Biochemistry – midterm period LIPIDS

The term “lipid” refers to both true fats and fat-like substances. These are biomolecules that are insoluble in
water but are soluble in organic solvents.

Another important function of lipids is the storage energy in the form of body fat. The brain and nerve tissues
are rich in certain lipids, a fact that indicates the importance of these compounds in the maintenance of life.

Five groups of lipids:

1. Fats and oils 2. Waxes 3. compound lipids
4. Sterols and steroids 5. Derived lipids General objectives:
At the end of the experiment, the student must be able to:
1. Familiarize himself/herself with the different classes of lipids; and
2. Identify each kind of lipid based on the chemical properties of its hydrolyzed products.
a. Spotting effect
All the lipids are greasy in nature. Therefore the test may be taken as group test for lipids.
PRINCIPLE: the oil does not wet the paper
b. Solubility

This test is based on the property of solubility of lipids in organic

solvents and insolubility in water.

PRINCIPLE: The oil will float on water because of lesser specific gravity.

c. Test for unsaturation (bromine water test)

PRINCIPLE: All neutral fats contain glycerides of some unsaturated fatty acids. These unsaturated fatty acids
become saturated by taking up iodine.

This test identifies the level of saturation and the number of bonds an oil, fat, or lipid has.

Visible result: Colorless

d. Acrolein test

It is used to detect the presence of glycerol or fat. When fat is treated strongly in the presence of a dehydrating
agent like potassium bisulphate (KHSO4), the glycerol portion of the molecule is dehydrated to form an
unsaturated aldehyde, acrolein that has a pungent irritating odor.

e. Extraction of brain lipids

Brain submerged in Ether

Extracts ceramides, sphingomyelins, choline glycerophospholipids, ethanolamine glycerophospholipids and

phosphatidylserines.

Not all lipids are soluble in Ether, therefore separates polar and non polar lipids
Molisch test

Principle: Molisch test is Specific for all carbohydrates – To differentiate between Proteins & Amino Acids (-
ve), and Carbohydrates (+ve).

This test is based on the fact that pentoses and hexoses are dehydrated by conc. H2SO4 acid to form furfural
or hydroxymethylfurfural, respectively.

1. Molisch test

Result: Reddish Violet ring at the interface - Any Carbohydrate No change - Non carbohydrate

2. Ninhydrin test

Principle: This test is due to a reaction between a amino group of free amino acid and ninhydrin. Ninhydrin is
a powerful oxidizing agent and its presence, amino acid undergo oxidative deamination liberating ammonia,
CO2, a corresponding aldehyde and reduced form of ninhydrin ( hydrindantin). The NH3 formed from a amino
group reacts with another molecule of ninhydrin and is reduced product
( hydrindatin) to give a blue substance diketohydrin ( Ruhemanns complex).

Positive compound: Phosphatidylserine

Visible result: blue violet or dark blue solution

3. Soda lime test

test for amino and amino group// test for acidity and basicity

Soda lime positive compound: Sphingomyelin and phosphatidylserine

Principle: deamination and acid-base reaction

Visible result:
- Red litmus -> Blue - Blue Litmus -> Blue

4. Ammonium molybdate test

It detects the presence of phosphate group Test positive compound: Phospholipids

Principle: Hydrolysis and double decomposition Visible results:

yellow precipitate (ammonium phosphomolybdate) insoluble in nitric acid

lecithin (yellow orange) Pigs brain (yellow)

5. Liebermann – burchard test

This test is used for the detection of cholesterol. The formation of a green or green-blue color after a few
minutes is positive.

Lieberman–Burchard is a reagent used in a colorimetric test to detect cholesterol, which gives a deep green
color.
 Chapter 4: Carbohydrates

Carbohydrates (which contain the elements carbon, hydrogen, and oxygen) form a class
of organccompounds that includes sugars, starches, and cellulose. Carbohydrates are
now defined
as polyhydroxyaldehydes or polyhydroxyketones, or substances that yields this
compounds on hydrolysis.

Occurrence and Functions of Carbohydrates

• Most of the matter in plants, except water, is carbohydrate material.

oCellulose: structural element

oStarch/glycogen: energy reservoir
▪small amount in human body
oPlant products are source of carbohydrates
▪average human diet contains 2/3 of carbohydrates
•Carbohydrates serve as the major source of energy and as a reserve
food material.
•Carbohydrates form part of the structural framework of DNA and RNA
molecules.
•Carbohydrates are involved in cell-cell interaction.

Classification of Carbohydrates

Carbohydrates are classified on the basis of molecular size as

monosaccharides, disaccharides, oligosaccharides, and
polysaccharides.

• Monosaccharide
o Contain single polyhydroxy aldehyde or ketone unit
o They can’t be broken down into simpler substances by hydrolysis (reaction with
water) reactions
o Contains 3-7 C atoms
o 5 and 6 carbon species are more common
o Water soluble white crystalline solids
• Oligosaccharides
o Contains ~2-10 monosaccharide units - covalently bonded to each other
o Disaccharides (contain 2 monosaccharide units) more common - crystalline water
soluble substances
o Table sugar (sucrose) and milk sugar (lactose)
 o - are common disaccharides
o Upon hydrolysis they produce monosaccharide
o In human body associated with proteins and lipids for structural and regulatory
functions
• Polysaccharides
o Contains many monosaccharide units covalently bonded
o Polymers: May contain 100s of 1000s of monosaccharide units

Reactions of Monosaccharides
•Five important reactions of monosaccharides: –Oxidation to acidic sugars

•Oxidation can yield three different types of acidic sugars depending on the type of
oxidizing agent used:

•Weak oxidizing agents such as Benedict’s solutions oxidize the aldehyde end to give an
aldonic acid.

•A reducing sugar is a carbohydrate that gives a positive test with Benedict’s solutions.

– Reduction to sugar alcohols

• Reduction to sugar alcohols: The carbonyl group in a monosaccharide (either an aldose
or a ketose) is reduced to a hydroxyl group using hydrogen as the reducing agent.

•Sorbitol - used as moisturizing agents in foods and cosmetics and as a sweetening agent in
chewing gum

–Glycoside formation

•Glycoside formation: are hemiacetals, they react with alcohols to form acetals:

•Monosaccharide acetals are called glycoside

–Phosphate ester formation

•Phosphate esters of various monosaccharides are stable and play important roles in the
metabolism of carbohydrates

–Amino sugar formation

•Amino sugars and their N-acetyl derivatives are important building blocks of
polysaccharides such as chitin

Disaccharides
 •Two monosaccharides can react to form disaccharide

•One monosaccharide act as a hemiacetal and other as alcohol

o Cellobiose is produced as an hydrolysis of the polysaccharide cellulose:

▪Cellobiose contains two b - D-glucose monosaccharide units linked through a b (1—4)

glycosidic linkage.

oMaltose is digested easily by humans because we have enzymes that can break a (1-4)
linkages but not b (1-4) linkages of cellobiose. Therefore cellobiose cannot be digested by
humans.

Lactose is made up of b-D-galactose unit and a b-D- glucose unit joined by a b(1-4)
glycosidic linkage

Lactose intolerance: a condition in which people enzyme lactase needed to

hydrolyze lactose to galactose and glucose.

Lactase hydrolyzes b(1-4) glycosidic linkages.

▪ Deficiency of lactase can be caused by a genetic defect, physiological decline

with age, or by injuries to intestinal
mucosa.
▪ When lactose is undigested it attracts water causing fullness, discomfort,
cramping, nausea, and diarrhea.
Bacterial fermentation of the lactose further along the intestinal
tract produces acid (lactic acid) and gas, adding to the discomfort.

o Any monosaccharide units bonded with glycosidic linkages

o Two types:
▪- Linear and branched, homo- and hetero- polysaccharides

Storage Polyssaccharides

o Starch
sucrose (table sugar): The most abundant of all disaccharides and found in
plants.

.
 General Characteristics of Polysaccharides

• Polymers
▪ A storage polysaccharide is a polysaccharide that is a storage form for
monosaccharides and is used as an energy source in cells.

o Amylopectin:

O
▪ Branched chain polymer - 80 - 85 % of the starch a (1 4) glycosidic bond for
O
straight chain and a (1 6) for branch

o Glycogen

▪ Humans and animals storage polysaccharide

▪ Contains only glucose units