 Alkaloids

INTRODUCTION –
One of the largest groups of chemicals produced by plants is the alkaloids. Many of these
metabolic by-products are derived from amino acids and include an enormous number of
bitter, nitrogenous compounds. More than 10,000 different alkaloids have been discovered
in species from over 300 plant families. Alkaloids often contain one or more rings of carbon
atoms, usually with a nitrogen atom in the ring.
The term “alkaloid” (alkali-like) is commonly used to designate basic heterocyclic nitrogenous
compounds of plant origin that are physiologically active.
Alkaloids are derived from plant sources, they are basic, they contain one or more nitrogen
atoms (usually in a heterocyclic ring) and they usually have a marked physiological action.
Alkaloids are cyclic organic compounds containing nitrogen in a negative state of oxidation with
limited distribution among living organisms.
E.g., atropine, caffeine, morphine, nicotine, quinine, and strychnine.
Ergot alkaloids a group of chemically related alkaloids either derived from ergot or synthesized; 
some cause ergotism while others are medicinal.
Vinca alkaloids alkaloids produced by the common periwinkle plant (Vinca rosea); two, vincristi
ne and vinblastine, are used as antineoplastic agents.
Biosources
Alkaloids are present in varying quantities, such as:
1) More in dicots than in monocot plants.
2) Lesser common in lower plants.
3) Apocynaceae, Rubiaceae, Solanaceae, and Papaveraceae families have abundance of alkaloids, while
Rosaceae and Labiatae families are free from alkaloids

Function of alkaloids: -The purpose of existence of alkaloids in plants i.e. their functions in
plants is uncertain. There are various views by different authorities, such as they are of no
importance and may be regarded as byproducts of plant metabolism. They may act as
reservoirs for protein synthesis. They may act as protective substances against the animal or
insect attacks. Like hormones, they may function as plant stimulants or regulators in activities
like growth, metabolism and reproduction. Or they may function as detoxicating agents by
methylating, condensing, and cyclizing the compounds whose accumulation might otherwise
cause damage to the plant.
Properties of Alkaloids:-
Physical Properties:
1- Most alkaloids contain oxygen; those compounds are usually colorless crystals at ambient
conditions. 
2- Oxygen-free alkaloids, such as nicotine or coniine, are typically volatile, colorless, oily liquids.
3- Some alkaloids are colored, like berberine (yellow) and sanguinary (orange).
4- Most alkaloids have a bitter taste. It is believed that plants evolved the ability to produce
these bitter substances, many of which are poisonous, in order to protect themselves from
animals; however, animals in turn evolved the ability to detoxify alkaloids.
Chemical Properties:
1- Most alkaloids are weak bases, but some are amphoteric, for example theo bromine and
theophylline.
2- Most alkaloids are poorly soluble in water but readily dissolve in organic solvents, such as
diethyl ether, chloroform and 1,2-dichloroethane. However, caffeine dissolves well in boiling
water. 
3- With acids, alkaloids form salts of various strengths. Those salts are usually soluble in water
and alcohol and poorly soluble in most organic solvents. Exceptions include scopolamine hydro
bromide, which is soluble in organic solvents and water-soluble quinine sulfate.

There are three main types of alkaloids:

(1) True alkaloids : Having heterocyclic ring with nitrogen and derived from amino acids.
For example: Atropine

(2) Proto alkaloids: It does not have heterocyclic ring with nitrogen and derived from amino
acids. For example Colchicine, Ephedrine etc

(3) Pseudo Alkaloids: Having heterocyclic ring with nitrogen but not derived from amino
acids.For example : Caffeine

On the basis of chemical nature:

Non heterocyclic alkaloids:

1. True and proto alkaloids (Amino acid derived alkaloids)

a. Ornithine and Lysine derived alkaloids

i. Pyrolidine alkaloids e.x. Nicotine

ii. Tropane alkaloids e.x. Atropine, Hyoscine

iii. Pyrolizidine alkaloids e.x. Alkaloids of Borage and Symphytum

iv. Quinolizidine alkaloids e.x. Lupanine and sparteine

v. Indolizidine alkaloids e.x. Castanospermine (Anti-HIV)

vi. Piperidine alkaloids e.x. Lobeline, Piperine, pelleterine and coniine

b. Phenylalanine and tyrosine derived alkaloids

i. Phenyl ethyl amine alkaloids e.x. Ephedrine

ii. Isoquinoline alkaloids e.x. Papaverine

iii. Aporphine alkaloids e.x. Apomorphine

iv. Morphinans e.x. Morphine, codine, thebain

c. Tryptophan derived alkaloids

i. Tryptamine and carbolines e.x. Muscarine, serotonine, harmine and harmaline

ii. Indolines e.x. Neostigmine and physiostigmine

iii. Ergolines e.x. Lysergic acid derivative ergot alkaloids

iv. Monoterpenoid indole e.x. vinca, cinchona, nux-vomica, rauwolfia and camptotheca
alkaloids.

d. Histidine derived alkaloids e.x. Pilocarpine

2. Pseudo alkaloids (non amino acid derived alkaloids)

a. Alkaloids derived from terpene metabolism

i. Mono and sesque terpene alkaloids e.g. alkaloids of Nymphea sp.

ii. Diterpene alkaloids e.g. alkaloids of Aconite sp.

iii. Steroidal alkaloids e.g. alkaloids of Solanum and Veratrum sp. and kurchi alkaloids.

b. Purine bases e.g. caffeine and theine

c. Miscellaneous alkaloids like peptide alkaloids of Ziziphus sp. and maytansinoids.

action of pupil

DRAW THESE STRUCTURES

Forms of Alkaloids

1) They may occur in the form of free bases.

2) They may occur in the form of salts with organic acids, e.g., oxalic acid and acetic acid.

3) They may occur in the form of salts with inorganic acids, e.g.,HCl andH2SO4.

4) They may occur in the form of salts with specials acids, e.g., meconic acid in opium and quinic
acid in cinchona.

5) They may occur in the form of glycosides, e.g., solanine in solanum.

Functions in Plants
1) Their bitterness and toxic nature provide protection against herbivores.

2) They are the waste products of detoxification.

3) Being the major source of nitrogen, they can cure nitrogen deficiency.

4) They also aid in regulating growth in some metabolic systems.

5) Those containing a sugar moiety may be utilised as a source of energy in deficiency of

carbon dioxide assimilation.

Extraction of Alkaloids:

 There is no single method of extracting alkaloids from natural raw materials due to the
structural diversity of alkaloids.

 Plants contain several alkaloids, firstly their mixture is extracted with suitable solvent
and then individual alkaloids are separated.

 Mostly alkaloids are present in the plant material in the form of salts of organic acids.

 An extraction of alkaloids involves following methods:

In general, alkaloids may be extracted by any of the following three well -defined and
widely accepted processes:

1) Stas-Otto Process:

Alkaloid extraction is based on its basicity and solubility. Extraction is basically carried
out by Stas-Otto process in which the moistened drug is treated with alkali to set free the
base existing in salt form and then the free base is separated with an organic solvent.
DRAW THIS STUCTURE

2. Kippenberger’s Process: This process is performed as follows:

i) The powdered and sieved plant substance is first and foremost digested with solution
of tannin in glycerol at a co nstant temperature of 40ºC for 48 hours.

ii) The resulting mixture is heated to 50ºC to help in the complete coagulation of
proteinous substance.

iii) Then the mixture is cooled to ambient temperature and filtered.

iv) The resulting filtrate is thoroughly shaken with petroleum ether to get rid of fa ulty
material, and the last traces of petroleum ether are removed from the extract by heating
either on a water bath or exposing to IR lamp.

v) The fat-free crude plant extract is subsequently acidified and shaken with chloroform
to remove the bulk of alkaloids.
 vi) The resulting residual extract may still contain narceine and curarine.

vii) However, narceine and morphine may be isolated by passing freshly generated CO2
directly into hydroxide into their corresponding carbonate which is then ultimately
subjected to solvent extraction using alcohol and chloroform mixture.

viii) Finally, the third alkaloid, curarine may be extracted by agitation with mixture of
equal volume of ether and chloroform.

ix) A combination of Kippenberger‘s process and Stas -otto process by its application to
the final alcoholic extract obtained by the latter process is found to give a better
separation of alkaloids.

3. Steam Distillation:

This method is specially employed for volatile liquid alkaloids like coniine, nicotine, and
sparteine. This process is not suitable for high molecular weight alkaloids.

Method A: It involves followings steps:

(a) The powdered plant material moistened with water and then mixed with lime that combine
with acids, tannins and phenolic substances and sets alkaloids free.

(b) Extraction process is then carried out by treating with organic solvents such as ether or
petroleum spirit.

(c) The concentrated organic liquid is shaken with aqueous acid and allowed to separate. An
alkaloids salts are now in aqueous liquid.

Method B : it involves following steps:

(a)The powdered plant material extracted with water or aqueous alcohol containing dilute acid.

(b)Unwanted materials and pigments are removed by shaking with chloroform or other organic
solvents.

(c)Then the free alkaloids are precipitated by the addition of excess of sodium bicarbonate or
ammonia and separated by filtration or by extraction with organic solvents.
 The separation and purification of alkaloid mixture sometimes may be done by fractional
precipitation or by fractional crystallization. Chromatographic technique is mainly suitable for
separation alkaloids from extract which are in small quantity.

Purification of Crude Extract of Alkaloids Different alkaloidal crude extracts are purified
by different ways:

1) Direct Crystallisation from Solvent: This simple isolation method is not used for complex
mixtures.

2) Steam Distillation: This method is used for volatile liquid alkaloids like coniine, sparteine ,
and nicotine. However, high molecular weight alkaloids cannot be isolated by this technique.

3) Chromatographic Techniques: This method is ideal for separating majority of plant

alkaloids. The different chromatography techniques such as TLC, HPTLC, column, gas, liquid
and ion-exchange chromatography, etc. are used for separating individual alkaloids from
complex mixtures.

4) Gradient p H Technique: Though alkaloids are basic in nature, there are variations in the
extent of basicity of various alkaloids of the same plant. Depending on this character, the crude
alkaloidal mixture is dissolved in 2% tartaric acid solution and extracted with benzene so that the
first fraction contains neutral and or very weakly basic alkaloids. The pH of aqueous solution is
increased gradually by 0.5 increments up to pH 9 and extraction is carried out at each pH level
with benzene , ether, or chloroform. By this way, the alkaloids with different basicity are
extracted. Strongly basic alkaloids are extracted at the end.

Chemical Test for Alkaloids: The chemical test are performed from neutral or slightly acidic
solution of drug following type of chemical test given by alkaloids are-

TEST FOR ALKALOIDS:

(a) Dragendroff’s Test: To 2-3 ml.filtrate add few drops Dragendroff’s Reagent. Reddish brown
colored precipitate is formed.

Preparation of Dragendroff’s Reagent: Boil 14 g. of sodium iodide with 5.2 gm. Basic bismuth
carbonate in 50 ml. Glacial acetic acid for a few minutes. Allow to stand overnight and filter off the
precipitate of sodium acetate crystals. To 40 ml. of the red brown filtrate and add 160 ml. of ethyl
acetate and 1 ml. Water. Preserve the stock solution in ambered colored bottle. When needed. Add
20 ml of acetic acid to 10 mL. Of this solution and make up to 100 ml.with water.
(b) Hager’s Test: To 2-3 ml. filtrate with few drops Hager’s Reagent gives yellow Colored precipitate
is formed.

Preparation of Hager’s Reagent: A saturated aqueous solution of picric acid used for detection of
alkaloids.

(c) Mayer’s Test: To 2-3 ml. filtrate with few drops Mayer’s Reagent gives cream Colored
precipitate is formed.

Preparation of Mayer’s Reagent: Dissolve 1.36 gm. of mercuric chloride in 60 ml.distilled water. (A)
Dissolve 5 gm. of potassium iodide in 20 ml. distilled water. (B). Mix (A) and (B) and adjust the
volume to 100 ml. with distilled Water

(d) Wagner’s Test: To 2-3 ml. filtrate with few drops Wagner’s Reagent gives Reddish brown
colored precipitate is formed.

Preparation of Wagner’s Reagent: Dissolve 1.27 gm. of iodine and 2 gm. of potassium iodide in 5
ml. of water and make up to the volume to 100 ml. with distilled water.

(e) Tannic Acid Test: To 2-3 ml. filtrate with few drops tannic acid solution gives Buff colored
precipitate is formed.

ISOLATION OF ALKALOIDS: Method -

Powdered crude drug

Defatted by using light petroleum

Defatted Powdered Drug

Moisten & make alkaline by Na2CO3/K2CO3/
NH3/Ca(OH)2

Free Alkaloids in crude powdered drug

Extract with organic solvents like
CHCl3/Ether/Methylene Chloride
Extract (inorganic solvent)
Conc.the extract under vacuum.
Shake the conc.extract with successive quantities of acid like dil H 2SO4

Aqueous Acidic Solution (Alkaloidal Salt) Residual organic fractions

Make Alkaline Extract alkaloids with an
immiscible solvents

It contains pigments fats (if present) & very weak bases

Or CHCl3 soluble alkaloid sulphate
Residual Aq. fraction
Organic solution of Alkaloid bases
Remove Solvent
Crude alkaloid Mixture
Commercial Applications
The commercial applications of alkaloids are given below:

1) As Medicines: In a few serious disorders like hearth -failure, cancer, blood pressure, etc ., alkaloids
serve as lifesaving drugs. Digoxin is used in heart failure and it also improves heart functioning and
maintains life. Vincristine obtained from vinca is used as an anticancer agent. Ephedrine derived
from ephedra is used in blood pressure. Codeine is used as a cough suppressant. Ergot alkaloids
relieve migraine pain.
2) As Insect Repellents and Pesticides: Pyrethrin is an insect repellent and is added in mosquito coils,
mosquito repellents, and pesticides also.
3) For Research and Scientific Study : Alkaloids are used in research and scientific study because of
their specific effects on body. For e xample, atropine causes pupil dilation. For confirming whether a
new substance has similar or opposite effects, it is compared with atropine. Hence, atropine is used
as a standard for comparison.
4) To Catch Animals: Alkaloids were used earlier for capturing heavy animals (like deer, elephants,
etc.) by making them immobile.
DRAW THESE TABLES
2.TERPENOIDS
Introduction
The volatile and aromatic substances present in plants and giving them their characteristic
fragrance or odour are terpenoids. Generally, most of the terpenoids are a group o f
compounds present naturally in plants with some exceptions (i.e., obtained from other sources)
including civet musk, etc. In plants, terpenoids are present in leaves and fruits of higher plants,
conifers, citrus plants, and other plants. Volatile liquid, turpentine obtained from pine trees
yields terpene on isolation.

The term terpene represents the hydrocarbons (C5H8)n , while terpenoids include the
hydrocarbons, as well as their oxygenated derivatives.
Terpenes and terpenoids are found in all volatile oils of plant and animal origin.
During their formation, the isoprene units are linked in head and tail fashion. The number of
units incorporated into a particular terpene serve as a basisi of their classification. Diterpenes
tend to be most abundant in Lamiaceae family and have microbial and antiviral properties.
Carotenoids are another group of compounds formed by the tetraterpenoids. The term terpene
was coined to be used initially for the mixture of isom eric hydrocarbons (molecular formula C
10H16) present in the essential oils yielded from tissues and sap of trees and plants.

Properties
 Since terpenoids occur only in the volatile oils, they are normally colourless liquids or solids
with pleasant smell.
 They are insoluble in water , soluble in alcohol , organic solvent and fixed oils.
 They readily volatilise in steam.
 Most of them are optically active.
 Terpenoids normally contain one or more double bonds and form additive compounds with
halogens, nitrosyl chloride, nitrosyl bromide.
 They oxidised by oxidising agents.
Chemical Classes
(C5H8)n is the general formula for most of the natural occurring terpenoid
 Based on the number of rings present in the structure, each class of terpenoids are further divided
into the following sub-classes:

1) Acyclic Terpenoids: They contain open structure.

2) Monocyclic Terpenoids: They contain one ring in the structure.

3) Bicyclic Terpenoids: They contain two rings in the structure.

4) Tricyclic Terpenoids: They contain three rings in the structure.

5) Tetracyclic Terpenoids: They contain four rings in the structure.

Chemical test
Salkowski test- To each 0.5g of the extract was added 2ml of chloroform. Concentrated
sulphuric acid was carefully added to form a layer. A reddish brown colouration of the interface
indicated the presence of terpenoids.
Qualitative Analysis
The qualitative analysis of terpenoids is done by performing the following tests:
1) Noller’s Test: The sample to be analysed is mixed with tin and thionyl chloride and subjected
to warmth. Pink colouration appears if tri -terpenoids are present.
2) Test for Terpenoids: In a test tube, the plant extract (0.5gm) is taken along with chloroform
(2ml). Conc. H 2SO4 (5ml) is added carefully in the tube in such a manner that it forms a
separate layer. Appearance of reddish brown colouration at the junction of two layers confirms
the presence of terpenoids.
Quantitative Analysis
The terpenoids are quantitatively analysed by extracting them from a crude drug and then
separating or isolating by the following three steps:
1) Extracting terpenoids and essential oils,
2) Separating essential oils, and
3) Isolating terpenoids from essential oils.

3. GLYCOSIDES
Glycosides are the organic compound obtained from plants or animal sources which undergoes
enzymatic hydrolysis or acids hydrolysis to give one or more sugar moiety and non-sugar
moiety. The former part is called glycon and latter portion is called as aglycon or genin.
Chemically glycosides are acetals or sugar ethers which are formed by interaction of hydroxyl
group. Sugars involved in glycosides are rhamnose,fructose, galactose, glucose, arabinose,
ribose etc.
classification of glycosides is based on the chemical nature of the aglycone moiety The most
acceptable present in them, namely:
(i) Anthracene glycosides
(ii) Phenol glycosides
(iii) Steroid glycosides
(iv) Flavonoid glycosides
(v) Coumarin and Furanocoumarin glycosides
(vi) Cyonogenetic glycosides
(vii) Thioglycosides
(viii) Saponin glycosides
(ix) Aldehyde glycosides
(x) Bitter glycosides
(xi) Miscellaneous glycosides

Biogenesis
Formation of glycosides involves two steps:
1) The first step involves the reactions resulting in the formation of aglycones, and
DRAW THE REACTION

2) The second step involves the metabolic pathway inv olving coupling of aglycones with
sugar moiety. To synthesise O-glycosides found in plant cells, the nucleotide glycoside
(such as UDP -glucose) and the alcoholic or phenolic group of the second moiety
(aglycones) interact with each other. The other naturally occurring glycosides are C-
glycosides (involving the linkage is through carbon), N -glycosides (linkage through
nitrogen), or S-glycosides (linkage through sulphur).

DRAW THE REACTION

 The glycosides may contain either of the types of sugars:
1) Monosaccharides (rhamnose, glucose, and fructose), or
2) Deoxysugars (digitoxose or cymarose) as in cardiac glycosides.

 ON THE BASIS OF GLYCON PORTION

 EXAMPLES-GLYCOSIDES WITH GLUCOSE
 Rhamnosides with rhamnose
 Fructosides with fructose
 Galactosides with galactose.

ON THE BASIS OF THERAPEUTIC ACTIVITY OF AGLYCON PORTION

 CARDIOTONIC
 ANTIEMATIC
 ANTIARTHMIC
 ANTIRHEUMATIC

BASED ON THE LINKAGE OF AGLYCON PORTION

C GLYCOSIDE
 -glycosides isolated from different plant genus and charcterized spectroscopically are:
Carminic acid, Aloin, Scoparin, Saponarin,’.
O-GLYCOSIDE
A type of carbohydrate-proteincovalentlinkagebetween a serine or
threoninehydroxylsidechainamideand a sugar;type II linkage
Examples-Senna and rhubarb
S GLYCOSIDES

 Example-sinigirnfrom black mustard.

N GLYCOSIDES

 Example-nucleosides.

ISOLATION OF GLYCOSIDES:

 The dried plant material is rendered into a moderately coarse powder.

 The powder is then extracted in a soxhlet apparatus with aqueous-ethanol.
 The non-glycoside impurities which get extracted along with glycosides are removed by
precipitating them with lead acetate solution.
  The excess of lead acetate is then removed by passing hydrogen sulphide gas through
the extract.
 Lead gets precipitated as lead sulphite which is filtered out.
 The filtrate contain the glycoside.
 The glycoside can be obtained by removal of the solvent under reduced pressure or any
other suitable procedure.
 Further purification of the isolated glycosides is done by column chromatography.

Chemical tests of glycosides

Glycosides are the compounds with organic molecules having attached glucose or any mono-
oligo saccharide unit. Usually, these are crystalline or amorphous solids; optically active, soluble
in water and alcohol but insoluble in organic solvents like ether, chloroform and benzene etc.
Generally, aqueous or alcoholic extracts of crude drugs are tested with specific reagents for
presence of various types of glycosides.
 Borntrager’s test: To 1 gm of drug add 5-10 ml of dilute HCl boil on water bath for 10
min and filter. Filtrate was extracted with CCI/ benzene and add equal amount of
ammonia solution to filtrate and shake. Formation ofpink or red colour in ammonical
layer due to presence of anthraquinone moiety.
 Modified Borntrager’s test:To 1 gm of drug, add 5 m1 dilute HCl followed by 5 ml ferric
Chloride (5% w/v). Boil for 10 min on water bath, cool and filter, filtrate was extracted
With carbon tetrachloride or benzene and add equal volume of ammonia solution,
formation of pink to red colour due to presence ofanthraquinone moiety. This is used C-
type ofanthraquinone glycosides.

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4. VOLATILE OILS
Volatile oils are odorous volatile principles of plant and animal source, evaporate when exposed
to air at ordinary temperature, and hence known as volatile or etheral oils. These represent
essence of active constituents of the plant and hence also known as essential oils
Chemically, they are derived from terpenes and their oxygenated compounds. They are made
up of isoprene units (C6H8) and are usually mono-, sesqui and diterpenes with empirical
formulae as C5H3, C10H16, C15H24 and C20H32 respectively. Volatile oils are soluble in alcohol, ether
and other lipid solvents and practically insoluble in water. They are usually lighter than water.
They possess characteristic odours and they have high refractive index. Most of them are
optically active. They are secreted in special structures such as duct, cell, schizogenous or
lysigenous glands, trichomes, etc. They are commonly found in the species of Labiatae,
Rutaceae, Piperaceae, Zingiberaceae, Umbelliferae, Myrtaceae and Lauraceae. They are present
in entire plant or in any part of the plant.
Chemistry
The volatile oils chemically comprise of the following:
1) Terpene Hydrocarbons
i) Monoterpenes
a) These compounds are present in almost all volatile oils, and have a 10 carbon atom structure
(derived from two isoprene units ) and not less than one double bond.
b) These compounds react to air and heat sources. Due to this , citrus oils do not last long, since
the content of monoterpene hydrocarbons is high in them and they react quickly with air and
get easily oxidised.
ii) Sesquiterpenes

a) These compounds contain 15 carbon atoms and exhibit complex pharmacological actions.

b) These compounds show anti-inflammatory and anti -allergic properties. Farnesene is ano ther
sesquiterpene that occurs in chamomile, rose, and other floral oils.

2) Oxygenated Compounds
i) Phenols
a) Generally, a carbon side chain is present in phenols found in volatile oils. Examples of such
compounds are thymol, eugenol, and carvacrol. These compounds show antiseptic, anti
-bacterial, disinfectant and stimulating therapeutic properties.
c) They show antiseptic properties, but are also considered as irritants for skin and muc ous
membrane, e.g., cinnamon and clove oil can cause severe skin reactions
ii) Alcohols
a) Monoterpene Alcohols
These alcohols exhibit good antiseptic, anti -viral and anti -fungal properties, and produce very
less side effects like skin irritation or toxicity. They also show a stimulating effect.
b) Sesquiterpene Alcohols
These alcohols are rarely present in the volatile oils, but bisabolol in German chamomile exhibit
properties like liver and glandular stimulant, anti-allergen, and anti-inflammatory.
3) Aldehydes
These compounds show anti -fungal, anti -inflammatory, disinfectant, sedative yet stimulating
therapeutic properties. These compounds impart the citrus -like smell in melissa, lemongrass ,
and citronella. These properties of aldehydes are utilised in aromatherapy when the volatile oil
is used in low dilution, i.e., around 1%.
4) Ketones
i) In thujone (found in thuja ), wormwood oil , and picocamphone , the ketones present can be
toxic; however, these co mpounds also exhibit therapeutic effects as they facilitate mucus
secretion and tissue regeneration

Chemical tests for Volatile Oils:

Presence of volatile oil in the natural drugs can be detected by the following tests:
1. To the thin section of the drug, add alcoholic solution of Sudan Ill. Red colour obtained
by globules indicates the presence of volatile oil.
2. To the thin section of the drug, add a drop of tincture alkane. Red colour indicates the
Presence of volatile oil.
CLASSIFICATION OF VOLATILE OILS
Volatile oils are classified on the basis of functional groups
Classification of volatile oil
Groups Drugs
Hydrocarbons Turpentine oil

Alcohols Peppermint oil, Pudina, Sandalwood oil, etc.

Aldehydes Cymbopogon sp., Lemongrass oil, Cinnamon,Cassia, and Saffron
Ketones Camphor, Caraway and Dill, Jatamansi, Fennel, etc.
Phenols Clove, Ajowan, Tulsi, etc.
Phenolic ethers Nutmeg, Calamus, etc.
Oxides Eucalyptus, Cardamom, and Chenopodium oil
Esters Valerian, Rosemary oil, Garlic, Gaultheria oil, etc.

Biosources

Volatile oils are present in the following plant parts:

1) Glandular Hair: Mint stems and leaves (Labiatae).

2) Mesophyll: Eucalyptus leaves.

3) Modified: Piperaceae and Parenchyma.

4) Vitale: Anise (Umbelliferae).

5) Lysigenous: Gossypium species.

6) Schizogenous Gland: Pinaccae and Rutaceae.

7) All Tissues: Conifers.

8) Petals: Rose.

9) Bark: Cinnamon.

10) Rind: Orange.

11) Hear Wood: Sandalwood.

EXTRACTION OF VOLATILE OILS- Volatile oils are prepared by means of several techniques and
those techniques are discussed below:

Extraction by Distillation-

The distillation is carried out either by water or steam. The volatile oils from fresh materials are
separated by hydro distillation, and volatile oils from air dried parts are separated by steam
distillation. However it is better to use fresh materials in either case.

Extraction by Scarification-

This method is used for the preparation of oil of lemon, oil of orange, and oil of bergamot. The
two principal methods of scarification are the sponge and the ecuelle method.

(a) Sponge Process:

In this process the contents of the fruit are removed after making longitudinal or transverse cut,
and the peel is been immersed in water for a short period of time. Then it is ready for
expression. The operator takes a sponge in one hand and with the other presses the softener
peel against the sponge, so that the oil glands burst open and the sponge absorbs the exuded
oil, which is transferred to a collecting vessel. The turbid liquid consisting of oil and water is
allowed to stand for a short time, whereupon the oil separates from water and is collected. The
whole of the above process is carried out in cool, darkened rooms to minimize the harmful
effects of heat and light on the oil.

(b) Ecuelle Process: In this process, the rinds are ruptured mechanically using numerous
pointed projections with a rotary movement and the oil is collected.
Extraction by Non-Volatile Solvent

A nonvolatile solvent, for example, a fine quality of either lard or olive oil, is used in this
process. After saturation with the floral oil the lard or olive oil is sometimes used as a flavouring
base for the preparation of pomades, brilliantine, etc., or converted to a triple extract. In the
latter instance the lard or oil is agitated with two or three successive portions of alcohol, which
dissolve the odorous substances. The mixed alcoholic solutions so obtained constitute the‘triple
extract’ of commerce.

There are three chief methods that come under this; they are enfleurage, maceration and a
spraying process.

(a)Enfleurage:

In this a fatty layer is prepared using lard and the flower petals are spreaded over it, after the
imbibitions is over the fatty layer is replaced with fresh petals. After the saturation of fatty layer
the odorous principles are removed by treating with alcohol and a triple extract then prepared.
When oil is used as a solvent the flowers are placed on an oil-soaked cloth supported by a metal
grid enclosed in a frame. Fresh flowers are added as required, and finally the oil is expressed
from the cloths. It may then be used as perfumed oil, or extracted with alcohol to produce a
triple extract.

(b) Maceration:

This is also used to extract the volatile matters of flowers. The lard or oil is heated over a water
bath, a charge of flowers added and the mixture stirred continuously for some time. The
exhausted flowers are removed, pressed, the expressed fluid returned to the hot fat, fresh
flowers, added and the process continued until defined weights of flowers and solvent have
been used. Again, a triple extract is prepared by extracting the perfumed lard or oil with
alcohol.

(c) Spraying:

In this process a current of warm air is sprayed through a column of the flowers. Then oil or
melted fat is sprayed over this oil-laden air which absorbs and dissolves most of the perfume,
the collected oil or fat is then extracted with alcohol as described above.

Extraction by Volatile Solvent

In this the flowers are extracted by using the solvent light petroleum and the latter is distilled
off at a low temperature, leaving behind the volatile oil.
Therapeutic Uses

Volatile oils are employed as flavouring and perfuming agents in pharmaceutical

products, foods, beverages, and in cosmetics.

They serve as medicinal agents for therapeutic effects like:

1) Carminative (e.g., umbelliferous fruits),

2) Anthelmintic (e.g., chenopodium oil),

3) Diuretics (e.g., juniper),

4) Antiseptic (e.g., eucalyptus),

5) Counter irritant (e.g., oil of wintergreen),

6) Local anaesthetic (e.g., clove),

7) Sedative (e.g., jatamansi),

8) Local irritant (e.g., turpentine),

9) Insect repellent (e.g., citronella), and

10) Source of vitamin A (e.g., lemongrass).

TANNINS

The term tannin was first applied by Seguin in 1976 to denote substances present in plant
extracts which are able to combine with proteins of animal hides, prevent their putrefaction
and convert them into leather.
Tannins are one of the most widely occurring groups of natural substances in different
families of higher plants. These secondary metabolites are present in solution form in the cell
sap and also in distinct vacuoles. They have been known since long time as the astringent
substances, having the capacity to combine with tissue proteins and precipitate them. They
are, therefore, used in medicines or allied purposes or as mild antiseptics, in treatment of
diarrhoea, and to check small haemorrhages. Commercially, they find extensive application in
leather industries, when the skins of animals are treated with tannins to prevent the
putrefaction.
Tannins form colloidal solutions with water and are non-crystalline substances. In solution,
 they show acidic reactions due to phenols. They are also soluble in alcohol, glycerine, dilute
alkalies, but practically insoluble in organic solvents except acetone. Tannins exhibit some
specific chemical reactions.
Tannins are defined as, ‗Complex substances that occur as mixtures of
polyphenols that are very difficult to separate since they do not crystallise‘.

Definition:-
“Chemically, they contain the mixture of complex organic substances in which polyphenols
are present, generally with o-dihydroxy or o-trihydroxy groups on a phenyl ring.”
Normally, they have high molecular weight and unlike alkaloids are devoid of nitrogen.

Classification of Tannins:-

The tannins are broadly classified into three groups based on complexity of their chemical
nature and according to their behaviour on dry distillation –
(1) Hydrolysable Tannins : As the name indicates, these tannins are hydrolysed by acid or
enzymes quickly and the products of hydrolysis are gallic acid or ellagic acid. On dry
distillation, gallic acid and other components get converted to pyrogallol. They respond to
ferric chloride solution, producing blue colour. The examples of hydrolysable tannins are
gallotannin in nutgall, rhubarb, amla, clove and chestnut; ellagitannin from oak, myrobalans
and pomegranate bark.

(2) Condensed Tannins : They are also called as non-hydrolysable tannins, phlobatannins or
proanthocyanidins. They are much resistant to hydrolysis. They are related to flavonoid
pigments, because they are formed via derivatives of flavones, like catechin or flavan-3-ol or
flavan-3, 4-diols. Unlike the hydrolysable tannins, on treatment with enzymes or mineral
acids, they are polymerised or decomposed into red coloured substances called
phlobaphenes, which are insoluble in water and indicate the typical brownish- red colour of
many plants and drugs. On dry distillation they yield catechol. Tannins with ferric chloride
produce brownish-green colour.
They are distributed in different parts of plants. The green tea and hamamelis leaves,
cinchona, cinnamon and wild cherry bark, male fern rhizome, cocoa, cola and areca seeds,
pale and black catechu contain these types oftannins.
(3) Pseudotannins : This is not as a separate group of tannins, but may be treated as sub
group because they do not obey to goldbeater’s skin test and are low molecule weight
compounds. Chlorogenic acid in coffee and nuxvomica, ipecacuanhic acid in ipecacuanha and
catechins in cocoa are examples of pseudotannins.
The detection test for chlorogenic acid is carries out by extracting the drug with water and
treating this extract with ammonia solution, followed by exposure to air, which leads slowly
 to formation of greencolour.
Biosources
Rhubarb, red rose petals, bearberry leaves, Chinese galls, Turkish gall, hamamelis, che stnut,
and maple are rich in gallotannins. Pomegranate rind, pomegranate bark, myroba lans,
eucalyptus leaves, some Australian kinds , and oak bark are rich in ellagitannins. Some drugs,
e.g., tea, ham amelis bark and leaves contain both hydrolysable and condensed tannins.

CHARACTERISTICS OF TANNINS
1. Tannins are colloidal solutions with water.
2. Non crystalline substance.
3. Soluble in water (exception of some high molecular
weight structures), alcohol, dilute alkali, and glycerin.
4. Sparingly soluble in ethyl acetate.
5. Insoluble in organic solvents, except acetone.
6. Molecular weight ranging from 500 to >20,000.
7. Oligomeric compounds with multiple structure unit with free phenolic groups.
8. Can bind with proteins and form insoluble or soluble tannin—protein complexes
ISOLATION OF TANNINS

 Both hydrolysable and condensed tannins are highly soluble in water and alcohol but
insoluble in organic solvents such as solvent ether, chloroform, and benzene.
 Tannin compounds can be easily extracted by water or alcohol.
 The general method for the extraction of tannic acid from various galls is either with
water-saturated ether, or with mixture of water, alcohol, and ether.
 In such cases, free acids such as Gallic and ellagic acid go along with ether, whereas true
tannin gets extracted in water.
 If the drug consists of chlorophyll or pigment, it may be removed by ether.
 After extraction, the aqueous and ethereal layers are separately concentrated, dried,
and subjected to further isolation and purification using various separation techniques
of chromatography.

Chemical tests for Tannins :-

(1) Gelatin test : To a solution of tannin, aqueous solution of gelatin and sodium chloride are
 added. A white buff coloured precipitate isformed.

(2) Goldbeater’s skin test : A small piece of goldbeater skin (membrane prepared from the
intestine of an ox) is soaked in 20% hydrochloric acid, ringed with distilled water and placed
in a solution of tannin for 5 minutes. The skin piece is washed with distilled water and kept in
a solution of ferrous sulphate. A brown or black colour is produced on the skin due presence
oftannins.

(3) Phenazone test : A mixture of aqueous extract of a drug and sodium acid phosphate is
heated and cooled and filtered. A solution of phenazone is added to the filtrate. A bulky
coloured precipitate isformed.

(4) Match stick test (Catechin test) : A match stick is dipped in aqueous plant extract, dried
near burner and moistened with concentrated hydrochloric acid. On warming near flame, the
matchstick wood turns pink or red due to formation ofphloroglucinol.

(5) Chlorogenic acid test : An extract of chlorogenic acid containing drug is treated with
aqueous ammonia. A green colour is formed on exposure toair.

(6) Vanillin-hydrochloric acid test : Sample solution and added vanillin- hydrochloric acid
reagent (Vanillin 1 gm, alcohol 10 ml, concentrated hydrochloric acid 10 ml). A pink or red
colour is formed due to formation ofphloroglucinol.

Therapeutic Uses
The medicinal uses of tannins are:
1) Styptics: Tannin-containing drugs precipitate proteins and have been used traditionally
as styptics and internally for the protection of inflamed surface of mouth and throat.

2) Anti-diarrhoeal: They act as anti -diarrhoeal and have been employed as antidotes in
poisoning by heavy metals, alkaloids, glycosides. In Western medicine, their use
declined after World War -II when it was found that absorbed tannic acid can cause
severe liver necrosis.

3) Anti-Tumour: They exhibit a strong anti -tumour activity. Ellagitannins monomer units
having galloyl groups at the C -2 and C -3 positions on the glucose core exhibit anti-
tumour activity.

4) Antidote: They are used as antidote s in alkaloid poison ing. They form insoluble and
non -toxic tannates with alkaloids. Only dilute solution is applied for this purpose.
1. RESINS
The hydrocarbon secretion of various pla nts, especially the coniferous trees is known as the
resin which is considered valuable because of its chemical constituents and various uses like
adhesives, varnishes, perfumes, incense, nail polish, etc. Apart from this, resins are also used as
a raw material for synthesizing organic compounds. The fossilised resins act as a source of
amber.
Resins are amorphous products of complex chemical nature. These are amorphous mixtures of
essential oils, oxygenated products of terpene and carboxylic acids found as exudations from
the trunk of various trees. They contain large number of carbon atoms. Chemically, they
contain organic acids, alcohols, esters and neural resins.

Properties
 They are transparent or translucent solids, semi-solids or liquid substances.
 Most of the resins are heavier than water.
 They are insoluble in water, but soluble in alcohol, volatile oils, fixed oils.
 They are hard, electrically non-conductive and combustible masses.
 When heated, they soften and ultimately melt.
They are transparent or translucent solids, semi-solids or liquid substances containing large
number of carbon atoms. Most of the resins are heavier than water. They are insoluble in
water, but soluble in alcohol, volatile oils, fixed oils, chloral hydrate and non-polar organic
solvents like benzene or ether. They are hard, electrically non-conductive and combustible
masses. When heated, they soften and ultimately melt. They are usually formed in
schizogenous or schizolysigenous cavities or ducts as end products of metabolism.
Chemically, they contain organic acids, alcohols, esters, and neutral resins. Depending upon the
type of the constituents of the resin.
They are further classified as: (a) Acid resin, (b) Ester resin, and (c) Resin alcohols.
1. Acid resins: Following are few examples of this type of resins alongwith their acids;
colophony (abietic acid), safetid (sandracolic acid), copaiba (copaivic and oxycopaivic acids),
myrrh (commiphoric acid) and shellac (alleuritic acid).
2. Ester resins: This group contains esters as the chief constituents of the resins, e.g.
benzoin and storax. Benzoin contains coniferyl benzoate and storax contains cinnamyl
cinnamate.
 3. Resin alcohols The contents are the complex alcohols of high molecular weight. They are
either found in free state or as esters. The examples are balsam of peru with
peruresinotannol, gurjan balsam with gurjuresinol and guaiacum resin with guaic-resinol.

Biogenesis
Resins can be obtained from carbohydrate produced via photosynthesis. These
carbohydrates breakdown into pyruvate products (simpler compou nds), from which
terpenoid and phenolic compounds can be synthesised via different metabolic pathways

Quantitative Analysis
Balsam and gum resin s can be estimated on the basis of quantity by the
determination of the following:
1) Acid value,
2) Ester value (direct or indirect),
3) Saponification value, or resin value and gum value,
4) Percentage of moisture,
5) Ash,
6) Percentage soluble in alcohol,
7) Percentage insoluble in alcohol,
8) Specific gravity, and
9) Percentage soluble in other solvents.

ISOLATION Methods

Method – A Powdered drug

1. Extract the resin with alcohol
2. Filter
3. Concentrate extract an excess of water, shake Resins get precipitate.
Method – B Powdered drug containing oleo-resin, percolate the powdered drug with non-
polar solvent (e.g. acetone, chloroform)

IDENTIFICATION TEST
Three types:
1.Physical test Such as solubility, taste, odor and examination of powder under the
microscope
2.Chemical test – For the identification of resins Acid Value Saponification Value Iodine
value
 3. Specific chemical test – For specific constituents such as Cinnamic acid, Benzoic acid in
Benzoin, Tolu balsam and Peru Balsam

CHEMICAL TESTS
HCl TEST: HCl Formation of pink color Presence of resins
FeCl3 TEST:-Fecl3 Greenish blue color Presence of resins