PBC20403 Pharmacognosy

Pharmacognosy Laboratory Manual

Introduction
Instructions for Students

Students shall read the points given below for understanding theoretical concepts and
practical applications.
1. Students should wear white lab coat, mask, and gloves before entering into the laboratory.
2. Students should keep their belongings in locker which are not required during practical like
bag, extra files, etc.
3. Students should always carry Laboratory Manual, rough notebook and practical
requirements without exception.
4. Listen carefully to the lecture given by teacher about importance of subject, skills to be
developed, information about equipment, instruments, procedure, method of continuous
assessment, tentative plan of working laboratory, and total amount of work to be done.
5. Students should perform the practical only at the place which allocated to her. (No change
can be done without permission of subject teacher)
6. Students shall undergo study visit of laboratory for types of equipment, instruments,
material to be used, before performing experiment.
7. Read write up of each experiment to be performed, a day in advance.
8. Organize the work in the group and make a record of all observations.
9. Understand the purpose of experiment and its practical applications.
10. Write the answer of the questions allotted by teacher during practical hours if possible or
afterwards, but immediately.
11. Students should not hesitate to ask any difficulty faced during conduct of practical.
12. Students shall study all the questions given in the laboratory manual and practice to write
the answers to these questions.
13. Students shall develop maintenance skill as expected by the industries.
14. Students should develop the habits of pocket discussion, group discussion related to the
experiments so that exchanges of knowledge, skills could take place.
15. Students shall attempt to develop related hands-on -skills and gain confidence.
16. Students shall focus on development of skills rather than theoretical or codified
knowledge.
17. Students shall visit nearby workshops, workstation, industries, technical exhibitions, trade
fair etc. even not included in the lab manual. In short, students should have exposure to the
area of work right in the student's hood.
18. Students shall insist for the completion of recommended laboratory work, industrial
visits, answers to the given questions, etc.
19. Students shall develop habits of evolving more ideas, innovations skills etc. than included
in the scope of the manual.
20. Students shall develop technical magazines, proceedings of the seminars, refers websites
related to the scope of the subjects and update their knowledge and skills.
21. Students should develop the habit of not to depend totally on the teachers but to develop
self- learning techniques.
22. Students should develop the habit to react with the teacher without hesitation with respect
to the academics involved.

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23. Students should develop the habit to submit the practical exercise continuously and
progressively on the scheduled dates and should get the assessment done.
24. Students should be well prepared while submitting the write up of the experiments. This
will develop the continuity of the studies and he will not be over loaded at the end of the
term.
25. Students should clean the platform before leaving the laboratory.

Mark Distribution

Marks Submission
Practical Reports 10% Next week Friday
(5 lab reports) after the practical
Project 10% As indicated
schedule

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Plant Morphology and Microscopic Characteristics

Experiment No. 1

PART A: Plant Morphology

PART 1-Leaves:
Consists of:
-Blade (= lamina): Flat expanded area.
-Petiole (= stalk): Stalk that connects leaf blade to stem, and transports materials
-Stipule: Is an outgrowth of the lower zone of a young leaf, part of the leaf base

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Part 2-Stem

Types of stem
A) Over ground: Normal types in majority of plants
B) Underground stem
1. Bulb, in which the shoot consists of very short vertical stem (bearing roots below) and
fleshy storage leaves (e.g. Onions).
2. Corm, in which the shoot consists mostly of vertical storage stem(e.g Colcicum)
3. Rhizome, in which the stem is horizontal and underground (e.g. Zingiber officinale,
Ginger).
4. Tuber, which consists of a thick, underground storage stem, usually not upright, (e.g.,
Solanum tuberosum, Potato)

Part 3-Flowers
The typical flower consists of four sets of flower parts arranged on a short swollen structure
called receptacle. The four parts from outside are:
Sepals (collectively called calyx) are the outermost organs below the petals
Petals (collectively called corolla) are the showy part of most flowers. In some flowers, the
petals are green and are called sepaloid.
Stamens (collectively called androecium) are the male sexual organ of the flower. A stamen
consists of an anther which contains the pollen, supported by a thin filament.
Pistils, which are often called carpels, (collectively called gynoecium) are the female sexual
organ of the flower which are usually vase-like in appearance

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Part 4: Seeds and Fruits

In case of fruits and seeds, generally T.S. of various parts are observed under microscope e.g.
in case of umbelliferrous fruits like fennel etc. Thus in case of fruit of seed drugs, separate
sectioning technique is required for individual drug. Sometimes, longitudinal sections cut
through the center of the seed of the fruit has been found useful in investigation.

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PART B. Plant cell inclusions by microscopy

Background
The use of herbs as medicine is the oldest form of healthcare known to humanity and has
been used in all cultures throughout history. Early humans recognized their dependence on
nature for a healthy life and since that time humanity has depended on the diversity of plant
resources for medicine to cure myriads of ailments. Primitive people learned by trial and
error to distinguish useful plants with beneficial effects from those that were toxic or inactive,
and also which combinations or processing methods had to be used to gain consistent and
optimal results.

Nowadays, microscopical examination is one of the most important tools in herbal drug
identification. Some structures are very important in this process and sometimes called “Key
elements”. The most important procedures that should be observed while studying the
powder microscopy of herbs are:

PART 1: Preparation of Sample for Sectioning

1.1: STEM

Put the selected stem sample in test tube and add water so that the sample remains submerge.
Boil the sample in water over a bunsen flame for a few minutes. In case of leaf, this step is
NOT necessary.

1:2 LEAF

In case of a leaf drug, cut a part of the leaf passing

through the midrib. This cut off portion may or may not
be boiled (Boil if dry for softening). Since the lamina
of a leaf is very thin, section cutting is difficult. The
surface area of the surface to be cut has to be increased.
This is done by embedding the sample in a block of
pith (potato cube). A cubical portion of a pith is cut-off
and used as showed:

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PART 2: SECTION CUTTING TECHNIQUE

SECTION OF A STEM, ROOT, STOLON

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Transverse section (T.S.)

Transverse section is obtained by cutting along the radial place of cyclindrical portion of the
stem/ root/ stolon and perpendicular to the long axis.

Longitudinal Section

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PART 3- Staining and Mounting sections

A transverse section of required part of ingredient was taken on a glass slide to which are
added a few drops of safranin for 1 minute then washed immediately with water 3-4 times to
remove excess of safranin.

Then mounted with glycerin. Care however is to be taken to avoid air bubbles and to see that
there is sufficient glycerine under the cover slip. Excess of glycerine outside the cover slip is
to be withdrawn using a blotting paper.
Then the anatomy of the transverse section were observed and identified under microscope
(10xX10x).

PART 4- Observation of Calcium Oxalate crystals by Pizzoloto Method

1. Take uniformly thin section of crude plant.

2. Treat them with 2 N acetic acid for about 15 min.
3. Remove and treat them with 1% solution of silver nitrate in 15% hydrogen peroxide
for about 15 min (at 22oC) .
4. Remove the sections and wash them with distilled water.
5. Counterstain the sectionswith 2% safranin for 1 to 3 minutes.
6. Following usual technique, mount the sections and observe under microscope.
Calcium oxalate crystal appear clack against red background.

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SUMMARY OF THE REPORT:

PLANT MORPHOLOGY AND INCUSIONS

SYNONYMS
BIOLOGICAL SOURCES:
MACROSCOPY: (Draw and
identify the parts of the plant)

Organoleptic Characteristics: Color: Odor: Taste:

MEDICINAL USES:
MICROSCOPY:

CALCIUM OXALATE
TEST: (Draw the Crystals
identified)
CHEMICAL
CONSTITUENTS

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