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Leaf Constant

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1K views

Leaf Constant

Uploaded by

Samriti
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
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Leaf constants

(Microscopic evaluation of
crude drugs)
Leaf constant
1. Stomatal number
2. Stomatal Index
3. Palisade number
4. Vein islet number
5. Lycopodium spore method
1.Stomatal number :-

• It can be defined as the average number of


stomata present in per square mm area of
epidermis cell
• stomatal number can be determined by Treating
the lamina of the leaves with chloral hydrate
and then part of the epidermal surface is
removed carefully and observed under
microscope
• PROCEDURE :-
Place leaf fragment of about 5×5 mm in size in a test tube
containing about 5ml of chloral hydrate solution

Heat in a boiling water bath for about 15 minutes or until the


fragments become transparent

Transfer fragment to a microscopic slide and prepare the


Mount the lower epidermis uppermost, in chloral hydrate
solution
And put a small drop of glycerol ethanol solution on one side of the cover
glass to prevent the preparation from drying

Examine with a 40x objective and 6x eye piece , to which a microscopical


drawing apparatus is attached

Mark on the drawing paper a cross (×) for each stomata, and calculate the
average no of stomata Per square mm

Ex: Datura innoxia :141


2.Stomatal index:
• The stomatal index is the percentage of the
number of stomata formed by the total number of
epidermal cells, including the stomata, each stoma
being counted as One cell
• Procedure :-
• Place the leaf fragments of about 5×5mm in size in a test
tube containing about 5ml of chloral hydrate solution
• Heat in a water boiling bath for about 15minutes or until
the fragments become transparent
• Transfer a fragment to a microscopic slide and prepare
the mount ,the lower the epidermis uppermost in chloral
hydrate solution
• Put a small drop of glycerol ethanol solution on one side
of the cover glass to prevent the preparation from drying
Mathematical expression:

Stomatal index :

Where,
Where S = the number of stomata in a given area of leaf; and
E = the number of epidermal cells (including trichomes) in
the same area of leaf.
3. Palisade Ratio :-
Palisade ratio is the average number of palisade
cell under one epidermal cell
For ex :
Digitalis purpurea - 3.7 to 4.2
Datura stramonium – 4 to 7
Procedure:
• Place leaf fragments of about 5 × 5 in size in test
tube containing about 5 ml of chloral hydrates
solution
• Heat in a water bath for 15 minutes or until the
fragment become transparent
• Transfer a fragment to a microscopic slide and prepare the
mount of the uppermost epidermis in chloral hydrate solution
• And put a small drop of glycerol solution on one side of the
cover glass to prevent the preparation from drying
• Examine with 40 X objective and 6x eye piece to which a
microscopical drawing apparatus is attached
• Trace four adjacent epidermal cells on paper. focus gently
downward to bring the palisade into view and trace sufficient
palisade cells to cover the area of the outlines of four epidermal
cells
Calculate average number of palisade cells beneath
one epidermal cell; dividing the Count by 4
• This is the palisade ratio
4. Vein islet number :
Vein-islets present in per square millimeter is
known as “Vein-Islet number”.
Procedure:
• Take pieces of leaf lamina with an area of not
less than 4 square millimeters from the central
portion of the lamina, excluding the midrib and
the margin of the leaf.
• Clear the pieces of lamina by heating in a test
tube containing chloral hydrate solution on a
boiling water-bath for 30 to 60 minutes until
clear and prepare a mount in glycerol-solution
or, if desired, stain with safranin solution and
prepare the mount in Canada balsam.
• Place the stage micrometer on the microscope stage and
examine with 4x objective and a 6x eye piece.
• Draw a line representing 2 mm on a sheet of paper by
means of a microscopical drawing apparatus and construct
a square on the line representing an area of 4 square
millimeters
• Move the paper so that the square is seen in the centre of
the field of the eyepiece.
• Place the slide with the cleared leaf piece on the
microscope stage and draw in the veins and veinlets
included within the square, completing the outlines of
those vein-islets which overlap two adjacent sides of the
square.
• Count the number of vein-islets within the square
including those overlapping on two adjacent sides and
excluding those intersected by the other two sides
The result obtained is the number of vein-islets in 4 square
millimeters. For each sample of leaf, make no fewer than three
determinations and calculate the average number of vein-islets per
square millimeter.
Examples: Cassia angustifolia: 19 – 23; Digitalis purpurea: 2 – 5.5;
Cassia acutifolia: 25-30.
5. Lycopodium spore method :
This method is use to identify the crude drugs when the
chemical and physical methods are inapplicable.
This method is also useful to detect the adulteration
present in the crude drugs containing starch grains.
Examples: Adulterated drug containing starch can be
determined by counting the number of starch grains per
mg and calculating the amount from the known number of
starch grains per mg of the pure starch. The percentage
purity of an authentic powdered ginger is calculated using
the following equation
Where,
N = Number of characteristic structures (starch grain) in 25
fields.
W = Weight in mg of lycopodium taken.
S = Number of lycopodium spores in the same 25 fields.
M = Weight in mg of the sample, calculated on the basis of
sample dried at 105°C.
P = 2,86,000 in case of ginger starch grains powder.

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