Lipids

Def.: → Heterogenous water insoluble compounds, such as fats, oil, steroids, waxes and other related compounds.
Used for:
1) Production of fat soluble vitamins, steroids and bile acids 4) Cell membrane composition
2) Palatable food 5) Energy production
3) Thermal insulator

Classification of lipids

Compound lipids Simple lipids Derived lipids

Fat and oils (Neutral fats) Waxes

Fatty acids with glycerol (sugar alcohol) Fatty acids with long carbon chain alcohols

Simple lipids:
 Esterification of fatty acids with alcohols to form either Neutral fats (short chain alcohols) or waxes (long chain alcohols)
 Neutral fats are also called Triglycerides or Tri-acylglycerol and include:
a- Fats b-Oils
Solid at room temperature Liquid at room temperature
Contain saturated fatty acids Contain unsaturated fatty acids

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 Glycerol
1) Its trihydric alcohol . 4) Enters in all cosmetic preparations.
2) Soluble in water and insoluble in fat solvents. 5) Used in treatment of glaucoma.
3) When heated with H2SO4 gives acrolein.

Classified in to: Fatty Acids (F.As)

1) According to number of atoms 2) According to the degree of saturation

a) Odd numbered fatty b) Even numbered fatty a) Saturated Fatty Acid b) Unsaturated Fatty Acid
acid propionic acid acid Palmatic acid Short chain Mono unsaturated
valeric acid stearic acid Long chain Poly unsaturated

a) Saturated Fatty Acids

 They contain no double bonds
 They are classified into short chain and long chain fatty acid

A- Short chain fatty acids B- Long chain fatty acids

Contain 2-6 C atoms Contain 7-10 C atoms  Contain more than 10 C atoms
 volatile  Non-volatile  Non-volatile, solid at room temperature
 liquid at room temperature  solid at room  Soluble in fat solvents
 EX: temperature  EX:
 Acetic acid  Soluble in fat solvents  Palmatic acid (16C)
 Propionic acid  Stearic acid (18C)
 Butyric acid  Lignoceric acid (24C)
 Valeric acid

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 b) Unsaturated Fatty Acids
 They contain double bonds and classified into:
Monounsaturated F.As Polyunsaturated F.As
 They are non essential fatty acids  They are essential fatty acids → due to inability of the body
 They have one double bond & produced by several tissues to form more than one double bond
as liver  They contain more than one double bond & present in
 Ex. vegetable oil (Corn, Olive Oil)
 Palmitoleic acid (16:1 ∆9 ).  Ex.
 Oleic acid (18:1 ∆9 ). Linoleic acid (18 : 2 ∆𝟗 ,𝟏𝟐 )
Linolenic acid (18 : 3 ∆𝟗 ,𝟏𝟐 ,𝟏𝟓 )
Arachidonic acid (20 : 4 ∆𝟓 ,𝟖 ,𝟏𝟏 ,𝟏𝟒 )
Eicosapentanoic acid (20 : 5 ∆𝟓 ,𝟖 ,𝟏𝟏 ,𝟏𝟒 ,𝟏𝟕 )

 Importance of essential fatty acids:

1) Essential for growth and fertility.
2) Act as lipotropic factors (Prevent fatty liver).
3) Enter in the formation of phospholipids in cell
membrane.
4) Prevent atherosclerosis by forming esters with
cholesterol.
5) Precursor of Eicosanoids.

Note:
Timnodonoic acid is unsaturated fatty acid contain 5 double bonds present in cod liver oil.

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W3 and w6 fatty acids:
Omega 3 Omega 6
 Fish, olive, garlic  Vegetable oil, dairy, eggs, fast food, chicken and beef
 Reduction of inflammation  Decrease risk factor of diabetes
 Increase vision adequacy  Esterification of cholesterol

 Improve mental function  Lower risk of heart diseases

 First unsaturated bond at omega carbon number 3  First unsaturated bond at omega carbon number 6
 Linolenic and eicosapantoic  Linoleic and arachidonic

Eicosanoids

• They are a group of hormone-like compounds produced by many cells in the body.

• They derived from polyunsaturated fatty acids as arachidonic acids that contain 20 carbons (eicosa).
• The eicosanoids including prostaglandins, thromboxanes and leukotrienes.
• They control cellular function in response to injury or inflammation.

Importance of eicosanoids:
1. Eicosanoids participate in the inflammatory response that occurs after infection or injury.
2. It controls bleeding through the formation of blood clots.
3. In inflammation can produce symptoms as pain, swelling & fever.
4. Eicosanoids also regulate smooth muscle contraction (particularly in the intestine and uterus).

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5. They increase water and sodium excretion by the kidney and are involved in regulating blood pressure
6. They are also involved in regulating bronchoconstriction and bronchodilation

Formation of eicosanoids:
• The most abundant and the most common precursor of the eicosanoids is arachidonic acid ( a polyunsaturated fatty acid with
20 carbons and 4 double bonds )

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 Properties of neutral fats
I) Physical properties

1. Freshly prepared fats and oils are odorless, colorless, and tasteless.
2. Melting point depends on

a) the presence and absence of double bonds

b) the length of fatty acids.

3. Rancidity: → unpleasant odor & taste resulted from the effect of moisture, light, oxygen, temperature & lipase enzyme produced
by some bacteria
Types of rancidity:
a) Oxidative rancidity: Oxidation of fatty acids by oxygen, moisture and temperature to form aldhydes , ketones and peroxides.
b) Hydrolytic rancidity: Oxidation of fatty acids by bacterial lipase and moisture, temperature to form volatile fatty acids (bad odor).

Effect of rancidity:
1. Unpleasant odor and taste of fat. 2. The products of rancidity are toxic.
3. Destruction of fat-soluble vitamins (A, D, E, and K). 4. Irritation of gastric and intestinal mucosa.

Detection of rancidity: Rancid fats and oils could be detected by their changes in odor, taste and increase in iodine number.

Prevention of rancidity:
1. keeping the food in cold place (refrigerator) and avoid light, moisture.
2. Using antioxidants.

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 II- Chemical properties

1. Oxidation:

• Exposure of fat and oil to oxygen leads to oxidation of double bond, which present in unsaturated fatty acids.
• This resulted in production of aldehydes , ketones, peroxides and volatile fatty acids

2. Hydrogenation:

• Hydrogen is usually added to double bond present in unsaturated fatty acids in presence of Nickel forming corresponding
saturated fatty acids
• Vegetable oils when hydrogenated yield solid products (margarine). These are now widely used as substitutes to natural
fat.
• Margarine contains neither fat soluble vitamins nor essential fatty acids

3. Hydrolysis:

• This means breakdown of lipids by addition water in the presence of lipase enzyme, acids or alkali.
• Acid and enzymatic hydrolysis of neutral fat (triglycerides) produces glycerol and three fatty acids.

4. Addition reactions (halogenations):

• Halogens like iodine can be added at the double bonds of unsaturated fatty acids in triglycerides and give halogenated
saturated derivatives.

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 II- Compound lipids (Conjugated lipids)

• They contain beside alcohol and fatty acids other substances or prosthetic group

A- Phospholipids
 They are compound lipids containing phosphorus and they include:

I. Glycerophospholipids:

1- Phosphatidic acid. 2- Cardiolipin. 3- Plasmalogen

4- Lecithin. 5-Cephaline (phosphtidyl ethanolamine, serine or inositol)

1. Phosphatidic acid: 2. Cardiolipin: (diphosphatidyl glycerol )

act as an intermediate compound in synthesis of
triacylglycerols & phosphoglycerol .
Not found in a great quantity in tissues.
On hydrolysis yield glycerol, fatty acids and phosphoric acid.
It is an important phospholipid of mitochondrial membrane.
In which two phosphatidic acids are joined by a molecule of
glycerol.
These phospholipids are particularly rich in the polyunsaturated
fatty acids especially linoleic acid.

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3. Lecithin (Phosphatidyl choline): 4. Cephalin (Phosphatidyl ethanolamine, Serine or Inositol)
Formed from glycerol, two fatty acids , phosphoric acid and base.
 Formed from glycerol, two fatty acids, phosphoric acid & The base in is ethanolamine, serine, Inositol.
choline
 The first fatty acid is saturated fatty acid, while the
second one is
unsaturated fatty acid.
 lecithine present in egg yolk, liver, brain and plasma.

It can be hydrolyzed by lecithinase enzyme present in snake

venom causing removal of the unsaturated fatty acid
producing lysolecithin which causes haemolysis of R.B.Cs.

Dipalmitoyl lecithin is a very effective surface-active agent

and a
major constituent of the surfactant preventing collapse
Its absence from the lungs of infant causes
respiratory distress syndrome.

Phosphatidyl ethanolamine Present in nervous tissues and

activating coagulation factors.

Phosphatidyl Inositol (lipositol) in cell membrane of different

organs, has a role in cell signaling and act as 2nd messenger.
Phosphatidyl serine in cell membranes, has a role in cell
signaling.
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5- Plasmalogen:
These compounds constitute as much as 10% of the phospholipids of brain and muscle.
Structurally, the plasmalogen resemble phosphatidyl ethanolamine
but an ether link on the carbon-1 instead of the ester link.

II- Sphingophospholipids:
Sphingomyelin:

Found in large quantities in brain and nerve tissue

Sphingolipids do not have a glycerol backbone, they are formed from sphingosine alcohol (18 C) derived from serine & palmitic
acid.

Ceramides formed from sphingosine by attaching a fatty acid.

sphingomyelins are then formed by attaching choline (base) on ceramide.

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 B- Glycolipids (Cerebrosides)
They are compound lipids containing carbohydrates.
They are widely distributed in every tissue of the body, particularly in nervous tissues such as brain
They are formed from Sphingosine, sugar (usually β-galactose) and fatty acid.

Cerebrosides are differing according to the type of fatty acids present:

1. Kerasin → contains lignoceric acid (24 carbon atoms).
2. Cerebron → contains cerebronic acid (hydroxy lignoceric acid)
3. Nervon → contains nervonic acid (unsaturated lignoceric acid)
4. Hydroxy nervon → contains hydroxy nervonic acid (hydroxy unsaturated lignoceric acid)

Gaucher’s disease
Inability to break sphingolipids due to deficiency of 𝛽 −glucocerebrosidase and galactocerebrosidase that would result in
accumulation of sphingolipids (glycolipids) in liver and spleen causing hepatomegaly and splenomegaly (large liver and spleen).

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 C- Sulpholipids
These are sulphate ester of kerasin or cerebron
The sulphate group is usually attached to OH group of C3 of galactose
They are present in the testis, brain and muscle.

D- Gangliosides
Glycolipids containing Sphingosine with glucose or galactose and bound with N-acetyl galactoseamine and sialic acid.
Present in ganglia
E- Lipoproteins
► They are compound lipids containing proteins, present in cell membrane, mitochondria, cytoplasmic particles and blood
plasma

Structure of lipoproteins
The lipid fraction
1- Triglycerides. 2- Phospholipids. 3- Cholesterol (free or esterified).
4- Fat-soluble vitamins. 5- Steroid hormones

The protein fraction

1- -globulin: -lipoprotein.
2- β-globulin: β-lipoprotein.

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The increase in protein fraction would result in the increase in the density of lipoprotein and vice versa for example 60% in
HDL and 1-2% in chylmicron

Methods of separation of lipoproteins

I- Separation by ultracentrifugation:
• Means floatation of in sodium chloride solution.
• This method depends on as the proportion of lipids to proteins in lipoproteins increases, the density decreases.
By ultracentrifugation lipoproteins classified into 4 types:
1. Chylomicrons.
2. Very low density lipoproteins (VLDL or pre-β-lipoproteins).
3, Low-density lipoproteins (LDL or β-lipoproteins).
4. High-density lipoproteins (HDL or α-lipoproteins).

II. Separation by electrophoresis:

Electrophoresis means migration of charged particle in an electric field (anode or cathode)
1. Chylomicron → the predominant lipid is triglycerides absorbed from small intestine
2. Pre-β-ipoproteins(VLDL) → the predominant lipid is triglycerides synthesized in liver.
3. β-lipoproteins(LDL) → the predominant lipid is cholesterol. (They transport 60% of plasma cholesterol)
4. α-lipoproteins(HDL) → the predominant lipid is phospholipids . (Also transport 40% of plasma cholesterol)

Note:
 HDL is the lipoprotein with the fastest electrophoretic mobility and the lowest triglyceride content.
 LDL is a lipoprotein associated with high incidence of coronary atherosclerosis.
 HDL is a lipoprotein inversely related to the incidence of coronary artherosclerosis.

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 III- Derived lipids
► They are substances derived from the hydrolysis of simple and compound lipids.

► Derived lipids include: 1. Fatty acids 2. Alcohols 3. Steroids.

Steroids
• They are compounds containing steroid nucleus (perhydro- cyclopentano-phenanthrene)
• The steroid nucleus is formed from four fused rings of 19 carbon atoms
• The steroids are differing from each other as regards the nature of side chain, which is usually attached to 𝒄𝟏𝟕

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• The most important steroids are:
1- Sterols. 2- Sex hormones → Testosterone , Estrogen, Progesterone
3- Adrenal cortical hormones → Glucocorticoids , Mineralocorticoids 4- Bile acids.
5- Vitamin D. 6- Cardiac glycosides.

1- Sterols ex: cholesterol and ergosterol.

A- Cholesterol B- Ergosterol
► Cholesterol is animal sterol ► It is a plant sterol isolated from ergot and yeast.
► It has two sources ► It has exogenous source (from diet)
1) exogenous (from diet)
2) endogenous sources (tissue biosynthesis).
► It contains 27 carbon atoms & there is a OH group at C3 and ► It contains 28 carbon atoms; there is a OH group at C3 and two
a double bond between 𝐶5 − 𝐶6 double bond between 𝐶5 − 𝐶6 and 𝐶22 − 𝐶23

side chain = 8C

► It is the precursor of bile acids, adrenal cortical hormones, ► It is the precursor of vitamin 𝑫𝟐 by irradiation with ultraviolet
sex hormones, vitamin 𝑫𝟑 and cardiac glycosides. light.
► It is present in blood in a concentration 150-250 mg/dl
1) 70% are present as cholesterol ester
(cholesterol combined with fatty acids)
2) 30% are present as free cholesterol

► Cholesterol is partly excreted in bile and if its concentration

increases, it may form gallstone.

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 Chemical properties of cholesterol
1-Cholesterol crystals:

Observation and result:

• They appear as four sided plates with notched angels

2-Color reactions of cholesterol:

• This test is carried out with dry reagents on a dry test tube.

Liebermann's test
Procedure:

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Observation & result:
• The solution becomes red then blue then changes gradually into bluish green.
• It is cholesterol.

Salkowski 's test

Procedure:

Observation & result:

• The upper chloroform layer becomes red and the lower layer becomes yellow with green fluorescence.

2- Sex hormones:

3- Adrenal cortical hormones:

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 4- Bile acids
• There are four types of bile acids:
1. Cholic acid. 3. Deoxycholic acid.
2. Chenocholic acid 4. Lithocholic acid.

• These bile acids are conjugated with glycine or taurine (derived from cystiene) giving glycocholic or taurocholic acids
• These acids are conjugated with sodium or potassium giving sodium or potassium glycocholate or taurocholate (Bile salts).

• Greater than 95% of the bile salts are reabsorbed in the ileum and return to the liver via the enterohepatic circulation.
• Less than 5% of the bile salts entering the gut are excreted in the feces each day.

5- Cardiac glycosides

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 Oxidative stress
Definition:

A metabolic condition where the generated free radicals exceeds the antioxidant defense system of the body which would result on a serious
damages for the animal body.

Free radical:

is an atom which contain single unpaired electron on the outer shell generated from a leakage in mitochondrial system , which is highly reactive
causing oxidative damage through stealing electron from other atom to be stable which transfer this atom to free radical and so on.

Free radical subdivided into reactive oxygen species and reactive nitrogen species.

Anti oxidants:

Are reducing agents which prevent other chemical compounds from oxidation and become less effective from environmental factors

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 Reactive oxygen species (ROS)

 Chemically highly reactive compounds containing a radical oxygen perform certain physiological function.

 However during any environmental stress it increase causing severe damages such as superoxide radical (O2.), hydroxyl radical (OH. ) , hydrogen
peroxide (H2O2).

Generation of ROS

Major Sources of Reactive Oxygen Species:

1) formed during phagocytosis and inflammation:
a) the response of infectious agent stimulates the immune system to produce inflammatory cells stimulate consumption of oxygen
b) generation of free radical due to the action of antimicrobial defense system intend to destroy invading microorganisms
2) Hydrogen peroxide and lipid peroxide are generated as an intermediate for the synthesis of many eicosanoids
3) radiation, pollution increase the formation of free radicals
4) the leakage of electrons from electron transport system forming superoxide radical

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Effect of free radicals on cellular components:

 lipid degradation occurs, forming such products as Malondialdehyde

 Malondialdehyde appears in the blood and urine and is used as an indicator of free radical damage.

Reactive nitrogen species

 They are highly reactive chemical compounds generated from nitric oxide (NO)
 Reactive nitrogen species act together with (ROS) to result in cellular damage, causing nitrosative stress
 Nitric oxide (NO)
At low concentrations → it functions physiologically as 1) neurotransmitter
. 2) hormone that causes vasodilatation (control blood pressure)

At high concentrations → it combines with O2 or with superoxide to form additional reactive and toxic species containing both nitrogen and
oxygen (RNS) → involved in 1) neurodegenerative diseases such as Parkinson's disease.
2) Chronic inflammatory diseases such as rheumatoid arthritis.
 Nitric oxide (NO) is synthesized from arginine by nitric oxide synthase.
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Generation of RNS

• NO + O2·− → ONOO− (peroxynitrite)

• NO + NO2 → N2O3
• ONOO− + H+ → ONOOH (peroxynitrous acid)
• ONOO− + CO2 → ONOOCO2− (nitrosoperoxy carbonate)

Cellular defenses against oxygen toxicity

[Antioxidant]
 These are substances used to control and reduce lipid peroxidation both in human and in nature.

Types of antioxidants:

1) Some chemicals used as food additives. Like propyl gallate and others. Sodium benzoate

2) Some vitamins: Like vitamin E, vitamin C and B-carotene (precursor of vitamin A).

3) Some hormones: Estrogen and melatonin.

4) Some enzymes: Catalase, superoxide dismutase (SOD), glutathione peroxidase, glutathione reductase, etc.

Mechanism of antioxidant enzymes

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