Qualitative Tests for Lipids Procedure Test for ester 10 drops of the eluates were placed in separate test tubes. 0.5 mL ethanol:1-butanol (3:1) was added followed by sequentially adding two drops of 2 M NH 2 OH HCL and 3 M NaOH and was mixed. The samples were allowed to stand for 5 minutes. Then 2 drops of 6 M HCl and 1 drop 5% FeCl 3 ● 6H 2 O in 0.1 M HCl were added and later were mixed. The samples with esters produced a burgundy color. Test for Gycerol (Acrolein test) A pinch amount of KHSO 4 was added to 10 drops of the eluate in a test tubes. The test tubes were then heated in a boiling water bath and the odor produced was noted. The odor was burn fat which indicates the presence of glycerol. Test for Glycerol (Kraut’s Test) 3 mL of Kraut’s reagent was added to 10 drops of the eluate in a test tube. It was later warmed and observed if there were any changes. Test for Cholesterol (Liebermann- Burchard Test) 10 drops eluate were placed in a test tube. 0.25 mL dichloromethane was added and 6 drops of acetic anhydride was added with 2 drops of concentrated H 2 SO 4 and were mixed. A greenish color was produced after a few minutes which indicates the presence of cholesterol. Test for Lipid Unsaturation with Br 2 10 drops eluate were placed in a test tube. 3 mL dicholoromethane was added and mixed. Under a fume hood, 5% Br 2 was added dropwise in dicholoromethane into the test tube. It was shake after each addition until a reddish brown color persisted. The number of drops of 5% Br 2 in dichloromethane was recorded.
Test for ester10 drops of the eluates were placed in separate
test tubes. 0.5 mL ethanol:1-butanol (3:1) was added followed by
sequentially adding two drops of 2 M NH2OH HCL and 3 M NaOH and was
mixed. The samples were allowed to stand for 5 minutes. Then 2
drops of 6 M HCl and 1 drop 5% FeCl3 6H2O in 0.1 M HCl were added
and later were mixed. The samples with esters produced a burgundy
color.
Test for Gycerol (Acrolein test)A pinch amount of KHSO4 was
added to 10 drops of the eluate in a test tubes. The test tubes
were then heated in a boiling water bath and the odor produced was
noted. The odor was burn fat which indicates the presence of
glycerol.
Test for Glycerol (Krauts Test)3 mL of Krauts reagent was added
to 10 drops of the eluate in a test tube. It was later warmed and
observed if there were any changes.
Test for Cholesterol (Liebermann- Burchard Test)10 drops eluate
were placed in a test tube. 0.25 mL dichloromethane was added and 6
drops of acetic anhydride was added with 2 drops of concentrated
H2SO4 and were mixed. A greenish color was produced after a few
minutes which indicates the presence of cholesterol.
Test for Lipid Unsaturation with Br210 drops eluate were placed
in a test tube. 3 mL dicholoromethane was added and mixed. Under a
fume hood, 5% Br2 was added dropwise in dicholoromethane into the
test tube. It was shake after each addition until a reddish brown
color persisted. The number of drops of 5% Br2 in dichloromethane
was recorded.
