BIOCHEM | GROUP 4

LIPIDS
- Fatty, waxy, or oily substances are known as lipids. They are soluble in organic
solvents but insoluble in polar solvents like water.

- Long-chained carboxylic acids called fatty acids produce the RCOO-onion and
H+ when they ionize. Soaps are created when fatty acids and NaOH combine.

- Soaps are the alkali metal salts of fatty acids that typically have between 10 and
18 carbon atoms. At one end of them is a long, non-polar hydrocarbon chain that
dissolves in oil, and at the other is a water soluble carboxylate ion.

- By introducing bromine together with CCI4, one may ascertain the degree of
unsaturation of lipids. Double bonds are formed between the free bromine and
the carbons. The decolorization of additional bromine indicates the existence of
unsaturation.

OBJECTIVES
● To prepare some physical and chemical properties of lipids.
● To prepare soap
● Compare the properties of soap with detergents.

SAFETY RULES INSIDE AND DURING THE LABORATORY EXPERIMENT

1. Wear appropriate PPE
- Laboratory Gown
- Protective Goggle
- Gloves
- Hairnet
2. No Horseplay
- No Eating, Drinking, and Applying Cosmetics inside the LAB.
- Never engage in horseplay or practical jokes. This is when injuries occur.
3. Act Responsibly Inside the Laboratory
- Inspect the laboratory apparatus before using it.
- Wash your hands before and after the experiment
- Maintain the noise inside the laboratory
4. Minimize All Chemical Exposure
- Avoid skin and eye contact with chemicals; Minimize all chemical exposures.
- Do not work with hazardous chemicals or processes when alone in the
laboratory.
MATERIALS
● Tubes (10pcs)
● Test Tube Rack
● Test Tube Holder
● Test Tube Brush
● Bunsen Burner
● Pipette and Aspirator
● Graduated Cylinder
● Droppers
● 250 ml Erlenmeyers flask
● Platform Balance
● 150 ml Beaker
● Watch Glass
● Water Bath
● Tripod
● Wire Gauge
● Stirring Rod
● 600 ml Beaker
● Funnel
● Filter Paper
● Spatula
● Evaporating Dish
● Hot Plate
● Molder for the Prepared Soap
● pH paper
● Large Test Tube
● Olive Oil
● Coconut Oil
● Cholesterol
● Oleic Acid
● Corn Oil
● Peanut Oil
● Stearic Water
● Linseed Oil
● Bromine in CCI4
● CHC13
● Acetic Anhydride
● Conc. H2SO4
● Powdered Detergent
 ● Distilled Water
● 0.5% Calcium Chloride
● 0.5% Magnesium Chloride
● NaOH Pellets

PROCEDURES
I. A. Test For Unsaturation
- Olive oil, oleic acid, stearic acid, peanut oil, corn oil, linseed oil and coconut oil in
seven separate test tubes. Add bromine in CC14, shaking it until a reddish brown
color persist.Take care to prevent cross contamination of samples. Record the
number of bromine drops consumed.

B. Liebermann-Burchard Test
1. Using a clean dry test tube, place a few grains of cholesterol in 3 ml anhydrous
chloroform.
2. Then add 10 to 20 drops of acetic anhydride and then carefully add 2 drops of
concentrated H2SO4.
3. Gently mix and observe the formation of a lilac color that develops gradually,
turning to blue, and then finally to an emerald green color.

II. Preparation of Soap

1. Weigh 10 g of coconut oil in a beaker.
2. Mix with 20 ml of 10% NaOH solution, cover with a watch glass, and heat the
mixture in a boiling water bath, stirring continuously in one direction until
saponification is complete. Test for saponification: shake one or two drops of the
mixture with 5 ml of warm water if the mixture produces. Plenty of suds, and no
oily globules, separate on standing, saponification is complete.
3. Pour the mixture into a mold and allow to set.
4. Rinse out the remaining soap from the container with 50 ml of distilled water and
set aside for Part III.

III. Comparison of the Properties of Soap and Detergents

1. Prepare a solution of 1 g of a powdered detergent in 50 ml of distilled water.
2. Test the pH of your soap solution and detergent solution by adding
phenolphthalein. Record your result in the report sheet.
3. Pour 10 ml of your soap solution in two labeled test tubes. Add 2 ml of 0.5%
CaCl2, solution in test tube 1; and 2 ml of 0.5% MgCl2 to the second tube.
Repeat procedure 3, but this time, Use Detergent solution instead of the soap.
4. Fill a test tube with about 10 ml of warm water. Add 1 ml of cottonseed oil,
stopper and shake. Observe. Now, add 1 ml of soap solution to the tube, stopper
and shake again. Observe again. Repeat procedure 4 using detergent instead of
soap or observed in a similar manner.

- Allow the setup to stand for 30-45 minutes or until the solvent reaches the 2 cm
mark from the upper edge. DO NOT MOVE THE SETUP AS THE PAPER IS
TAPED AND SUSPENDED IN THE JAR. Remove the paper from the jar and
mark the positions of the different dye components and the position of solvent
before it dries up completely.