CHEMISTRY OF LIPIDS

Definition of lipids: Lipids are heterogeneous group of compounds that are highly insoluble in water, but are soluble in non-polar solvents like Benzene, Chloroform, ether, alcohol e.t.c. Biological Roles of Lipids Lipids perform several important functions 1. Lipids are the concentrated fuel reserves of the body (Triglycerides) 2. Lipids involves in the construction of all biological membranes. 3. Lipids are the sources of the fat soluble vitamins (A, D, E, K) 4. Some lipids acts as cellular metabolic regulators (steroid hormones and prostaglandins) 5. Lipids act as the thermal insulators and maintain the body temperature 6. Lipids act as electrical insulators and help the neuronal transmissions. 7. Lipids make diet palatable (tasty) CLASSIFICATION OF LIPIDS

1. FATTY ACIDS Fatty Acids are carboxylic acids with aliphatic hydrocarbon side chain. These are the simplest Lipids. Fatty acids mainly occur in the esterified form as major constituents of various lipids, they are also present in free form Based on presence of single or double covalent bonds fatty acids are divided in to Saturated Fatty acids: Do not contain double bonds Unsatur(ated Fatty acids: Contain double bonds Mono unsaturated fatty acids: Contains one single double bond Poly unsaturated fatty acids: Contains two or more double bonds Numbering of carbon atoms in Fatty acids There are different types of numbering systems 10 9 8 7 6 5 4 3 2 1 Delta numbering system CH3 CH CH =CH2 CH2 CH2 CH2 CH2 CH2 COOH Greek letters 1 2 3 4 5 6 7 8 9 10 Omega numbering system Counting carbons with Greek alphabets The first CH2 adjacent to the carboxyl group is named as , and so on The last carbon with CH3 group is called as omega () carbon Delta ( ) system:

Carbons are numbered from COOH end to CH3 Carboxyl carbon (COOH) is counted as 1, and carbon with CH3 as the last Above fatty acid is 10: 1 or 10:1;7

(Total 10 carbons: 1 double bond between 7&8 carbons) Omega () system: Carbons are numbered from CH3 to COOH CH3 is counted as 1 and COOH as the last
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Above fatty acid is 10:1 7 (10 carbons: 1 double bond between 3&4carbons) Length of Fatty acids: Short chain fatty acids: Medium chain Fatty acids: Long chain Fatty acids: with less than 6 carbons with 8-14 carbons with 16-24 carbons

List of Important saturated fatty acids Common name Numbering Acetic acid Propionic acid Butyric Acid Valeric Acid Caproic acid Capric acid Palmitic acid Stearic acid Arachadic acid 2:0 3:0 4:0 5:0 6:0 10:0 16:0 18:0 20:0

List of Important Unsaturated Fatty acids Common name Palmitoleic acid Oleic acid Linoleic acid Linolenic acid Arachidonic Acid -ngmbering cis16: 1; 9 cis18:1;9 All cis18: 2;9,12 All cis18:3;9,12,15 -gnirebmgn 16:1 7 18:19 18:2 6,9 18:3 3,6, 9

All cis20:4; 5,8,11,14 20: 4 6,9,12,15

Even and Odd Carbon Fatty acids: Most of the fatty acids that occur in natural lipids are even carbons. Most common even carbon fatty acids: Palmitic acid (16C) and Stearic acid (18C) Most common odd carbon fatty acids: Propionic acid (3C) and valeric acid (5C) Isomerism in unsaturated fatty acids Unsaturated fatty acids shows geometric isomerism depending on the orientation of the groups around the double bond Cis configuration: Similar groups are present on the same side of the double bond.

Trans configuration: Similar groups are present on the opposite side of the double bond.

Essential fatty acids: The fatty acids that cannot be synthesized by the body and therefore should be supplied in the diet are known as essential fatty acids. Essential fatty acids are chemically poly unsaturated fatty acids. They are Linoleic aicd ( 18:2;9,12) Linolenic acid( 18:3; 9,12,15) Arachidonic acid (20:4;5,8,11,14) becomes essential when its precursor linoleic acid is not supplied in diet.
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Essential fatty acids are present in vegetable oils, and fish oils Functions of essential fatty acids: Normal growth of the body Formation of cell membrane Transport of cholesterol Formation of lipoproteins Formation of eco sanoids Protects against atherosclerosis Non essential fatty acids: These can be synthesized by the body These are saturated and mono unsaturated fatty acids Synthesized from acetyl-CoA which is derived from glucose oxidation

2. SIMPLE LIPIDS
Esters of Fatty acids with alcohol There are two types of simple lipids i. Triacylglycerols (Fats & Oils), ii. Waxes

i. Triacylglycerols (Fats & Oils) Esters of 3 fatty acids with a glycerol COOH group of fatty acids condenses with OH of glycerol and forms ester link

Triacylglycerol = 3 fatty acids are esterified to glycerol Also known as Triglycerides and neutral fats Insoluble in water and soluble in non-polar solvents They are fuel reserves of animal, stored in the adipose tissue The fat stored in normal men 20% women 25% of the body weight Triglycerides are not found in biological membranes. Triglyceride up on hydrolysis gives diacylglycerols and monoacylglycerols Based on the type of fatty acids present triglycerides are divided in to Simple triglycerides: Contains the same type of fatty acids at all the three carbons Mixed triglycerides: Contains 2 or 3 different types of fatty acids Mixed triglycerides are more common in nature. Oil: triglyceride with one or more unsaturated fatty acids exists as liquid at room temperature termed as oil Plant triglycerides are having higher content of unsaturated fatty acids compared to that of animals. Hence animal fat will be solid and plant fat is liquid at room temperature

Properties of Triglycerides: a) Hydrolysis:

Triglyceride +3 H2O

Lipase

3 Fatty acids + Glycerol

This reaction helps the digestion and absorption of the fat from the intestine Also helps the mobilization of fat from the adipose tissue. b) Saponification:

Triglyceride + 3 NaOH

3 Fatty acidsNa (Soap) + Glycerol

The alkali can be sodium hydroxide (NaOH), or Potassium Hydroxide (KOH). In the intestine soaps are formed with bile salts known as emulsification
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Emulsification process helps the easy absorption of fats from the intestine c) Rancidity: Oxidation of fat due to air, moisture, light, Bacteria is known as rancidity Oxidation takes place at double bonds of the free fatty acids Fats will be spoiled due to rancidity, the taste and smell will be unpleasant Steps and types in rancidity Hydrolytic rancidity: Triglycerides are hydrolysed by bacterial lipase to produce glycerol + free fatty acids Free fatty acids undergo further oxidative and ketonic rancidity. Hydrolysis increases in presence of moisture and warm temperature Oxidative rancidity: oxidation at double bonds forms peroxides

Ketonic rancidity: peroxides at double bond decomposes to form aldehydes & ketones

Enhancers of rancidity: Oxidizing agents like nascent oxygen (O), Pb2+, Cu2+ , Fe2+ (Heme), Lipoxygenase Detection of rancidity: Copper acetate reacts with free fatty acids Prevention of rancidity: Prevention of fat for the exposure to light and moisture Addition of antioxidants like Vit-C, E and molecules with SH group (cystine)

ii) Waxes: Esters of long chain fatty acids with long chain monohydric alcohols.

Eg: Bee wax, sebaceous secretion, cuticles of leaves Waxes are not hydrolysed by lipase, hence animals cannot digest the wax Waxes are solids at room temperature, highly hydrophobic

3. COMPLEX OR COMPOUND LIPIDS Alcohol + Fatty acid + Some other molecule There are three major types of complex (compound) lipids are present i. Phospho Lipids ii. Glyco Lipids iii. Lipoproteins

They are further divided in to i. Phospho Lipids A) Glycero Phospho Lipids a) Phosphatidic acid b) Lecithin c) Cephalin d) Phosphotidyl Inositol e) Phosphotidyl serine f) Plasmalogen g) Cardio Lipin ii. Glyco Lipids ( Glycosphingo lipids) a) Cerebrosides b) Gangliosides iii. Lipoproteins
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B) Phospho Sphingo lipids a) Sphingomylin

i. Phospho Lipids: These lipids contain alcohol + Fatty acids + phosphoric acid. There are two classes of phospho lipids A) Glycerophospholipids (Phosphoglycerides): contain glycerol as the alcohol B) Sphingophospholipids: Contains shingosine as the alcohol.

A) Glycerophospholipids (Phosphoglycerides): a) Phosphatidic acid: Glycerol esterified with a Saturated fatty acid at carbon 1, Unsaturated fatty acid at its carbon 2 Phosphoric acid at carbon 3

b) Lecithins ( Phosphotidyl Choline) Choline attached to phosphate of phosphatidic acid Carbon:1 = Saturated fatty acid Carbon: 2 = Unsaturated fatty acid Carbon 3: phosphoric acid + choline

Functions of lecithin Construction of cell membrane Formation of lecithin cholesterol esters Lecithin + Cholesterol Lecithin cholesterol acyltransferase (LCAT) Cholesterol ester (Cholesterol-Fatty acid) + Lysolecithin

Cholesterol esters are taken to liver, mixed in bile and excreted Absence of lecithin increases cholesterol in the blood, leads to atherosclerosis
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 Lecithin in bile prevents formation of gall stones (cholesterolstones) Storage of choline: lecithin acts as store for choline Choline involves in neurotransmissions (forms acetyl choline) Choline prevents accumulation of fat in liver (lipotropicfactor) Choline gives methyl group for methylation reactions Dipalmitoyl lecithin: Is a surfactant (decreases surfacetension) found in lungs and prevents the adherence of inner surfaces of lungs due to the surface tension dipalmitoyl lecithin is less produced in premature infants causing respiratory failure this condition is known as respiratory distress syndrome. c) Cephalin ( Phosphotidyl Ethanolamine): Ethanolamine attached to phosphatidic acid is termed as cephalin or phosphatidyl ethanolamine Involves in blood coagulation mechanism

d) Lipositol (Phosphotidyl Inositol) : Inositol attached to the phosphatidic acid. This is an important component of cell membranes. Acts as a second messenger in Ca2+ dependent hormonal actions

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e) phosphotidyl Serine The aminoacid serine is attached to phosphatidic acid found in brain, blood and other tissues

f) Plasmalogens Fatty aldehyde attached at C1 of glycerophospholipids by ether link known as plasmalogens.

Lecithin with fatty aldehyde at C1 = phosphatidal choline Cephalin with fatty aldehyde at C1 = phosphatidal ethanolamine Present in brain and muscle g) Cardiolipin: Two molecules of phosphotidic acids attached to a glycerol through their phosphoric acid groups is called cardiolipin. Cardiolipin is an important component of inner mitochndrial membrane.

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B) Sphingo Phospho Lipids: Sphingo lipids contain sphingosine as alcohol, they do not contain glycerol Sphingo lipids are found in brain and nervous tissue.

a) Ceramide: Fatty acid + Sphingosine Fatty acid attached to Sphingosine through amide link in ceramide

b) Sphingomyelin: Fatty acid+ Sphingosine+ Phosphric acid+ Choline Choline is attached to the phosphate of ceramide through ester link Present in nervous tissue and brain

Niemann picks disease Accumulation of sphingomyelin in liver,spleen and brain Caused by the deficiency of an enzyme sphingomyelinase Clinical findings are enlargement of liver & spleen, and mental retardation

Functions of Phospholipids: Construction of cell membranes and regulates the cell permeability. Helps the absorption of fat from the intestine, Involves in the synthesis of lipoproteins Acts as surfactants, eg: Dipalmitoyl lecithin in lungs.
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Prevents the accumulation of fat in liver Involves in cholesterol metabolism Cephalin participates in blood clotting. Phosphatidyl inositol participates in signal transduction across the cell membrane.

ii. Glycolipids: Present in cell membranes and nervous tissue, Contains sphingosine + fatty caid + carbohydrate, There are two important classes of glycolipids. a) Cerebrosides a) Cerebrosides: These are the simplest glycolipids contains Sphingosine + Fatty acid (ceramide) + sugar (galctose or glucose). Present in brain, nervous tissue Acts as insulators for neurotransmissions b) Gagliosides

Gauchers disease: Cerebrosides accumulate in liver, spleen and bone marrow. Because of the deficiency of enzyme glucocerebrosidase Spleen and liver will be enlarged

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b) Gangliosides: These are complex glycosphingolipids Ceramide attached to N-acetyl neuramnic acid (sialic acid) and other mono saccharides. The important gangliosides present in the brain are GM1, GM2, GD, GT Also present in membrane receptors Where: G= Ganglioside; D= Di sialic acid residues M= Mono sialic acid residue T=Tri sialic acid residues

Tay sachs disease: accumulation of GM2 gangliosides in brain, spleen Because of deficiency of an enzyme hexosaminidase A Leads to mental retardation, blindness and muscular weakness

iii. Lipoproteins: = Lipid + apolipoprotein Lipids combined with proteins are termed as lipoproteins The proteins present in lipoproteins are known as apolipoproteins Apolipoproteins are mostly globulins in nature Lipids present in lipoproteins are triglycerides, phospholipids, cholesterol, cholesterol esters and free fatty acids Lipoprotein helps the transport of lipids in the blood, and in and out of the cells Lipoproteins involves in the construction of cell membrane If lipid concentration increases the density of lipoprotein decreases If the lipid concentration decreases the density of lipoprotein increases
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Classification of lipoproteins based on centrifugation Lipoproteins can be separated by centrifugation based on their relative densities They are mainly divided in to 4 classes Type Chylomicrons VLDL IDL LDL Lipids present Important Apolipoproteins present apo-A-I,A-II,C-I,C-II apo-B-100,C-I,C-II,E apo-B-100,C-I, C-II,E apo-B-100,C-I, C-II,E

Triglycerides Triglycerides, cholesterol Less TG & cholesterol Cholesterol Cholesterol esters Phospholipids Cholesterol HDL Cholesterol esters Phospholipids Free fatty acids-Albumin Free fatty acids

apo- A-I, A-II,E,D

Albumin

Functions of apo-lipoproteins Transport of lipids (gives hydrophilic covering for lipids) Binds membrane receptors
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Regulates enzyme activities (Activates /inhibits) Electrophoretic separation of lipoproteins Proteins present in the lipoproteins shows charge (ve & +ve) During electrophoresis lipoproteins will separate based on their charge Centrifugation CM VLDL LDL HDL FFA-albumin Electrophoresis CM LDL ( lipoproteins) VLDL (pre lipoproteins) HDL ( lipoproteins) Free fatty acids-Albumin Separation based on charge

Separation based on density

4. DERIVED LIPIDS

Hydrolysis or chemical modification of simple or complex lipids gives derived lipids Some of important derived lipids are Fatty acids Ecosanoids Glycerol Steroids & Sterols (Cholesterol, Vitamin-D, Bile acids & Salts, Sex hormones)

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i) ECOSANOIDS Cyclic compounds that are derived from arachidonic acid (20 C poly unsaturated) Note: Ecosa = 20 Ecosanoids acts like local hormones They are derived in to two types A) Prostanoids Prostaglandins (PG) Prostacyclines (PGI) Thromboxanes (Tx) A) Prostanoids a) Prostaglandins There are four main groups of prostaglandins are present A, B, E, and F Recently some more groups are identified like, C,D,H,G,I,X Causes vasodilatation, and contraction of uterus & intestine, b) Prostacyclines Causes vasodilatation Prevents platelet aggregation c) Thromboxanes Causes vasoconstriction Enhances platelet aggregation B) Leukotriens (LT)

B) Leukotriens Are present in leukocytes Causes chemo taxis of leukocytes towards the site of inflammation

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ii. GLYCEROL Contains 3 carbons, all with alcoholic groups (poly hydric alcohol) Derived from hydrolysis of triglycerides Can be converted to glucose in a process named gluconeogenisis Used to prepare nitroglycerine the medicine used to dilation of coronary artery Used to prepare soaps, creams and lotions for dry skin

iii. STEROIDS & STEROLS Compounds containing steroid nucleus (cyclopentano perhydro phenanthrene ring) Steroid nucleus contains 16 carbons making 4 rings Methyl side chains (carbons 19 , 18) are attached to carbons 10 , 13 respectively

Sterols: When steroids contains one or more hydroxyl groups then it is called as sterol One of the important biological sterol in animal systems is cholesterol. Other important sterols include, Bile acids, Vitamin-D, Sex Hormones, Adrino cortical hormones.

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a) Cholesterol:

Structure: Steroid nucleus attached with an aliphatic side chain at carbon 17 Aliphatic side chain contains 8 carbons; hence cholesterol contains total 27 carbons Hydroxyl group (-OH) is present at carbon 3 Double bond between carbons 5-6 Cholesterol contains five methyl (CH3) groups ( C18, C19, C21, C26, C27).

Properties and functions of cholesterol Cholesterol is a yellowish crystalline solid in nature Cholesterol is the most abundant animal sterol Cholesterol is found only in animals. Cholesterol is present cell membranes (increases fluidity of cell membrane) Cholesterol is a poor conductor of heat and electricity hence functions as insulator for nervous tissue. Cholesterol involves in the synthesis of Bile acids, steroid hormones and Vitamin-D.

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b) Vitamin D: Is synthesized from cholesterol in animals

Ergosterol (Fungi) U.V light

Cholesterol (Animals) U.V light (In skin)

Vitamin D2 (ergocalciferol)

Vitamin D3 (cholecalciferol)

By enzyme 25,Hydroxylase in Intestine 25,hydroxy cholecalciferol (Calcidiol) 1- Hydroxylase in Kidneys

1,25 dihydroxy cholecalciferol (Clacitriol) is the active form of vitamin-D

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c) Bile Acids:

Primary Bile Acids:

Cholic acid ,

Cheno dioxy cholic acid

These are synthesized in the liver from cholesterol.

Secondary Bile acids: Deoxy cholic acid, Litho cholic acid. In the intestine primary bile acids will be converted to secondary bile acids by the intestinal bacteria.

Conjugates of bile acids & Bile salts:

Conjugates of bile acids: Primary bile acids complexes with glycine and taurine Eg: Glyco cholic acid, Tauro cholic acid Glyco cheno dioxy cholic acid, Tauro chenodioxy cholic acid

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Bile salts: Sodium salts of conjugated bile acids Eg: sodium glycocholate, Sodium tauro cholate

Functions of bile salts Helps in digestion and absorption of lipids from the intestine (emulsification) Stimulates liver to secrete more bile (choleretic effect) Prevents formation of cholesterol stones

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d) Steroid Hormones: There are two major classes of steroid hormones All the steroid hormones are derived from cholesterol i) Sex Hormones: a) Estrogens (Female sex hormones) Estrone Estradiol Estriol b) Progestins (Female sex hormones) Progesteron c) Androgens (Male sex hormones) Testosterone Dihydrotestosterone i) Sex hormones: a) Estrogen: Is a female sex hormone There are three types of estrogens Estrone (E1), Estradiol (E2), Estriol (E3) Estradiol (E2) is the most active estrogen Structure: Ring A is unsaturated -OH group at C3 Hydroxyl group at C17 Methyl group (C18) at C13 Site of production: Females: Ovary, Placenta, Adrenal cortex (minor amounts) Males: Testes, Adrenal cortex (minor amounts) Functions: Development of female secondary sexual characters Anabolic effect on bones and cartilage
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ii) Adrinocortical hormones a) Gluco corticoids Corticosterone Cortisol Dehydrocorticosterone Cortisone b) Mineralo corticoids Aldosterone 11-deoxy corticosterone

Contraception (prevents pregnancy) b) Progesterone: Is a female sex hormone Ketone group at C3 Double bond between C4,C5 Methyl groups (C19&C18) at C10 & C13respectively Site of production: Females: Ovary, Placenta, Adrenal cortex Males: Adrenal cortex Functions: Prepares uterus for implantation of ovum Stabilizes pregnancy Inhibits milk production during late pregnancy Antagonizes estrogen function

c) Androgens: Are male sex hormones Testosterone: Ketone group at C3 Double bond between C4 &C5 Methyl groups at C10,C13 -OH at C17 Dihydro testosterone is testosterone without double bond between C4 & C5 Dihydrotestosterone is the active form in tissues Site of production: Males: Testes, Adrenal cortex, Females: Adrenal cortex Functions: Development of male secondary sexual characters
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Stimulates spermatogenesis, Shows anabolic effect on proteins ii. Adrenal cortical hormones: Adrenal cortical hormones are mainly divided in to two types Are produced from adrenal cortex

a) Glucocorticoids: Corticosterone, Dehydrocorticosterone, Cortisol , Cortisone They regulates the carbohydrates, Fat and protein metabolisms Shows catabolic effect on fat and protein metabolisms Shows anabolic effect on carbohydrate metabolism

b) Mineralcorticoids: Aldosterone, 11-deoxy corticosterone Regulates the mineral metabolism Acts on kidneys stimulates secretion of K+, H+ and reabsorption of Na+

All adrenal cortical hormones are derived by slight modifications of corticosterone Corticosterone structure Keotne group at C3 Double bond between C4& C5 -OH at C11 Methyl groups at C10,C13 Ketol group at C17

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5. FREE RADICALS AND ANTIOXIDANTS

Free Radical: a molecule or atom that contains one or more unpaired electrons, and is capable of independent existence.

Different types of free radicals produced in the body O O2 OHHOOROO(singlet oxygen / nascent oxygen) (Super Oxide radical) ( Hydroxyl Radical) ( Hydro Peroxy Radical) (Lipid Peroxy Radical)

Free radicals are formed during the normal metabolism in presence of oxygen or metal ions ( Cu ++, Fe ++) they are highly reactive. Free radicals can generate new free radicals by chain reaction Free radicals damages the cell membranes and biomolecules

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Lipid Peroxidation Free radicals oxidizes the poly unsaturated fatty acids and forms lipid peroxides Lipid peroxides are free radicals, they further oxidizes other poly unsaturated fatty acids Lipid peroxidation takes place in three steps (phases)

Initiation Phase: Hydroxyl Radical (OH) removes hydrogen atom from polyunsaturated fatty acids (RH) forming fatty acyl radical ( R) RH + OH R + H2O

Propagation Phase: Fatty acyl radical will be converted to peroxy radical ( ROO) in presence of oxygen R + O2 ROO

Peroxy Radical ( ROO-) removes Hydrogen atom from another poly unsaturated fatty acid and becomes lipid hydroperoxide ( ROOH) ROO + RH ROOH + R

Termination Phase: When all the poly unsaturated fatty acids are oxidized lipid peroxidation stops Lipid peroxidation in body destroys cell membranes and leads to atherosclerosis and cardiovascular diseases

Toxic effects of free radicals Damage of cell membranes, and biomolecules leads to Atherosclerosis, myocardial infarction, cancers, aging, rheumatoid arthritis, etc

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Antioxidants: The molecules that can stop or slow down the oxidation of lipids and other biomolecules are termed as anti oxidants.

i) Natural anti oxidants: These are of two types:

a) Preventive antioxidants: Reduces the rate of chain initiation (first step of lipid peroxidation) Eg: Catalase, Glutathione peroxidase, selenium EDTA( Ethylene Diamine Tetra Acetate) DTPA ( Diethylene Triamine Penta Acetate)

b) Chain Breaking Anti Oxidants: They stops the chain elongation, (second step of the lipid peroxidation) eg: Super oxide desmutase ( SOD) Vitamin-E, Vitamin-C

ii. Artificial antioxidants : These are chemically synthesized Added in lipid based foods to prevent the oxidation of lipids ( Rancidity) in the food. Eg: Propyl Gallate, Butylated Hydroxy anisole (BHA) Butylated Hydroxy Toluene (BHT)

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