UNIT III

Fats and Oils

In pharmaceutical organic chemistry, the study of fats and oils is crucial as they serve various purposes in
pharmaceutical formulations. These substances can act as excipients, solvents, carriers, or even active
pharmaceutical ingredients (APIs) themselves.

Fats and oils are called triglycerides because they are esters composed of three fatty acid units
joined to glycerol, a trihydroxy alcohol:

Below is an outline of what a chapter on fats and oils might cover in pharmaceutical organic chemistry:

Introduction to Fats and Oils:

Definition of fats and oils.

Classification based on saturation (saturated, unsaturated, polyunsaturated).
Chemical structure and properties.
Sources and Extraction:

Natural sources of fats and oils (animal and plant sources).

Extraction methods (mechanical pressing, solvent extraction, supercritical fluid extraction).
Composition:
Triacylglycerol structure.
Fatty acids (saturated, unsaturated, omega-3, omega-6).
Other components (phospholipids, sterols, tocopherols).
Physical Properties:

Melting point, boiling point, and viscosity.

Solid fat content (SFC) and iodine value.
Relationship between fatty acid composition and physical properties.
Chemical Properties:

Hydrolysis (acidic, basic, enzymatic).

Oxidation (rancidity, oxidative stability).
Saponification and esterification reactions.
Role in Pharmaceutical Formulations:

Excipients in oral dosage forms (tablets, capsules, emulsions).

Solvents and carriers in topical formulations (creams, ointments).
Lipid-based drug delivery systems (liposomes, lipid nanoparticles).
Active pharmaceutical ingredients (omega-3 fatty acids, essential fatty acids).

Quality Control and Analysis:

Methods for determination of fatty acid composition (gas chromatography, high-performance liquid
chromatography).
Analysis of oxidative stability (peroxide value, anisidine value, TOTOX value).
Testing for impurities (heavy metals, pesticides, solvent residues).
Regulatory Considerations:

Pharmacopoeial standards for fats and oils (USP, BP, EP).

Good Manufacturing Practices (GMP) for production and testing.
Regulatory requirements for labeling and documentation.
Applications

Novel extraction techniques (green extraction, ultrasound-assisted extraction).

Development of lipid-based drug delivery systems for enhanced bioavailability.
Exploration of functional lipids for therapeutic applications.

Physical Characteristics of Oils and Fats

Pure fats and oils have no taste, no smell, and no colour. Some of them have distinct colours, smells, and
flavours that we are familiar with because of outside molecules that are absorbed by these lipids and lipid
soluble. For instance, the pigment carotene gives butter its yellow colour; bacteria in the maturing cream
from which butter is created produce the chemicals diacetyl and 3-hydroxy-2-butanone, which give butter
its flavour.
Oils and fats are diverse substances with varying physical characteristics, largely influenced by their
chemical composition and structure. Here are some key physical characteristics of oils and fats:
1. State at Room Temperature:
● Oils are typically liquid at room temperature, while fats are solid or semi-solid. This
difference is primarily due to the saturation level of fatty acids present. Saturated fats tend
to be solid at room temperature, while unsaturated fats are usually liquid.
2. Melting Point:
● The melting point of a fat or oil depends on the types of fatty acids present. Saturated fats
have higher melting points compared to unsaturated fats. Fats with a higher proportion of
saturated fatty acids will have a higher melting point. For example, coconut oil, which is
high in saturated fats, has a relatively high melting point, while oils high in unsaturated fats,
like olive oil, have lower melting points.
3. Viscosity:
● Oils generally have lower viscosity compared to fats. This means that oils flow more easily
than fats at a given temperature. Viscosity is influenced by factors such as fatty acid
composition, temperature, and processing methods.
4. Cloud Point:
● The cloud point is the temperature at which a clear oil becomes cloudy or begins to solidify
due to the formation of crystals. This phenomenon occurs because some fats and oils
contain saturated fatty acids that crystallize at specific temperatures. The cloud point varies
depending on the composition of the oil or fat.
5. Color:
 ● Oils and fats can vary widely in color, ranging from clear or pale yellow to deep green or
amber, depending on factors such as the source of the oil, processing methods, and
presence of pigments or impurities.
6. Odor and Flavor:
● Oils and fats may have distinct odors and flavors influenced by their source, processing
methods, and storage conditions. For example, olive oil may have a fruity or grassy aroma,
while coconut oil has a characteristic coconut scent.
7. Density:
● The density of oils and fats varies depending on factors such as composition and
temperature. Generally, oils are less dense than water, while fats are denser. This property
can affect their behavior in culinary applications and industrial processes.
8. Transparency:
● The transparency of oils and fats can vary from clear to opaque, depending on factors such
as purity, processing methods, and the presence of impurities or particles.
Chemical Characteristics of Oils and Fats
1. Fatty acids that are free: These are individual fatty acid molecules that are not bound to glycerol
or other molecules. They can be found in various biological processes such as energy metabolism
and membrane synthesis.
2. Triacylglycerol, or triglyceride: This refers to a type of lipid molecule composed of three fatty
acid chains esterified to a glycerol backbone. Triacylglycerols serve as the primary storage form of
fatty acids in organisms, providing a concentrated source of energy.
3. Lipids phosphorylated: This term refers to lipids that have undergone phosphorylation, a process
where a phosphate group is added to the molecule. Phosphorylated lipids, such as phospholipids,
are crucial components of cell membranes and play roles in cellular signalling and metabolism.
4. Lipids: This term encompasses a broad category of biomolecules that are water-insoluble and
include fatty acids, triglycerides, phospholipids, steroids, and other compounds. Lipids play vital
roles in cellular structure, energy storage, signalling, and insulation.
5. Anabolic agents: While not exclusively related to lipids, anabolic agents are substances or
processes that promote the synthesis of complex molecules from simpler ones, typically associated
with growth, repair, and maintenance of tissues. Lipids can serve as building blocks for anabolism,
contributing to processes such as cell membrane formation and energy storage.

Triglycerides
Triglycerides are molecules that are related to glycerol and consist of three fatty acid units. These
molecules make up fats and oils. Triglycerides, also known as triacylglycerols, are indeed composed of
glycerol and three fatty acid molecules. In these molecules, the three fatty acids are esterified to the three
hydroxyl (-OH) groups of a glycerol molecule, forming ester bonds.

Triglycerides are the main components of both fats and oils, which are collectively referred to as lipids. The
distinction between fats and oils lies primarily in their physical state at room temperature: fats are typically
solid, while oils are liquid. This difference in state is largely determined by the types of fatty acids present
in the triglyceride molecules, with saturated fats tending to be solid and unsaturated fats typically being
liquid.

Triglycerides serve important functions in living organisms, including energy storage, insulation, and
protection of organs. They are stored in specialized cells called adipocytes in animals and in seeds and fruits
in plants. When energy is needed, triglycerides can be hydrolyzed into glycerol and fatty acids, which can
then be metabolized to produce ATP (adenosine triphosphate), the primary energy currency of cells.

Triglycerides can exist in different forms based on the fatty acids they contain and how those fatty acids are
attached to the glycerol molecule. Here's a breakdown:
1. Simple Triglycerides: In a simple triglyceride, all three fatty acid chains attached to the glycerol
molecule are identical. These are relatively uncommon in nature but can be synthesized in
laboratory settings.
2. Mixed Triglycerides: Most triglycerides found in natural fats and oils are mixed triglycerides. This
means that the three fatty acid chains attached to the glycerol molecule can be different from one
another. Mixed triglycerides are more prevalent in biological systems and are found in a variety of
dietary fats and oils.
physical state of triglycerides (whether solid or liquid) and their composition, particularly in terms of
saturation with fatty acids.
1. Solid vs. Liquid Triglycerides: Triglycerides that are solid at room temperature are commonly
referred to as fats, while those that are liquid are typically called oils. This distinction is largely
determined by the composition of fatty acids within the triglyceride molecule.
2. Composition and Melting Points: The degree of unsaturation and the length of carbon chains in
the constituent fatty acids influence the melting points of triglycerides. Saturated fats, with no
double bonds between carbon atoms, tend to have higher melting points and are solid at room
temperature. In contrast, unsaturated fats, which contain one or more double bonds, have lower
melting points and are liquid at room temperature.
3. Sources of Fats: Animal-derived fats, such as lard and tallow, tend to be solid due to their high
content of saturated fatty acids. In contrast, plant-based oils, like olive oil and linseed oil, are
typically liquid because they contain a higher proportion of unsaturated fatty acids.
4. Role in the Body: Fats serve as a storage form of energy in the body, providing a reserve of fuel
that can be accessed when needed. When required, fats are broken down into their constituent fatty
acids and glycerol, which can then be metabolized for energy production. The liver can convert
glycerol into glucose through a process called gluconeogenesis, providing an additional source of
energy.

Distinguish between oils and fats.

outlines the key differences between fats and oils based on their physical characteristics and composition.
1. State at Room Temperature:
● Fats are solids or semisolids at room temperature.

● Oils are liquids at room temperature.

 2. Composition of Fatty Acids:
● Fats contain a large amount of saturated fatty acids, such as stearic and palmitic acids.

● Oils contain a large amount of unsaturated fatty acids, such as oleic acid.
3. Melting Point:
● Fats melt at high temperatures.

● Oils melt at low temperatures.

4. Presence of Double Bonds:
● Fats do not contain double bonds in their fatty acid chains.

● Oils have double bonds in their fatty acid chains.

5. Stability:
● Fats are more stable than oils.

● Oils are less stable due to the presence of unsaturated fatty acids, which are prone to
oxidation.
6. Source:
● Fats are commonly found in animal sources.

● Oils are predominantly derived from vegetable sources, although some oils can also come
from animal sources (e.g., fish oil).

Fats oils

Fats are solids or semisolids Oils are liquids at room temperature

at room temperature

Fats contains large amount of saturated fatty Oils contains large amount of unsaturated
acids e.g. stearic and palmitic acids

fatty acids e.g. oleic acid

Fats melt at high temperature Melt at low temperature

Fats do not contain double bonds Oils have double bonds

Fats are more stable Oils are less stable

Fats are animal fats Oils are vegetable fats

Pure fats and oils have no taste, no smell, and no colour. Some of them have distinct colours, smells, and
flavours that we are familiar with because of outside molecules that are absorbed by these lipids and lipid
soluble. For instance, the pigment carotene gives butter its yellow colour; bacteria in the maturing cream
from which butter is created produce the chemicals diacetyl and 3-hydroxy-2-butanone, which give butter
its flavour.
With densities of roughly 0.8 g/cm3, fats and oils have a lower density than water. They act as great
insulators for the body, limiting the loss of heat via the skin since they are poor conductors of heat and
electricity.
Chemical composition

1. Basic concepts: All types of fats share a common basic concept, consisting of glycerol and fatty
acids.
2. Glycerides: Fats are primarily composed of glycerides, with triglycerides being the predominant
form. Triglycerides consist of three fatty acid molecules bonded to a glycerol molecule.
3. Ester Bonds: The carboxyl end (-COOH) of each fatty acid molecule reacts with one of the
hydroxyl groups (-OH) of the glycerol molecule. This reaction forms ester bonds between the fatty
acids and glycerol, resulting in three fatty acid chains per triglyceride molecule.
4. Dehydration Synthesis: During esterification, water molecules are removed from the reactants,
and the carbon atoms of the fatty acids are joined to the glycerol molecule by oxygen atoms,
forming ester bonds. This process is also known as dehydration synthesis.
5. Esterification: The process of forming ester bonds between fatty acids and glycerol is called
esterification. This process results in the formation of triglycerides, which are esters of glycerol and
fatty acids.
6. Molecular Structure: If the angles and kinks in the fatty acid chains were straightened out, the
triglyceride molecule would resemble a capital letter "E," with glycerol as the central vertical line
and the fatty acid chains as the horizontal lines.
In a fat molecule, each fatty acid is represented by a horizontal line, while the vertical line
connecting them represents the glycerol backbone. This arrangement forms ester bonds between the
fatty acids and glycerol.
Fatty Acid Chain Length: Fatty acids consist of a chain of carbon atoms arranged in a zigzag
pattern, with hydrogen atoms attached. The chain length of fatty acids increases with the number of
carbon atoms, and longer chains have higher melting points due to increased intermolecular forces
of attraction, such as van der Waals forces.
Degree of Saturation: The presence and type of double bonds in the carbon chain of fatty acids
determine their classification. Saturated fatty acids (SFA) lack double bonds, monounsaturated fatty
acids (MUFA) have one double bond, and polyunsaturated fatty acids (PUFA) have multiple double
bonds.
Effect on Membrane Fluidity: The length and saturation of fatty acids influence the
configuration and fluidity of cell membranes. Shorter chain fatty acids and those with
higher unsaturation contribute to more flexible membranes, as they are less viscous
and rigid.
Biological Impact: The fluidity of cell membranes plays a crucial role in various
biological processes, affecting membrane permeability, receptor function, and signal
transduction pathways.
Various chemical reactions of fats and oils
1. Hydrolysis

The hydrolysis of fats and oils in the presence of a base makes soap and is known as
saponification. Double bonds present in unsaturated triglycerides can be hydrogenated to convert
oils (liquid) into margarine (solid).

Hydrolysis can also be done by heating fat with water under pressure. Alkaline hydrolysis of fats
produces salts of fatty acids called as soaps and hence this reaction is also known as
saponification. Common soaps are the mixture of sodium salts of ‘C’ atoms (12 atoms) and higher
fatty acids. Soap molecules have both lipophillic (lipid loving) and hydrophilic (water loving)
group. The lipophillic group dissolves oils while hydrophilic portion dissolves water. Soap
molecules on dissolution in water forms micelle. Hydrolysis can be done in three ways-

a) Hydrolysis by water: Fats undergoes hydrolysis in presence of water at 443K and 6-8
atmospheric pressure. Zinc oxide is used as catalyst.
b) Hydrolysis by enzymes: hydrolysis of fats and oils can be done by adding enzyme lipase to an
emulsion of fat in water.

c) Hydrolysis by acids: mineral acids cause hydrolysis of fats. For this mixture of sulphonic acids
which are obtained by sulphonation of mixture of oleic acid and benzene.

The above three ways gives glycerol and fatty acids as a product of hydrolysis of fats while
alkaline hydrolysis of fats gives glycerol and soap which are used as cleansing agent. The
cleansing property of soap depends upon the ability to form emulsion with fat soluble materials.

2. Hydrogenation
Oils have large amount of unsaturated portion in the form of glycerides. When hydrogen is passed
through oils under pressure and by using catalyst at high temperature oils gets converted into solid
fats. This process is known as hardening of oils. By hydrogenation, unsaturated acid part of oil
gets reduced into saturated part and hence liquid oil gets converted into semi-solid fat.
Hydrogenation is carried out in a closed container in the presence of finely powdered catalyst
(0.05 - 0.2% of nickel) at temperature as high as 150-200 oC. The catalyst is usually removed by
filtration. During hydrogenation process a proportion of the cis double bonds are isomerized to
trans double bonds and there is also migration of double bonds. The hydrogenation process has
made it possible to extend the food uses of a number of vegetable oils and marine oils whose
melting points are too low.
3. Hydrogenolysis:
This is a cleavage reaction in which fat or oil molecule is treated with excess of hydrogen under
pressure in presence of copper-chromium catalyst. In this reaction fat or oil gets splits up into
glycerol and higher aliphatic alcohols.

4. Saponification:
Saponification is a process that involves conversion of fat, oil or lipid into soap and glycerol by
the action of heat in the presence of aqueous alkali (e.g. NaOH). Soaps are salts of fatty acids and
fatty acids are monocarboxylic acids that have long carbon chains (at least 10) e.g. sodium
palmitate.
5. Rancidification:
Rancidification is the process of complete or incomplete oxidation or hydrolysis of fats and oils
when exposed to air, light, or moisture or by bacterial action, resulting in unpleasant taste and
odor. Rancidity reactions may be due to hydrolysis of ester bonds (hydrolytic rancidity) or due to
oxidation of unsaturated fatty acids (oxidative rancidity). Rancidity occurs by the following ways-
a) Oxidation of unsaturated fatty acids: in presence of light and moisture, small amount of
unsaturated acids present in fats/oils gets oxidized by air to form peroxides which further break
down into aldehydes having unpleasant smell and taste. Saturated fatty acids do not get rancid.
This problem can be checked by adding small quantity of phenolic substances which act as
antioxidant.

b) Enzymatic hydrolysis: Due to presence of micro-organisms fats gets hydrolyzed by enzymes

to produce fatty acids having sour taste and unpleasant odour. For example butter gets rancid due
to production of butyric acid in this manner.

c) β-oxidation of saturated fatty acids: fats having saturated fatty acids undergo ketone
rancidity. Saturated acids undergo β-oxidation to form keto acids which gives carbon dioxide to
form ketones having unpleasant odour.

6. Drying oils:
A drying oil is the oil that hardens to a tough, solid film after a period of exposure to air. The oil
hardens through a chemical reaction in which the components crosslink (and hence, polymerize)
by the action of oxygen (not through the evaporation of water or other solvents). Drying oils are a
key component of oil paint and some varnishes. Some commonly used drying oils include linseed
oil, tung oil, poppy seed oil, perilla oil, and walnut oil. Drying oils (wild rose oil, linseed oil,
wheat oil) contain more than 50% of polyunsaturated acids. They are quickly absorbed and leave
no greasy layer on oily skin. Their light consistency makes them a good make-up primer.

The "drying", hardening, or, more properly, curing of oils is the result of autoxidation, the addition
of oxygen to an organic compound and the subsequent crosslinking. This process begins with an
oxygen molecule (O2) in the air inserting into carbon-hydrogen (C-H) bonds adjacent to one of the
double bonds within the unsaturated fatty acid. The resulting hydroperoxides are susceptible to
crosslinking reactions. Bonds form between neighboring fatty acid chains, resulting in a polymer
network, often visible by formation of a skin-like film on samples. This polymerization results in
stable films that, while somewhat elastic, do not flow or deform readily. Diene-containing fatty
acid derivatives, such as those derived from linoleic acid, are especially prone to this reaction
because they generate pentadienyl radicals. Monounsaturated
 fatty acids, such as oleic acid, are slower to undergo drying because the allylic radical
intermediates are less stable.

Oils depending upon their exposure to light and air can be classified as-

a) Non-drying oils: These oils on exposure to light and long storage get rancid. Oils get
decomposed into glycerol and fatty acids (saturated and unsaturated). The unsaturated acids gets
oxidised into aldehydes and acids with lesser carbon atoms in the molecule. The saturated acids
get decomposed by enzymes to form ketones. For example olive oil, almond oil, Babassu oil,
Baobab oil, Coconut oil, Peanut oil and Tiger Nut Oil.

b) Drying oils: They form a solid elastic film. A good drying oil dries within 4-5 hours. For
example Linseed oil.

c) Semi-drying oils: It is the oil which partially hardens when exposed to air. This is as opposed
to a drying oil, which hardens completely, or a non-drying oil, which does not harden at all. Oils
with an iodine number of 115-130 are considered semi-drying. Semi-drying oils contain 20%-
50% of polyunsaturated acids. They include: sweet almond oil, apricot seed oil, Cottonseed oil,
Sesame oil and Grape seed oil.

Analysis of fats and oil

Purity and composition of oil depends upon the degree of unsaturation, acidity on hydrolysis, and
its molecular weight. A variety of physical tests have been performed on fats and oils to determine
their composition and quality. Some of the common tests are as follows and these tests are also
known as analytical constants.

a) Acid value: The Acid value is the number, which expresses in milligrams, the amount of
potassium hydroxide necessary to neutralize the free acids present in one gram of the substance
(fats or oil).

Procedure: Weigh about 10g of the substance being examined, in an iodine flask. Prepare 50ml
mixture of equal volumes of ethanol (95 per cent) and ether, add 0.5 ml phenolphthalein solution
and titrate it against 0.1 N aqueous potassium hydroxide (KOH) solution until the solution
remains faintly pink after shaking for 30 seconds. Calculate the acid value from the following
equation.

Acid value = 5.61×VKOH/W

Where, VKOH = Volume of potassium hydroxide solution used (ml)

W = the weight of the fat or oil being examined (gm).

Significance: Acid value indicates the degree of rancidity of the given fat. High acid values arise
in rancified oils. Rancidification is the decomposition of fats and other lipids by hydrolysis and/or
oxidation. Oxidation primarily occurs with unsaturated fats by a free radical-mediated process,
which is responsible for producing the unpleasant and obnoxious odours and flavours.
Rancidification can be reduced (but not completely eliminated) by storing the fats and oils at low
temperature and away from light [2, 8, 9, 15, 16].

b) Saponification value: Saponification value is defined as the number of milligrams of

potassium hydroxide required to completely saponify one gram of fat or oil.

Procedure: Weigh about 2 g of the substance being examined in an iodine flask with reflex
condenser. Add 25 ml of 0.5 ethanolic potassium hydroxide solution and boil under reflux on
water bath for 30 minutes. Remove the condenser and add 1 ml of phenolphthalein solution and
titrate immediately with 0.5 M Hydrochloric acid. Note the reading as ‘A’. Repeat the operation
omitting the substance being examined. Note the reading as ‘B’. Calculate the saponification value
from the following equation,

Saponification value = 28.05 × (B-A)/W

Where, W = weight of substance (gm).

B = ml of HCl used for blank titration

A = ml of 0.5 N HCl used for titration

Significance: The saponification value is used primarily as an identification aid to detects

adulteration with unsaponifiable matter. It is also used to determine the extent of compounding
(fats and oils added to improve oiliness) in a lubricant [10, 11, 15, 16].
c) Ester value: The Ester value is the number of milligrams of potassium hydroxide required to
react with esters present in 1 g of fat or oil. The difference between saponification value and acid
value is called Ester value [2, 14, 15, 16].

Ester value = Saponification value – Acid value

d) Iodine value: Iodine value is defined as the number of grams of iodine taken up by 100 gm of
fat or oil or it is the number of grams of iodine which will combine with 100 gm of the fat or oil.
1) Hubl’s method: Fat or oil sample is dissolved in carbon tetrachloride and is treated with
excess of standard solution of ethanolic iodine in presence of mercuric chloride. Unused iodine is
then calculated by titration with standard sodium thiosulphate solution.

2) Wij’s method: This method uses iodine monochloride in acetic acid in presence of iodine.
Iodine monochloride readily combines with the double bonds present in fat and oil. The unreacted
iodine is then calculated by the addition of potassium iodide and titration with standard solution of
sodium thiosulphste using starch as indicator.

Calculate the Iodine value from the following equation:

Iodine value = 1. 269 × (b-a)/W
Where, W = weigh of fat or oil taken.
b = reading of actual titration
a = reading of blank titration
Significance: Iodine value gives us an idea about the proportion of unsaturated fatty acids presents
both in free and combined forms of esters. Susceptibility of rancidity increases for the oils or fats
having higher iodine values. Iodine value helps to indicate the composition of complex mixture, as
well as pure substances. It may be determined by any of the following methods [2, 12, 13, 15, 16].

e) Acetyl value: It is defined as the number of milligrams of potassium hydroxide required to

neutralise acetic acid produced by the saponification of one gram of completely acetylated fat or
oil. It helps in determining the number of alcoholic groups present in oil or fat.
Procedure: to the given sample add 5 ml of acetic anhydride-pyridine mixture (1:7). Add 5
ml of water. Put on a water bath for about 30 minutes then cool it. Titrate with 0.5N KOH
using phenolphthalein as an indicator.
Acetyl value = E ×4.3/ A
Where, A = Weight of sample acetylated (gm)
E = Acidity equivalent
f) Reichert-Meissl Value (R.M. Value): It is defined as the number of ml of 0.1N KOH
solution required to neutralise the water soluble steam or to neutralise the distillate of 5 gm
of hydrolysed fat or oil. It is an indicator of how much volatile fatty acid can be extracted
from fat through saponification. It is a measure of the volatile fatty acid residues present in a
given fat or oil.
Procedure: To the 10 gm of sample add an excess of 0.1N NaOH solution in order to
completely saponify the fat. The solution is then acidified with dil.H 2SO4 and is undergo
steam distillation. The distillate containing the volatile acid is then titrated with 0.1N KOH
solution using phenolphthalein as an indicator.
R.M. value = 1.10 (T1-T2)
Where, T1 = volume of 0.1N KOH used for the
titration T2 = volume of 0.1N KOH used for
blank titration
Significance: R.M. value is useful for testing the purity of the butter and desi ghee which
may contain a high amount of glycerides of butyric acid and other steam volatile fatty acid
residues. For e.g. Adulterated butter has low R.M. value than that of pure butter. This R.M.
value number is an indicator of non-fat compounds in edible fats like butter and ghee. Hence,
it helps in determining the purity of ghee and butter .