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y To

be able to cite and explain the different properties of lipids y To able to explain the process of saponification. y To able to explain each test to characterize lipids


a. Solubility Test

Cottonseed oil

Dist. Water


It is insoluble because water is Polar thus it wont react with Cottonseed oil which is Nonpolar Even though Ethyl Alcohol is Polar yet it is soluble in Cottonseed is because it is weak polar solvent.

Ethyl Alcohol


Cottonseed Oil



Even though Ethyl is Polar yet it is soluble in Cottonseed is because it is a weak polar solvent Chloroform is also non polar and makes Cottonseed oil soluble in it. Cottonseed oil is insoluble in Hydrcholric acid beacause it can easily be separated when mixed with other solutions that have dipoles.



5% HCl


5% NaOH


NaOH is a strong polar thus making Cottonseed oil insoluble in it.

Cottonseed Oil h Ethyl Alcohol


Ethyl Alcohol did not evaporate and it is insoluble in Cottonseed Oil Ether evaporated and it is soluble in Cottonseed Oil because is a non polar.

Cottonseed Oil h Ether

The solubility test is used to determine if the substance (cottonseed oil) is soluble in the given solvents such as benzene, ether, ethyl alcohol, 5% HCl, 5% NaOH and distilled water. The cottonseed oil is a polyunsaturated fatty acid and it is made up of different kinds of acid, such as: Linoleic acid, Oleic acid, Stearic acid, Palmitic acid, Myristic acid. 54% of the cottonseed oil is made up of linoleic acid which makes the oil a lipid.

b. Test for Unsaturation

Oleic Acid Palmitic Acid

Reagents Added
Bromine in CCl4 Chloroform Hanus Iodine

Formed a colorless solution. The color changed into pink after 15 seconds

Oleic Acid

Chloroform Hanus Iodine

The color changed into pink after 15 seconds

Cottonseed oil

Chloroform Hanus Iodine

The color changed into light pink after 14 seconds

c. Acrolein Test
Test Solution
Glycerol Cotton Seed Oil

Reagents Added
KHSO4 (s) KHSO4 (s)

Pungent Pungent Smell

The acrolein test is used to detect glycerol as indicated by the formation of propenal or acrolein that has the pungent odor.The glycerol was heated using the dehydrating agent, the KHSO4 to form the acrolein as an unsaturated aldehyde

d. Rancidity Test
Test Solution Fresh Coconut Oil Reagents Added Phenolphthalein Methyl Orange Litmus Paper Blue Red Result No Reaction No Reaction


Rancid coconut Oil

Phenolphthalein Methyl Orange Litmus Paper Blue Red

No Reaction No Reaction

- Neutral

e. Saponification & Properties of Soap

Name of Test E. Saponification F. Properties of Soap Salting-out Soap solution NaCl A solid soap was formed and there was formation of suds when it was mixed with water Formation of insoluble white precipitate. Both had a formation of widely spread insoluble precipitate an insoluble liquid layer was formed (cloudy) Test Solution Coconut oil Reagents Added 10% KOH Results Soap produced

Formation of Fatty Soap solution Acids Insoluble Soaps Soap solution

10 % hydrochloric acid 5% CaCl2 5% MgCl2


5% CaCl2 5% MgCl2

The alkaline hydrolysis of fats is called saponification; a process that yields to glycerol and the salt of the fatty acid.1This involves the reaction of a strong base with the triglyceride. The base that was used for the saponification process in the experiment is potassium hydroxide. The sodium salt of the fatty acid is obtained instead of the fatty acid as one of its products. Even if there is a formation of a fatty acid during the hydrolysis, the KOH will neutralize it to form the soap. The soap molecule is composed of a hydrophilic and a hydrophobic end. Its long nonpolar hydrocarbon chain is the hydrophobic end (water-hating) while the other end is the highly polar carboxylate salt which is hydrophilic (water-loving). In order to put the soap out of the solution, NaCl was dissolved in the solution. It is used to purify the soap. There were still other by-products that were formed during the process of saponification and these by-products are soluble in saltwater. When these by-products are already dissolved in the saltwater, the soap comes out of the solution. The formation of a white mass in the solution is the soap. There is a need to make sure that there is no more NaCl that is being dissolved in the soap solution to make sure that the by-products are already separated from the soap itself.

g. Liebermann-Buchard Test LiebermannTest Solutions Cholesterol Reagents Added chloroform acetic anhydride conc. sulfuric acid Results Initially blue then turned blue-green

There is a presence of sterol/unsaturated steroids In cholesterol. The Liebermann- Burchard is used to detect cholesterol and it produces a deep green color. The change is gradual because when the reagents were added, the color became pink then changed to blue-purple and finally to deep green. A water molecule is removed from the cholesterol molecule when it is reacted to conc. sulfuric acid. After the removal of the water molecule, it is now oxidized to form 3,5-cholestadiene. The product is then converted to a polymer containing a chromospheres which results in the green color.

h. Emulsifying Action
Test Solutions 2% albumin Reagents Added Cholesterol Cholesterol h Lecithin Result Turbidity and emulsion Turbidity and Emulsion

Emulsion is the mixture of two or more liquids in which one is present as droplets, of microscopic or ultramicroscopic size, distributed throughout the other. Emulsions are formed from the component liquids either spontaneously or, more often, by mechanical means, such as agitation, provided that the liquids that are mixed have no (or a very limited) mutual solubility. Stable emulsions can be destroyed by inactivating or destroying the emulsifying agente.g., by adding appropriate third substances or also by freezing or heating.


Why are fatty acids insoluble in water? Fatty acids are insoluble in water because they are composed mostly of hydrocarbon chains which are insoluble in water. The only one that is soluble in water is the carboxyl group attached to it. The longer the chain of the fatty acid is, the more it gets insoluble in water.

2. Explain why the cis-form is the predominant configuration of unsaturated fatty acids? Unsaturated fats tend to have a predominant configuration of cisform instead of trans since most unsaturated fats are liquid at room temperature, and the said configuration causes the unsaturated acid to be in liquid state.

The double bond is, in the normal cis configuration, asymmetric and so forces a kink bend into the carbon chain. As a result the unsaturated fatty acids are unable to pack so closely together, or crystallize as readily as straight-chain saturated fatty acids.

3. Why is the acrolein test a general test for fats?

It is because when a fat is heated strongly in the presence of a dehydrating agent such as KHSO4, the glycerol portion of the molecule is dehydrated to form the unsaturated aldehyde, acrolein (CH2=CH-CHO), which has the peculiar odor of burnt grease. A sample is heated with potassium bisulfate, and acrolein is released if the test is positive. That's why Acrolein test is a test for the presence of glycerin or fats.

4. What type of rancidity occurs in vegetable shortenings? How can it be prevented?

The type of rancidity which occurs in vegetable shortenings is autoxidation which is oxidation requiring only oxygen. It is a radical-chain process that converts an R-H group to R-O-O-H group, the hydroperoxide. The hydroperoxides are unstable and, under biological conditions, degrade to short-chain aldehydes and carboxylic acids with unpleasant rancid smells.

Antioxidants are often added to fatcontaining foods in order to retard the development of rancidity due to oxidation. Natural antioxidants include flavonoids, polyphenols, ascorbic acid and tocopherols. Synthetic antioxidants include butylated hydroxyanisole, butylated hydroxytoluene and ethoxyquin. Additionally, rancidification can be decreased, but not completely eliminated, by storing oils in a place with little exposure to oxygen or free radicals, low temperature, and away from light. Light and heat warp the structure of fat, and greatly accelerate its reaction with oxygen.

5. Explain the cleansing action of detergents.

6. Write the structure of the parent compound of cholesterol.

7. Explain the cooperative solvent effect of lecithin and albumin. y Lecithin is an emulsifier that has a variety of ingredients. One of its active components, the lipids consists of long chain polar lipids. Its emulsifying properties results from the combination of a hydrophilic and a hydrophobic group which helps in the dispersion of oil into aqueous solution or vice versa, creating the emulsion. Albumins are water-soluble proteins with massive polar molecules. The two having the same effect because both of them have polar molecules makes emulsion easier to happen.


Lipids are fat-like substances thus, they constitute a large heterogenous group of unrelated physiological and chemical substances classified together which also observes the rule of thumb like disolves like. The carboxyl group (-COOH), a polar end of the fatty acid contributes to the polarity of the lipids. Lipids are nonpolar because of the hydrocarbon chains, thus making them insoluble in polar solvents like water. The saturation and unsaturation can also be distinguished through the bonds in their hydrocarbon chains, which may be classified as saturated, monosaturated, disaturated and polysaturated.

Rancidification is also associated to lipids, Rancidification is the chemical decomposition of fats, oils and other lipids. Rancidity is the state of being rancid, having a rancid scent . It has two types, one is Hydrolytic rancidity which occurs when water breaks larger compounds into smaller ones. And the other is Oxidatile rancidity which the double bond of an unsaturated fatty avid reacts chemically with oxygen to result to two or more short molecules. Saponification is a process that yields glycerol and the salt of the fatty acid, in short the alkaline hydrolysis of fats. This involves the reaction of a base which is potassium hydroxide.