Joycelyn Alag BS Chemistry 3 PROPERTIES of LIPIDS

Chem 161L -1:30-4:30 March 17, 2014

I. Objectives: 1. To give the operational definition lipids 2. To differentiate the following lipids in terms of their structural formulas and chemical test 3. To differentitate water-soluble and fat-soluble vitamins and give examples with their corresponding functions. 4. To determine the effect of heat on the activity of vitamin C 5. To identify the physiological functions of triglycerides, cholesterol, and phosphoglycerides II. Materials and Apparatus: 24 test tubes in rack, two 400-mL beakers, Bunsen Burner, test tube holder, wire gayze, thermometer, clay flame shield, stain test tube, litmus paper Cholesterol, lecithin, linseed oil, coconut oil, vitamin E., vitamin C, chloroform, distilled water, acetic anhydride, concentrated sulfuric acid, 1% iodine in chloroform, red litmus papers, concentrated nitric acid, ammonium molybdate solution, glycerol, juice with a high vitamin C content, 0.01M iodine solution, 1% starch solution, 20% sodium hydroxide III. Procedure & Observations: 1. Solubility 6 test tubes were prepared with 2-mL of water and dissolve in one microspatula each of : 1. Cholesterol, 2. Lecithin, 3. Vitamin C, 4. Vitamin E, 5. Linseed oil, 6 . coconut oil. The solvent was then replaced with chloroform. Cholesterol Lecithin Vitamin C Vitamin E Linseed oil Coconut oil Water x x x x x Chloroform x

2. Detection of Cholesterol a. Salkowski Test - one microspatula was dissolved in a three milliliter chloroform. Three milliliter of concentrated sulfuric acid was carefully introduce. Three colored layers were observed: red on top, then orange and yellow. b. Liebermann-Burchard Test - one microspatula of cholesterol was dissolved in three milliliter chloroform anf one milliliter of acetic anhydride. Concentrated sulfuric acid was added drop by drop in the mixture. Dark green solution was observed after five minutes. 3. Chemical composition of Lecithin a. Detection of Choline two milliliter of sodium hydroxide was added to one microspatula of lecithin, then heated. One red litmus paper was placed on the mouth during heating. Light yellow solution with solids, fishy smell, no change in red litmus paper were observed. b. Detection of Phosphorus- one microspatula of lecithin was completely dissolved in concentrated nitric acid and sulfuric acid, them warm maintained at 60 C. ten drops of ammonium molybdate was added and warm again. Dark fumes,foam formation, brown precipitate were observed. 4. Test for Unsaturation Five different test tubes containing: 1. One microspatula of cholesterol dissolved in one milliliter chloroform, 2. One microspatula of lecithin dissolved in one milliliter chlororform, 3.one milliliter of coconut oil, 4. One milliliter of linseed oil, 5. One milliliter of distilled water.five drops of 1% iodine in chloroform was added to each tube. T1- cholesterol Red solution T2- lecithin Orange solution T3- coco oil Red orange solution T4-linseed oil Yellow layer at the top of brown layer at the bottom T5- water Red violet at the top and light yellow at the bottom 5. Acrolein Test Three drops of coco oil with 0.2 g of potassium bisulfate was mixed and heated cautiously. Burnt grease smell was observed. .6.Heat Destruction of Vitamin C Three different test tubes containing five milliliter of juice having high vitamin C content and ten milliliter of water to each. In a water bath, tube 1 contents were boiled for 1-10 minutes, tube 2 contents boiled for 30 minutes while tube 3 contents were set aside. Both tubes were cooled in running water. Then, one milliliter of acetic acid and five drops of starch indicator was added to all tubes. Iodine solution was added to all until solution turns deep blue.

T1 T2 T3

Number of iodine drops 14 15 1

IV. Conclusion Solubility Not all vitamins are lipophilic vitamins. Vitamin C is hydrophilic and this is not a lipid. Cholesterol shows a positive result in Salkowkis Test and Libermanns Test. Detection of Cholesterol Salkowkis Test. When conc. Sulfuric acid is added to a chloroform solution of cholesterol, the chloroform layer shows a red to orange color. Liebermann-Burchard Test. Cholesterol produces a characteristic gree color when it is mixed with the Liebermann Burchard reagent. Chemical composition of Choline Detection of Choline. The released choline decomposes to ethylene glycol and trimethylamine. It has a characteristic odour Detection of P. Phosphate contained in lecithin is released during alkaline hydrolysis. In reaction with ammonium molybdate (VI), it creates ammonium phosphomolybdate, which colors the yellow solution. Test for Unsaturation. Fatty acids in animal fats are usuallu saturated, whereas those in vegetable oils are generally unsaturated. Halogen I will add across the double bonds and thus the decoloriation of an iodine will indicate the presence of unsaturated fatty acid. The more saturated, the paler the solution. Acrolein Test. When glycerol is heated with potassium bisulphate or conc, sulfuric acid, dehydration occurs and aldehyde acrolein formed which has a sharp irritating odor, burnt grease. Heat Destruction of Vitamin C. The solution that was exposed to heat at longer time need only less number of drops of iodine because Vitamin C was destroyed and only less of it remained on it. Thats why the solution that was not heated needs the highest numbers of drops of iodine.

Theoretical Background Lipids are a class of biological molecules that are insoluble in water and soluble in nonpolar solvents. Lipids are essential components of every living cell. Lipids are classified into: simple lipids - they are esters of fatty acids with various alcohol, complex lipids they contain (in additional to fatty acids and alcohols) additional groups as phosphates, carbohydrates and protein. In solubility, oil dissolves in the organic solvent chloroform but not in water because oil contains long hydrocarbon tails which are hydrophobic in nature. Fatty acids are nonpolar, it does not dissolve in water but dissolves in chloroform because it is non polar. Salkowkis Test is a test for cholesterol. Salkowski reagent is aqueous ferric chloride to which is added a slug of concentrated sulfuric acid. It is specific for indoles, like cholesterol. the highly acid reagent causes two indole molecules to join together at the 2-m and 2'-positions. The Fe(III) chelates the two N atoms from the resulting condensation compound to give a variety of hues of colors based on substituents on the indoles. Liebermann-Burchard Test is used in the estimation of blood cholesterol. LiebermannBurchard reagent is a mixture of acetic anhydride and sulfuric acid. The change in color may be gradual, initially pink, then blue-purple and finally deep green. Detection of Choline. .In a highly alkaline reaction, the hydrolysis of the ester bond takes place between choline and the phosphate residue of lecithin. Equation:

Detection of P. Alkaline hydrolysis releases fatty acids in the form of potassium salts (soaps), reduce the surface tension of water Equation:

Iodine test is used for distinguishing saturated and unsaturated fatty acid as well as between fats and oils. Equation:

Acrolein test responds to glycerol free or linked as an ester. Equation:

Heat Destruction of Vitamin C. The presence of Vitamin C can be detected with iodine using starch indicator