EVALUATION OF CRUDE DRUGS

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Evaluation of crude drugs:
Naureen Shehzadi Department of Pharmaceutical Sciences, Faculty of Pharmacy, Superior University Evaluation of crude drugs

introduction:
A natural substance is regarded as food if it is used for filling up the stomach daily without experiencing any deleterious effects. But, it may
be referred to a drug if it results in changing a pathological/disease state of an individual or animal to a normal/physiological state without
producing any side effects in a specific dose. introduction

Why needed?:
The evaluation of crude drug is necessary because of three reason; 1. Biochemical variations in the crude drug 2. Deterioration due to
treatment and storage 3. Substitution and adulteration, as a result of carelessness, ignorance or fraud Why needed?

Drug evaluation:
Evaluation of crude drug ensures its identification and determination of its quality and purity. These are needed to establish whether or not a
substance qualifies to be a drug or food or eliminated from general use. Drug evaluation

Identification:
In every country, there is national herbarium where most of the native plants are preserved. Identification of the crude drug is usually done
by; A qualified, specialized and experienced personnel (Botanist, Herbal specialist) Comparison with the standard/authenticated sample
specimens Identification

quality:
It refers to intrinsic value of the crude drug i.e. nature and amount of active/medicinal principles. These active principles may be; Alkaloids
Glycosides Carbohydrates Volatile oils Fixed oils Tannins etc. quality

quality:
To maintain quality of the crude drugs, it is necessary to; Select proper source of crude drugs (either wild or cultivated) Collect the crude
drugs at appropriate time Collect the required parts of the plants Preparation of collected drugs by proper cleaning and drying Properly
preserve the crude drugs in order to avoid contamination due to microbes and against moisture, heat, air and light. quality

purity:
Purity of the crude drug can be achieved by; Proper identification Quality assurance purity

Morphology and morphography:

The study of the form of crude drugs is called morphology and determination of the form of crude drugs is called morphography.
Morphology and morphography

Types of drug evaluation:

Types of drug evaluation

PowerPoint Presentation:
ORGANOLEPTIC EVALUATION OF THE CRUDE DRUGS

DEFINITION:
DEFINITION It refers to the evaluation of crude drugs using organs of sense. The study includes; Macroscopic or external appearance Color
Odor Taste Sounds of its fractures etc….

characteristics:
characteristics The macroscopic features of the crude drugs include; Size Shape External color Odor Taste Fractures and internal color

size:
size The size of crude drug encompasses measurements of; Length Breadth Diameter It may be measured in mm or cm.

shape:
shape The crude drug may be having different shapes e.g. Crude drugs Shape Sarsaparilla root Cylindrical Podophyllum Sub-cylindrical
Aconite Conical Jalap Ovoid Calumba Disk

External color:
External color The crude drugs may acquire a variety of colors. The color of the crude drugs vary from white to yellowish gray, yellowish
brown, reddish orange or brownish black. The fruits and seed crude drugs have different colors whilst barks posses brownish gray to
brownish black color. The internal and external color of the barks varies; internal color is little lighter than the outer one.

Odor of crude drugs:

Odor of crude drugs The crude drugs may be odorless or pose an odor which may be; Distinct Indistinct General terms for describing odor
are; Aromatic Balsamic Spicy Fruity Pungent etc….

TASTE of crude drugs:

TASTE of crude drugs The crude drugs may posses a characteristic taste or may be tasteless. The different terms used to define taste of a
crude drug are; General terms used in describing taste are: 1- Acid (sour) 2- Saccharine ( sweet) 3- Saline (salty) 4- Alkaline 5- Bitter 6-
Tasteless

TASTE of crude drugs:

TASTE of crude drugs Taste Examples Mucilaginous Starches Oily Linseed oil Astringent Pomegranate, catechu Pungent Ginger Acrid
Coca Nauseous Ipeca

Fractures and internal color:

Fractures and internal color Fractures and internal color of crude drugs refer to the external markings that are characteristics of a particular
drug and color after breaking the crude drug. Internal color of a crude drug is usually lighter compared to the external colour. Example is
turmeric, ginger etc…..

PowerPoint Presentation:
Determine basic shapes of a leaf. Determine internal and external color of 15 plant drugs. Key components in a national herbarium Write the
characteristic odor of 10 crude drugs. Write taste of 5 animal drugs and 5 mineral drugs. CLASS ACTIVITY

PowerPoint Presentation:
PHYSICAL EVALUATION OF THE CRUDE DRUGS

Physical evaluation:
Physical evaluation The physical evaluation of the crude drugs is accomplished by the determination of various physical
characteristics/parameters by using various physico-chemical techniques. In crude plant evaluation, physical methods are used to determine;
Solubility Specific gravity Optical rotation Viscosity

Physical evaluation:
Physical evaluation Refractive index Melting point Water content Degree of fiber elasticity Ash values Extractive values Foreign organic
matter etc…….

solubility:
solubility I n this evaluation, the specific behavior of the drugs towards solvents is taken into consideration. Crude drugs Characteristic
solvents Colophony Light petroleum Peru B alsam Chloral hydrate Castor oil Light petroleum (half to the volume of castor oil) Alkaloidal
bases Organic solvents Alkaloidal salts Polar solvents

Optical rotation:
Optical rotation Certain drug substances may have capability of rotating the plane polarized light to specific orientation (+, dextrorotatory
and -, levorotatory). Substances that rotate the plane polarized light to the right/clockwise are dextrorotatory (d) or (+). Substances that
rotate the plane polarized light to the left/anti-clockwise are levorotatory (l) or (-). Measuring optical rotation helps in determining; Whether
a substance is optically active or not Purity Quality Strength Crude drugs Optical rotation Eucalyptus 0 ° – (+)10° Honey (+)3 ° - (-)15°

Refractive index:
Refractive index Refractive index is the property of a material that changes the speed of light, computed as the ratio of the speed of light in a
vacuum to the speed of light through the material. n = c/v Where; n = Refractive index c = Speed of light in vacuum v = Speed of light in the
substance Its measure gives an idea of; Identification of a substance Purity Concentration Crude drugs Refractive index Castor oil 1.4758-
1.527 Clove oil 1.527-1.535

Specific gravity:
Specific gravity It is also referred to as the “relative density”. It is the ratio of the mass of a solid or liquid to the mass of an equal volume of
distilled water at 4°C or of a gas to an equal volume of air or hydrogen under prescribed condition of temperature or pressure. Its
measurement gives the idea of floatability of the drug. Specific gravity > 1: Substance sinks Specific gravity < 1: Substance floats Crude
drugs Specific gravity Cottonseed oil 0.88-0.93 Coconut oil 0.925 Castor oil 0.95

viscosity:
viscosity It refers to the resistance of a liquid to flow. This resistance acts against the motion of any solid object through the fluid and also
against the motion of a fluid itself past stationary surfaces. It is a constant value.

Melting point:
Melting point It refers to a temperature at which it changes state from solid to liquid. Plant constituents have very sharp and constant melting
points. Crude drugs Melting point (°C) Beeswax 62-65 Wool fat 34-44 Agar 85

Moisture content:
Moisture content The moisture content of a crude drug is responsible for decomposition of crude drug due to chemical change or microbial
attack. It is necessary to determine and control the moisture content of the crude drug. It is determined by heating the drug at 105°C in an
oven to a constant weight. Crude drugs Moisture content Digitalis Not more than 5% w/w Ergot Not more than 8% w/w

Ash values:
Ash values Determination of ash values is useful for detecting low-grade products, exhausted drugs and excess of sandy or earthy material.
Different ash values are calculated for such purposes e.g. Total ash Acid-insoluble ash Water soluble ash Sulphated ash

Ash values:
Ash values Total ash It is useful for detection of the crude drug mixed with various minerals e.g. sand, soil, calcium oxalate, chalk powder
or other drugs with different inorganic contents to improve their appearance. The maximum temperature used for total ash is not more than
450°C. It is because of the fact that at higher temperatures, volatile alkali chlorides may be lost.

Ash values:
Ash values Acid-insoluble ash Acid-insoluble ash means ash insoluble in dilute hydrochloric acid. Its value is higher than total ash. The
majority of crude drugs contain calcium oxalate whose quantity varies frequently. Therefore, total ash of a crude drug vary within wide
limits for specimen of genuine drug e.g . for Rhubarb, total ash ranges from 8-40% and in this case , determination of acid-insoluble content
is more preferable.

Ash values:
Ash values Acid-insoluble ash The calcium oxide formed due to incinerated oxalate is soluble in hydrochloric acid. The insoluble as is then
weighed. In this way, excessive earthy matter that is usually present in root, rhizome and leaves can be determined. Water-soluble ash It is
used to detect the presence of material which is exhausted by water .

Ash values:
Ash values Sulphated ash This ash is produced after treating the drug with sulphuric acid to get sulphate salts. percentage ash is calculated
with reference to air-dried sample. Temperature used for the procedure is more than 600°C. Crude drugs Ash values Ginger (acid-insoluble)
3% Ginger (total ash ) 16%

Extract values:
Extract values Extracts of crude drugs are obtained with different solvents. These extract give approximate measures of their chemical
contents. Various solvents are used according to the type of constituents to be analyzed. Water soluble extracts are used for determination of
crude drugs containing glycosides, tannins and mucilage and alcohol-soluble extracts are used for determination of resins, certain glycosides
whilst ether-soluble extracts are used for drugs containg fats and volatile principles.

Foreign organic matter:

Foreign organic matter The parts of the organs or organs other than the required are called foreign organic matter. These may include;
Insects Moulds Earthy material Animal excreta Each crude drug has its own limits for presence of foreign matter e.g . garlic and saffron
should not contain more than 2%

PowerPoint Presentation:
CHEMICAL EVALUATION OF THE CRUDE DRUGS

INTRODUCTION:
INTRODUCTION

Chemical tests:
Chemical tests These are carried out using various chemical reagents to identify the nature and quantity of chemical constituents of crude
drugs. Chemical tests may be; Qualitative Quantitative

Qualitative chemical tests:

Qualitative chemical tests Qualitative chemical tests involve identification tests for various phytochemical constituents e.g. alkaloids,
glycosides, tannins etc. these tests provide information regarding nature of active principle. Examples include; Molisch's test for
carbohydrate drugs e.g. Glucose, Sucrose etc. Iodine test for identification of starches Spot test for identification of fixed oils

Quantitative chemical tests:

Quantitative chemical tests These tests give the value or amount of active constituent in the crude drug. Examples include; Saponification
values for lipids (number of milligrams of potassium hydroxide required to saponify 1g of fat under the conditions specified. It gives a
measure of molecular weight of the crude drug.) Ester value for volatile oils (number of mg of potassium hydroxide required to saponify the
esters in 1.0 g of the substance) Acid number for resins

CHEMICAL ASSAYS:
CHEMICAL ASSAYS These tests give an approximate value of total phytochemical constituents in a crude drug. Examples include; Total
alkaloid in belladonna leaf Total alkaloid in ipecacuanha Total resinous principle in jalap Total vitamins in cod liver oil

Instrumental techniques:
Instrumental techniques These techniques identify the chemical group in phytoconstituents using chromatography and spectroscopy
techniques.

PowerPoint Presentation:
Write chemical assay method of any crude drug from official book Write down at least 2 identification tests for carbohydrates, alkaloids,
glycosides, fixed oils and tannins Prepare a list of the approximate value of phytochemical constituents in 5 crude drugs e.g. turmeric,
fennel, caraway, cardamom and cinnamon Write a method for quantification of active principle of crude drug using UV-visible
spectrophotometer Class activity

PowerPoint Presentation:
BIOLOGICAL EVALUATION OF THE CRUDE DRUGS

DEFINITION:
DEFINITION It refers to evaluation of therapeutic or pharmacological and toxicological activity of the whole crude drug or its active
principle by using animals or microbes. Biological evaluation determines; Therapeutic activity of drug or active principle Potency Toxicity

importance:
importance There are several method by which the plant or extract can be evaluated for pharmacological activity. These are a method of
choice when the drug cannot be assayed satisfactorily by chemical and physical methods. These are also necessary to carry out since the
crude drug is considered important only if it exerts significant biological effects.

types:
types

method:
method These assays are conducted by determining the amount of drug of known potency required to produce a definite effect on suitable
test animals or organs under standard conditions. Reference standards are used in procedures to minimize errors. Toxicity studies are
performed in suitable animal models to decide the lethal dose and effective dose of crude drugs.

Animal models:
Animal models Drugs Animal models Vaccines Mice Vasopressin injection Guinea pigs Oxytocin injection Domestic chickens Digitalis
glycosides Pigeons Depressor activity and mydriatic drugs Cats Anthelmintic drugs Worms Eye preparations Rabbits GIT and cardiac drugs
Dogs Next to the animal drugs, the studies are carried out in humans and it is not necessary that same results be expressed by the human
beings.

Microbiological studies:
Microbiological studies Microbiological assays are carried out to determine anti-microbial activity of various drugs. Various methods
include; Agar diffusion methods Turbidimetric methods Disc-diffusion methods Yeasts, moulds and living bacteria are used for assaying
vitamins. Living bacterial strains are used for testing antimicrobials.

Agar diffusion method:

Agar diffusion method

Turbidimetric method:
Turbidimetric method

Disc-diffusion method:
Disc-diffusion method

PowerPoint Presentation:
MICROSCOPIC EVALUATION OF THE CRUDE DRUGS

DEFINITION:
DEFINITION This evaluation is based on microscopic examination of whole, certain parts or powdered crude drug. Microscopy techniques
may be simple or complex. The powdered crude material can be observed using only chloral hydrate (simple technique) but the presence of
starch in that crude material is detected using iodine tincture. The starch will be stained blue and other components of the sample will not be
stained.

importance:
importance Microscopic evaluation of the crude drug is indispensable in; Initial identification of herbs Identification of small fragments of
crude or powdered drugs Detection of adulterants (insects, animal feacal matter, moulds, fungi) Identification of plant by its characteristic
tissue features

types:
types

Identification of crude drug using qualitative microscopy:

Identification of crude drug using qualitative microscopy Every plant possess a characteristic tissue structure that can be demonstrated
through study of; Tissue arrangement Cell walls structure Cell contents (calcium oxalate crystals, fibers, parenchyma etc.) Configuration
(when properly mounted on the slide) Reagents (Lignin stains red or pink with a drop of phloroglucinol and concentrated hydrochloric acid;
Mucilage are stained pink with ruthenium red; Starches stained blue with iodine solutions) Media

example:
example Surinam Quassia is recognized from the other herbs due to following characteristic features that are observed microscopically
Absence of calcium oxalate crystals Presence of crystal fibres Presence of uniseriate medullary rays

Microscopic linear measurements:

Microscopic linear measurements T hese include identification of crude drug by measurements like; Size of starch grains (e.g. diameter of
starch grain in cassia bark distinguishes from cinnamon) Size of the stomata Diameter of phloem fibers (e.g. detection of cassia in
cinnamon) Width of the vessel (e.g. detection of clove stalks in powdered cloves) Length and width of fibres

Determination of leaf constants:

Determination of leaf constants

Stomatal index:
Stomatal index It is the percentage which the numbers of stomata form to the total number of epidermal cells (each stomata being counted as
one cell). S Here; S = Number of stomata per unit area E = Number of epidermal cells in the same unit area.
Vein-islet number:
Vein-islet number It is defined as the number of vein islets per square mm of leaf surface midway between the midrib and the margin. This
value is constant for a given specie of a plant and used as a characteristic for the identification of allied specie.

Veinlet termination number:

Veinlet termination number It is defined as the number of veinlet termination per square mm of leaf surface midway between the midrib and
the margin (unbroken leaf part is required). A vein termination is the ultimate free termination of the veinlet. Hall and Melville in 1951,
determined veinlet termination number of Indian and Alexandrian Senna which was quite different for each other.

Palisade ratio:
Palisade ratio It is the average number of the palisade cells beneath each epidermal cell. This ratio can be determined with powdered drug.

Quantitative microscopy:
Quantitative microscopy This is an important technique employed in identification of crude drug when physicochemical methods are
inapplicable. The powdered crude drugs with well-defined particles that can be counted or the objects with measurable diameter and
thickness under suitable magnification are calculated using lycopodium spore method. Adulterated starchy drugs can be determined by
counting the number of starch grains per mg and calculating the amount from the known umber of starch grains per mg of the pure
starch/starchy material.

EVALUATION OF CRUDE DRUGS

1. 1. K.Sudheer Kumar, Assistant professor. Dept.of Pharmacognosy Chilkur Balaji college of Pharmacy
Hyderabad. E-mail:sudheer.y2k8@gmail.com EVALUATION OF CRUDE DRUGS K.Sudheer Kumar,
Assistant professor. Dept.of Pharmacognosy Chilkur Balaji college of Pharmacy Hyderabad. E-
mail:sudheer.y2k8@gmail.com
2. 2. INTRODUCTION Crud drugs : Vegetable or animal drugs that consist of natural substances that have
undergone only the processes of collection and drying. Natural substances: 1- Plant origin: leaves,
flowers, seeds and barks. Or vegetable saps, extracts and secretions. 2- Animal origin: whole animals,
glands or organs, extracts and secretions. INTRODUCTION Crud drugs : Vegetable or animal drugs
that consist of natural substances that have undergone only the processes of collection and drying.
Natural substances: 1- Plant origin: leaves, flowers, seeds and barks. Or vegetable saps, extracts and
secretions. 2- Animal origin: whole animals, glands or organs, extracts and secretions.
3. 3. Purity – the extent of foreign organic material present in a crude drug. • Importance of evaluation of
crude drugs: • Determination of Biochemical variation in the drugs • Identification of deterioration due
treatment and storage • Repoting Substitution and adulteration, as result of carelessness, ignorance and
fraud Quality – the quantity of the active constituents present.  Identity – identification of biological
source of the drug.  Purity – the extent of foreign organic material present in a crude drug. •
Importance of evaluation of crude drugs: • Determination of Biochemical variation in the drugs •
Identification of deterioration due treatment and storage • Repoting Substitution and adulteration, as
result of carelessness, ignorance and fraud Drug evaluation may be defined as the determination of
identity, purity and quality of a drug.  Quality – the quantity of the active constituents present.  Identity
– identification of biological source of the drug. Drug evaluation may be defined as the determination of
identity, purity and quality of a drug.
4. 4. METHODS OF DRUG EVALUATION The evaluation of a drug is drug done by studying its various
properties. The various properties are: (1) Organoleptic evaluation (2) Microscopic evaluation (3)
Physical evaluation (4) Chemical evaluation (5) Analytical evaluation (6) Biological evaluation
METHODS OF DRUG EVALUATION The evaluation of a drug is drug done by studying its various
properties. The various properties are: (1) Organoleptic evaluation (2) Microscopic evaluation (3)
Physical evaluation (4) Chemical evaluation (5) Analytical evaluation (6) Biological evaluation
5. 5. 1.Organoleptic (Morphological) Evaluation • This refers to drug evaluation by means of organs of
sense and includes other sensory organs like color, odour, taste ,size ,shape and texture. • It includes
the study of morphology and other sensory characters. S.NO: CHARACHTER DRUG EXAMPLES.NO:
CHARACHTER DRUG EXAMPLE 1 Brown colour Cinnamon 2 Aromatic odour Umbelliferous fruits 3
Sweet taste Liquorice 4 Fractured surface Cinchona 5 Wavy shape Rauwolifia 6 7 to 8mm width 25 to
60 mm length (size) Senna leaf
6. 6. (a) Study of Morphology • It includes the visual examination of drug. S.NO PART OF DRUG EXAPLE
1 BARK KURCHI 2 UNDERGROUND TURMERIC,ZINGER 3 LEAVES DIGITALIS 4 FLOWERS
SAFFRON 5 FRUITS FENNEL5 FRUITS FENNEL 6 SEEDS NUX-VOMICA 7 RESIN ASAFOETIDA 8
WOOD SANDAL WOOD 9 GUMS ACACIA 10 ENTIRE DRUG ERGOT
7. 7. 1- Shape and size. Flowers: Floral parts: stigmas, corollas, anther, ovary, receptacle. Leaves and
leaflets: Length, width, apex, margin, base, venation, the texture of the leaf and the hairs in upper and
lower surface. The feel of the surface described as soft, hairy smooth. Bark: The barks occur in three
shapes: •Flat or curved pieces. • Single quill. •Double quills. ii- Barks have two surfaces, an outer and
inner. iii- The inner surface is usually lighter in color than the outer surface 2- Odor and taste. Odor: 1-
distinct 2- indistinct aromatic-balsamic,- spicy 1- Shape and size. Flowers: Floral parts: stigmas,
corollas, anther, ovary, receptacle. Leaves and leaflets: Length, width, apex, margin, base, venation, the
 texture of the leaf and the hairs in upper and lower surface. The feel of the surface described as soft,
hairy smooth. Bark: The barks occur in three shapes: •Flat or curved pieces. • Single quill. •Double
quills. ii- Barks have two surfaces, an outer and inner. iii- The inner surface is usually lighter in color
than the outer surface 2- Odor and taste. Odor: 1- distinct 2- indistinct aromatic-balsamic,- spicy
8. 8. Taste: 1) Acidic (sour) 2) Saccharine (sweet): indicates sugar or sugar like substances 3) e.g.,
liquorice. 4) Saline (salty) 5) Alkaline 6) Bitter: indicates presence of substances such as bitter principle
7) e.g., glycoside, alkaloids. 8) Tasteless 9) Distinctive sensations to the tongue I. Mucilaginous and oily
(soft feeling) e.g., linseed. II.Astringent indicates presence of tannin. III.Pungent (warm biting sensation)
e.g., ginger. IV.Acrid (irritant sensation) e.g., Aconite, coca. V.Nauseous (those tending to excite
vomiting), Ipecac. Taste: 1) Acidic (sour) 2) Saccharine (sweet): indicates sugar or sugar like
substances 3) e.g., liquorice. 4) Saline (salty) 5) Alkaline 6) Bitter: indicates presence of substances
such as bitter principle 7) e.g., glycoside, alkaloids. 8) Tasteless 9) Distinctive sensations to the tongue
I. Mucilaginous and oily (soft feeling) e.g., linseed. II.Astringent indicates presence of tannin. III.Pungent
(warm biting sensation) e.g., ginger. IV.Acrid (irritant sensation) e.g., Aconite, coca. V.Nauseous (those
tending to excite vomiting), Ipecac.
9. 9. 3- Color and external markings. I. 1- White: e.g., starch, II. 2- Pale yellow:e.g., ginger,squill,white
pepper. III. 3- Deep yellow: e.g., peeled liquorice. IV. 4- Light pale brown e.g., nux-vomica, fennel. V. 5-
Dark brown: e.g., cloves buds. VI. 6- Dark reddish brown: cinchona. VII.7- Red: (brick red). e.g.,
cinnamon bark inner portion VIII.8- Pale green e.g., lobelia. IX. 9- Greenish brown: most of the leaf
herbs. 3- Color and external markings. I. 1- White: e.g., starch, II. 2- Pale yellow:e.g., ginger,squill,white
pepper. III. 3- Deep yellow: e.g., peeled liquorice. IV. 4- Light pale brown e.g., nux-vomica, fennel. V. 5-
Dark brown: e.g., cloves buds. VI. 6- Dark reddish brown: cinchona. VII.7- Red: (brick red). e.g.,
cinnamon bark inner portion VIII.8- Pale green e.g., lobelia. IX. 9- Greenish brown: most of the leaf
herbs.
10. 10. 2. Microscopic or Anatomical Evaluation • This method allows a more detailed examination of a drug
and it can be used to identify organized drugs by their known histological characters. • Before
examination through a microscope the material must be suitably prepared. • This can be done by
powdering, cutting thin sections of the drug or preparing a macerate. 2. Microscopic or Anatomical
Evaluation • This method allows a more detailed examination of a drug and it can be used to identify
organized drugs by their known histological characters. • Before examination through a microscope the
material must be suitably prepared. • This can be done by powdering, cutting thin sections of the drug or
preparing a macerate.
11. 11. 1. Palisade Ratio 2. Stomatal Number 3. Stomatal Index 4. Stomata 5. Vein-islet Number 6. Vein-
termination Number 7. Trichomes or plant hairs 8. Calcium oxalate crystals Quantitative Microscopy
1.Lycopodium spore method 1. Palisade Ratio 2. Stomatal Number 3. Stomatal Index 4. Stomata 5.
Vein-islet Number 6. Vein-termination Number 7. Trichomes or plant hairs 8. Calcium oxalate crystals
Quantitative Microscopy 1.Lycopodium spore method
12. 12. 1.Palisade ratio: • It represents the average number of palisade cells beneath one epidermal cell,
using four continuous epidermal cells for the count. • It is determined from powdered drugs with the help
of camera-Lucida. • Examples: Atropa belladona – 05-70 Adhatoda vasica –5.5-6.5 Cassia angustifolia
–5.5-10.0 upper,4.0-7.4 lower(senna) Digitalis lanata –2.5-6.5 1.Palisade ratio: • It represents the
average number of palisade cells beneath one epidermal cell, using four continuous epidermal cells for
the count. • It is determined from powdered drugs with the help of camera-Lucida. • Examples: Atropa
belladona – 05-70 Adhatoda vasica –5.5-6.5 Cassia angustifolia –5.5-10.0 upper,4.0-7.4 lower(senna)
Digitalis lanata –2.5-6.5
13. 13. 2.Stomatal Number: • The average number of stomata present per square millimeter of the
epidermis is known as stomatal number Examples: a)Atropa belladonna upper epidermis---07-10 lower
epidermis---77-115 b)Datura metel upper epidermis---147-160 lower epidermis---200-209 c)Ocimum
sanctum upper epidermis---64-72 lower epidermis---175-250 2.Stomatal Number: • The average number
of stomata present per square millimeter of the epidermis is known as stomatal number Examples:
a)Atropa belladonna upper epidermis---07-10 lower epidermis---77-115 b)Datura metel upper epidermis-
--147-160 lower epidermis---200-209 c)Ocimum sanctum upper epidermis---64-72 lower epidermis---
175-250
14. 14. 3.Stomatal Index: • It is the percentage proportion of the number of stomata to the total number of
epidermal cells. • Stomatal number varies considerably with the age of the leaf but stomatal index is
relatively constant for a given species. • Stomatal index calculated by • S.I = Example: a)Atropa
belladonna upper epidermis--- nil lower epidermis---20.2-23-0 3.Stomatal Index: • It is the percentage
proportion of the number of stomata to the total number of epidermal cells. • Stomatal number varies
considerably with the age of the leaf but stomatal index is relatively constant for a given species. •
Stomatal index calculated by • S.I = Example: a)Atropa belladonna upper epidermis--- nil lower
epidermis---20.2-23-0 S E+S S.I = Stomatal index S= Number of stomata per unit area E=Number of
epidermal cells in the same unit area
15. 15. 4.Stomata: (primary and important function is gaseous exchange) a minute epidermal opening
present on arial parts of plants, Stomata consists of central pore ,two kidney shaped similar cells(guard
cells) & varying number of subsidiary cells. Epidermis of leaf shows different characteristics
e.g.cuticle,stomata,trichomes, Types of stoma: 1.Moss type 2.Gymnospermous type 3.Gramineous type
4.Dicoteyledonous---it is having diagnostic significance and classified based on form of arrangement of
 subsidiary cells. 4.Stomata: (primary and important function is gaseous exchange) a minute epidermal
opening present on arial parts of plants, Stomata consists of central pore ,two kidney shaped similar
cells(guard cells) & varying number of subsidiary cells. Epidermis of leaf shows different characteristics
e.g.cuticle,stomata,trichomes, Types of stoma: 1.Moss type 2.Gymnospermous type 3.Gramineous type
4.Dicoteyledonous---it is having diagnostic significance and classified based on form of arrangement of
subsidiary cells.
16. 16. a) Paracytic or rubiaceous or parallel-cell stomata: in this stomata two guard cells covered by two
subsidiary cells, e.g. senna b) Diacytic or caryophyllaceous or cross-celled stomata: in this stomata the
guard cells are covered by two subsidiary cells on right angle to that of stomata e.g. peppermint
c)Anisocytic or cruciferous or unequal-celled stomata: in this stomata number of guard cells is two but
covered by three subsidiary cells and in that one is small in size with other two e.g.Datura d) Anomocytic
or ranunculaceous or irregular- celled:in this type stoma is surrounded by varying number of subsidiary
cells. e.g. digitalis e)Actinocytic or radiate-celled stomata: two guard cells are surrounded by radiating
subsidiary cells. a) Paracytic or rubiaceous or parallel-cell stomata: in this stomata two guard cells
covered by two subsidiary cells, e.g. senna b) Diacytic or caryophyllaceous or cross-celled stomata: in
this stomata the guard cells are covered by two subsidiary cells on right angle to that of stomata e.g.
peppermint c)Anisocytic or cruciferous or unequal-celled stomata: in this stomata number of guard cells
is two but covered by three subsidiary cells and in that one is small in size with other two e.g.Datura d)
Anomocytic or ranunculaceous or irregular- celled:in this type stoma is surrounded by varying number of
subsidiary cells. e.g. digitalis e)Actinocytic or radiate-celled stomata: two guard cells are surrounded by
radiating subsidiary cells.
17. 17. 5.Vein-islet Number: • Vein-islet number is defined as the number of vein-islets per sq.mm. of leaf
surface. S.NO NAME OF DRUG Vein-islet Range 1 Andrograohis paniculata 9-12 2 Bacopa monniera
6-13 3 Cannabis sativa 18-243 Cannabis sativa 18-24 4 Digitalis purpurea 2.5-3 5 Eucalpytus globules
8-13.5 6.Vien-termination Number: It is defined as the number of veinlet terminations per.sq. mm of the
leaf surface between midrib and margin.
18. 18. 8.Calcium oxalate crystals: Several cell contents present in vegetable drugs. the inorganic crystalline
compounds by virtue of their specific shapes can be utilized for the identification of herbal drugs. due to
this reason they are called as diagnostic characters of the plant. 1.Cubical (cube shape)
e.g,senna,Glycyrrhiza. 2.Rhombic (diamond shape) e.g., 3.Tetragonal e.g,onion. 4.Mono clinic(all three
axes are un equal) e.g,Gall. 5.Acicular (long slender, pointed , budles) e.g, Squill, Cinnamon. 6.Rosettes
–clusters (aggregation of crystals) e.g, Clove,Arjuna. 7.Microsphenoidal (minute in structures) e.g,
Henbane. 8.Calcium oxalate crystals: Several cell contents present in vegetable drugs. the inorganic
crystalline compounds by virtue of their specific shapes can be utilized for the identification of herbal
drugs. due to this reason they are called as diagnostic characters of the plant. 1.Cubical (cube shape)
e.g,senna,Glycyrrhiza. 2.Rhombic (diamond shape) e.g., 3.Tetragonal e.g,onion. 4.Mono clinic(all three
axes are un equal) e.g,Gall. 5.Acicular (long slender, pointed , budles) e.g, Squill, Cinnamon. 6.Rosettes
–clusters (aggregation of crystals) e.g, Clove,Arjuna. 7.Microsphenoidal (minute in structures) e.g,
Henbane.
19. 19. 7.Trichomes or plant hairs Trichomes are the tubular elongated or glandular outgrowth of the
epidermal cells Trichomes are also called as plant hairs.trichomes consists of two parts root and
body.trichomes present in most of plant parts and are function less but some times perform secretory
function. Depending up on the structure and the number of cells present in trichomes,they are classified
in to following. 1.Covering Trichomes 2.Glandular Trichomes 3.Hydathode or special Trichomes
7.Trichomes or plant hairs Trichomes are the tubular elongated or glandular outgrowth of the epidermal
cells Trichomes are also called as plant hairs.trichomes consists of two parts root and body.trichomes
present in most of plant parts and are function less but some times perform secretory function.
Depending up on the structure and the number of cells present in trichomes,they are classified in to
following. 1.Covering Trichomes 2.Glandular Trichomes 3.Hydathode or special Trichomes
20. 20. The most basic terms used are glabrous—lacking hairs— and pubescent— having hairs. Details are
provided by: a. glabrous, glabrate – lacking hairs or trichomes; surface smooth b. hirsute – coarsely
hairy c. hispid – having bristly hairs d. articulate – simple pluricellular-uniseriate hairs e. downy – having
an almost wool-like covering of long hairs f. pilose – pubescent with long, straight, soft, spreading or
erect hairs g. puberulent – minutely pubescent; having fine, short, usually curly, hairs h. pubescent –
bearing hairs or trichomes of any type i. strigillose – minutely strigose j. strigose – having straight hairs
all pointing in more or less the same direction as along a margin or midrib k. tomentellous – minutely
tomentose l. tomentose – covered with dense, matted, woolly hairs m. villosulous – minutely villous n.
villous – having long, soft hairs, often curved, but not matted The most basic terms used are glabrous—
lacking hairs— and pubescent— having hairs. Details are provided by: a. glabrous, glabrate – lacking
hairs or trichomes; surface smooth b. hirsute – coarsely hairy c. hispid – having bristly hairs d. articulate
– simple pluricellular-uniseriate hairs e. downy – having an almost wool-like covering of long hairs f.
pilose – pubescent with long, straight, soft, spreading or erect hairs g. puberulent – minutely pubescent;
having fine, short, usually curly, hairs h. pubescent – bearing hairs or trichomes of any type i. strigillose
– minutely strigose j. strigose – having straight hairs all pointing in more or less the same direction as
along a margin or midrib k. tomentellous – minutely tomentose l. tomentose – covered with dense,
matted, woolly hairs m. villosulous – minutely villous n. villous – having long, soft hairs, often curved, but
not matted
21. 21. Quantitative microscopy Lycopodium spore method: it is used when especially chemical and other
methods of evaluation of drugs fails to determine quality. Lycopodium spores are very characterized in
shape and appearance and uniform in size(25μm) on avg,94000 spores present/mg of lycopodium
powder . it consists of 1.well defined particles which may be counted. 2.Single layered cells or tissues
the area of which may be traced under suitable magnification and actual area calculated 3.The objects
of uniform thickness, the length of which can be measured, and actual area calculated. Quantitative
microscopy Lycopodium spore method: it is used when especially chemical and other methods of
evaluation of drugs fails to determine quality. Lycopodium spores are very characterized in shape and
appearance and uniform in size(25μm) on avg,94000 spores present/mg of lycopodium powder . it
consists of 1.well defined particles which may be counted. 2.Single layered cells or tissues the area of
which may be traced under suitable magnification and actual area calculated 3.The objects of uniform
thickness, the length of which can be measured, and actual area calculated.
22. 22. The percentage purity of an authentic ginger powder calculated as follows N X W X 94,000 X 100
N= NUMBER OF CHARACTERISTIC STRUCTURES(STRACH GRAINS) IN 26 FIELDS W=WEIGHT
IN mg OF LYCOPOSIUM TAKEN S=NUMBER OF LYCOPODIUM SPORES IN THE SAME 25 FIELDS
M=WEIGHT IN mg OF SAMPLE CALCULATED ON BASIS OF DRIED SAMPLE AT 105 C P=2,86,000
IN CASSE OF GINGER STARCH GRAIN POWDER S x M x P = % Purity of drug The percentage purity
of an authentic ginger powder calculated as follows N X W X 94,000 X 100 N= NUMBER OF
CHARACTERISTIC STRUCTURES(STRACH GRAINS) IN 26 FIELDS W=WEIGHT IN mg OF
LYCOPOSIUM TAKEN S=NUMBER OF LYCOPODIUM SPORES IN THE SAME 25 FIELDS
M=WEIGHT IN mg OF SAMPLE CALCULATED ON BASIS OF DRIED SAMPLE AT 105 C P=2,86,000
IN CASSE OF GINGER STARCH GRAIN POWDER
23. 23. 3. Physical Evaluation • Physical contents such as elasticity in fibres, viscosity of drugs containing
gums, swelling factor for mucilage containing materials, froth number of saponin drugs, congealing point
of volatile and fixed oils, melting and boiling points and water contents are some important parameters
used in the evaluation of drugs. • Ultraviolet light is also used for determing the fluorescence of extracts
of some drugs. 3. Physical Evaluation • Physical contents such as elasticity in fibres, viscosity of drugs
containing gums, swelling factor for mucilage containing materials, froth number of saponin drugs,
congealing point of volatile and fixed oils, melting and boiling points and water contents are some
important parameters used in the evaluation of drugs. • Ultraviolet light is also used for determing the
fluorescence of extracts of some drugs.
24. 24. • Physical constants are extensively applied to the active principles of drugs, such as alkaloids,
volatile oils, fixed oils etc. A few of them are I. Moisture Content II. Viscosity III. Melting point IV.
Solubility V. Optical Rotation VI.Refractive Index VII.Ash values VIII.Extractive values IX.Volatile oil
Content X.Foreign organic matter XI.swelling factor • Physical constants are extensively applied to the
active principles of drugs, such as alkaloids, volatile oils, fixed oils etc. A few of them are I. Moisture
Content II. Viscosity III. Melting point IV. Solubility V. Optical Rotation VI.Refractive Index VII.Ash values
VIII.Extractive values IX.Volatile oil Content X.Foreign organic matter XI.swelling factor
25. 25. I. Moisture Content: • Presence of moisture in a crude drug can lead to its deterioration due to either
activation of certain enzymes or growth of microbes. • Moisture content can be determined by heating
the drug at 150⁰C in an oven to a constant weight and calculating the loss of weight. I. Moisture Content:
• Presence of moisture in a crude drug can lead to its deterioration due to either activation of certain
enzymes or growth of microbes. • Moisture content can be determined by heating the drug at 150⁰C in
an oven to a constant weight and calculating the loss of weight. S.NO DRUGS MOISTURE CONTENT
W/W 1. Aloes Not more than 10 2. Digitalis Not more than 5 3. Starch Not more than 15
26. 26. II.Viscosity: • Viscosity of a liquid is constant at a given temperature and is an index of its
composition. • Hence, it is used as a means of standardising liquid drugs. i)Liquid paraffin-kinematic
viscosity not less than 64-centistokes at 37.8° ii)Pyroxylin-kinematic viscosity,1100-2450 centistokes
II.Viscosity: • Viscosity of a liquid is constant at a given temperature and is an index of its composition. •
Hence, it is used as a means of standardising liquid drugs. i)Liquid paraffin-kinematic viscosity not less
than 64-centistokes at 37.8° ii)Pyroxylin-kinematic viscosity,1100-2450 centistokes
27. 27. III.Melting Point: • It is one of the parameters to judge the purity of crude drugs containing lipids as
constituents. They may of animal or plant origin and contain fixed oils, fats and waxes. The purity of the
following crude drugs can be ascertained by determining their melting points in the range shown against
each of them. III.Melting Point: • It is one of the parameters to judge the purity of crude drugs containing
lipids as constituents. They may of animal or plant origin and contain fixed oils, fats and waxes. The
purity of the following crude drugs can be ascertained by determining their melting points in the range
shown against each of them. S,NO DRUGS MELTING POINT (°C) 1 COLOPHONY 75-85 2 BEES WAX
62-65 3 WOOL FAT 34-44
28. 28. IV.Solubility : The presence of adulterant in a drug could be indicated by solubility studies S.NO
DRUG SOLUBILITY 1 Castor oil Soluble in 3 volumes of alcohol 2 Balsam of Peru Soluble in chloral
hydrate solution 2 Balsam of Peru Soluble in chloral hydrate solution 3 Asafoetida Soluble in carbon
disulphide 4 Alkaloid bases Soluble in chloroform 5 colophony Soluble in light petroleum
29. 29. V.Optical Rotation: • Many substances of biological origin, having a chiral centre, can rotate the
plane of polarised light either to right(dextro rotatory)or to the left(laevo). The extent of rotation is
expressed in degrees, plus(+) indicating rotation to the right and minus(-) indication rotation in the left.
Such compound are optically active and hence called optical rotation. V.Optical Rotation: • Many
 substances of biological origin, having a chiral centre, can rotate the plane of polarised light either to
right(dextro rotatory)or to the left(laevo). The extent of rotation is expressed in degrees, plus(+)
indicating rotation to the right and minus(-) indication rotation in the left. Such compound are optically
active and hence called optical rotation. S.NO Drugs Angles of Optical Rotation 1. Caraway oil + 75° to
+80° 2. Clove oil 0° to +6.0° 3. Honey +3° to -15°
30. 30. VI.Refractive Index: When a ray of light passes from one medium to another medium of different
density, it is bent from its original path. Thus, the ration of velocity of light in vaccum to its velocity in the
substance is said to the Refractive index of the second medium. It is measured by means of
refractometer. RI of a compound varies with the wavelength of the incident light, temperature and
pressure. VI.Refractive Index: When a ray of light passes from one medium to another medium of
different density, it is bent from its original path. Thus, the ration of velocity of light in vaccum to its
velocity in the substance is said to the Refractive index of the second medium. It is measured by means
of refractometer. RI of a compound varies with the wavelength of the incident light, temperature and
pressure. S.NO DRUGS REFRACTIVE INDEX 1 Arachis oil 1.4678 to 10470 2 Castor oil 104758 to
10527 3 Clove oil 1.527 to 10535
31. 31. VII.Ash values The residue remaining after incineration is the ash content of the drug.( inorganic
salts of carbonates, phosphates, silicates of sodium, potassium, calcium and magnesium) is known as
ash content. Ash value is a criterion to judge the identity OR purity of the crude drug The residue
remaining after incineration is the ash content of the drug.( inorganic salts of carbonates, phosphates,
silicates of sodium, potassium, calcium and magnesium) is known as ash content. Ash value is a
criterion to judge the identity OR purity of the crude drug S.NO Drugs total ash(% w/w) acid insoluble
ash % (w/w) 1 Agar - 1.00 2 Bael 03.5 - 3 Cannabis 15.0 5.00 4 Gelatin 03.6 - 5 Valerin 12.0 -
32. 32. TYPES OFASH VALUES 1.Total ash value Useful for detecting low grade products, exhausted
products, excess of sandy and earthy matter with drug. 2.Acid insoluble ash value Used for the
determination of earthy matter present on roots, rhizomes, and also on the leaves, Crude drugs contain
calcium oxalate crystals the amount may varies depending on the environmental conditions.
3.Sulphated ash value Used for the detection of low grade products. 4. Water soluble ash value Water
soluble ash value Used to detect either material exhausted by water or not ( Tea leaves, Ginger
rhizomes). 1.Total ash value Useful for detecting low grade products, exhausted products, excess of
sandy and earthy matter with drug. 2.Acid insoluble ash value Used for the determination of earthy
matter present on roots, rhizomes, and also on the leaves, Crude drugs contain calcium oxalate crystals
the amount may varies depending on the environmental conditions. 3.Sulphated ash value Used for the
detection of low grade products. 4. Water soluble ash value Water soluble ash value Used to detect
either material exhausted by water or not ( Tea leaves, Ginger rhizomes).
33. 33. Determination Total ash value 1.Weigh accurately about 3gms of the powdered drug in a tared silica
crucible 2.Incinerate the powdered drug by gradually increasing the heat until free from carbon and cool.
Keep it in desiccators 3. Weigh the ash and calculate the % of the total ash with reference to the air
dried sample 1.Weigh accurately about 3gms of the powdered drug in a tared silica crucible 2.Incinerate
the powdered drug by gradually increasing the heat until free from carbon and cool. Keep it in
desiccators 3. Weigh the ash and calculate the % of the total ash with reference to the air dried sample
34. 34. 1. Boil the total ash obtained as above for 5 minutes with 25ml of dilute HCL 2.Filter and collect the
insoluble matter on the ashless filter paper , wash the filter paper with hot water, ignite in tared crucible,
cool and kept in desiccators 3.Weigh the residue and calculate the acid insoluble ash of the drug
Determination of Acid insoluble ash value 1. Boil the total ash obtained as above for 5 minutes with 25ml
of dilute HCL 2.Filter and collect the insoluble matter on the ashless filter paper , wash the filter paper
with hot water, ignite in tared crucible, cool and kept in desiccators 3.Weigh the residue and calculate
the acid insoluble ash of the drug
35. 35. VIII.Extractive values In crude drugs, sometimes the active chemical constitutes cannot be
determined by normal procedures. In such cases, water, alcohol or ether soluble extractive values are
determined for evaluation of such drugs. Significances : 1.Useful for the evaluation especially when the
constituents of the drugs can not be readily estimated by any other means 2.It also helps to indicate the
nature of chemical constituents present in the drug 3. Also helps in the identification of adulterants
VIII.Extractive values In crude drugs, sometimes the active chemical constitutes cannot be determined
by normal procedures. In such cases, water, alcohol or ether soluble extractive values are determined
for evaluation of such drugs. Significances : 1.Useful for the evaluation especially when the constituents
of the drugs can not be readily estimated by any other means 2.It also helps to indicate the nature of
chemical constituents present in the drug 3. Also helps in the identification of adulterants
36. 36. Types of extractive values A. water soluble extractive values B. Alcohol soluble extractive values C.
Ether soluble extractive values Types of extractive values A. water soluble extractive values B. Alcohol
soluble extractive values C. Ether soluble extractive values
37. 37. Determination of water soluble extractive value 1. Macerate about 5gm of the accurately weighed
coarse powder with 100ml of chloroform water in a 100ml volumetric flask for 24 hours . 2. Shake
frequently for first 6 hours 3. Filter rapidly through filter paper and evaporate 25ml of water extract to
dryness in a tared flat-bottomed shallow dish. 4. Dry the residue at 105 and weigh. Keep it in a
desiccators 5. Dry the extract to constant weight ,finally , calculate the % W/W of Water soluble
extractive value with reference to the air dried drug. Determination of water soluble extractive value 1.
Macerate about 5gm of the accurately weighed coarse powder with 100ml of chloroform water in a
 100ml volumetric flask for 24 hours . 2. Shake frequently for first 6 hours 3. Filter rapidly through filter
paper and evaporate 25ml of water extract to dryness in a tared flat-bottomed shallow dish. 4. Dry the
residue at 105 and weigh. Keep it in a desiccators 5. Dry the extract to constant weight ,finally ,
calculate the % W/W of Water soluble extractive value with reference to the air dried drug.
38. 38. A. Water soluble extractive value Water soluble extractive value is applied for the drugs which
contain water soluble constituents such as tannins, sugars, plant acids and mucilage. S.NO DRUG
WATER SOLUBLE EXTRACTIVE (% W/W) S.NO DRUG WATER SOLUBLE EXTRACTIVE (% W/W) 1
Aloe Vera NLT 25.0 2 Linseed NLT 20.0 3 Senna leaves NLT 30.0 4 Ginger NLT 10.0 5 Glycyrrhiza NLT
20.0 NLT= Not less than
39. 39. Determination of Alcohol soluble extractive values 1. Macerate about 5gm of the accurately weighed
coarse powder with 100ml of 90% alcohol in a 100ml stoppered flask for 24 hours . 2. Shake frequently
for first 6 hours 3. Filter rapidly through filter paper and collect the filtrate evaporate 25ml of alcohol
extract to dryness in a tared flat-bottomed shallow dish. 4. Dry the residue at 105 and weigh. Keep it in a
desiccators 5. Dry the extract to constant weight ,finally , calculate the % w/w of alcohol soluble
extractive value with reference to the air dried drug. Determination of Alcohol soluble extractive values
1. Macerate about 5gm of the accurately weighed coarse powder with 100ml of 90% alcohol in a 100ml
stoppered flask for 24 hours . 2. Shake frequently for first 6 hours 3. Filter rapidly through filter paper
and collect the filtrate evaporate 25ml of alcohol extract to dryness in a tared flat-bottomed shallow dish.
4. Dry the residue at 105 and weigh. Keep it in a desiccators 5. Dry the extract to constant weight ,finally
, calculate the % w/w of alcohol soluble extractive value with reference to the air dried drug.
40. 40. B.Alcohol soluble extractive value Alcohol soluble extractive value is applied for the drugs which
contain alcohol soluble constituents such as tannins, resins and alkaloids .Official method for the assay
of myrrh & asafoetida. Generally,95% ethyl alcohol is used for determination of Alcohol soluble
extractive. B.Alcohol soluble extractive value Alcohol soluble extractive value is applied for the drugs
which contain alcohol soluble constituents such as tannins, resins and alkaloids .Official method for the
assay of myrrh & asafoetida. Generally,95% ethyl alcohol is used for determination of Alcohol soluble
extractive. S.NO DRUG Alcohol soluble extractive. (% W/W) 1 Aloe vera NLT 10.0 2 Benzoin NLT 90.0
3 Asafoetida NLT 50.0 4 Ginger NLT 04.0 5 Myrrh NLT 24.0 NLT= Not less than
41. 41. C.Ether soluble extractive value Ether soluble extractive value is applied for the extraction of volatile
oils, fixed oils and resins. 1.Volatile ether soluble extractive value –(volatile oil) 2.Non volatile ether
soluble extractive value –(resin, fixed oils, coloring matter) C.Ether soluble extractive value Ether soluble
extractive value is applied for the extraction of volatile oils, fixed oils and resins. 1.Volatile ether soluble
extractive value –(volatile oil) 2.Non volatile ether soluble extractive value –(resin, fixed oils, coloring
matter) S.NO DRUGS LIMIT FOR NON-VOLATILE ETHER SOLUBLE EXTRACTIVES(% W/W) 1
CAPSICUM NLT 12.0 2 MALE FERN NLT 01.0 3 LINSEED NLT 25.0 NLT= Not less than
42. 42. IX.Volatile oil content: Efficiency of several drugs is due to their odorous principle (volatile oils).Such
crude drugs are standardized on the basis of their volatile oil contents. Weighed quantity of the drug is
boiled with water in a round bottomed flask fitted with clevenger apparatus. The distillate collected is
graduated into volatile oil. The amount thus obtained is recorded from the tube. IX.Volatile oil content:
Efficiency of several drugs is due to their odorous principle (volatile oils).Such crude drugs are
standardized on the basis of their volatile oil contents. Weighed quantity of the drug is boiled with water
in a round bottomed flask fitted with clevenger apparatus. The distillate collected is graduated into
volatile oil. The amount thus obtained is recorded from the tube. S.NO DRUGS VOLATILE OIL
CONTENT (% W/W 1 CARAWAY NLT 2.5 2 CLOVE NLT 15.0 3 FRESH LEMON PEEL NLT 205 4
FENNEL NLT 1.4 5 DILL NLT 205 NLT= Not less than
43. 43. The physical or Physico chemical parameters useful in quality profile of a crude drug evaluation
The maximum limit for the foreign organic matter is defined in the monograph of crude drug. If it
exceeds the limits, deterioration in quality of the drug takes place.  The parts of the organ or organs
other than those named in the definition and description of the drug are defined as foreign organic
matter.  The physical or Physico chemical parameters useful in quality profile of a crude drug
evaluation x.Foreign organic matter:  The maximum limit for the foreign organic matter is defined in the
monograph of crude drug. If it exceeds the limits, deterioration in quality of the drug takes place.  The
parts of the organ or organs other than those named in the definition and description of the drug are
defined as foreign organic matter. x.Foreign organic matter:
44. 44. XI.Swelling Factor: Significances : Useful in the evaluation of crude drugs containing mucilage
Useful for the detection of purity of the crude drug Determination 1. Transfer 1 gm of the seeds to a
25ml stoppered cylinder 2. Fill up to the 20ml mark on the cylinder with water. Agitate gently and
occasionally during 24 hours and allowed to stand 3.Measure the volume occupied by the swollen seeds
XI.Swelling Factor: Significances : Useful in the evaluation of crude drugs containing mucilage Useful for
the detection of purity of the crude drug Determination 1. Transfer 1 gm of the seeds to a 25ml
stoppered cylinder 2. Fill up to the 20ml mark on the cylinder with water. Agitate gently and occasionally
during 24 hours and allowed to stand 3.Measure the volume occupied by the swollen seeds
45. 45. 4. Chemical Evaluation • Determination of the active constituent in a drug by chemical tests is
referred to as chemical evaluation. • The following are various methods of chemical evaluation 1.
Instrumental methods 2. Chemical tests 3. Individual constituent chemical tests 4. Micro chemical tests
4. Chemical Evaluation • Determination of the active constituent in a drug by chemical tests is referred
 to as chemical evaluation. • The following are various methods of chemical evaluation 1. Instrumental
methods 2. Chemical tests 3. Individual constituent chemical tests 4. Micro chemical tests
46. 46. 1.Instrumental methods: They make use of various instruments for evaluation like colorimetry,
flourimetry spectrophotometry etc. 2.Chemical constants tests: These are like acid value, iodine value
and ester value etc are used for the identification of fixed oils and fats. 3.Individual chemical tests:
These are the tests which are used for identifying particular drugs. 4.Microchemical tests: These are the
tests which are carried on slides. Example: Euginol in clove oil is precipitated as potassium euginate
crystals. 1.Instrumental methods: They make use of various instruments for evaluation like colorimetry,
flourimetry spectrophotometry etc. 2.Chemical constants tests: These are like acid value, iodine value
and ester value etc are used for the identification of fixed oils and fats. 3.Individual chemical tests:
These are the tests which are used for identifying particular drugs. 4.Microchemical tests: These are the
tests which are carried on slides. Example: Euginol in clove oil is precipitated as potassium euginate
crystals.
47. 47. These extracts along with positive and negative controls were tested for the presence of active
phytochemicals viz: tannins, alkaloids, phytosterols, triterpenoids, falvonoids, cardiac glycosides,
anthroquinone glycosides, saponins, carbohydrates, proteins, amino acids and fixed oilsExtract
obtained using petroleum ether, chloroform, ethanol and water was prepared using the respective
solvent. Method for chemical evaluation & These extracts along with positive and negative controls
were tested for the presence of active phytochemicals viz: tannins, alkaloids, phytosterols, triterpenoids,
falvonoids, cardiac glycosides, anthroquinone glycosides, saponins, carbohydrates, proteins, amino
acids and fixed oilsExtract obtained using petroleum ether, chloroform, ethanol and water was
prepared using the respective solvent. fats following standard methods Method for chemical evaluation
& fats following standard methods
48. 48. I. Tannins 1. Ferric chloride Test: Added a few drops of 5% ferric chloride solution to 2 ml of the test
solution. Formation of blue color indicated the presence of hydrolysable tannins. 2. Gelatin Test: Added
five drops of 1% gelatin containing 10% sodium chloride to 1 ml of the test solution. Formation of white
precipitates confirmed the test. II. Alkaloids Approximately 50 mg of extract was dissolved in 5 ml of
distilled water. Further 2M hydrochloric acid was added until an acid reaction occurred and filtered. The
filtrate was tested for the presence of alkaloids as detailed below I. Tannins 1. Ferric chloride Test:
Added a few drops of 5% ferric chloride solution to 2 ml of the test solution. Formation of blue color
indicated the presence of hydrolysable tannins. 2. Gelatin Test: Added five drops of 1% gelatin
containing 10% sodium chloride to 1 ml of the test solution. Formation of white precipitates confirmed
the test. II. Alkaloids Approximately 50 mg of extract was dissolved in 5 ml of distilled water. Further 2M
hydrochloric acid was added until an acid reaction occurred and filtered. The filtrate was tested for the
presence of alkaloids as detailed below
49. 49. 1.Dragendorff’s Test: To 2 ml of the filtrate was added 1 ml of Dragendorff’s reagent. Formation of
orange or reddish brown precipitate indicated the test as positive. 2.Mayer’s Test: To 1 ml of test
solution or filtrate was added a drop or two of the Mayer’s reagent. white or a creamy precipitate
confirmed the test as positive. 3.Hager’s Test: To 1 ml of test solution or filtrate, a drop or two of Hager’s
reagent formation of yellow precipitate indicated the test as positive. 4.Wagner Test: Two drops of
Wagner’s reagent was added to 1ml of the test solution. The formation of yellow or brown precipitate
confirmed the test as positive for alkaloids. 1.Dragendorff’s Test: To 2 ml of the filtrate was added 1 ml
of Dragendorff’s reagent. Formation of orange or reddish brown precipitate indicated the test as positive.
2.Mayer’s Test: To 1 ml of test solution or filtrate was added a drop or two of the Mayer’s reagent. white
or a creamy precipitate confirmed the test as positive. 3.Hager’s Test: To 1 ml of test solution or filtrate,
a drop or two of Hager’s reagent formation of yellow precipitate indicated the test as positive. 4.Wagner
Test: Two drops of Wagner’s reagent was added to 1ml of the test solution. The formation of yellow or
brown precipitate confirmed the test as positive for alkaloids.
50. 50. III. Phytosterols 1. Liebermann-Burchard’s Test: The extract (2 mg) was dissolved in 2 ml of acetic
anhydride, heated to boiling, cooled and then 1 ml of concentrated sulfuric acid was added.A brown ring
formation at the junction and the turning of the upper layer to dark green color confirmed the test for the
presence of phytosterols. IV. Triterpenoids 1. Salkowski Test: Approximately 2 mg of dry extract was
shaken with 1 ml of chloroform and a few drops of concentrated sulfuric acid were added.A red brown
color formed at the interface indicated the test as positive for triterpenoids. III. Phytosterols 1.
Liebermann-Burchard’s Test: The extract (2 mg) was dissolved in 2 ml of acetic anhydride, heated to
boiling, cooled and then 1 ml of concentrated sulfuric acid was added.A brown ring formation at the
junction and the turning of the upper layer to dark green color confirmed the test for the presence of
phytosterols. IV. Triterpenoids 1. Salkowski Test: Approximately 2 mg of dry extract was shaken with 1
ml of chloroform and a few drops of concentrated sulfuric acid were added.A red brown color formed at
the interface indicated the test as positive for triterpenoids.
51. 51. V. Flavonoids 1.Shinoda test:A few magnesium turnings and 5 drops of concentrated hydrochloric
acid was added drop wise to 1 ml of test solution. A pink, scarlet, crimson red or occasionally green to
blue color appeared after few minutes confirmed the test. 2. Alkaline reagent test: Addition of 5 drops of
5% sodium hydroxide to 1 ml of the test solution resulted an increase in the intensity of the yellow color
which became colorless on addition of a few drops of 2 M hydrochloric acid which indicated the
presence of falvonoids. 3.Lead acetate test: A few drops of 10% lead acetate added to 1ml of the test
solution resulted in the formation of yellow precipitate confirmed the presence of falvonoids. V.
 Flavonoids 1.Shinoda test:A few magnesium turnings and 5 drops of concentrated hydrochloric acid was
added drop wise to 1 ml of test solution. A pink, scarlet, crimson red or occasionally green to blue color
appeared after few minutes confirmed the test. 2. Alkaline reagent test: Addition of 5 drops of 5%
sodium hydroxide to 1 ml of the test solution resulted an increase in the intensity of the yellow color
which became colorless on addition of a few drops of 2 M hydrochloric acid which indicated the
presence of falvonoids. 3.Lead acetate test: A few drops of 10% lead acetate added to 1ml of the test
solution resulted in the formation of yellow precipitate confirmed the presence of falvonoids.
52. 52. VI. Saponins 1.Foam Test: 5 ml of the test solution taken in a test tube was shaken well for five
minutes. Formation of stable foam confirmed the test. 2. Olive oil test: - Added a few drops of olive oil to
2ml of the test solution and shaken well. The formation of a soluble emulsion confirmed the test. VII.
Cardiac glycosides 1.Keller -Killiani test: Added 0.4 ml of glacial acetic acid and a few drops of 5% ferric
chloride solution to a little of dry extract. Further 0.5 ml of concentrated sulfuric acid was added .The
formation of blue color in acetic acid layer confirmed the test. VI. Saponins 1.Foam Test: 5 ml of the test
solution taken in a test tube was shaken well for five minutes. Formation of stable foam confirmed the
test. 2. Olive oil test: - Added a few drops of olive oil to 2ml of the test solution and shaken well. The
formation of a soluble emulsion confirmed the test. VII. Cardiac glycosides 1.Keller -Killiani test: Added
0.4 ml of glacial acetic acid and a few drops of 5% ferric chloride solution to a little of dry extract. Further
0.5 ml of concentrated sulfuric acid was added .The formation of blue color in acetic acid layer
confirmed the test.
53. 53. VIII. Test for carbohydrates 1.Molisch’s test: To 1 ml of test solution added a few drops of 1 % alpha-
napthol and 2-3 ml concentrated sulfuric acid. The reddish violet or purple ring formed at the junction of
two liquids confirmed the test. 2. Barfoed’s test: 2ml of reagent was added to 2 ml of the test solution,
mixed & kept a in boiling water bath for 1 min. Red precipitate formed indicates the presence of
monosaccharide's. 3.Seliwanoffs test: To 3 ml of Seliwanoffs reagent was added to 1 ml of the test
sample and heated on a water bath for one minute. The formation of rose red color confirmed
carbohydrates VIII. Test for carbohydrates 1.Molisch’s test: To 1 ml of test solution added a few drops of
1 % alpha-napthol and 2-3 ml concentrated sulfuric acid. The reddish violet or purple ring formed at the
junction of two liquids confirmed the test. 2. Barfoed’s test: 2ml of reagent was added to 2 ml of the test
solution, mixed & kept a in boiling water bath for 1 min. Red precipitate formed indicates the presence of
monosaccharide's. 3.Seliwanoffs test: To 3 ml of Seliwanoffs reagent was added to 1 ml of the test
sample and heated on a water bath for one minute. The formation of rose red color confirmed
carbohydrates
54. 54. 4. Fehlings test: Dissolved 2 mg dry extract in 1 ml of distilled water and added 1ml of
Fehling’s(A+B) solution, shooked and heated on a water bath for 10 minutes. The brick red precipitate
formed confirmed the test IX. Anthraquinone glycosides Hydroxyanthraquinone Test To 1 ml of the
extract, added a few drops of 10% potassium hydroxide solution. The Formation of red color confirmed
the test. X. Test for proteins 1. Biuret test: To 2 ml of the test solution added 5 drops of 1% copper
sulphate solution and 2 ml of 10% NaOH .Mix thoroughly. Formation of purple or violet color confirmed
proteins. 4. Fehlings test: Dissolved 2 mg dry extract in 1 ml of distilled water and added 1ml of
Fehling’s(A+B) solution, shooked and heated on a water bath for 10 minutes. The brick red precipitate
formed confirmed the test IX. Anthraquinone glycosides Hydroxyanthraquinone Test To 1 ml of the
extract, added a few drops of 10% potassium hydroxide solution. The Formation of red color confirmed
the test. X. Test for proteins 1. Biuret test: To 2 ml of the test solution added 5 drops of 1% copper
sulphate solution and 2 ml of 10% NaOH .Mix thoroughly. Formation of purple or violet color confirmed
proteins.
55. 55. XI. Test for amino acids 1. Millon’s test: Added 5 drops of millons reagent to 1 ml of test solution and
heated on a water bath for 10 min, cooled and added 1% sodium nitrite solution. Appearance of red
color confirmed the test. XII. Fats and fixed oils To 5 drops of the sample was added 1 ml of 1% copper
sulphate solution and a few drops of 10% sodium hydroxide. The formation of a clear blue solution
confirmed the test. XI. Test for amino acids 1. Millon’s test: Added 5 drops of millons reagent to 1 ml of
test solution and heated on a water bath for 10 min, cooled and added 1% sodium nitrite solution.
Appearance of red color confirmed the test. XII. Fats and fixed oils To 5 drops of the sample was added
1 ml of 1% copper sulphate solution and a few drops of 10% sodium hydroxide. The formation of a clear
blue solution confirmed the test.
56. 56. 5.Analytical evaluation Chromatographic techniques: a)TLC-Thin layer chromatography b)HPTLC-
High performance thin layer chromatography c)HPLC-High performance/pressure liquid chromatography
d)GLC-Gas chromatography e)CC-column chromatography f)Gel permeation chromatography g)Affinity
chromatography 5.Analytical evaluation Chromatographic techniques: a)TLC-Thin layer chromatography
b)HPTLC-High performance thin layer chromatography c)HPLC-High performance/pressure liquid
chromatography d)GLC-Gas chromatography e)CC-column chromatography f)Gel permeation
chromatography g)Affinity chromatography
57. 57. Spectrophptometric methods: i) UV- Ultra violet /visible spectroscopy ii)IR-Infra Red spectroscopy iii)
Fluorescence analysis iv) NMR-nuclear magnetic resonance spectroscopy v) MS-Mass spectroscopy vi)
X-ray diffraction vii) RIA-radio immuno assay Spectrophptometric methods: i) UV- Ultra violet /visible
spectroscopy ii)IR-Infra Red spectroscopy iii) Fluorescence analysis iv) NMR-nuclear magnetic
resonance spectroscopy v) MS-Mass spectroscopy vi) X-ray diffraction vii) RIA-radio immuno assay
58. 58. TLC/HPTLC are micro analytical techniques used for determination of natural products Advantages
:simple in operation and rapid The Rf value vary depend on the pirity,nature,of substance, composition
of solvent and impurities  TLC is useful to analyse Alkaloids, Glycosides like all bio- constituents 
After development of chromatography spots are revealed by spraying with suitable detecting agent 
Adsorbent silica gel G/C coated to a thickness of minutes and used.  TLC/HPTLC are micro analytical
techniques used for determination of natural products Advantages :simple in operation and rapid
Chromatographic techniques: a)TLC-Thin layer chromatography Principle :Adsorption  The Rf value
vary depend on the pirity,nature,of substance, composition of solvent and impurities  TLC is useful to
analyse Alkaloids, Glycosides like all bio- constituents  After development of chromatography spots
are revealed by spraying with suitable detecting agent  Adsorbent silica gel G/C coated to a thickness
of minutes and used. Chromatographic techniques: a)TLC-Thin layer chromatography Principle
:Adsorption