INTRODUCTION

The various molecules found in all life forms are called organic compounds because they are
produced by organisms. Organic molecules are carbon based and range from small to enormous
in size. These large molecules are composed of simpler, smaller molecules known as monomers
that linked together into long chains or polymers. There are four major classes of important
biological organic compounds with specific functions.

OBJECTIVE

Explain the specific laboratory tests used to identify reducing sugars, starch, lipids and proteins

DETERMINING THE PRESENCE OF REDUCING SUGARS

APPARATUS/MATERIAL

Test tubes, test tube rack, marker pen, distilled water, glucose solution, sucrose solution, starch
solution, Benedicts reagent, label stickers

PROCEDURE

1. From the bottom of 4 clean test tubes, 1cm and 3 cm was marked. The top end of the test
tube was labelled 1, 2, 3, 4.
1. Each test tube was filled to the 1 cm mark with one of the following
Distilled water
Glucose solution
Sucrose solution
Starch solution
2. Each test tube was filled with 2cm Benedicts reagent and was swirled to mixed.
3. All the test tube was placed in a test tube rack for 3 minutes.
4. The colour change was observed. Data was recorded in a table.


RESULT/DATA


Test tube Observation (colour change) Conclusion
1


No changes Absent
2


Blue
-
Brick red precipitate
Present
3


No changes Absent

4


Blue
-
Greenish blue
Present


POST LAB QUESTIONS

1. Which test tube contains reducing sugars ?

Test tube 2 and test tube 4

2. Name the functional group present in carbohydrate molecule that can produce positive
reaction with Benedicts reaction.

Carbonyl C = O


3. If onion juice mixed with Benedicts reagent gives a mustard yellow colour, in what form is
sugar stored in onions ?

In a form of reducing sugar , Fructose



DETERMINING THE PRESENCE OF STARCH


APPARATUS/MATERIAL

Test tube, test tube rack, marker pen, microscope, distilled water, glucose solution, sucrose
solution, white tile, fresh potato, razor blade, clean microscope slides, coverslip, label stickers,
starch solution

PROCEDURE

2. From the bottom of 4 clean test tubes, 1cm was marked. The top end of the test tube was
labelled 1, 2, 3, 4.
3. Each test tube was filled to the 1 cm mark with one of the following :

Distilled water
Glucose solution
Sucrose solution
Starch solution

4. 3 drops of iodine solution was added to each test tube and swirled to mixed. The colour of
each test tube was recorded.
5. The contents of the test tube was discarded and rinsed properly. Excess water was shake out
from the test tubes. The next exercise used them.
6. A razor blade was used to cut a small, thin, slice of fresh potato. The potato slice should be
transparent.
7. The potato slice was placed onto a clean microscope slide and wet mount was prepared. The
potato slice was observed under low and high power.
8. The coverslip was removed and a few drops of iodine solution was added. The coverslip was
replaced, iodine to penetrate was waited for a few minute and the potato was observed again
under low and high power. The observation was draw.



RESULT AND DATA


Test tube Observation (colour changes) Conclusion
1


No changes Absent
2


No changes Absent
3


No changes Absent
4


Yellow
-
Blue black
Present


POST LAB QUESTIONS

1. Draw the potato slide observed under a microscope.













2. Does a blue black colour present in any of the carbohydrates solutions beside starch ?

No

3. Is iodine test suitable to distinguish starch from sucrose ? Explain why

Yes because iodine dissolved in an aqueous solution of potassium iodide, reacts with
the starch producing a purple black colour. The colour can be detected visually with
concentrations of iodine as low as 0.00002M at 20C. However the intensity of the colour
decreases with increasing temperature and with the presence of water-miscible, organic
solvents such as ethanol. Also the test cannot be done at very low pHs due to the hydrolysis
of the starch under these conditions
















DETERMINING THE PRESENCE OF PROTEINS


APPARATUS/MATERIAL

Test tube, test tube rack, small ruler, marker pen, distilled water, label stickers, honey solution,
corn oil, egg white solution, 10 NaOH solution, 1 CuS

PROCEDURE

1. 4 test tubes was marked at 2cm and 4cm measured from the bottom of the test tube. The top
end of test tube was labelled as 1, 2, 3, 4.
2. Each test tube was filled up to the 2cm mark with one of the following :

Distilled water (tube 1)
Honey solution (tube 2)
Corn oil (tube 3)
Egg white solution (tube 4)

3. 10 sodium hydroxide (NaoH) solution was added to each test tube up to the 4cm mark.
4. 5 drops of copper sulfate (CuS

) solution was added to each test tube and swirled to mix.

The tubes was examined for the appearance of any violet colour and result was recorded.
5. The contens of the test tubes was discareded, washed and rinsed with detergent properly.
Excess water was shake out from the test tube. Test tube was used for the next exercise.









RESULT/DATA


Test tube Observation (colour change) Conclusion
1


No change Absent
2


Form 2 layer
Upper layer : cloudly yellow
Bottom layer : yellow
Absent
3


Form 2 layer
Upper layer : cloudly yellow
Bottom layer : colourless
with blue precipitate
Absent
4


Violet Present


POST LAB QUESTIONS

1. Which test tube contains protein ?

Test tube 4

2. Name the chemical bond in protein that can react with biuret reagent producing a violet
colour.

Peptide bond







DETERMINING THE PRESENCE OF LIPIDS


APPARATUS/MATERIAL

Test tube, test tube rack, Sudan III dye, marker pen, small ruler, egg white solution, honey
solution, corn oil, distilled water, label stickers

PROCEDURE

1. 4 test tubes was measured at 2 cm from the bottom of the tube. The top end of test tube was
labelled as 1, 2, 3, 4.
2. Each test tube was filled up to the 2cm mark with one of the following :

Distilled water (tube 1)
Honey solution (tube 2)
Corn oil (tube 3)
Egg white solution (tube 4)

3. 10 drops of Sudan III dye was added to each test tube.
4. The tube was swirled to mix. The tube was examined for result. Result was recorded.
5. The contents of all tubes was discarded in the container designated for Sudan III disposal.
Sudan III was ensure not to pour it down the sink drain. Test tube was cleaned thoroughly.








RESULT/DATA


Test tube Observation (colour change) Conclusion
1


Light orange Absent
2


Light brown Absent
3


Light orange with fat droplet Present
4


Milky orange Absent


POST LAB QUESTIONS

1. Which test tube contain lipids ?

Test tube 3

2. Which test tube as a control ?

Test tube 1





DISCUSSION

From the experiment, we can know identify suitable experiment for testing organic compounds
such as carbohydrates, starch, lipids and proteins. Benedicts reagent is used to determining the
presence of reducing sugars that is carbohydrates. Benedict's reagent is used as a test for the
presence of reducing sugars. This includes all monosaccharides and many disaccharides,
including lactose and maltose. A positive test with Benedict's reagent is shown by a colour
change from clear blue to a brick-red precipitate. Glucose solution shows the solution have
reducing sugars by changing its colour blue to brick red precipitate. Iodine is used to determining
the presence of starch in four test tube. Test tube 1 act as control. Test tube 4 show the presence
of starch by changing its colour yellow to blue black. The Iodine test is used to test for the
presence of starch. Iodine solution iodine dissolved in an aqueous solution of potassium
iodide reacts with the starch producing a purple black colour. Next, Copper sulphate,
Cus

solution was used to determining the presence of proteins. Test tube 4 show changes its
colour to violet that is egg white yolk. Copper(II) sulfate is also used in the Biuret reagent to test
for proteins. Moreover, Sudan II was used to determining the presence of lipids. Sudan III
changes colour of corn oil to light orange with fat droplet in it. Sudan dyes have high affinity to
fats, therefore they are used to demonstrate triglycerides, lipids, and lipoproteins.















CONCLUSION

As a conclusion for this experiment, the specific laboratory test was used to identify reducing
sugars, starch, lipids and protein. After using some chemical subtances such as Cus

solution,
Sudan III and Iodine , result show that different kind of subtances have different type of
compounds. The test tube must be shaken to get better result and all of the test tube must be
labelled to avoid error in data. For reducing sugars, carbohydrate may be classified as
monosaccharides, disaccharides or polysaccharides. Some carbohydrates called reducing sugars
have an aldehyde functional group as part of their molecular structure which make them react
with Benedicts reagent when heated. For starch, Iodine reacts with starch resulting in a dark,
blue black colour that is a positive test for starch. The reaction occurs because starch is a coiled
polysaccharide made of repeated glucose molecules. For protein, protein molecules contain
multiple amino acid molecules linked together in long chain. The amino acid are linked by
peptide bonds that react with copper sulfate that is used in the Biuret test producing violet colour.
For lipids, lipis are nonpolar compounds composed of a glycerol molecule bonded to three fatty
acid molecules. Solubility of Sudan III dye can be used as a positive test for lipids.