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REFERANCE:
2. K. S. Ladhha, herbal drug microscopy ,yucca publication house ,first edition ,march 2003,
page no. 1 to7
MICROSCOPY:
A microscope (micro= small, and scope =to view) is an optical instrument consisting of lens or
combination of lenses. It helps in magnification (enlargement) of the image of an object, which is
too small to be viewed by the naked eye.
There are three principal kinds of microscope: Simple, Dissecting and compound.
SIMPLE MICROSCOPE:
It consists of single lens or magnifying glass, fixed on the suitable frame to view any object e.g. a
hand lens. It is useful where only a low magnification required e.g. for a examination of external
characteristics of crud drugs. Here the magnification obtained is approximately two times.
DISSECTING MICROSCOPE:
This is nothing but the simple microscope with additional features of stage. On which a dissection
can be carried out a suitable hand test for convenience and a mirror to focus the light on the object.
The lens can be raised or lowered by rack and pinion arrangement or moved horizontally for proper
focusing. The magnification in this case is about five times.
COMPOUND MICROSCOPE:
It consists of two sets of lenses. Here one set of lenses of short focal length is used to produce an
enlarge image of an illuminated object at a short distance, which is further enlarged by the second set
of lenses placed appropriately.
The stage : A horizontal shelf with graduated mechanical slide holder with X and Y movement for
holding the slide to be examined .The stage bears a hole in the center for transmitting light reflected
up by the mirror.
The mirror: Situated below the stage, reflects light upward through the hole in the stage. The mirror
is usually double faced. The plane face is for initial light intensity and the concave for concentrating
the light on the object.
The Diaphragm: Situated in between the hole on the stage and mirror, regulates the amount of light
reflected by the mirror.
The Body Tube: A cylinder holding a draw tube and moves up and down vertically above the hole
in the stage. The tube is raised or lowered by the coarse adjustment and is use for finding the focus.
The fine adjustment which on being turned produces a very slow motion of the entire framework
which holds the body tube and is used for exact focusing of the higher power lenses.
The ocular or Eye piece: Is to be inserted into the upper end of the draw tube. It consists of 2 plane
convex lenses, the lower and large collective or field lens increasing the field of vision and the upper
and smaller eye lens. Ocular enlarges the image formed by the objective. Midway between the field
lens and eye lens is perforated diaphragm which cut out edge rays from the image, determining the
size of the field of view. Oculars are designated usually with magnification number as 5X, 10X, 15X
etc. or by figures which represent focal length.
The objectives: Are fitted in to the bottom of the body tube or nose piece. Each of these consists of
a system of 2, 3 or more lenses. Objectives, like oculars are usually designated by magnification
numbers as 10, 45 etc. or by focal length. The smaller the number of focal length, the greater is its
magnifying power. If only two objectives accompany your microscope, the lower power objective is
shorter in length. Objective enlarges the object and projects them in the direction of eyepiece.
Magnification and field view: Microscopes are usually fitted with two objectives of 16mm and
4mm, two or three eyepieces and condenser. Different combination of eyepiece and objectives give
different magnification and field of view as indicated in the following table.
When using the microscope, it is useful to have the knowledge about the size of the field of view,
e.g. using 4mm objective and 5X eyepiece, field of view is approximately 0.42mm or 420µ.
However, accurate measurements are made with eyepiece micrometer or Camera Lucida. Objectives
are of two types, dry lenses and immersion lenses. The lens is called dry one, if an air space is
present between the tip of objective and the object, if the liquid is present the lens is called an
immersion lens (water immersion lens, if it is water or oil immersion if it is oil). Some microscopes
are fitted with a nosepiece capable of carrying 2, 3 or 4 objectives, which may be adjusted into place
at the lower end of the body tube. Others have a condenser to concentrate the light upon the object to
be examined. When using the condenser use only plan mirror.
Place the microscope on the table with the arm or pillar nearest to you.
Turn the lowest power objective into position into position which find light by looking into
ocular (eyepiece) and at the same time turn the mirror at such an angle that it reflects light from the
window or lamp up through the hole in the stage to objective.
Place the prepared slide in the slide holder on the stage in the horizontal position with the object
right in the center of the hole through which light is reflected from the mirror.
Make the lower power objective quite close to the slide by turning the adjustment down. Then
while looking through the eyepiece, move the coarse adjustment upward until the object is seen
distinctly. The object if not under the lens may now be brought into the field by moving the X-Y
movement very slowly while looking through the eyepiece. Then slowly turn the fine adjustment to
improve the focus.
Regulate the quantity of light with the iris diaphragm to improve the clarity of the field.
To focus with the light power objective, first find the object with the low power and arrange in
centre of the field. Raise the low power objective by means of coarse adjustment. Then turn the high
power objective into position and lower until the objective front lens nearly touches the cover glass.
A slight movement of the fine adjustment should show the object clearly. Never try to focus with
the high power objective while looking through the eyepiece because of the danger of cover glass
and damaging the tissue lying underneath. It may also cause damage to the delicately mounted
lenses.
Care of Microscope:
While carrying the microscope from one place to another, hold it firmly by the arm in an erect
position so that the ocular which is fitted loosely in the drawtube may not fall out.
Do not allow the dry object or the liquid in the draw tube, may not fall out.
Do not touch the ocular or stand with cleaning cloth soaked by reagents.
Raise the body tube sufficiently while changing from low power to high power objective to avoid
damage to the objective or the mounts.
If the lens is soiled it may be cleaned with clean cloth wetted with few drops of Xylol.
Never observe objects without putting a cover glass.
For removing slide from the stage, first raise the body tube and slowly slide it out of stage.
The slide must be prepared in such a manner that the stage is never wetted with any solvent or
reagent.
When not in use, microscope must be kept covered.
Histological techniques:
For obtaining satisfactory result of the following items are needed:
1. A sharp shaving blade for sectioning
2. A scalpel
3. Dissecting needle
4. Forceps
5. Watch glasses
6. A sufficient number of Microscopic slide
7. Cover slips
8. A 0 no. Camel hair brush
9. A piece of clean cloth
10. A record book
11. Pencil, pen etc.
The preparation of plant material from microscopic examination and the reagents used are described
below. Preparation of material for section cutting:
The drug available for pharmacognostic study are generally in the dry condition. The necessity tales
soaking of these drugs to soften them sufficiently, which permit easy sectioning of drug. The
duration of soaking depends up on nature of tissue of drug.
Chloroform water prevents microbial growth. Drugs which are available in fresh form like Datura,
Eucalyptus, Ginger, Neem , Vasaka etc. should be kept in water to avoid cell shrinkage by loss of
water.
SECTIONING:
The material to be sectioned is held between the thumb and for finger of the hand. Using a sharp
razor held in right hand, thin sections are made by drawing the razor across the object in quick
succession, sliding over the forefinger with the edge of forefinger pointing towards you. Transfer the
section cut, into water kept in watch glass with the help of Camel’s hair brush. In case of tender and
flexible material such as fresh leaf, the section can be taken conveniently by placing it between the
two flat surfaces of pith. A pith is usually a piece of potato( about 3×1×1cm ) in which a
longitudinal slit of two cm deep is made into which the material to be sectioned is placed is placed
and the section are taken as described above.
1. Tranverse section (TS): Is made in horizontal place at right angle to the long axis of
material.
2. Radial- longitudinal section (RLS): is made in a longitudinal plane parallel to the long axis
of the material along its radius.
3. Tangential-longitudinal section (TLS): is made in longitudinal plane parallel to the long
axis of the material and to the tangent.
In addition to the above three a surface view can be obtained by pealing of outer most layer e.g.
epidermal area of the leaves or the cork layer of the bark.
Techniques Of mounting:
From the watch glass containing the section transfer section to the center of glass slide, put 2-3 drops
of chloral hydrate solution on it, heat the slide very gently by passing it to and fro over low flame
when bubble start to appear stop heating, add a drop of glycerin- water solution to avoid drying of
the preparation and crystallization of chloral hydrate and place the cover glass carefully. To avoid air
bubbles getting entrapped, the cover glass is allowed to stand on one side and gently squeeze the
liquid between the cover glass and slide. Chloral hydrate is good clearing agent, which dissolves
starch, proteins, chlorophyll, resins and volatile oils which causes expansion of shrunken cells. The
method describe above is useful for observation of calcium oxalate crystals, as chloral hydrate does
not dissolve calcium oxalate.
For staining the preparation after treatment with chloral hydrate, add a drop of phloroglucinol
reagent on the section, allow it to evaporate and then add then add a drop of hydrochloric acid and
wait for 2 min. Then add then mounted in water. Their presence can be conforming by staining with
a iodine solution. The powder of crude drugs can also be mounted describe above for identification
of various characteristics.
Reagent Used:
Chloral hydrate solution: Dissolve 25gm of chloral hydrate in 10ml water.Glycerin- water solution:
Prepare by adding 1:1 glycerin and distilled water. Iodine water: Add as much iodine to distilled
water as it well dissolves.Phloroglucinol solution: Dissolve 1 gm of Phloroglucinol in 50 ml of 95%
alcohol. Use this solution within.Conc. Hydrochloric acid.
MICROMETRY:
Micrometry is the determination of particles with the help of microscope. The accessories required
for the of measurement are eyepiece micrometer and stage micrometer.
I. Stage Micrometer: It is a slide .It contains a standard scale length of 1mm which is divided into
100 divisions.
or
II. Eyepiece Micrometer: It is a circle of glass with scale attached on the surface. It is suitable for
insertion inside the ocular and used during the operation of measurement. A convenient form is a
linear scale of 1mm divided into 100 divisions.
CALCULATION:
1division of stage micrometer = 0.01mm = 10µ
If, 6division of eyepiece micrometer = 4 division of stage micrometer.
The value of one division of eyepiece micrometer is calculated as follows-
1division of stage = 0.01mm=10µ
4division of stage =40micron
Now, 6 division of eyepiece micrometer = 4 division of stage
=40µ
Now, to measure any object under consideration, place the prepared slide on the stage. See how
many lines are required.
Swift Ives Camera Lucida and Abbe Model are the commonly used instrument to trace
the image of an object under microscope on the paper. The image of object under the microscope
can be traced on paper with the help of swift- Ives camera lucida and a drawing board , whole
inclination can be adjusted . The camera lucida fixed over the eyepiece of the microscope. Figure
shows path of light from the object passing directly to the observer's eye through an opening in the
silvered surface of the left hand prism. At the same time light from the drawing paper and pencil is
reflected by the right hand prism and by the silvered surface, so the pencil appears superimposed on
the object enabling it to be traced on the paper. Other type of camera lucida working on same
principle is also available. The abbe drawing apparatus is another form of apparatus, which can be
used to trace the image of an object without any inclination in the board. It utilizes a plane mirror
carried on a side arm, instead of the adjustable prism, with the mirror at 450 to the bench surface.
Camera lucida or drawing ocular is useful for tracing a magnified image of the object under
microscopical studies with proper adjustments of camera lucida and illumination. It is possible to see
simultaneously the drawing paper, the pencil point and the object under microscope and it is then
easy to trace the require cutlines. This is much quicker and more accurate than the most skilled free
hand drawing, but it requires the subsequent addition of details by free hand.
Swift Ives, camera lucida and abbe model are commonly used instruments. Swift Ives, camera
lucida consist of a prism fitted over the microscope should be in a vertical position and microscope.
Lamp should be carefully adjusted, so that illumination on the drawing paper, placed at the site of
the microscope is equal to that on the mounted preparation. The drawing pare should be supported
drawing board and if necessary tilt it at correct angle to avoid distortion. Test the equal illumination
on all sides of drawing board. By placing a stage micrometer on stage of the microscope and tracing
its division by co-inciding the image of tip of pencil and graduation of stage micrometer. If division
drawn are not equal, then the angle of tilted board is readjusted, until equality is obtained. Remove
the stage micrometer and place a mounted slide with specific drug preparation on stage of
microscope, after drawing of stage micrometer scale on drawing sheet, the distance between 2 point
is measured with the help of scale. For measurement of object, replace the stage micrometer with
object under study without disturbing the adjustment. Trace the outline of object same as that of
stage micrometer. Co-incide the tip of pencil in margin of object finish the drawing. Measure the
dimension of object. By this arrangement, the circle over the eyepiece comes just above the cornea
of eye and camera lucida. The field of view is not in anyway reduced and changed. All that can be
seen directly through the eyepiece perfectly disciplined camera lucida, while the drawing being
viewed directly on drawing sheet but not drawn on field.
In practice the drawing board should be adjusted at particular angle approx 45 0 and the image
accurately focused without the eyepiece. The camera is slide on eyepiece and pushed down more or
less until the microscopical image is seen directly and illumination of the field is equal throughout.
The drawing paper is placed on the table, immediately under the camera. The observer will then see
the microscopical image projected on paper. At the same time waving the pencil point directly the
whole pupil of eyepiece is available for both images (object and drawing board) the diaphragm on
the apparatus being considerably larger than the pupil so it may be necessary to balance the
illumination either by subdividing the light in the microscope or by increasing it on drawing paper. It
will generally be found that when the object is ion luminious field the light on the object may be
advantageously subdivided by glass around it or similar measures. The eye may be removed as
required from the camera to minimize the parallax produced because of camera lucida and change in
intensity of illumination.The drawing paper should be kept at a distance of distinct vision according
to change in camera lucida and the person who is studying that object.
b. Abbe Model:
The Abbe drawing apparatus is another form of apparatus, which can be used to trace the image of
an object without any inclination in the board. It utilizes a place mirror carried on the side arm,
instead of adjustable prism, with the mirror at 45˚ to the bence surface.
EXPERIMENT NO: 2 DATE:
REFERENCE:
STARCH.
a. BOTENICAL SOURCE-
Potato starch.
Simple granules, irregularly ovoid or spherical and sub spherical 30-100µ in diameter;
hilum-eccentric; striations well marked and concentric.
Rice starch.
Compound granules (aggregation of large number of simple granules)up to 150
component granules,2 to 10µ,polyhedral with sharp, hilum minute central point, rarely
conticuous ,striations absent
b. CONSTITUENTS:
c. STARCH USES:
d. SUBSTITUENTS:
Tapioca starch or cassava starch from rhizomes of Manihot utilissiama and Manihot
aipi, fam Euphortbiaceae.
Sago starch from metroxylan sagu,M.rumphii ,Fam palmae.
Sweet potato starch
Brazilian arrowroot starch
RESULT:
EXPERIMENT NO: 3 DATE:
REFERENCE:
Dr. K.r. khandelwal, practical pharmacogonosy,nirali publication,pg no. 9.1-9.2
Crystals of calcium oxalate occurs in many plants. This are form by reaction of
calcium salt (which have been absorbed from the soil) with oxalic acid(by product of
protein and another metabolic process). Among the various cell content, calcium
oxalate crystals of different type are found in different organs of the plant. They may
be present in almost all parts of plant
Crystals belonging to the tetragonal system have three axis at right angle to each other,
two of these axis are equal in length, the third being of different length.
Crystals belonging to the monoclinic system have three axis of unequal length, two of
which are obliquely inclined to each other and other two are right angles to these two
Usually Calcium Oxalate crystals are described according to their general form and size.
1] Solitery(single) crystals- Usually in the form of prism, occurs as sharp angular bodies.eg:
Quassia, Senna.
2] Twin Crystals- Two prisms are united.eg: Hyoscyamus.
3] Rosette Aggregates- It consists of numorous small prism appears like rosette or star.eg: Rhuhab,
Senna.
4] Columnar Crystals- Elongated prisms.eg: Quillaia Saponaria.
5] Acicular or needle shape crystals- Single needle shape crystals usually found scattered in
parenchymatous cells of cinnamon bark etc.
6] Microcrystals (sandy crystals or microsphenoidals)- these are arrow-shaped minute completely
filling the cells in which they are occur.eg: Belludonna, Tobacco etc.
7] Crystals Fibers - These are superimposed parenchyma cells each of which contains single prism
or a rosette aggregate.
Significance:
-They give protection to the plant against birds and animals.
-They have great diagnostic value.
-Presence of or absence of crystals useful in identification of crude drugs.
-Helps in identification of adultrants.
RESULT:
EXPERIMENT NO: 4 DATE:
REFERENCE:
Functions of trichomes: Trichomes or hairs are adapted to many different purposes. A dense
covering of Trichomes prevents the damage by insects and the clogging of stomata due to
accumulation of dust. Trichomes perform the function of secreting volatile oil.
Types of Trichomes:
A) Covering Trichomes
a) Unicellular Trichomes
B) Glandular Trichomes
REFERENCE:
THEORY:
Stomata are minute openings usually found in the epidermis of the leave as in digitalis,senna etc. or
in young green stems as in ephedra, in flower as in clove and in fruit as in fennel , orange as orange
peel. This opening are surrounded with a pair of kidney shaped cells called guard cells. The term
“STOMA” is often applied to the stomatal apparatus which consist of slit like opening along with the
guard cells.The epidermal cells surrounding the guard cells are called neighbouring cells or
subsidiary cells.This in many cases as in digitalis etc. resemble the other epidermal cells, but in large
number of plants they differ in size, arrangement and shape from the other epidermal cells. On the
basis of the characteristic of the guard cells and subsidiary cells, Stamatas can be classified as:
Stomata perform the function of gaseous exchange and transpiration in the plants body. They are
most abundant in the lower epidermis of a dorsiventral leaf and less abundant on the upper in
isobilateral leaf, stomata remain confined to the upper epidermis alone, in submerged leaves no
stoma is present.In buchu and neem ,stomata are present only on the lower surface, while in case of
bellodona,datura,senna etc. stomata are present on both the surface.
The distribution of stoma shows great variation between upper and lower epidermis. In desert plants
and in those showing xerophytic adaptations eg.ephedra, agave, oleander etc.stomata are situated in
grooves or pits in the stem or leaves. This is special adaptation to reduce excessive evaporation as
the stoma sunken in the pits are protected from gusts of wind.
PROCEDURE:
1. Take a small amount of powder of a given sample in a test tube, add chlorhydrate and mix
well.
2. Take the mixture on watch glass and mix properly again.
3. Take this mixture on a slide add a drop of water with the help of brush.
4. Put the coverslip without any air bubble.
5. Observe under microscope at 10x and 45x respectively.
RESULT:
EXPERIMENT NO: 6 DATE:
REFERENCE:
1. Practical Pharmacognosy, C.K. Kokate, 5th edition, Vallabh Prakashan, pg. no. 159,160.
Waring blender, centrifuge, stirrer, shaking sieves, oven, distilled water, potatoes.
PROCEDURE:
1. Wash potatoes thoroughly with water to remove adhering soil and earthly matter and reduce
to fine slurry with water in a blender.
2. Pass the slurry through shaking sieves in order to remove the cell debris and other impurities.
3. Allow the milky liquid to settle down. Decant the supernatant liquid. Wash starch 2-3 times
with distilled water with constant stirring.
4. Centrifuge the milky liquid, dry it in oven at a low temperature and powder.
The yield of starch is approximately 10 percent and it gives blue colour with weak iodine solution.
RESULT
EXPERIMENT NO: 7 DATE:
REFERENCE:
THEORY:
EXTRACTIVE VALUES
Alcohol is an ideal solvent for extractive of various chemicals like tannins, resin etc. Therefore this
method is frequently employed to determine the approx resin content of drug. It is also used as an
official method for assay in case of myrrh and asafetida .Generally, 95% ethyl alcohol is used for
determination of alcohol soluble extractive. In some diluted alcohol may also be used, depending
upon solubility of the constituent of crude drug.
PROCEDURE:
Weigh about 4 gm of the coarsely powdered drug in a weighing bottle and transfer in to a dry 250 ml
conical flask.
Fill a 100 ml graduated flask to the delivery mark with the solvent (90% alcohol). Wash out the
weighing bottle and pour the washing with the remainder of the solvent into the conical flask.
Cork the flask and set aside for 24 hours shaking frequently (maceration)
Filter into 50 ml cylinder. When sufficient filtrate has collected, transfer 25 ml of the filtrate to a
weighed thin porcelain dish, as used for the ash values determination
Evaporate to dryness on a water bath and complete the drying in an oven at 105 0 C for 6 hours
CALCULATION:
X = ------ x ------
= -------%
For Liquorice ,
Wt. of Petri dish =
Wt. of extract =
X= ------ x ------
------
RESULT:
From the above observation the alcoholic extractive value Ginger and Liquorice powder was found
to be -------- % w/w and -------- % w/w respectively.
EXPERIMENT NO: 8 DATE:
REFERENCE:
THEORY:
Extractive value:
This method is applied to drug which contain water soluble active constituents of crude drugs, such
as tannins, sugar, plant acids, mucilage, and glycosides.
PROCEDURE:
Weigh about 4 gm of the coarsely powdered drug in a weighing bottle and transfer in to a dry 250 ml
conical flask.
Fill a 100 ml graduated flask to the delivery mark with the solvent (chloroform water).
Chloroform acts as preservative. Wash out the weighing bottle and pour the washing with the
remainder of the solvent into the conical flask.
Cork the flask and set aside for 24 hours shaking frequently (maceration)
Filter into 50 ml cylinder. When sufficient filtrate has collected, transfer 25 ml of the filtrate to a
weighed thin porcelain dish.
Evaporate to dryness on a water bath and complete the drying in an oven at 105 0 C for 6 hours
CALCULATION:
= ______g
X =------ x ------
_____
X = ______% w/w
______ml = ______g
______ml = y
y = ------ x ------
_____
y = ______%w/w
RESULT:
From above observation, the water soluble extractive value of Ginger and Liquorice was fond to be
______% w/w and ______% w/w respectively.
EXPERIMENT NO: 9 DATE:
REFERENCE:
1. Herbal drug microscopy by T.N vasudevan and K.S ulka publishing house pg no 20
2. Practical pharmacognosy Techniques and experiments by Dr. K.R. khandelwal, nirali
publication pg no 24.1 to 24,2.
THEORY –
Stomatal no – stomatal number is the average no of stomata per sq.mm of the epidermis.It is more
significant in evaluation of leaf drug.
Stomatal index – stomal index is the percentage of epidermal cell of leaf which have been
converted into stomata.
T= S * 100
T+S
STANDARD VALUE
A. Stomatal number
PROCEDURE:
Stomatal number
1. Clear the piece of the leaf by boiling with chloral hydrate solutionor alternatively with
chlorinated soda , peel out upper and lower epidermis. Separetly by means of forceps keep it on slide
and mount in glycerin water.
2. Arrange the camera lucida and drawing board for making the drawing to scale.
3. Draw a square of 1 mm by means of stage micrometer.
4. Place the slide with clean leaf ( epidermis ) on the stage trace, the epidermal cell and stomata
on the paper.
5. Count the no of stomata present in the area of 1 sq.mm include the cell if atleast half of its
area ;ies within the square
6. Record the result for each of ten fields and calculate the average number of stomata/ sq.mm
Stomatal index :
Step no 1, 2, 3, 4 are similar as mentioned in the determination of stomatal no.
7. Count the no of stomata, also the number of epidermal cells in each field.
8. Calculate the stomatal index using the above formula.
9. Determine the value of upper and lower surface ( epidermis ) separetly, for determination of
average index , not less than 10
RESULT :
EXPERIMENT NO: 10 DATE:
REFERENCE:
THEORY:
vein islate is the area of photosynthetic tissue and circulated by ultimate division of the conducting
strands.ion
vein termination NO:- It is the no of vein termination per sq.mm of leaf structure.
Significance
Vein islet and vein termination no. can be used for the identification of plants and can be used as a
tool for standardistion of crude drug to prevent the use of an adultrated drug.
PROCEDURE:
Take senna leaflet and to a leaf prepare a specimen by cutting the leaf from midrib to margin
( near about 2-4mm2 ).
Take these pieces in a test tube containing a porcelain piece.
Add chloral hydrate (1ml) and few drops of glycerin water, attach cotton plug wetted with
water to the test tube. Heat the test tube till bubble appears.
Then remove out the chloral hydrate with tap water.
Repeat this treatment 2-3 times till the specimen turns colorless (Pale yellow).
Keep it on slide and mount with glycerin water.
Arrange a camera lucida and drawing board for making the drawing to scale.
Draw the scale with the help of stage micrometer.
Trace out the veins which are seen from the eyepiece, completing the outlines of those islet .
Select the particular area ( square or rectangle) in a drawing and count the number of vein
islets and vein termination within that area.
Then calculate vein islet number and vein termination number in 1sq.mm area.
RESULT:
Vein islet number of senna leaflet was found to be ------- per square mm area.
Vein termination number of senna islet was found to be ------ per square mm area.
EXPERIMENT NO: 11 DATE:
THEORY:
PROCEDURE:
AIM: To determine number of stone cells in Cinnamon powder by lycopodium spore method.
REFERENCE:
THEORY:
It is an important analytical technique for powdered drugs, especially when chemical and other
methods of evaluations of crude drugs fails as accurate measures of quality. It is inexpensive with
official status. Lycopodium spores are very characteristic in shape and appearance and exceptionally
uniform in size (25µm) on an average 94,000 spores per mg of powdered lycopodium are present.
1. Well defined particles which may be counted eg.: starch grains or pollen grains.
2. Single layered cells or tissues, the area of which may be traced under suitable magnification
and actual area calculated or
3. The object of uniform thickness, the length of which can be measured under suitable
magnification and actual area calculated.
The percentage purity of an authentic powdered ginger is calculated using the following equation,
Lycopodium spore method can be used for evaluation of powdered kurchi,clove, ginger, cardamom,
nutmeg, umbeliferous fruits, etc.
STONE CELLS
Scleroids, stone cells or sclerenchymatous occurs in the parenchyma of many barks. These cells are
parenchymatous elements and may be rounded, polyhedral or prismatic, they have lignified walls
and the lumen may vary from a narrow, branching, slit like hollow to a fairly large, sub-rectangular
cavity. The walls commonly show striations and are performed by tubular pits, which are often
branched. The external opening of the pits appear as small, circular or irregular pores dotted over the
surface view. The value of these cells for diagnostic purposes is illustrated by the fact that they are
absent from frangula bark, but are present in the very similar cascara bark, they are few in quillaia
and very numerous in cinnamon and cassia.
PROCEDURE:
OBSERVATION:
Table no. 1
Field. 1 2 3 4 5
Lycopodium spore
Stone cell
Table no. 2
Field. 1 2 3 4 5
Lycopodium spore
Stone cell
Table no. 3
Field. 1 2 3 4 5
Lycopodium spore
Stone cell
Table no. 4
Field. 1 2 3 4 5
Lycopodium spore
Stone cell
Table no.5
Sr. no. 1 2 3 4 5
Lycopodium spore
Stone cell
CALCULATION
Total number of lycopodium spores in 25 fields = x=
Total number of stone cells in 25 fields = y =
If _x_ no of lycopodium spores have _y_ no of stone cells in 25 fields.
Therefore,
Percentage of number of stone cells = ______ x 100
in given sample
= ________ %
RESULT:
From the above observation, percentage of number of stone cells in given sample of kurchi powder
was found to be _______ %
EXPERIMENT NO: 13 DATE:
REFERENCE :
2.Biren Shah, A.K.Seth , Textbook of pharmacognosy and phytochemistry , second edition , page no.
119.
THEORY :
Principle: The ash content of a crude drug is generally taken to be the residue remaining after
incineration like carbonates, phosphates and silicates of sodium, potassium, calcium and magnesium
etc. It usually represents the inorganic salts naturally occurring in the drug and adhering to it, but it
may also include inorganic matter added for the purpose of adulteration so for the determination
different types of ash value are used in detection of crude drugs like total ash , acid-insoluble ash,
water soluble ash and sulphate ash . And this standards have been established for a number of
official drugs.
Significance: Ash values are helpful in determining the quality and purity of crude drugs, especially
in powder form. And it is also useful for detecting low- grade product, exhausted drug, and excess of
sandy or earthy matter.
Defination :
The total ash is the residue remaining after incineration. The acid insoluble ash is the part of the total
ash which is insoluble in diluted hydrochloric acid.
Procedure :
Incinerate about 2 to 3 g accurately weighed, of the ground drug in An tared platinum or silica dish
at a temperature not exceeding 450º until free from carbon, cool and weigh. If a carbon free ash
cannot be obtained in this way, exhaust the charred mass with hot water, collect the residue on an
ashless filter paper, incinerate the residue and filter paper, add the filtrate, evaporate to dryness, and
ignite at a temperature not exceeding 450º. Calculate the percentage of ash with reference to the air-
dried drug.
Significance :
Total ash value is useful in detecting the crude drugs that are mixed ith various minerals substances
like sand, soil,calcium oxalate,chalk powder or other drugs with different inorganic contents to
improve their appearance .
AIM : To determine Acid soluble ash value, water soluble ash value of given Sample.
REFERENCE :
1. C.K.Kokate,Practical pharmacognosy, fifth edition , vallabh prakashan, page no. 123,124.
2. Biren Shah, A.K.Seth , Rextbook of pharmacognosy and phytochemistry , second edition , page
no. 119.
Definition: The acid insoluble ash is the part of the total ash which is insoluble in diluted
hydrochloric acid.
Procedure: Boil the ash obtained for 5 minutes with 25 ml of dilute hydrochloric acid; collect the
insoluble matter in a Gooch crucible, or on an ashless filter paper, wash with hot water and ignite to
constant weight. Calculate the percentage of acid-insoluble ash with reference to the air dried drug.
Significance: Used for determination of earthy matter present on roots, rhizomes and also on leaves.
Crude drugs carry calcium oxalate crystals the amount may vary depending on environmental
conditions
Example;
Agar – NMT 1%
Amla – NMT 2%
Bael- NMT 1%
REFERENCE:
Practical Pharmacognosy, Dr. K.R. Khandelwal and Dr. Vrunda K. Sethi, 24th edition, Nirali
Prakashan, pg. no. 27.1, 27.11, 27.17, 27.20.
Practical Pharmacognosy, C.K. Kokate, 5th edition, Vallabh Prakashan, pg. no. 183-186.
REFERENCE:
1. Textbook of Pharmacognosy by C.K. Koakate, S.B. Gokhale, Nirali Prakashan, 43rd edition,
pg no. 34-36
2. Textbook of Pharmacognosy and Phytochemistry by Vinod Rangari, pg no.
THEORY:
Unorganized drugs:
These are derived from plants or animals by some process of extraction and followed by purification,
if necessary.
Few unorganized crude drugs are commonly describes by their physical characters only. The
description is as below:
The non uniform physical and chemical nature of these substances makes them difficult to define.
They are obtained from plant as well as from animal sources. The resins of animals sources is shellac
or lac which finds number of applications in pharmaceutical industry.
Oleo resins:
When the natural plants resins are accompanied with volatile oils in homogeneous form they are
known as oleo resins. Canada balsam and capaiba are suitable examples of oleo resins.
Balsams:
Aromatic resinous substances of plant origin containing balsamic acids are known as balsams.
Neither Canada balsam nor balsam of Capaiba contains any balsamic acids and hence, they are not
balsams in real sense. The examples are balsam of tolu, benzoin, storax and peru.
These are the combinations of volatile oils, gums and resins. Sometimes they also contain ether
substance like enzyme e.g. Myrrh and asafetida.
Dried juices:
Theses juices are obtained from fleshy leaves (aloes) of from stems of trees. In all cases incisions are
made to respective part of plants and juice coming out is collected and dried.
Latices:
The latex is product contained in special secretory tissues of certain plants. It is usually a white
aqueous suspension wherein microscopically small particles of oily globules are suspended. These
natural suspension of milky consistency may contain proteins, sugars, minerals and alkaloidal salt in
the true solution, whereas gums, starch and resins I n the suspended form.
Extracts:
The extracts covered under crude drugs differ from galenical extracts. The extract of pharmacognostic
origin consists of extracting the parts of the plant with water followed by concentration, while
pharmaceutical preparations known as extracts are prepared by using alcoholic solutions and
adjusting the products is as standard strength.
DRUGS BIOLOGICAL PHYSICAL CHEMICAL IDENTIFICATION USES
SOURCE CHARACTERISTICS CONSTITUENTS TESTS
AGAR Dried Strips: colorless, Carbohydrate: -Boil agar with Bulk laxative,
(Agar-agar, gelatinous slender, translucent, Polysaccharides water→ forms stiff pharmaceutical aid,
Japanese substance, lustrous, 4 mm wide i.e. agarose and jelly on cooling. in the preparation of
Isinglass) obtained from Bands: yellowish, 4 agaropectin. culture media.
Gelidium cm wide. -Agar solution +
amansii, G. Sheets: 45-60cm long ruthenium red →
cartilagineum, and 10-15 cm wide. pink color.
Flakes or course
G. Pristodes,
powder: grayish -Agar solution (hot)
Gracilaria
white, odorless + BaCl2 reagent→
confervoides,
Taste: mucilaginous white ppt.
P. capillacea Solubility: practically
and other insoluble in cold -Agar solution +
closely allied water, but swells to a Fehling’s solutions
members of gelatinous mass. + heat→ red ppt.
family Soluble in boiling -Agar + Iodine
Rhodophyceae. water. solution→ crimson
to brown color.
-Powder + 5%
aqueous caustic
potash→ canary
yellow color.
EXPERIMENT NO: 16 DATE:
1] SENNA Senna, Sona Obtained from leaf and pods Glycosides, Laxative, Supply
mukhi of Cassia angustifodia(fam- Antrancene- in habitual/
Leguminosae) Flavonides constipation
6] ISAPGOL Isphapgol Obtained from dried seeds Alkaloids, fixed To relieve gout,
of plantago ovate(fam- oils Epidermal cancer,
Isabghol plantagin-aceae Leukamia
(fam. Liliceae)
(fam. Liliaceae)