Unit 1 Quality control and Standardization (Evaluation Parameter) of Herbals

1. QUALITY CONTROL OF DRUGS OF
NATURAL ORIGIN
PREPARED BY
MRS. MEGHA S SHAH
1

2. Objective of Study
Confirmation of its identity
determination of its quality, purity
detection of nature of adulteration
2

3. ADULTERATION
Adulteration is a practice of substituting original crude drug partially
or whole with other similar looking substances but the latter is either
free from or inferior in chemical and therapeutic properties.
or
Adulteration in simple words is the debasement of an article.
or
Adulteration is broadly defined as admixture or substitution of original
or genuine article/ drug with inferior, defective or otherwise useless or
harmful substances.
3

4. ADULTRANT
The adulterant must be some material which in both cheap and available in
fairly large amounts.
4

5. TYPES OF ADULTRATION
1. Deliberate ( Intentional ) adulteration : Deliberate adulteration –
Are normally commercial mainly with the intention of enhancement
of profits.
2. Accidental ( In-deliberate) adulteration
5

6. REASON FOR ADULTRATION
1. Scarcity of the drug
2. The high price of the drug in the market,
eg: Clove, Cinnamon, Cardamom
3. It is very common with the contraband drugs
e.g. Opium
6

7. The term 'adulteration' or debasement of an article covers a number of conditions,
which may be deliberate or accidental.
Inferiority: is a natural substandard condition (e.g. where a crop is taken whose
natural constituent is below the minimum standard for that particular drug) which
can be avoided by more careful selection of the plant material.
Spoilage is a substandard condition produced by microbial or other pest infestation,
which makes a product unfit for consumption, which can be avoided by careful
attention to the drying and storage conditions.
7

8. Deterioration: is an impairment of the quality or value of an article due to
destruction or abstraction of valuable constituents by bad treatment or
aging or to the deliberate extraction of the constituents and the sale of the
residue as the original drugs.
Admixture: is the addition of one article to another through accident,
ignorance or carelessness e.g. inclusion of soil on an underground organ or
the co-collection of two similar species.
8

9. Sophistication: is the deliberate addition of spurious or inferior material
with intent to defraud; such materials are carefully produced and may
appear at first sight to be genuine e.g. powder ginger may be diluted with
starch with addition of little coloring material to give the correct shade of
yellow colour.
Substitution: is the addition of an entirely different article in place of that
which is required e.g. supply of cheap cottonseed oil in place of olive oil.
9

10. TYPES OF ADULTRATION/SUBSTITUTION OF
HERBAL DRUGS
1. Substitution with substandard Commercial Varieties The adulterants used here
may resemblance to the authentic drugs by morphological, chemical or therapeutic
characters, but are substandard in nature and hence cheaper in cost. This is rather a most
common practice of adulteration.
E.g. Strychnous nux blanda or S.Potatorum in place of S. Nux vomica
Capsicum minimum replaced by C.Annum
Arabian Senna (Cassia angustifolia ) and dog Senna (Cassia obovata ) have been used to adulterate
Indian Senna (Cassia senna)
Gentian substituted by kutki
Japanese ginger ( Zingiber mioga ) to adulterate medicinal ginger (Zingiber officinale).
10

11. 2. Adulteration by Artificially Manufactured Substitutes
It has been also observed that substances artificially prepared to resemble original
drug are used as substitutes. Generally this practice is followed for much costlier
drugs.
Compressed chicory in place of coffee,
paraffin wax made yellow coloured and substituted for beeswax,
properly cut and shaped basswood for nutmeg
11

12. 3. Substitution by Exhausted Drugs
 Here the same plant material is mixed which is having no active medicinal components
as they have already been extracted out. This practice is most common in case of volatile
oil containing materials like clove, fennel etc.,
 where the dried exhausted material resembles the same like original drug (similarly with
drugs like Cascara sagrada and ginger). Sometimes when coloring matters have been
extracted or removed during exhaustion, the residue is re-colored with artificial dyes as
is done with saffron and red rose petals.
12

13. 4. Substitution by Superficially Similar inferior drugs
These inferior drugs used may or may not be having any chemical or therapeutic value
as that of original natural drug. Due to their morphological resemblance to
authenticate drug, they are marketed as adulterants.
Belladona leaves are substituted with Ailanthus leaves,
saffron is admixed with dried flowers of Carthamus Tinctorius,
scented bdellium is used for myrrh,
mother cloves and clove stalks are mixed with clove and
beeswax is substituted by Japan wax
13

14. 5. Harmful adulterants
The waste from market are collected and admixed with authenticate drugs. This is particularly
noticed for liquids or unorganized drugs.
Example include piece of amber coloured glass in colophony,
limestone in asafetida,
lead shot in opium,
white oil in coconut oils,
cocoa butter mixed with stearin or paraffin
Rodent fecal matter to cardamom seed is very harmful adulterant.
14

15. 6. Adulteration of powders
Besides the entire drugs, the powdered forms are frequently found to be adulterated.
Some examples are
Dextrin in Ipecacuanha,
powdered liquorice or gentian admixed with powdered olive stones,
exhausted ginger powder in powdered colocynth or ginger,
red sanders wood in capsicum.
Powdered bark is frequently found to be adulterated with brick powder.
15

16. 7. Presence of Vegetative Matter from the Same Plant
 Sometimes, the other miniature plants growing along with medicinal plant are mixed
with drug due to their resembling colour, odour and in some cases constituents.
 For example liver warts and epiphytes growing in bark portion are mixed with
Cascara or Cinchona
 stems of portions are mixed along with leaf drugs like stramonium, lobelia and senna.
16

17.  Besides these common practices, sometimes other methods are employed
like use of synthetic chemical to enhance the natural character as in case
of addition of
 benzyl benzoate to balsam of peru,
 Citral to citrus oils like oil of lemon and orange oil
17

18. DRUG EVALUATION
OBJECTIVE TO STUDY DRUG EVALUATION
Biochemical variation in the drug
Deterioration due to treatment and storage
Substitution and adulteration, as a result of carelessness, ignorance or fraud
18

19. Organoleptic evaluation (Morphological evaluation) : Natural and degree of
evaluation of crude drugs has undergone a systematic change.
Microscopically evaluation : Used to identified the organized drugs by their known
histological character. Mostly used for qualitative evaluation.
Chemical Evaluation: method of estimating active constituents present in the crude drug
Physical evaluation: with advance in separation technique and instrumental analysis,
possible to perform analysis both of qualitative and quantitative
Pharmacological evaluation: Biological behavior of crude drug extracts constitute
pharmacological evaluation
19

20. ORGANOLEPTIC EVALUATION
 Organoleptic (or morphological) refers to evaluation by means of the organs
of sense and includes the macroscopic appearance of the drug, its colour,
Odour and taste, occasionally the sound or snap of its fracture and the feel of
the drug to touch.
For convenience of description the macroscopic characteristics of a drug may be
divided into following headings.
 Shape and size Colour
 Fracture External markings
 Odour and taste
20

21. SHAPE AND SIZE OF CRUDE DRUGS
Crude drugs can be of different shapes etc.
Cylindrical (Sarsaparilla)
Sub cylindrical (Podophyllum)
21

22. Conical (Aconite) Fusiform (Sweet potato) Ovoid (Cactus)
22

23. Pyriform (Jalap) Terrete (Orchid) Disc shape (Calumba)
23

24. Leaf
Types of leaf Apex Types of leaf Margin
24

25. Flowers
 A flower is a modified shoot meant for
production of seeds and it is built up on the
enlarged end of stem called thalamus.
 It consist of four basic parts
 Calyx
 Corolla
 Androceium
 Gynoecium
 The bunch of flower is called inflorescence
Types of inflorescene
25

26. 26

27. Seed
 A seed is a fertilized ovule.
 It consist of three parts. Coat, embryo
and endosperm
 Seed are characterized by the hilum, a
point of attachment of seed to stalk
 Microphyle, a minute opening for
absorption of water and
 the raphe, a longitudinal marking of
adherent stalk.
27

28. Fruits
 Fruits are matured ovary with or without associated parts.
1. Simple fruits: which develops from single ovary of single flower with or without other parts. (a)
dehiscent (b) indehiscent
2. Aggregate fruits: developed from polycarpellory apocarpus ovary. Each carpel forms a single
fruitlets. All fruitlets arise from a single flower and are attached on the same axis therefore termed as
aggregate fruits.
3. Composite fruits: developed from inflorescence. The peduncle, perianth or calyx and corolla as well
as ovular parts after maturity and ripening forms a fleshy fruit. The shape of fruits may be oblong,
ellipsoidal and globular. The example of fruit drugs are fennel, coriander, cardamom, dill, bael
28

29. SIZE OF CRUDE DRUGS
Size of crude drugs may vary because their length, diameter and width is different.
Size of crude drugs can be measured by vernier callipers in cm & mm.
COLOUR
External colour of crude drugs is mostly greyish brown and internal colour is
whitish yellow, external colour maybe yellowish grey, yellowish brown, reddish
orange to brown black. It is often more or less grey from clay dust.
29

30. FRACTURE
There are three types of fracture.
 Complete fracture : the drug breaks completely when pressure is applied.
 Incomplete fracture: the drug does not break completely when fracture is
applied.
 Short fracture: the drug breaks with a snap sound when pressure is applied.
30

31. Nature of fracture
Nature of fractured surface is as important as fracture.
 Even: smooth surface
 Uneven: irregularly broken surface
 Granular : grain like appearance
 Hard: a compact surface
 Horny: Horn like surface
 Powdered: powdery surface
 Resinous: smooth glossy surface
 Conchodial : a resinous surface having the surfaces curved in convex and concave form.
31

32. External markings
How the drug looks externally. They may be classified as
 Furrows: alternate ridges and valleys due to shrinkage of internal parts caused by drying
 Wrinkles: fine or delicate furrows
 Annulations: transverse ring like markings on exterior
 Fissures: splits extending into tissues
 Nodule: rounded outgrowth on surface
 Scars: produced due to injury or cut
32

33. ODOR
 It is a very sensitive criteria based on individual perception. Odor may be divided
into two main classes
(i) Distinct odor e.g. camphor and clove (ii) Indistinct odor
 depending upon the amount of volatile constituents the drug possesses.
The different odors may include
1. Aromatic odor
2. Alliaceous
3. Camphoraceous
4. Spicy etc.
33

34. Taste
Usually crude drugs cannot be tasted because some may be poisonous or may cause
allergic reactions. But some tastes which are known are:
 Sweet taste e.g. liquorice, honey
 Acidic/sour taste e.g. lemon
 Saline/salty taste e.g. kashar
 Acrid taste : unpleasant and irritating e.g. Jalap
34

35. 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.
35

36. Microscopic or Anatomical Evaluation
Microscopic
evaluation
Quantitative
Microscopy
Stomatal number
Stomatal index
Palisade ratio
Vein islet number
Vein termination
number
Qualitative
Microscopy
Stomata
Trichomes
Starch grains
Calcium oxalate crystals
Xylem vessels
36

37. Stomata
The stomata are minute pores which occur in the epidermis of the plants. Each
stoma remains surrounded by two kidneys or bean shaped epidermal cells the
guard cells.
The stomata may occur on any part of a plant except the roots. The epidermal
cells bordering the guard cells are called accessory cells or subsidiary cells.
Stomata is one of the best standard to identify the plant microscopically.
37

38. Function of stomata: Gaseous exchange and Tanspiration
The leaves of bryophytes and submerged leaves of aquatic parts do not contain stomata.
Mostly present in green parts of plants (mostly in leaves) and absent in roots.
They are present in stem (ephedra), flower (Clove) and fruits (fennel)
Abundantly present in dicot leaves.
In some cases present in upper surface of leaves while in other cases lower surface only
(coca and cherry).
In some cases present in both surface of leaves (Senna, Belladona, Datura)
38

39. Types of stoma
1.Moss type: Found on the apophysis of the areca and when mature posses guard cells which are united
by the breaking down of the dividing wall during growth. E.g funaria
2.Gymnospermous type : Characteristic of Graminaceae and cyperaceae and has guard cells which in
surface view are more or less dumb bell shaped and outline sub-rectangular.
3.Gramineous type: Have guard cell oval In transverse section and are place at an angle of about
45 with outer surface and have walls which are in part lignified. E.g savin
4.Dicoteyledonous: It is having diagnostic significance and classified based on form of
arrangement of subsidiary cells.
39

40. Various types of stomata
1. Paracytic stomata
2. Diacytic stomata
3. Anisocytic stomata
4. Anomocytic stomata
5. Actioncytic stomata
40

41.  Paracytic or rubiaceous or parallel-cell stomata: in this stomata two guard cells covered
by two subsidiary cells, e.g. senna
 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
 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
 Anomocytic or ranunculaceous or irregular- celled: In this type stoma is surrounded by
varying number of subsidiary cells. e.g. digitalis
 Actinocytic or radiate-celled stomata: two guard cells are surrounded by radiating
subsidiary cells.
41

42. Leaf constant
 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.
 Stomatal number: The average number of stomata present per square millimeter of the
epidermis is known as stomatal number
42

43. 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 = S/E+S*100
S.I = Stomatal index
S= Number of stomata per unit area
E=Number of epidermal cells in the same unit area
43

44.  Vein-islet Number: Vein-islet number is defined as the number of vein-islets per
sq.mm. of leaf surface.
 Vein-termination Number: It is defined as the number of veinlet terminations per.
sq. mm of the leaf surface between midrib and margin.
44

45. 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.
45

46. Trichomes
 Trichomes or plant hairs Trichomes are the tubular elongated or glandular outgrowth
of the epidermal cells.
 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
46

47.  Function of trichomes:
 Ion exchange
 Protective
 Secretion of essential oil
 Absorption of water
47

48. Trichomes
1. Unicellular Trichomess:
Vary From Small Papillose Outgrowth To
Large Robust Structure.
Linear, Thick Walled And Warty
Trichomes- Damiana
Linear Strongly Waved, Thick Walled
Trichomes- Yerba Santa
Large, Conical, Longitudinally Striated
Trichomes: Lobellia
Long, Tubular, Flattened And Twisted
Trichomes: Cotton
Lignified Trichomes: Nux Vomica,
Strophanthus
Short Conical Trichomes: Tea
Short, Conical, Warty Trichomes: Senna
Short, Sharp, Pointed, Curved, Conical
Trichomess-cannabis
48

49. 2. Multicellular Trichome
(A) Multicellular Unbranched Trichomes
(i) Uniseriate
Bicellular, Conical: Datura
Three Celled Long: Stramonium
Four To Five Celled Long: Belladona
(ii) Biseriate: Calendula Officinalis
(iii) Multiseriate: Euphorbia Pilulifera , Male
Fern
(B) Multicellulat Branched Trichomes:
(I)stellate: Hamamelis And Altheae
(II) Peltate: Leaves Of Eleagnus, Leaves And
Stem Of Croton Eleuteria
Candelabra (Uniseriate Branched Axis) :
Verbascum Thapsus And Rosemary
T-shaped Trichomes: Pyrethrum And
Artemisia
49

50. 1. Unicellular glandular trichomes: they do not possess
stalk.
2. Multicellur glandular trichome:
Uniseriate stalk with single spherical secreting cell at the
apex: Digitalis pupurea
Uniseriate mutlicellular stalk with single spherical cell at
the apex: Digitalis thapsi and belladona
Unicellular stalk and bicellular head: D. purpurea
Unicellular stalk and multicellular head: Hyoscyamus
Biseriate stalk and biseriate secreting head: santonica
Short stalk and secreting head formed of a rosette of club
shaped cell: Mentha species
Multiseriate cylindrical stalk and a capitate rosette of
secreting cells: cannabis
Glandular trichomes
50

51.  Hydathodes
 Hydathodes are glands of secretion or absorption developed in certain plants.
 They may consist of unicellular or multicellular hairs.
 They are most commonly found on the leaves of aquatic plants or herbaceous
plants growing in most places.
 They occur either at the tip or on the margins of the leaves.
 Each hydathodes is found in very intimate relation of a vein. E.g Piper betle
51

52. 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.
52

53.  The percentage purity of an authentic ginger powder calculated as follows
N x W x 94,000 x 100 = % Purity of drug
S x M x P
 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
53

54. Camera Lucida
 The principle of the Swift Ives camera lucida Light
from the viewed object is reflected twice as it passes
through a prism, so the object is perceived by the eye
as in nature, right side up. ... At the same time, the eye
receives light directly from the drawing paper.
 Magnified image of the object under the microscope
can be traced on paper.
 While using the instruments the illumination of both
object and paper must be titled at the correct angle to
avoid distoration.
54

55.  Swift-Ives camera lucida should be fit over the
eyepiece and the light from the object passes direct
to the observer’s eye through on opening in the
silvered surfaced of the left hand prison.
 At the same time light from the drawing paper
and pencil is reflected by the right hand hand
prism and by the silvered surface so that the pencil
appears superimposed on the object which can be
traced.
 For proper position palce a stage micrometer on
the microscope stage and trace its division on
paper.
55

56. Physical Evaluation
 Physical constant 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
are some important parameters used in the evaluation of drugs. Ultraviolet light is also used for
determining the fluorescence of extracts of some drugs.
I. Moisture Content (water content) II. Viscosity (viscosity of drug containing gum)
III. Melting point and boiling 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
56

57. 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.
57
S.NO Drugs total ash(% w/w) acid insoluble ash % (w/w)
1 Agar - 1.00
2 Bael 03.5
3 Cannabis 15.00 5.00
4 Gelatin 3.6 -
5. Valerin 12.0

58. TYPES OF ASH 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).
58

59. Determination Total ash value
 Weigh accurately about 3gms of the powdered drug in a tared silica crucible
 Incinerate the powdered drug by gradually increasing the heat until free from carbon and
cool. Keep it in desiccators .
 Weigh the ash and calculate the % of the total ash with reference to the air dried sample.
 Determination of Acid insoluble ash value 1. Boil the total ash obtained as above for 5
minutes with 25ml of dilute HCL
 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
 Weigh the residue and calculate the acid insoluble ash of the drug
59

60.  Determination of Water soluble ash value
 It is used to detect the presence of material exhausted by water.
 Boiled the total ash with 25 ml of water for 5 minutes.
 Filter through ashless filter paper.
 Wash the residue with hot water and ignite the tarred crucible by using nuffle
furnace, cool and keep in desiccators.
 Weight the ash obtained after incineration and calculate water soluble ash of
drug with reference to air dried sample.
60

61. 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
61

62. Types of extractive values
A. Water soluble extractive values
B. Alcohol soluble extractive values
C. Ether soluble extractive values
62

63. Water soluble extractive value
 Macerate about 5gm of the accurately weighed coarse powder with 100ml of
chloroform water in a 100ml volumetric flask for 24 hours .
 Shake frequently for first 6 hours
 Filter rapidly through filter paper and evaporate 25ml of water extract to dryness
in a tared flat-bottomed shallow dish.
 Dry the residue at 105 and weigh. Keep it in a desiccators
 Dry the extract to constant weight ,finally , calculate the % W/W of Water soluble
extractive value with reference to the air dried drug.
63

64.  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.
64
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

65. Alcohol soluble extractive values
Macerate about 5gm of the accurately weighed coarse powder with 100ml of 90% alcohol in a
100 ml stoppered flask for 24 hours .
Shake frequently for first 6 hours
Filter rapidly through filter paper and collect the filtrate evaporate 25ml of alcohol extract to
dryness in a tared flat-bottomed shallow dish.
Dry the residue at 105 and weigh. Keep it in a desiccators
Dry the extract to constant weight ,finally , calculate the % w/w of alcohol soluble extractive
value with reference to the air dried drug.
65

66.  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.
66
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

67. 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)
67
S.NO DRUGS LIMIT FOR NON-VOLATILE ETHER
SOLUBLE EXTRACTIVES(% W/W)
1 CAPSICUM NLT 12.0
2 MALE FERN NLT 1.0
3 LINSEED NLT 25.0

68. 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.
68

69. Volatile oil content
69
S.NO DRUGS VOLATILE OIL content
(%w/w)
1 CARAWAY NLT 2.5
2 CLOVE NLT 15.0
3 FRESH LEMON PEEL NLT 2.5
4 FENNEL NLT 1.4
5 DILL NLT 2.5

70. Swelling Factor
Significances :
Useful in the evaluation of crude drugs containing mucilage
Useful for the detection of purity of the crude drug Determination
Procedure:
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.
70

71. Foreign organic matter
I.P describes, foreign organic matter is the material consisting of any or all of the following
(i)parts of the organ from which the drug is derived other than the parts named in
definition and description or for which a limit is prescribed in the individual monograph.
(ii) The parts of the organ or organs other than those named in the definition and
description.
(iii) moulds, insects or other animal contamination
71

72. Foreign organic matter
 The maximum limit for the foreign organic matter for vegetable crude drugs. If it
exceeds the limits than the drug is declared substandard and deterioration in
quality of the drug takes place.
72
Drugs Foreign organic matter
Fennel NMT 2%
Caraway NMT 2%
Cardamom NMT 1%
dill NMT 2%

73. 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.
73
Sr no Drug Moisture content %W/W
1 Digitalis NMT 5
2 Balsam of peru NMT 4
3 Storax NMT 5
4 Tragacanth NMT 15

74. 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 standardizing liquid drugs.
i)Liquid paraffin-kinematic viscosity not less than 64-centistokes at 37.8°
ii) Pyroxylin-kinematic viscosity,1100-2450 centistokes
74

75. 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.
75
Sr No Drugs Melting Point (°C)
1 Colophony 75-85
2 Bees Wax 62-65
3 Wool Fat 34-44

76. Solubility
 The presence of adulterant in a drug could be indicated by solubility studies.
76
S.No Drug Solubility
1 Castor Oil Soluble In 3 Volumes Of Alcohol
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

77. 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.
77
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°

78. 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.
78
S.No Drugs Refractive Index
1 Arachis Oil 1.4678 To 10470
2 Castor Oil 104758 To 10527 3
4 Clove Oil 1.527 To 10535

79. 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 Constituent tests
3. Individual chemical tests
4. Micro chemical test
79

80. 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.
80

81. Method for chemical evaluation
 Extract obtained using petroleum ether, chloroform, ethanol and water prepared
using the respective solvent.
 These extracts along with positive and negative controls test for the presence of
active phytochemicals viz: tannins, alkaloids, phytosterols, triterpenoids,
flavonoids, cardiac glycosides, anthroquinone glycosides, saponins,
carbohydrates, proteins, amino acids and fixed oils & fats.
81

82. 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.
82

83. 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.
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.
83

84. Phytosterols
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.
Triterpenoids
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.
84

85. 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 flavonoids.
85

86. 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.
Cardiac glycosides
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.
86

87. 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.
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.
87

88. 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.
Test for proteins
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.
88

89. Test for amino acids
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.
89

90. 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
90

91. Spectrophotometric 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
91

92. TLC-Thin layer chromatography
Principle :Adsorption
Adsorbent silica gel G/C coated to a thickness of minutes and used.
After development of chromatography spots are revealed by spraying with suitable detecting
agent .
TLC is useful to analyse Alkaloids, Glycosides like all bio- constituents.
The Rf value vary depend on the pirity,nature,of substance, composition of solvent and
impurities.
TLC/HPTLC are micro analytical techniques used for determination of natural products
Advantages :simple in operation and rapid Chromatographic techniques
92

93. Thin layer chromatography(TLC)
has become increasingly popular for both qualitative and quantitative evaluation
of drugs.
Rf values refers to the ration of distance travelled by the solute to the distance
moved by the solvent on a thin layer adsorbent.
Rf = Distance travelled by the compound(solute)
Distance travelled by the solvent
93

94. HPTLC-High performance thin layer chromatography
 It is very useful qualitative/quantitative method for pharmaceutical analysis.
 HPTLC is a major advancement of TLC principle requiring shorter time better
resolution.
 HPTLC plates available in the form of pre coats  Silica gel-Gel with very small
particle size used as a stationary phase gives rapid separation with sensitivity .
 About 36 cm solvent front migration is sufficient to effect proper separation.
 HPTLC plates are produced from 4-5μm layer.
 About 7cm distance achieved in about 4 minutes.
94

95.  Sample preparation :HPTLC needs high concentration sample.
 Small amounts of sample need to apply, sample spot size 1 mm in diameter and sample
applied by capillaries.
95
S.NO COMPONENT MOBILE PHASE
1 Amino acids, alkaloids Butanol-acetic acid-water(4:5:1)
2 Cardiac glycoside Dichloromethane-methnol- formamide(8:2:1)
3 Flavonoids, coumarin glycosides acetate-methyl ketone-acetic acid- water(5:3:1:1)
4 Saponins Chloroform-methanol-water(7:4:1:)
5 Terpenes, essential oils ,sterols Hexane-acetone-(9:1)

96. HPLC-High performance/pressure liquid
chromatography
The term liquid chromatography used to refer to those methods in which the separation takes
place with packed column.(stationary)
A liquid mobile phase used eluent.
In HPLC mobile phase forced to column under high pressure
Derivatisation in HPLC undertaken to increase sensitivity of detection for a given compound.
Colum used in HPLC narrow (1 mm or less) flow rate of mobile phase is (100μl /min)
Advantages : most versatile ,safest. Uses :quality control of drugs like morhine, emetine, steroids
96

97. GLC-Gas liquid chromatography
GLC separates volatile substances by percolating a gas stream over a
stationary phase.
Principle :GLC works on partitioning Carrier gas used as mobile phase
(Nitrogen, Helium)
A film of a liquid spread over an inert solid. Acts as stationary phase.
GLC applied for
i. Assay of impurities
ii.Examination of volatile oils plant alkaloids.
97

98. CC-column chromatography
 Liquid chromatography in which mobile phase in form of liquid passes over the
stationary phase packed in a column.
 Colum is either a glass, metallic column. the column adsorption
chromatography is oldest and still practiced to day for extraction process.
98

99. Gel permeation chromatography/Size-exclusion chromatography
 Separation occurs not on the basis of adsorption /partition ,but on the effective
size of solutes present in solution for the separation purpose.
 Stationary phase used are cross linked polymers which give an open network
with large number of pores during flow large size particles can’t enters in to
pores hence excluded.
 Various types of gels are used sofgel, semi-rigid gels, rigid gels.
 Use :separates biomolecules, protiens, poly-peptides.
99

100. Affinity chromatography
 This technique is mainly used for the separation of protiens, enzymes, antigens,
antibodies.
 The adsorbent used is one of biological substance having a specific affinity for
other substance .
 These two substances are biologically interacting pairs such adsorbent is
attached to a porous stationary phase and placed in a column, when mixture
containing the other complement of adsorbent passed through stationary phase
100

101. UV- Ultra violet /visible spectroscopy
 Ultra violet –visible absorption techniques encompass analytical methods
based up on measurement of light absorption by substances in wave length
region from 190 to 900 nm 190-380 nm UV region and 380-900 nm visible
region.
 Applications :we can analyze variety of pharmaceutical phytoconstituents like
Lobeline-244 nm, Morphine-286 nm Antharaquinone 505 nm
Spectrophptometric methods
101

102. IR-Infra Red spectroscopy
 IR is the study of reflected, absorbed, transmitted radiant energy in region of
electromagnetic spectrum 0.8 to 500 nm
 It is divided in to three regions Near IR-12,500-400 cm-1, Mid IR-4000-400 cm-1
Far IR-400-20cm-1
 IR spectrophotometer can be divided in to single and double beam and Fourier
transform spectrophotometer(FTIR).
 Applications :Identification of drugs, polymorphs Raw materials, excipients.
102

103. Fluorescence analysis
 The organic molecules absorbs light usually over a specific range of wave length,
and many of them re-emits such radiation known as luminescence.
 The phenomina when the re-emission of absorbed light losts only when substance
receiving exiting rays, and called as fluorescence.
103
S.No Herbal Drug NATURE OF Fluorescencecharacters Under
UV Light
1 Cinchona Purple Blue
2 Rhubarb Violet
3 Quassia Whitish Blue

104. NMR-nuclear magnetic resonance spectroscopy
 NMR absorbs radio frequency radiation by substance held in a magnetic field. 
 Absorption results from interaction of radiation with magnetic moment of nuclei in
sample and it occurs at different frequencies for nuclei with chemically different
environment.
 Applications: NMR is imp tool in elucidation of molecular structure It is applicable
in identification of impurities. It reveals position of protons in a complex molecule
104

105. MS-Mass spectroscopy
 Mass spectrometry concerned with the electron ionisation, subsequent
fragmentation of molecules, determination of the mass to charge ratio (m/e)and
relative abundances of ions which are produced.
 Applications : It determines molecular weight of compound
 It helps in identification of drug constituents
105

106. X-ray diffraction
Many compounds are capable of crystallising in more than one type of crystal lattice at a
particular temperature and pressure.
The rate of phase transformation of a metastable polymorph to the stable one can be quite slow.
Polymorphs plays a very imp role in pharmaceutical science
RIA-radio immuno assayThis technique uses on antibody specific for the drug being assayed
and a labeled form of the same drug. The label may be particular radio-isotope, active-enzyme
or a C14 and iodine I125 commonly used isotopes in RIA. RIA is method of choice for
identification of cardiac glycoside, insulin.
106

107. Biological Evaluation
 It is employed when the drug cannot be evaluated satisfactorily by chemical and
physical methods.
 In this method, the response produced by the test drug on a living system is compared
with that of the stranded preparation.
 Such an activity is represented in units as International Units (I.U).
 Dose is termed as International units IU
 Digitalis 1IU=76mg of standard
 Vit-A 1IU=0.344 of standard
 Vit-D 1IU=0.025of standard
107

108. Indication of Biological Evaluation
 When the chemical nature of the drug is not known but is has an biological action.
 When chemical methods are not available.
 When the quantity of the drug is small and so it cannot be evaluated chemically.
 Drugs which have different chemical composition but same biological activity.Example: Cardiac
glycosides arte evaluated by this method on cats, frogs or pigeons.
SIGNIFICANCE
 1.The method is generally used when standardization is not done satisfactory by chemical or physical
methods
 2.When the quantity of the drug /sample are very less then the drugs are evaluated by biological
methods.
 3.These methods are performed on living animals, isolating living organ and tissue, animal
preparation and micro-organism ( Bioassay)
108

109.  METHODS OF STUDIES
 1)Toxic----animals are used
 2)Symptomatic-----animals are used
 3)Tissue-------isolated tissue is used 
To estimate potency of drug
With entire animal or with tissue
To conform therapeutic activity
109

110. EVALUATION OF HEAPATOPROTECTIVITY
Animals Used: Male And Female Albino Rats
Heapatotoxicity Indused By: Chemicals Industrial Pollutants, CCl4,
Drugs (Paracetamol, Rifampicin)
Parameters For Estimation
1.Physiological—hexobarbital Hypnosis
2.Bio-chemical—serum Estimation Enzymes Like SGOT (serum Glutamic Oxaloacetic
Transaminase) SGPT (Serum Glutamic Pyruvic Oxaloacetic Transaminase)
3.Blood Chelesterol, triglycerides Levels
4.Histopathological Methods (Liver Tissue Necrosis) For Testing Cultured Heaptocytes
Are Used For In-vitro Studies
110

111. Evaluation of Hypoglycemic activity
 deficiency of glucose in the bloodstream.
 Traditional Diabetic drugs –Momordica charantaka, Fenugreek, Gudmar.
 Diabetits is induced in animals by Alloxan & Streptazocin
 Alloxan cause necrosis of pancreatic islet-B cells which shows 180-250mg/ml fasting blood glucose
levels.
 Streptazocin cause formation of streptomycin they produce cytotoxic nitrourcido glucopyranose
which cause diabetes
 ANIMALS USED: Rabbits, Rats, Mice—(4 to 7 days)
 Dose :rats--80mg/kg ,mice—150mg/kg of streptazocin
single oral injection 140-180 mg/kg of alloxan for rabbits at marginal ear vein for 7 days
for rats and mice intraperitoneally 2 days
Insulin levels are noted by tests like RIA and ELISA.
111

112. Evaluation of Anti-inflammatory activity
Inflammation is caused by mechanical, infections, auto-immune
Types of inflammations: Rheumatoid arthritis, gout, dysmenorrhoea
PRINCIPLE: Anti-inflammatory activity is reduction of local edema induced in rat paw
by injecting irritant or inflammatory substance. Inflammation is induced by carrageenan
and croton oil.
Methods 1: Carrageenan is a mucopolysaccharide isolated from sea moss which induces
inflammation by giving through intraperitoneally saline in a dose of 0.1 ml, animal is
treated with herbal extract given orally (antagonist) Volume of paw is measured five times
with plethysmometer
Methods 2: Here albino rats or mice are used, edema is produced pina of ear with croton
oil(1 ml/ear) After induce, herbal extract is added to the same area. Edema is measured by
using vernier callipers and record the changes 0---no effect +ve---slight ++ve---pronounced
112

113. Evaluation of Anti fertility activity
Abortificient activity, contraceptives
 Traditional drugs like embelin from embilica and gossypol from gossypium produce
abortificient activity
 Types of Anti fertility evaluations
 In-females : Destruction of zygotes & prevention of ovulation
 In-males : estimation of Spermicidal activity & Anti-androgenic activity
113

114. Protocols for antispermatogenic activity
After acclimatization male rats are feed by herbal extract for 60 days
Between 12 day and 15 day & 60th day male rats are mated with female (morphological weight
of rat noted)
Histopathlogical study of sperm
 RESULT If No-fertilization b/w 12-15 days means functional sterility, after 56 days means
antispermatogenic.
114

115. Spermicidal activity (in-vitro)
Take human semen on a slide and add herbal extract and sorensens
phosphate buffer
Microscopic examination
Identification of Immobility means spermicidal
115

116. 116
Protocols for anti-fertility in female rats
Acclimatized 8-10 days ( I-group)
Vaginal smears are taken to measure cycle
Animals are feed by herbal extract
(oestrogenisity leads increase weight of uterus / cornifiction of vagina)
Further the rats are mate with males
(mating detected by smears)
Ovulation is absent due to contraceptive activity)
In case of fertilization ova are examined to see the transportation and implantation
Acclimatized 19 days ( II-group)
If weight gain contraception
If bleeding abortion

117. Testing of anti-ulcer activity
Causes of ulcer -- improper diet, alcohol consumption, stress, drugs (NSAIDS)
Traditional drugs: like liquorice, atropine, hyoscine and in less extent Gafaranate
extracted from cabbage juice shows anti ulcer effect
Chemical used to induce ulcer: Alcohol 1ml/kg orally
Aspirin -200mg/kg orally
Stress induced: animals are immobilized in cylindrical cage
117

118.  Animals are divided in to 3 groups
1.Those treated with normal saline
2.Those treated with ulcerogenic solution
3.Stress produced Animals are sacrificed after inducing
 All the groups are treated with herbal extracts and standard is treated
with ranitidine Stomach or duodenum is isolated Organs are opened to
know ulcer effect and gastric juice is measured and ulcer effect express in
table
118
0-no damage 0-absence
1.redness of mucosa 1.slight
2.Erosin of mucosa 2.One ulcer 5mm
length
3.ulceration 3.More than one
4.One above 5 mm
5.Total ulcerations

119. Evaluation of Neuropharmacological activity
 Testing of herbal drugs on CNS & ANS
 Drugs actions on CNS: cocaine, cannabis, morphine (stimulants, tranquilizers)
 METHODS FOR TESTING:
1. LOCOMOTAR ACTIVITY-----ACTIVITY CAGE (locomotion count is noted)
2. LOCOMOTAR CO-ORDINATION-----ROTATING ROD
3. PENTETRAZOLE CONVULSION IN MICE 80-120 mg/kg- intraperitonelly
a) time of onset of convulsions
b) number of convulsions
c) mortality rate
 4.BARBITURATE SLEEPING TIME stimulant or depressant effects on CNS
Phenobarbitone ----55mg/kg Hexobarbitone -----33mg/kg
119

120. Autonomic Nervous System (in-vitro studies )
1.Guinea pig ileum preparation: (non-specific anti-spasmodic activity)
Guinea pig ileum is useful for the evaluations of nerve mediated and directly stimulated
contractions because it contains non-adrenergic (cholinergic)nerves called as intrinsic
parasympathetic nerves, present between circular and longitudinal muscle layers.
120

121. 121
 Protocol
Food is withdrawn over night
Animal is killed
(suitable length of ileum is separated –1.5 to 2 cm long)
Kept in salt solution(physiological salt)
Jacketed organ bath at 32-37 c
Guinea pig ileum shows a response to
parasympathomimetics & parasympatholytics (ACH given to ileum)
Herbal drug is given

122.  Due to drug treatment if ACH type contractions are produced, persist even after giving a
dose of histamine H1-receptor antagonist, then it may be indicated that it is a cholinergic
drug. it can be verified by blocking effect with atropine.(it does not distinguish between
muscarinic and nicotinic receptor activity.
 If contractions caused by herbal drug are blocked by ganglion blocking agent then it may
be indicated that herbal drug is nicotinic receptor mechanism.
 Besides cholinergic activity the contractions occurred due to drug may be because of its
activity at other receptors like histamine.
122

123. 123
2.Isolated rabbit jejunum preparation
 With this study we can evaluate the nerve mediated and direct muscle stimulated adrenergic
effects
Rabbit jejunum is isolated along with mesenteric vessels as associated with sympathetic nerves
Mesentery vessels are carefully cut off & then isolated rabbit jejunum with periarterial nerve is
mounted in physiological solution in Jacketed organ bath 32-37c
Nerves are connected to a electrode and stimulated
If these are relaxed by herbal drug it may have noradrenalin effect or calcium channel blockdge
or potassium channel opening effect
If the action of drug is blocked by adrenergic receptor antagonist it may have effect on
adrenoreceptor.

124. 3. Rat. Phrenic nerve- diaphragm preparation
Phrenic diaphragm isolated from frontal part of right thoracic wall
Mount to phrenic nerve electrode
Muscle remains across platinum electrode
Drug shows effected neuromuscular junction
124

125. Mosquito repellence test
 Protection period offered by herbal extract against mosquito bite in terms of first bite.
3-6 days old, blood starved sucrose fed yellow fever mosquito (Aedes aegypti)
Human hand covered with snugly fitting polythene bag is introduced in a cage contains
about a thousand hungry mosquitoes, and allowed to bite on muslin cloth which is
treated wit herbal extract.
Number of bites received in 5 minutes counted
125

126. Anti insect activity
1) Larvicidal activity: Aedesaegyptic ---yellow mosquito in 100 ml of beaker,
Water Acetone extract of drug Larvae are feed with extract Estimation
parameter: mortality rate in 24 hours
2) Adulticidal activity: Tribolem castaneum---Red flour beetle Take Red
flour beetle in Petri dish in area of 10 cm Dried at 27 c and 60% RH Add
herbal extract, Mortality count after 24 hrs
126

127. Microbiological Assays
Drug substances suppress or influence the growth of micro organism are generally analyzed
by microbiological method.(Anti- biotics,Vitamins)
1.Cylindrical plate method Assay of antibiotics is based on measurement of the diameter of
microbial growth inhibition surrounding the cylinders containing various dilutions of test
compound which are placed on surface of solid nutrient medium .
2.Turbidimetric method Based on inhibition of microbial growth as indicated by
measurement of turbidity(transmittance) of suspension of a suitable micro-organism in a
fluid medium, to which have test compound. changes in transmittance is compared wit
standard known compound.
127

128. QUESTION BANK
1. Definition of Adulteration, substitution, Sophistication, deterioration, spoilage, admixture, Inferiority
2 marks
2. Enlist name of drug evaluation parameter 2 marks
3. Note on Different types of adulterants 5 marks
4. Briefly explain Morphological (Organoleptic ) evaluation parameter. 5 marks
5. Describe leaf constant parameter with suitable example. 5 Marks
6. Briefly describe Microscopical evaluation of crude drugs with suitable diagram. 10 marks
128

129. QUESTION BANK
7. Explain Chemical Evaluation method. ` 5 marks
8. Explain Biological evaluation method. 10 marks
9. Describe briefly Physical evaluation parameter for crude drugs. 5 marks
10. Significance of drug evaluation parameter. 2 marks
11. Describe Total ash, acid insoluble ash, water soluble ash with its significance.
5 marks
129

130. 130