Symbol of Chemicals

 Chloroform (CHCl₃)
 Hubl’s Reagent (I2 in HgCl2)
 Acetic Anhydride (C4H6O3)
 Concentrated Sulfuric Acid (H2SO4)
 Antimony Trichloride in Chloroform (SbCl3 in CHCl3)
 0.05% α-tochopherol in CHCl3
 N-Butanol (C₄H₁₀O)
 Concentrated Nitric Acid (HNO₃)
 Ammonium Molybdate ((NH4)2MoO4)

Schematic Diagram
Analysis of Lipids

l. Test for Unsaturation

- Prepare 4 test tubes for the following samples (olive and coconut oil, oleic and stearic
acid)
- Add 2 drops of the sample in separate test tubes
- Add 1 ml of Chloroform (CHCl₃) to the samples
- Add Hubl’s Reagent drop by drop then mixing the sample solution until the color of the
sample is the same as the color of the Hubl’s Reagant.
- Record the number of drops

II. Liebermann-Burchard or Acetic Anhydide Test

- Prepare 2 dry test tubes for the following samples (cholesterol and bile salt)
- Get a pinch of the test sample and put them separately
- Add 1ml Chloroform (CHCl₃) to dissolve the samples
- Add 5 drops of Acetic Anhydride (C4H6O3)
- Follow a drop of Concentrated Sulfuric Acid (H2SO4)
- Record Observations

III. Carr-Price Reaction

- Prepare 3 dry test tubes for the following samples (cod liver oil, coconut oil, olive oil)
- Put 2 drops of samples in separate test tubes
- Add 1ml Chloroform (CHCl₃) to the samples
- Prepare an Ice Bath and Let the solutions cool down
- Add 2 ml of Cold Saturated Chloroform Solution in SbCl 3 to the samples
 - Shake the solution
- Record observations and the subsequent fading.

IV. Modified Furter-Meyer Test

- Prepare 2 test tubes for the following samples (0.05% α-tocopherol in CHCl₃ and Vitamin
E)
- Add 1ml of the samples in the test tube
- Put 3.5ml of N-butyl alcohol
- Add in 0.5 Concentrated Nitric Acid (HNO₃)
- Mix the solution
- Place the test tubes in a water bath at 80° for 10 minutes
- Record Observations

V. Emulsification Test

- Prepare the following sample mixtures in separate test tubes

Test Tube 1: 1 ml coconut oil + 5 drops of 0.1% bile solution

Test Tube 2: 1 ml coconut oil + 5 drops H2O + a tiny crystal of cholesterol
Test Tube 3: 1 ml olive oil + 5 drops of 1% aqueous solution of lecithin

- Shake each sample very well and compare

- Examine a drop of each sample under a microscope right after shaking.
- Sketch your Observation
- Record Results

VI. Test for Phosphate

- A small amount of the lecithin will be incinerated in a porcelain crucible

- Let the residue cool down
- Add 5ml distilled water and mix
- Filter the mixture and place in a test tube
- To the 1ml Filtrate add 1ml of 5% Ammonium Molybdate ((NH4)2MoO4)
- Add in 3 drops of Concentrated Nitric Acid (HNO₃)
- Heat the solution to boil
- Record observations