LIPIDS

Isolation of lipids from brain tissue

Objectives
- To isolate cerebroside, cholesterol, and phospholipids from cow brain tissues
- To hypothesize the identity of the isolated precipitates
Definition of terms
Lipids

-a group of naturally occurring substances characterized by their insolubility in

water and their solubility in nonpolar organic solvents such as:

Chloroform
Ether
Hot Alcohol and;
Benzene
Fatty acids

-Molecules that are long chains of lipid-carboxylic acid found in fats and oils and in
cell membranes as a component of phospholipids and glycolipids.

Glycerol

-a three-carbon compound that contains three hydroxyl groups, one bound to each
carbon
Phospholipids

-Composed of a phosphate group, two alcohols, and one or two fatty acids

Cerebrosides

-a glycolipid that contains sphingosine and a fatty acid in addition to the sugar
moiety

Cholesterol

-a steroid that occurs in cell membranes; the precursor of other steroids

Phosphoglycerides

- a class of phospholipids, including lecithin and cephalin, consisting of a glycerol

backbone, two fatty acids, and a phosphorylated alcohol
Theory behind the experiment
Lipids

Classes of lipids

Principal lipids of the brain

Structure

Solubility

Extraction
Lipids
- Soluble in nonpolar organic solvents
- Insoluble in water
- Constitute the principal structural components of living cells (with proteins &
carbohydrates)
- Monomers: fatty acids and glycerol
Simple lipids - esters of fatty acids
- fats - esters of fatty acids and glycerol (mono,di, triglycerides)

- waxes - esters of long chain fatty acids and long chain alcohols
Compound lipids - esters of fatty acids and alcohol plus other groups

- phosphate, carbohydrate and nitrogeneous components

- Phospholipids - composed of a phosphate group, two alcohols, and one or two fatty acids
- Glycolipids - cerebrocides
- Proteins - lipoproteins and proteolipids
Derived lipids - hydrocarbon rings and a long hydrocarbon side
chain
-derivatives obtained by hydrolysis of simple and compound lipids

- fatty acids

-steroids - cholesterol
Principal lipids of the brain
- Cholesterol, cerebrosides, phosphoinosides, phosphosphingosides, and
phosphoglycerides
Structure, Solubility, and Extraction
Amph
General structure of a phosphoglyceride ipath
ic
Glycerol
part
Not soluble in Acetone
Soluble in ETHER, CHLOROFORM, BENZENE, HOT EtOH, & Carbon disulfide
GLYCOLIPIDS

de
Cerebrosi

ALMOST insoluble in ether

MORE soluble in acetone than phosphoglycerides.
CHOLESTEROL

Very high concentration in nervous tissues

Soluble in ETHER, CHLOROFORM, BENZENE, and HOT ALCOHOL

Discussion of results
Precipitate 1 - yellow(difficult to isolate)

Precipitate 2 - brownish yellow

Precipitate 3 - pale green

Discussion of Results
- abundant in the brain and nervous tissues
- able to extract minute amount
- difficulty in extracting Precipitate 1
- identity of of each lipid > compared and contrasted solubility in the solvents
and analyzed the procedure
Discussion of results
Cerebrosides: soluble in acetone & hot alcohol, insoluble in ether

Phospholipids: soluble in ether & hot alcohol, insoluble in acetone

Cholesterol: soluble in ether & hot alcohol

Conclusion
-Acetone, ether, and hot alcohol are used to isolate these lipids ( cerebroside,
phospholipid and cholesterol)

-We determined that Precipitate 1 is Cerebroside, They are soluble in acetone and
hot alcohol and insoluble in ether.

-Precipitate 2 is Phospholipid, They are soluble in ether and hot alcohol and
insoluble in acetone

-Cholesterol is the Precipitate 3, It is soluble in ether and hot alcohol

References
Biochemistry Laboratory Manual (2013 Edition)

Biochemistry by Campbell & Farrell