PHARMACOGNOSY (Y4S2) – BIOC 426

INTRODUCTION

Definition

Pharmacognosy is the formal study of natural products – plants, animals, minerals – used in
medicine. The word is derived from Greek pharmakon (drug) and gnosis (study). Natural
products have been part of medical practice for thousands of years. Drugs like opium, ginseng
and hyoscyamine have been part of medical practice for thousands of years.

Abroad spectrum of biological subjects encompass pharmacognosy e.g botany, ethnobotany,

medical anthropology, marine biology, microbiology, herbal medicine, chemistry,
biotechnology, phytochemistry, pharmacology, pharmaceutics, clinical pharmacy and
pharmacy practice.

Originally, nearly all medicines were derived from plants, hence in the beginning of the 19 th
century, pharmacognosy was all there was to the pharmacy profession. Eventually,
pharmacognosy became one of the biological disciplines taught in pharmacy schools and
colleges, which was concerned with the source and role of natural products utilized in
allopathic medicine. These natural products consisted mainly of plant derived materials and a
few from animals sources. Pharmacognosy was thus intimately related to the botany and
chemistry of the drugs and their therapeutic application.

History

 Since the beginning of civilization, people have used plants as medicine. Perhaps as
early as Neanderthal man, plants were believed to have healing powers.
 Evidence from mural inscriptions from ancient Egypt dating back to 3000 BC show
knowledge of the effect of medicinal plants.
 Clay tablets in the British Museum show that Sumerians had a form of herbal around
2500 BC.
 The earliest recorded uses are found in Babylon circa 1770 BC in the Code of
Hammurabi and,
 In ancient Egypt circa 1550 B.C. Ancient Egyptians believed medicinal plants to have
utility even in the afterlife of their pharaohs.
 At 660 BC, around 250 drugs were recognized by the Assyrians and some were
actively cultivated.
 Hippocrates was well acquainted with drugs-467 BC.
  Theophrastus, a pupil of Aristotle listed 500 plants known to him and distinguished
cinnamon from cassia.
The study of crude natural products in medicine, under the name pharmakognosie was first
developed in German-speaking areas of Europe, while other language areas often used the
older term materia medica taken from the works of Galen and Dioscorides.

 Galen was a Greek physician (circa-130-210 AD). The name galenical (below) is
derived from this physician’s name.
 Dioscorides’ Materia Medica, was compiled around 1334 and described medicinal
features of various plants in Arabic.

The term ‘Pharmacognosy’ was used for the 1st time by the Austrian physician Schmidt in
1811. In 1815 the German Anotheus Seydler in a work titled Analecta
Pharmacognostica..described pharmacognosy as… “the science which has the task to
learn everything about drugs originating from plants or animals in all aspects, except
the physiological effect , to describe them correctly and under a general vision connect
this knowledge.”

During the 19th century and the beginning of the 20th century, "pharmacognosy" was used to
define the ‘branch of medicine which deals with drugs in their crude, or unprepared form’.

Crude drugs;

the dried, unprepared material of plant, animal or mineral origin, used for medicine.

In this period, pharmacognosy was by far the most important pharmaceutical discipline, the
"mother" of all present day pharmaceutical disciplines. Microscopy was introduced for the
quality control of phytopharmaceuticals in the 19th century.

Synthesis of aspirin and its impact on position of pharmacognosy in colleges of pharmacy

Over a 100 years ago (1899), Aspirin -the first example of using nature as lead for a new
synthetic drug was introduced. This was the first signs for a new era.

Gradually, synthetic chemistry became more important for developing new drugs, and the
number of herbal preparations in the pharmacy were rapidly decreasing. The wish to have
pharmaceutical formulations with single pure compounds with well defined activities became the
in thing.

Pharmacognosy had great difficulties in holding its previous prominent position in the
pharmaceutical curriculum in the sixties and seventies.

Fortunately, some visionary pharmacognosists were able to open up new horizons.

The development of thin layer chromatography by the well known pharmacognosy
professor,Egon Stahl is probably the best example (Stahl, 1967). Also, pharmacognosists were
among the pioneers in the analysis of plant materials using gas and high pressure liquid
chromatographic techniques. For studies of the active compounds in plants, these
chromatographic methods became important tools. Moreover, in the seventies, the
spectrometric methods such as MS and NMR became commonplace for the pharmacognosists
in their search for new biologically active compounds in plants. In that period, developing
new drugs from plants was not as easy as with synthetic drugs, as the pharmacological test
systems, mainly in vivo animal experiments and in-vitro isolated organs, are not suited for
bioassay-guided fractionation of active compounds from plant extracts. That requires methods
that can readily measure large numbers of samples in a short time.

However, the structural diversity from synthetic chemistry will never match nature; a novel
active compound like paclitaxel, having 11 asymmetric carbons, will never be designed in a
synthetic laboratory.

As late as the beginning of the 20th century, the subject had developed mainly on the botanical
side, being particularly concerned with the description and identification of drugs (in western
pharmacopoeias) both in their whole state and in powder form. Such branches of
pharmacognosy are still of fundamental importance, particularly for pharmacopoeial
identification and quality control purposes. Rapid development in other areas has enormously
expanded the subject.

Although most pharmacognostic studies focus on plants and medicines derived from plants,
other types of organisms are also regarded as pharmacognostically interesting, in particular,
various types of microbes (bacteria, fungi, etc.), and, recently, various marine organisms.

The scope and practice of modern pharmacognosy

Modern isolation techniques, synthesis and pharmacological testing procedures have yielded
many purified substances most of which have been formulated into useful medicines.
However, this has not solved all the health care problems of the populace from the
pharmaceutical viewpoint. There are still many incurable diseases and diseases for which
the available medicines are either too toxic or very expensive. The search for alternative
sources of pharmaceuticals continues.

In recent years, there has been a reawakened scientific interest in the fundamental role plants
play in many cultures, including medicinal purposes. There is an increased interest in natural
products among the populace.
Even in Kenya, their popularity can be attested by the number of advertisements for herbal
products in the FM radios, the print media, matatus and on trees. Why?

 “Back to nature” fad

 Notion that “natural” drugs are safer than “synthetic” drugs
 Distrust or lack of confidence in conventional products (emergence of many incurable
diseases.
 High costs of conventional drugs.
 Very large parts of the population do not have access to desperately needed
pharmaceutical products.
*This increased prominence of herbal remedies requires additional contributions in
scientific investigation of the same.

What scientific investigation with regard to the natural products?

The following points are worthy to note:

 What is the mechanism of action of the natural product?

 What are the “active” compounds?
 Is there a biological/chemotaxonomic similarity between this and other natural
products?
 What evidence is there to substantiate or disprove the folkloric use of a product?
 Is the product safe?
 Are there interactions between prescription and natural products?
 What are the clinical implications of using “health foods’ and self-medicating with
natural products?
 In addition, what is the clinical impact of poisonous plants (and animals) in our
country?
 Medical community must be well informed about the use of all natural products,
since patients will continue to take them, natural products will remain an important part
of even the most advanced medical therapy.
 Thus proper medical treatment may not be achieved without the knowledge of the
potential hazards or benefits of natural products.
What should be done?

 There is need to investigate today’s herbal medicines in light of modern knowledge.

Products scoffed at in the past often were later widely accepted when a new
underlying mechanism was uncovered. (eg Ephedra tea – decongestant)
 We must weed out the ineffective natural drugs and focus on the potentially useful
ones.
 Many herbal products are presented without adequate quality control or support for
safety or effectiveness. Regulatory requirements should be introduced in Kenya in
order to control the quality of these products and to protect the public from quacks.
 There is need;
 To revert to the scientifically defined botanical standards once used to
establish what constitutes a legitimate, effective drug, it’s safe dose, it’s
appropriate method of administration.
 Data sheets and monographs giving descriptions, tests for identity and purity
and assays should then be compiled.
 WHO monographs and herbal phamacopoeias are examples of on- going work
in other countries.
 Such information can be useful instrument for promoting technical
advancement in Kenya. Cultivation of plants used in the formulation of these
medicines should be encouraged. Research is needed in the agronomy of
cultivated medicinal plants.
 Investigation of the economic, social and environmental impact of medicinal
plant cultivation and collection etc. is also necessary.
 Despite a rapidly expanding literature on phytochemistry and pharmacology, there
are literally thousands of plant drugs that have never been studied thoroughly. In
addition, only a small percentage of the total species (on land and in water) has been
examined. No doubt, there is a large field of future research in the plant and
 kingdom as source of new medicinal substances or as alternative sources of known
substances.
 In western countries, a large number of plants are already being investigated for their
possible pharmacological value as anti-inflammatory, hypotensive, hypoglycemic,
amoebicidal, anti-fertility, cytotoxic, antibiotic, antiparkinsonism, antimalarial, anti
– HIV agents. These are examples of biologically oriented screening programs.
 The westernization of areas with great wealth of knowledge concerning medicinal
plants (Africa, South America, Pacific countries) coupled with a rapid increase in
population may lead to extermination of plant species. Pharmacognosists in
collaboration with other scientists are needed to record local knowledge, then carry out
botanical, chemical, pharmacological and conservational studies before the
knowledge is lost forever.

Examples of drugs uncovered using search of early literature;

 Rauwolfia serpentina, the source of the antipsychotic and hypotensive alkaloids

reserpine, rescinnamine
 Cinchona spp., the source of the antimalarial quinine and the antiarrhythmic
agent quinidine.
 Papaver somniferum or opium, the source of the analgesics morphine and
codeine.
 Chondodendron tomentosum, the source of the curare muscle relaxant, d –
tubocurarine.
 Catharanthus roseus, the source of the antineoplastic alkaloid vinblastine and
antileukemic alkaloid, vincristine.
 Digitalis purpurea and D. lanata which have yielded digitoxigenin and digoxin.
 Atropa belladona (plus other Solanaceae) which are the sources of the
parasympatholytics atropine, scopolamine and hyoscyamine
Biotechnological procedures
 The sudden interest in a plant used in traditional medicine by the world at large may
exhaust local wild sources.
  It becomes necessary to research into the cultivation or artificial propagation by cell
culture of such species as well as other biotechnological procedures.
 Already, basic research into ways of culturing isolated cells and tissues of medicinal
plants for the large scale production of biologically active compounds is being
undertaken by pharmacognosists and plant physiologists.
 Production costs by this approach are relatively high. However, cell culturing offers
an alternative route to the production of drugs that have not yet been synthesized
cheaply.
 In addition, basic info on biosynthetic pathways can be elucidated which may help
researchers duplicate the synthesis in the lab.,
 Example of plants which have been cultivated with some degree of success include:
Digitalis species, Mentha species (mints), Dioscorea species and Papaver somniferum

Products from the sea-

 The disciplines of pharmacognosy, botany, zoology, chemistry, oceanography,

pharmacology and others have collaborated in approaching the resolution of medical
problems by developing products from the sea.
 Apart from searching for drugs from the sea, public health implications of ingestion
of or envenomation by marine toxins is of concern to pharmacognosists.
 A few long used classic drugs from marine sources include agar from the sea weed
Gelidium, Gracilaria and Hypnea spp; cod liver oil from cod fish Gadus morrhua;
protamine sulphate from the sperm or mature testes of salmon; the alginates from
the spp of Fucus seaweed; carrageenan from Chondrus; etc.
 These represent drugs in the older phamacopoeias, we can expect more exotic and
unique biodynamic molecules to be discovered and developed in the future.
 Ziconotide-a parentrally administered analgesic- is a more recent drug developed from
toxins of a marine snail, Conus magus.

Drug abuse-
The drug abuse problem is of concern to pharmacognosy because many abusable compounds
are naturally derived, e.g. opium derivatives like morphine and heroin,
tetrahydrocannabinal, hallucinogenic mushrooms etc

Multidisplinary approach in the study of natural drugs

 A comprehensive understanding of medicinal plants and other natural sources of drugs

inevitably requires a multidisciplinary approach by varied disciplines such as:
commerce, botany, agriculture, horticulture, chemistry, enzymology, genetics,
quality control, pharmacology etc.
 Modern pharmacognosy seeks to embrace these fields in a unified whole for the better
understanding and utilization of medicinal plants and other natural products.

 The main thrust of the discipline of pharmacognosy is to uncover new molecules from
nature that may be of therapeutic value.
 The history, commerce, evaluation of natural product pharmaceuticals, classification of crude
drugs, major phytochemical classes (carbohydrates, alkaloids etc.) are also important in the
study of pharmacognosy.
 Biotechnology
 Quality control of herbal medicines
 Clinical aspects
 Antibiotics
 Poisonous plants
 Drugs of abuse
 Allergens
 Pesticides
 Food additives
 Nutraceuticals
 Medicinal plants used to treat animal diseases
 Marine sources of drugs etc.
 Therapeutic drugs derived from natural products.
 Herb/food and drug interactions
Contemporary study of pharmacognosy

-can be divided into the fields of:

 Medical ethnobotany: the study of the traditional use of plants for medicinal purposes
 Ethnopharmacology: the study of the pharmacological qualities of traditional medicinal
substances
 Phytotherapy (the medicinal use of plant extracts).
 Phytochemistry, the study of chemicals derived from plants (including the identification of
new drug candidates derived from plant sources).
 Zoopharmacognosy, the process by which animals self-medicate, by selecting and using
plants, soils, and insects to treat and prevent disease.
 Marine pharmacognosy,the study of chemicals derived from marine organisms.
 Veterinary pharmacognosy,the study of medicines derived from plants used for
management of animal diseases.

Classification of vegetable drugs for study

1. Alphabetical classification –

Latin or vernacular names are arranged in alphabetical order as in pharmacopoeias, Handbook

of medicinal herbs(Duke J A-2002) etc

2. Taxonomic – Botanical classification in which drugs are arranged according to the plants
from which they are obtained – e.g. example showing the systematic position of peppermint,
Mentha piperita:

 Kingdom Plant
 Division Angiosperm
 Class Dicotyledonaeae
 Subclass Sympetalae
 Order Tubiflorae
 (Suborder) Verbenineae
 Family Labiatae (Lamiaceae)
 (Subfamily) Stachydoideae
 (Tribe) Satureieae
  Genus Mentha
 Species Mentha piperita Linnaeus (Pepperment)
 Varieties Mentha piperita var. vulgaris Sole (Black Peppermint)
Mentha piperita var. officinalis Sole (White Peppermint)

3. Morphological approach-e.g. into:

i. Organized drugs – flowers, fruits, seeds, herbs, entire organisms and herbs, wood,
bark, rhizomes, roots.
ii. Unorganized drugs – dried lattices, extracts, gums, resins, oils, fats, waxes. The
identification of powdered drugs is often based on micro morphological characters.
E g in Jackson and Snowdon (1990).Atlas of microscopy of medicinal plants,
culinary herbs and spices. Bellhaven Press, London.; Wallis T E
(1967).Pharmacognosy 5th edn Churchill Livingstone, London.
4. Chemical or Biogenetic classification – Which is based on the important chemical
constituents (SECONDARY Metabolites) such as alkaloids, glycosides, volatile oils etc or their
biosynthetic pathways. Note that a particular drug may possess a number of active
principles belonging to different phytochemical groups. This is a popular method of study
when teaching of pharmacognosy is phytochemically biased. eg in Dewick P M (2010)
Medicinal Natural Products, a Biosynthetic Approach 3nd edn. John Wiley & Sons Ltd,
Chichester, UK.

5. Pharmacological or Therapeutic – Where classification is according to the

pharmacological action of their most important constituent or their therapeutic use. E.g in
Heinrich M, et al (2012) Pharmacognosy and Phytotherapy. Churchill Livingstone, London.

Geographical sources & Distributions

 There are two factors which determine the commercial geographical sources of a drug i.e.
Suitability of the plant to a particular environment
 Economic factors associated with the production of a drug in a particular area.
Many plants grow equally well in numerous localities having similar climates; and as economic
conditions change in one area, so the collection or cultivation of a drug plant may move in
accordance.

Developing countries may also start the cultivation of medicinal plants, and in this respect South
America, India and S.E. Asian countries have been particularly active. Cinnamon, traditionally
produced in Sri Lanka, has been introduced to the Seychelles as a commercial crop, with so
much success that the plant has now become a weed! Cinnamon also grows well in West Africa
but is not commercially utilized there.

It must be remembered, however, that a plant may grow well in different situations but fail to
produce the same constituents (e.g. cinchonas growing at low altitude and in the plains).

The commercial cultivation of belladonna, stramonium, hyoscyamus and valerian in England

has long been uneconomic and material is now imported from Eastern Europe, largely via
Germany.

Environmental Conditions

a)Temperature

Temperature is a major factor controlling the development and metabolism of plants. Although
each species has become adapted to its own natural environment, plants are frequently able to
exist in a considerable range of temperature. Many tropical and subtropical plants will grow in
temperate regions during summer months, but lack frost resistance to withstand the winter. In
general, the highest temperatures are experienced near the Equator, but as the
temperature falls about 1°C for every 200 m of elevation, it is possible in, say, Jamaica to
have a tropical climate on the coast and a temperate one in the mountains. The formation of
volatile oils appears to be enhanced at higher temperatures, although very hot days may lead to
an excess physical loss of oil.
b)Rainfall
The important effects of rainfall on vegetation must be considered in relation to the annual
rainfall, its distribution throughout the year, its effect on humidity and its effect coupled with the
water-holding properties of the soil.
Continuous rain can lead to a loss of water-soluble substances from leaves and roots by leaching;
this is known to apply to some plants producing alkaloids, glycosides and even volatile oils.
This could account for low yields of some active constituents in wet seasons from plants whose
general condition appears to be good.
With Cassia angustifolia (Tinnevelly senna) it has been shown that short-term drought increases
the concentration of sennosides A+B but in the longer term causes loss of leaf biomass.

c) Day-length and radiation characteristics

Plants vary much in both the amount and intensity of the light which they require. In the wild
state the plant will be found where its shade requirements are met, and under cultivation similar
shade must be provided. In certain cases research has shown that light is a factor which helps to
determine the amount of glycosides or alkaloids produced.
With belladonna, stramonium and Cinchona ledgeriana full sunshine gives a higher content of
alkaloids than does shade.
It has been shown that under long-day conditions peppermint leaves contain menthone, menthol
and traces of menthofuran.

d)Altitude
The coconut palm needs a maritime climate and the sugar cane is a lowland plant. Conversely,
tea, cocoa, coffee, medicinal rhubarb, tragacanth and cinchona require elevation. In the case of
Cinchona succirubra the plants grow well at low levels but produce practically no alkaloids. The
bitter constituents of Gentiana lutea increase with altitude, whereas the alkaloids of Aconitum
napellus and Lobelia inflata and the oil content of thyme and peppermint decrease.
Pyrethrum gives the best yields of flower-heads and pyrethrins at high altitudes on, or near, the
Equator. It is therefore produced in East Africa and north-west South America.

CULTIVATED AND WILD PLANTS

Certain drugs are now obtained almost exclusively from cultivated plants. These include
cardamoms, Indian hemp, ginger, and peppermint and spearmint for oil production. Others
include Ceylon cinnamon, linseed, fennel, cinchona and opium. In other cases both wild and
cultivated plants are used. Some plants have been cultivated from time immemorial (e.g. flax,
opium poppy and coca). Others are now grown because supplies of the wild plants are
insufficient to meet the demand or because, owing to sparse distribution or inaccessibility,
collection is difficult.
Collection
Drugs may be collected from wild or cultivated plants, and the task may be undertaken by casual,
unskilled native labour (e.g. ipecacuanha) or by skilled workers in a highly scientific manner
(e.g. digitalis, belladonna and cinchona).
In the USA the explosive demand for some herbs has led to concern over wholesale
uncontrolled collection, so-called wildcrafting, resulting in the over-harvesting of such plants as
Panax quinquefolium, Polygala senega, Echinacea spp. and Cimicifuga racemose (black
cohosh).
Elsewhere Prunus africana (pygeum bark) found from Nigeria to Madagascar, Rauwolfia
serpentina from India,and Turnera diffusa (damiana) from Mexico are other examples of over-
exploitation.
A strategy for the sustained harvesting of Camptotheca acuminate (Nyssaceae), the source of the
anticancer drug camptothecin.
The alkaloid is accumulated in young leaves and by their repeated removal axillary bud
outgrowth is stimulated giving an increased harvestable amount of camptothecin in a non-
destructive manner. Studies showed that camptothecin accumulates primarily in the glandular
trichomes of the leaves and stems with overall variation among Camptotheca species and
varieties,and significantly, according to tissue ages and seasons. Details of the two best strains
for cultivation are given.
The season at which each drug is collected is usually a matter of considerable importance, as the
amount, and sometimes the nature, of the active constituents is not constant throughout the year.
This applies, for example, to the collection of podophyllum, ephedra, rhubarb, wild cherry and
aconite.
Rhubarb is reported to contain no anthraquinone derivatives in winter but anthranols which, on
the arrival of warmer weather, are converted by oxidation into anthraquinones; also the contents
of C-glycosides, O-glycosides and free anthraquinones in the developing shoots and leaves of
Rhamnus purshiana fluctuate markedly throughout the year.
Generally speaking, leaves are collected as the flowers are beginning to open, flowers just before
they are fully expanded, and underground organs as the aerial parts die down. Leaves, flowers
and fruits should not be collected when covered with dew or rain.

Drying
If enzymic action is to be encouraged, slow drying at a moderate temperature is necessary.
Examples of this will be found under ‘Orris Rhizome’, ‘Vanilla Pods’, ‘Cocoa Seeds’ and
‘Gentian Root.’
If enzymic action is not desired, drying should take place as soon as possible after collection.
Drugs containing volatile oils are liable to lose their aroma if not dried or if the oil is not distilled
from them immediately, and all moist drugs are liable to develop mould.
The duration of the drying process varies from a few hours to many weeks, and in the case of
open-air drying depends very largely on the weather. In suitable climates open-air drying is used
for such drugs as clove, colocynth, cardamom and cinnamon. Even in warm and dry climates
arrangements have to be made for getting the drug under the cover of sheds or tarpaulins at night
or during wet weather. For drying in sheds the drugs may be suspended in bundles from the roof,
threaded on strings, as in the case of Chinese rhubarb, or, more commonly, placed on trays made
of sacking or tinned wire-netting. Papers spread on a wooden framework are also used,
particularly for fruits from which it is desired to collect the seeds.
Drying by artificial heat is more rapid than open-air drying and is often necessary in tropical
countries (e.g. West Africa, where the humidity is very high, and Honduras for drying cardamom
fruits). In Europe continuous belt driers are used for large crops such as digitalis.
Rapid drying helps flowers and leaves to retain their colour and aromatic drugs their aroma, but
the temperature used in each case must be governed by the constituents and the physical nature
of the drug.
As a general rule, leaves, herbs and flowers may be dried between 20 and 40°C, and barks and
roots between 30 and 65°C

Storage
-The large-scale storage of drugs is a considerable undertaking. Except in a few cases, such as
cascara bark, long storage, although often unavoidable, is not to be recommended.
- Drugs such as Indian hemp and sarsaparilla deteriorate even when carefully stored.
- It has been reported that the content of taxol in Taxus baccata leaves and extracts stored at
room temperature for one year decreased by 30–40% and 70–80% respectively; storage in a
freezer and out of direct sunlight produced no adverse deterioration.
-Similarly the alkamides of the popular immune-stimulant herb Echinacea purpurea decrease
rapidly on storage.
Drugs stored in the usual containers—sacks, bales, wooden cases, cardboard boxes and paper
bags—reabsorb about 10–12% or more of moisture. They are then termed ‘air-dry’Plastic sacks
will effectively seal the contents.
The combined effects of moisture and temperature on humidity and the subsequent water-
condensation when the temperature falls, must be considered in drug storage.
Drugs such as digitalis and Indian hemp should never be allowed to become air-dry or they lose
a considerable part of their activity. They may be kept in sealed containers with a dehydrating
agent.

QUALITY CONTROL

Standards Applicable To Crude Drugs

There are a number of standards, numerical in nature, which can be applied to the evaluation of
crude drugs either in the whole or the powdered condition.
i)Sampling
Before a consignment of a drug can be evaluated, a sample must be drawn for analysis;
considerable care must be exercised to ensure that this sample is truly representative.
ii)Preliminary examination
In the case of whole drugs the macroscopical and sensory characters are usually sufficient to
enable the drug to be identified. The general appearance of the sample will often indicate
whether it is likely to comply with such standards as percentage of seed in colocynth, of ash in
valerian or of matter insoluble in alcohol in asafetida.
iii) matter
The difficulty of obtaining vegetable drugs in an entirely pure condition is fully recognized, and
pharmacopoeias contain statements as to the percentage of other parts of the plant or of other
organic matter which may be permitted e.g. cascara bark should be less than 1% Foreign matter.
iv)Moisture content
Moisture content must be determined to avoid deterioration of the herbal drug.
A large number of methods are now available for moisture determination, many being employed
in industries unrelated to pharmacy.
v)Loss on drying.
This is employed in the BP/EP and USP.
Although the loss in weight, in the samples so tested, principally is due to water, small amounts
of other volatile materials will also contribute to the weight loss. For materials (digitalis,
hamamelis leaf, yarrow, hawthorn berries, starch, aloes, fibres) which contain little volatile
material, direct drying (100–105°C) to constant weight can be employed.
Can be done using:-
a. Chemical method e.g. Karl Fischer
b. Spectroscopic methods e.g. NMR
c. Electrometric methods e.g. conductivity method
vi) Extractive values
The determination of water-soluble or ethanol-soluble extractive is used as a means of evaluating
drugs the constituents of which are not readily estimated by other means.
Vii) Ash values
When vegetable drugs are incinerated, they leave an inorganic ash which in the case of many
drugs (e.g. rhubarb) varies within fairly wide limits and is therefore of little value for purposes of
evaluation.
Viii) Other applicable standards
 Crude fibre
 Determination of volatile oil
 Tannin content
 Bitterness value
 Microbial contamination
 Toxic residues
  Heavy metal accumulation

PHYTOCHEMICALS

Alkaloids

Definition.
A precise definition of the term 'alkaloid' (alkali-like) is somewhat difficult because there is no
clear-cut boundary between alkaloids and naturally occurring complex amines.
-Typical 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 on man or other animals.
-Typical alkaloids can therefore be defined as nitrogen containing compounds mainly of
plant origin with a complex molecular structure and manifesting significant
pharmacological activity.
- 'proto-alkaloid' or 'amino-alkaloid' is sometimes applied to compounds such a hordenine,
ephedrine and colchicine which lack one or more of the properties of typical alkaloids.
-In practice, those substances present in plants and giving the standard qualitative tests for
alkaloids are termed alkaloids, and frequently in plant surveys this evidence alone is used to
classify particular plant as 'alkaloid-containing'.

Distribution of alkaloids in Plants

 By the mid-1940s, about 800 alkaloids had been isolated. Through new isolation
techniques, this figure has increased to the order of 10 000.
 Occur in about 15% of all vascular terrestrial plants in more than 150 diff. plant
families.
 True alkaloids rarely occur in lower plants.
 In the fungi, the lysergic acid derivatives and the sulphur-containing alkaloids, e.g.
the gliotoxins, are the best known.
  Among the pteridophytes and gymnosperms the lycopodium and ephedra, Taxus
alkaloids respectively have medicinal interest.
 Alkaloid distribution in the angiosperms is uneven.
 The dicotyledon orders Salicales, Fagales, Cucurbitales and Oleales at present appear to
be alkaloid-free.
 Alkaloids are commonly found in the families Amaryllidaceae, Apocynaceae,
Chenopodiaceae, Lauraceae, Magnoliaceae, Berberidaceae. Menispermaceae,
Ranunculaceae, Papaveraceae, Fumariaceae, Leguminosae. (Subfamily Papilionaceae),
Rubiaceae, Apocynaccae, Loganiaceae, Rubiaceae, Boraginaceae, Convolvulaceae.
Solanaceae, Campanulaceae (subfamily Lobelioideae), Compositae (subfamily
Senecioneae).

 Hegnauer, who has made an intensive study of alkaloid distribution, while recognizing
the undoubted potential chemotaxonomic significance of this group, is cautious about
its use without due regard to all the other characters of the plant. Nevertheless it
continues to be a popular area of research.

 Alkaloids are not confined to specific organs and can be found in all parts e.g Datura, in
all parts of the plant, cinchona in the bark, aconite in the root, pepper in fruit, areca nut-
the seed, belladonna-the leaves, ipecacuanha- rhizome & roots, tobacco alk- the leaves
(though produced in the root & translocated into the leaves), papaveraceae alk- latex of
the fruit.

Examples of alkaloids and their (higher) plant family origin

a) Monocotyledons

 Amaryllidaceae e.g. genus Narcissus typical alkaloid;

galanthamine
 Liliaceae genus Colchicum colchicine

b) Dicotyledons

 Solanaceae Solanum tuberosum (potato) solanine

Datura stramonium
hyoscyamine

 Rubiaceae Cinchona ledgeriana quinine

 Papaveraceae Papaver somniferum morphine
  Apocynaceae Cathanthus roseus vinblastine,
vincristine
 Compositae genus Senecio senecionine

Distribution of alkaloids in animals

 Nearly 300 alkaloids belonging to more than 24 classes are known to occur in the
skins of amphibians along with other toxins. They include the potent neurotoxic
alkaloids of frogs of the genus Phyllobates, which are among some of the most
poisonous substances known.
 Other reptilian alkaloids are strongly antimicrobial.
 Alkaloids derived from mammals include ones of indole and isoquinoline classes along
with mammalian morphine.
 A few are found in both plants and animals;

General properties of alkaloids

 Most alkaloids are crystalline, often optically active, substances, which react with
acids to form salts.
 Often impart bitter taste to the tongue. Caution; not all bitter substances are alkaloids and
not all alkaloids are bitter!
 They may exist in the plant in the Free State, as salts, or as N-oxides.
 Salts are from acetic, malic, oxalic, tartaric, succinic, tannic, meconic, etc acids
 Most alkaloids contain oxygen in addition to the elements carbon, hydrogen and
nitrogen.
 A few such as coniine from hemlock and nicotine from tobacco, are oxygen free and
are volatile liquids. Some are amorphous gums.
 Alkaloids are mostly colorless, with some exceptions, e.g. berberine is yellow,
betanidin is red, sanguinarine salts are copper red.
Extraction of alkaloids
Extraction methods vary with scale and purpose of the operation and with the raw material. In
general, drug is powdered through milling or grinding. This allows for effective contact between
solvents and drug tissues. Sometimes the material needs to be defatted using pet ether.
Following are general processes that may be used;
Process.
Process applied assuming alkaloid exists in salt form. The plant material may be defatted with
pet ether esp if drug is in form of seed or leaf.
 Moisten powdered material with water and mix with lime or ammonium hydroxide..
Lime combines with acids, tannins, other phenolic substances. This sets free the
alkaloids.
Alk2 SO4 + Lime [Ca (HCO3)2; Ca(OH)2] /NH4OH Alk base + salt
 Extract the material with organic solvents such as ether or petroleum spirit to take up
the alkaloid base.
 The concentrated organic liquid is then shaken with aq. acid and allowed to separate.
Alkaloid salts are now in the aq. liquid while many impurities remain behind in the
organic liquid.

PHENOLS

Phenols are hydroxyl derivatives of aromatic carbons. First member of this class is phenol.

 Currently believed to constitute the largest group of plants 20 metabolites.

 Found in most classes of natural compounds.
 Aromatic.
 Range from simple structures –with one aromatic ring to complex polymers such as
tannins and lignins.
 Important as medicinal agents and in food industry as coloring, flavoring, aromatizers
and antioxidants.
 Biosynthetic origin of some involves shikmic acid pathway and some from acetate
condensation.

Phenolic groups of pharmaceutical interest

1. Simple phenolic cpds
2. Tannins
3. Coumarins and their glycs.
4. Anthraquinones and their glycs.
5. Napthoquinones & glycosides
6. Flavone and related flavonoid glycs flavonoid
7. Anthocyanidins and anthocyanins greatest gp
8. Lignans and lignins

Extraction and detection of Phenolic compounds

 Phenolic cpds embrace a wide range of natural substances possessing an aromatic ring
bearing one or more hydroxyl groups eg. Hydroquinone.
 Usually water soluble as they frequently occur combined with sugar as glycosides.
 Among the natural phenolic compounds, the flavonoid form the largest group.
Monocyclic phenols, phenylpropanoids and phenolic quinones (anthraquinones) also
exist in considerable numbers. The tannins are complex phenolic compounds.
 Plant phenols have the ability of complexing with proteins via hydrogen bonding
hence posing a challenge to the phytochemist during isolation procedures.
 Phenols are very susceptible to enzymatic oxidation and phenolic material maybe
lost during isolation procedures due to enzymatic action in the plant.
 Extraction of phenols from plants with boiling alcohol normally prevents enzymatic
oxidation from occurring.

Classic procedure for detecting simple phenols-

Intense green, purple, blue or black colors are given by solution when 1% aq. or alcoholic
FeCl3 is added. However, majority of phenolic compounds can be detected on chromatograms
by their colors or fluorescences in UV light. The colors are intensified or changed by fuming
the chromatogram with ammonia vapor (bathochromatic shift).

 Phenolic pigments are visibly colored and they are easily monitored during their
isolation and purification.
 Phenolic compounds are all aromatic, they all show intense absorption in UV
region of the spectrum. In addition, phenolic compounds characteristically exhibit
bathochromic shifts in their spectra in the presence of alkali.
 Spectral methods are therefore, especially important for their identification and
quantification analysis of phenols.
 TANNINS

Introduction-

 True tannins have molecular weights of about 1000-5000.

 Tannins are substances in plant extracts which combine with protein of animal hides
preventing them from putrefaction and converting them into leather- a practical
definition.
 Hence tannins are detected qualitatively by a tanning test-the goldbeaters skin test-
and determined quantitatively by its adsorption on standard hide powder.
 Low molecular weight phenols, often present with tannins, such as, gallic acid,
catechins and chlorogenic acid, are excluded in the definition above, and are called
‘pseudo tannins’.
 Commercial tannins are obtained from quebracho, wattle, chestnut and myrobalan
trees.
 Many tannins are glycosides.
 Because they combine with tissue proteins and precipitate them, they are used as
astringents, in medicines as mild antiseptics, in the treatment of diarrhea, and to
check minor hemorrhages, for the protection of inflamed surfaces of mouth and
throat. They have been used as antidotes in poisoning by heavy metals, alkaloids
and glycosides. Commercially, they find extensive application in the leather industry.
 Recent research has demonstrated antitumor and anti-HIV activities by some
members.

Currently, tannins are not considered as a specific phytochemical grp, but as polyphenols
illustrating particular aspects of gallic acid and flavan-3-ol phytochemistry.

Chemistry and distribution-

 Tannins occur widely in vascular plants. In angiosperms, they are particularly associated
with woody tissues.
 By definition, they have the ability to react with protein, forming stable water-
insoluble co-polymers.
 Industrially, tannins are substances of plant origin which because of their ability to
cross-link with protein are capable of transforming raw animal skins into leather.
 In the plant cell, tannins are located separately from the proteins and enzymes of the
cytoplasm but when tissue is damaged, e.g, when animals feed, the tannin reaction
may occur, making the protein less accessible to the digestive juices of the animal.
 Plant tissues high in tannins are, in fact avoided by most feeders, because of the
astringent taste they impart.
 Tannins are classified into two grps based on complexity of their chemical structure and
according to their behavior on distillation.
 These are the hydrolysable tannins and the condensed tannins.

Hydrolysable tannins-

 Hydrolysable tannins are mainly of two classes;

 The simplest being the galloyl glucose depside, in which a glucose core is
surrounded by five or more galloyl ester groups (gallotanins).
 In the second type, the core molecule is a dimer of gallic acid, namely
hexahydroxydiphenic acid, again with glucose attachment (ellagitannins).
 Hydrolysed by acids or enzymes quickly to yield gallic or ellagic acids.
 On dry distillation, gallic acid and other components get converted to pyrogallol.
 They produce blue color with FeCl3.
 Examples of hydrolysable tannins are gallitannins in clove, rhubarb, red rose petals,
bearberry leaves, hamamelis; and ellagitannins of oak bark, myrobalans (dried mature
fruit of Terminalia chebula- Combretaceae), pomegranate rind/bark, eucalyptus leaves.

Condensed tannins-

 Also known as proanthocyanidins/flavolans.

 Not readily hydrolysed to simpler molecules and they do not contain a sugar moiety.
 They are related to flavonoid pigments and have flavan-3-ol structures. The flavan
units are linked to each other by 4-8 or 6-8 carbon-carbon bonds.
 These tannins have between two and twenty flavan-3-ol units.
 The name proanthocyanidin is used alternatively for condensed tannins because on
treatment with hot acid, some of the carbon-carbon linking bonds are broken and
anthocyanidin monomers are released.
 On treatment with acids or enzymes, condensed tannins are converted into red
insoluble compounds known as phlobaphenes.
 Phlobaphenes give the characteristic red color to many drugs such as red cinchona
bark, which contain these phlobatannins and their decomposition products.
 On dry distillation, they yield catechol and hence these tannins are sometimes called
catechol tannins.
 Like catechol, their solutions turn green with FeCl3.
 Some drugs such as tea, hamamelis bark/leaves contain both hydrolysable and
condensed tannins.
 They are distributed in different parts of plants; e.g leaves-green tea, hamamelis; bark as
in cinchona, cinnamon, wild cherry, mango; roots and rhizome-krameria and male fern;
 seeds e.g in cocoa, kola, guarana, areca; fruits- cranberries, grapes (red wines),
hawthorn; flowers- lime and hawthorn; extracts and dried juices- catechu, acacia cutch,
eucalyptus kino- kino is the name of the plant gum produced by various plants and trees,
particularly Eucalyptus, in reaction to mechanical damage.

COUMARINS AND THEIR GLYCOSIDES.

 Coumarins are derivatives of benzo-α-pyrone- the lactone of O-hydroxy cinnamic acid.

 Coumarin is present either in free state and as glycosides, the former being the most
common. Aesculetin, umbelliferone, and scopoletin are common in plants both in the
free state and as glycosides.
 Not all are phenolic and are included with phenolic derivatives for convenience.
 Coumarin derivatives are present in the families like Leguminosae, Solanaceae,
Rubiaceae, Caprifoliaceae, Umbelliferae, Oleaceae, etc.
 Coumarin itself has been found in about 150 species belonging to over 30 different
families.
 Coumarin gives a characteristic odour- that of new-mown hay and occurs in many
Leguminosae such as sweet clover.
 In ammoniacal solution, coumarin compounds have a blue, blue- green or violet
fluorescence. The fluorescence is used for chromatographic visualization of the
compounds.

Furanocoumarins; e.g psoralen are closely related to the above and are formed by the fusion
of the furan ring to coumarin at either position 6 & 7 or 7 & 8.

 Furanocoumarins occur particularly in the Rutaceae and Umbelliferae.

 Furanocoumarins are responsible at least in part for the unpredictable effects on drug
availability resulting from the consumption of grapefruit juice.
 Two components of the juice – 6’, 7’-dihydroxybergamottin and FC26- inactivate
cytochrome P450 enzymes (specifically CYP3A4 and CYP3A5) resulting in an
increased oral bioavalability of various drugs used to treat cancer, hypertension,
heart disease and allergies. Unnamed constituents of the juice have also been shown to
activate the efflux pump controlling P-glycoprotein mediated drug transport which
secretes absorbed drugs back to the gut.
 In vitro studies have demonstrated reduced absorption of vinblastine, cyclosporine,
losartan, digoxin and fexofenadine.
 FLAVONOIDS

 Flavonoids are related to flavones.

 They are sap pigments and the actual color of the plant organ is determined by the pH of the
sap e.g blue color of the cornflower and the red roses is due to the same glycoside. Both of
these plants on hydrolysis yield cyaniding hydrochloride.
 Flavonoids are derived from flavones. There are about 10 classes of flavonoids.
 Mainly are water-soluble compounds
 They can be extracted with 70% ethanol and remain in the aqueous layer following
partition of this extract with petroleum ether.
 Flavonoids are usually present in the plant-bound to sugars as glycosides.
Anyone flavonoid aglycone may occur in a single plant in several glyosidic combinations.
For this reason, when analyzing flavonoids, it is better to examine the aglycones present in
hydrolysed plant extracts before considering the complexity of glycosides that maybe present
in the original extract.
 Flavonoids are present in plants as mixtures and it is very rare to find only a single
flavonoid component in a plant tissue.
 The colored flavonoids in flower petals are anthocyanins which are almost always
accompanied by colorless flavones or flavonols.
 Mixtures of anthocyanins are also the rule, particularly in the flowers of ornamental plants.

Properties of some Flavonoid classes

Flavonoid class Distribution Characteristic properties

Anthocyanins Scarlet red, mauve and blue Water-sol, visible max 515-
flower pigs. 545nm, mobile in Butanol,
GAA, water 41.5 (upper
layer)
flavonols Mainly colourless After acid hydrolysis, bright
widespread in leaves yellow spots in UV light, on
forestall spectral max 350-
386 nm
Flavones As flavonols After acid hydrolysis dull
absorbing
Brown spots on forestall
chromatograms spectral max
330-350nm

Forestal=acetic acid-conc HCl-water:30:3:10

Significance

 Their therapeutic usefulness was not immediately realized

 Recent researches have demonstrated their usefulness in drugs such as liquorice, chamomile
& ginkgo
 It is possible that many herbal remedies, whose constituents are yet unknown, will be shown
to contain active flavonoids
 Of 84 drugs in the BHP, vol 1991, 36 contain flavonoids though not necessarily as active
ingredient.
 Known for anti-inflammatory and anti-allergic, antithrombic, vasoprotective properties,
inhibition of tumor promotion, and as a protective for gastric mucosa.
 Some of the pharmacological properties can be explained on the basis of antioxidant
activity
 Some are tannin materials.

BIOSYNTHETIC PATHWAY FOR ACTIVE COMPOUNDS

Biosynthetic pathway for cardiogenic Glycosides

H
R COOH
C
CH
H
HN 2
Amino Acid

H
R COOH
C
N-Hydroxy amino acid CH
H
NHOH

H
R COOH
C
Aldoxime CH
H
NOH
Cytochrome - P450

H
R
Nitrile C