You are on page 1of 3

NINHYDRIN TEST ~Dissolve residue B in 2.0 ml of water. Add 1 ml of 0.1%ninhydrin reagent.

Mix thoroughly and heat in boiling water bath for two minutes. ~Note the color of the resulting solution. a. b. c. d. e. Pupose: test for amines including alpha amino group. Principle involved in the test: oxidative decarboxilation, deamination and condensation. Observations: blue-violet solution. Compound responsible for the visible result: blue-violet complex. Sample equation using a lipid sample that gave a positive result.

f.

Structural formulas of other lipids that will be tested positive in test.

glycine

Phosphatidyl-Serine alanine

Leucine

SODA LIME TEST ~Grind a pinch amount of soda lime and residue B. Transfer the mixture in a dry test tube and heat gently over a small flame. Test vapor by placing moistened litmus paper at the tip of the stirring rod. ~note the change in color of the stirring rods. a. b. c. d. e. f. Pupose: test for amino and imino group. Principle involved in the test: deamination. Reagent: NaOH and CaO Observations: R to B (lithmus paper) Compound responsible for the visible result: hydroxyl group (OH)- basic Sample equation using a lipid sample that gave a positive result:

Soda lime removes the amino group from an amino acid. Ammonia, the gas evolved during heating reacts with water in the moistened lithmus paper to form ammonium hydroxide (responsible for R to B change)

NH3 + H2O
Possible identity of residuge B

NH4 + + OH-

From the above observations, we can say that the following lipids may be in the residue: Sphingolipids, phosphatydylserine or any lipid with amino group.

g. Structural formulas of other lipids that will be tested positive in test.

sulfatides

LEIBERMAN BURCHARD TEST ~Prepare two clean and dry test tubes. In first tube, dissolve residue C in 1 ml methylene chloride. In second test tube, place 1ml vegetable oil. Add 5 drops of acetic anhydride followed by three drops of conc. sulfuric acid into both tubes. Mix thoroughly. ~Compare resulting intensity and color of solution. a. b. c. d. Pupose: Unsaturated steroids (cholesterol) Principle involved in the test: Oxidation Reagents: Acetic anhydride and sulphuric acid Observations: Residue C: emerald green Vegetable oil: blue green + VR: emerald green e. Compound responsible for the visible result: steroid nucleus f. Sample equation using a lipid sample that gave a positive result. Possible identity of residue C: unsaturated steroid cholesterol

steroid cholesterol
g. Structural formulas of other lipids that will be tested positive in test.

You might also like