EXPERIMENT 5: LIPID ISOLATION FROM BRAIN TISSUES

I. OBJECTIVES
a. To isolate lipids from brain tissue and determine which one is the cerebroside,
cholesterol, and the phospholipids
b. To test the presence of glycerol through acrolein’s test
c. To determine the unsaturated lipids by dropping Hanus reagent
d. To identify the presence of free phosphate in acidic solution by adding a molybdate to
the solution
e. To
f. To estimate the blood cholesterol through Lieberman-Burchard test
g. To detect the presence of Vitamin A by Carr-Price reacton
h. To determine tocopherols, Vitamin E, in food stuffs through Furter-Meyer test
i. To detect the carbohydrates present in lipids using Molisch Test
II. OPERATIONAL DEFINITION OF TERMS
a. Lipids: are marginally soluble (at best) in water but readily soluble in organic solvents,
such as chloroform or acetone.
b. Lecithin:
III. THEORIES:
a. Acrolein’s Test: The presence of glycerol grouping as in coconut oil can be detected by
the acrolein’s test wherein substances to be tested is heated with the dehydrating
agent. The unsaturated aldehyde, acrolein is produced. It is readily detected by its
characteristic acrid, an irritating odor.
b. Unsaturated lipids: undergo addition reaction with iodine. The relative unsaturation of
these substances can be…
c. The presence of free phosphate in acidic solution can be detected by adding a molybdate
to the solution. Equation illustrates the pertinent reaction between phosphate and
ammonium molybdate solution in the presence of nitric acid (HNO3).
HPO42–(aq) + 12MoO42–(aq) + 3 NH4+(aq) + 23 H3O+(aq) (NH4)3[P(Mo3O10)4] (yellow,s) + 35 H2O(l)

After a few minutes, the yellow ammonium molybdo-phosphate precipitates from the
reaction mixture. When lipids containing phosphate groups in their structures are added to
a strong acid solution such as the solution used here, the lipid hydrolyses, producing free
phosphate. The free phosphate then reacts as in Equation, forming a yellow precipitate

d. Emulsification Test:
e. Lieberman-Burchard or Acetic Anhydride Test: Cholesterol gives characteristics colored
condensation products with acetic anhydride and conc. Sulfuric acid following LB
reaction (deep blue, green solution)
f.
g. Carr-Price reaction-
h. Modified Furter-Meyer Test
i. Molisch Test
j.
IV. DISCUSSION OF RESULTS
V. CONCLUSIONS
a. How emulsifiers are able to mix polar and non-polar substances together?
i. Emulsifiers work by forming physical barriers that keep droplets from
coalescing. A type of surfactant, emulsifiers contain both a hydrophilic (water-
loving, or polar) head group and a hydrophobic (oil-loving, or non polar) tail.
Therefore, emulsifiers are attracted to both polar and nonpolar compounds.
When oil suspended in an aqueous phase, emulsifiers surrounds the oil droplet
with their nonpolar tails and extending into the oil, and their polar head groups
facing water and vice versa. In this way, emulsifiers lower the interfacial tension
between the oil and water phases, stabilizing the droplets and preventing thm
from merging (Cassiday, 2018).
b. Lecithin structure with it being an emulsifying agent
i. Lecithin is an emulsifier made up of about five smaller molecules. It has a
backbone of glycerol that bonds up to three other molecules. Two of the
bonded molecules are fatty acids—these are hydrophobic. They give lecithin a
structure familiar to fats, or lipids. The third substance attached to glycerol is
phosphoric acid that has an amino alcohol attached called choline. The
phosphate/amino alcohol end of lecithin is hydrophilic. Thus, it has both polar
and nonpolar molecule which emulsify with other molecules.
c. Lecithins in food products:
i. Vegetables and legumes like brussels sprouts, cabbage, cauliflower, beans, soy,
and most leafy veggies
ii. Daily products such as eggs, cheese, yoghurt, milk
d. Tocopherols contribute to the keeping qualities of fats
i. They act as an antioxidants, that is, a good reducing agent- so it reacts with
oxidizing agents before it can attack other biolecule.
i. Vitamin E (α-tocopherol) 1, is an efficient lipid soluble antioxidant that
functions as a ‘chain breaker’ during lipid peroxidation in cell membranes and
various lipid particles including low-density lipoprotein (LDL). It functions to
intercept lipid peroxyl radicals (LOO˙) and to terminate the lipid peroxidation
chain reactions

ii. LOO˙ + α-tocopherol–OH → LOOH + α-tocopherol–O˙

iv. The resultant tocopheroxyl radical is relatively stable and in normal
circumstances, insufficiently reactive to initiate lipid peroxidation itself, which
is an essential criterion of a good antioxidant.45–47 It should be noted that,
vitamin E exerts antioxidant effects by scavenging lipid peroxyl radicals in
vivo as well as in vitro systems. However, vitamin E is not an efficient
scavenger of ˙OH and alkoxyl radicals (˙OR) in vivo.48