide

Name: Macajelos, Krizly Marie C. Score:

Course, Year and Section: BSMT II - C Date: 08/29/2020
Instructor: Engr. Aida Pao Instructor’s Sign:
Group Number:

Experiment No. 1 Test for Carbohydrates

I. Objectives:
At the end of the period, the students should learn the characteristics and
classification of carbohydrates.

II. Theory:
Carbohydrates are generally considered as substances made up of carbon,
hydrogen and oxygen in which the proportion of hydrogen and oxygen is 2:1. It can be
defined also as aldehyde or ketone derivatives of polyhydric alcohols. Carbohydrates
are classified according to the number of saccharide groups contained in their
molecules. Like the monosaccharide- containing one saccharide group,
oligosaccharides- composed of more than one saccharide group and polysaccharides-
made of several saccharide group. The members of monosaccharide are trioses,
Tetroses, Pentoses, hexoses and Heptoses. In the oligosaccharides the members are
Disaccharides and Trisaccharides. For the polysaccharides, there are two groups: the
starch and the cellulose groups. Lower members are soluble in all proportions with
water and generally insoluble in 95% ethyl alcohol. The reason is that carbohydrates
are derived from polyhydric alcohol, so their molecules contains several hydroxyl (OH)
groups, thus they will not dissolve in 95% alcohol which is also saturated (OH) groups.
Some are soluble in cold water, like glycogen and inulin, but some like starch is
insoluble in cold water but is soluble in boiling water. Cellulose can be dissolve only in
special reagents. 2 Reactions of carbohydrates with strong acids will act on CHO’s
dehydrating them into furfural if a pentose. It is hydroxyl methyl furfural if hexose or
higher carbohydrates. This dehydration product then will act with the corresponding
reagent to form colored products of unknown nature. The production of the colored
product will sometimes serve as a specific test for some carbohydrates. Molisch’s
reagent is prepared by adding alpha-naphthol in ethyl alcohol.

III. Materials:
Fructose Glucose Lactose Sodium Chloride Ethyl Alcohol Starch
Sulfuric Acid Molisch’s Reagent Sucrose

IV. Apparatus:
 Test tube Test tube rack Stirring rod Graduated cylinder Dropper
Beaker

V. Procedure:
1. Solubility Test the solubility of fructose, lactose, sucrose and starch in the
solvent water, 10% sodium chloride solution and ethyl alcohol. Use only a pinch of the
solid to be tested in 2 ml solvent.

Solubility Water Sodium Chloride Alcohol

Fructose Slightly Insoluble Soluble Insoluble
Lactose Soluble Soluble Insoluble
Sucrose Soluble Slightly Insoluble Insoluble
Glucose Soluble Soluble Slightly Insoluble
Starch Insoluble Insoluble Insoluble

2. Dialysis Place 1 ml of carbohydrate solution in cellophane (ice candy wrapper)

and immersed in a beaker of considerable amount of distilled water. Set aside for 20
minutes. Transfer the dialysate into a test then slowly pour on the side of the inclined
test tube two drops of Molisch’s reagent and 3 ml of concentrated sulfuric acid.
Compare the intensity of color produced. Record your result in the data table.
Carbohydrate solution to be used:

Carbohydrates Solution Result with Molisch

0.1 M Glucose Solution Colorless, Green & Brown, Pink
0.1 M Sucrose Solution Colorless, Green & Light brown, Pink
0.1 M Starch Solution Colorless, Green & Dark Brown

3. Furfural Reaction of Carbohydrates Prepare four test tubes containing 1 ml of

0.2 M glucose, 1 ml of 0.2 M of sucrose, 1 ml of 0.2 M fructose and 1 ml of 0.05 M
starch, respectively. To each of the four solutions add two drops Molisch’s reagent. Mix
well. Slowly pour along the side of each tube 3 ml of concentrated sulfuric acid. Note the
color of the rings formed after sometime. The first tube serves as control. In some
cases, as in polysaccharides it may require a longer time to develop a result.

Carbohydrate Solution Result with Alpha Nophthol Solution

1 ml of 0.2 M Glucose Colorless, Green, Purple
1 ml of 0.2 M Sucrose Colorless, Fuchsia, Purple
1 ml of 0.2 M Starch Colorless, Fuchsia, Purple
1 ml of 0.2 M Fructose Colorless, Green, Purple
VI. Guide Questions:
1. Compare the relative sweetness of the sugar in this exercise.
- Fructose is the sweetest sugar, followed by Sucrose and Glucose.

2. Why does starch considered insoluble to cold water?

- Starch considered very low solubility in water it is because a polymer consisting
of many Glucose molecules joined by Glucosides bonds.

3. Name the three varieties of sugar.

- Glucose
- Fructose
- Galactose

4. Give the chemical formula of the following materials/ reagents.

- Glucose – C6H12O6
- Fructose – C2H2O6
- Sucrose – C12H22O11
- Starch – C6H10O5
- Sodium Chloride – NaCl
- Sulfuric Acid – H2SO4

VII. SUMMARY AND DETAILED CONCLUSIONS:

- Carbohydrates are generally considered as substances made up of carbon,
hydrogen and oxygen in which the proportion of hydrogen and oxygen is 2:1. It can be
defined also as aldehyde or ketone derivatives of polyhydric alcohols. Carbohydrates
are classified according to the number of saccharide groups contained in their
molecules. Like the monosaccharide- containing one saccharide group,
oligosaccharides- composed of more than one saccharide group and polysaccharides-
made of several saccharide group. The members of monosaccharide are trioses,
Tetroses, Pentoses, hexoses and Heptoses. In the oligosaccharides the members are
Disaccharides and Trisaccharides. For the polysaccharides, there are two groups: the
starch and the cellulose groups. The test includes Solubility of the solution to the Water,
Sodium Chloride and Alcohol. The second procedure is Carbohydrates solution with
Molisch Solution, and the last procedure is the Carbohydrate solution with the Alpha
Nophthol Reaction.