this slide is based on pharmacognosy

1. STUDY OF PHARMACOGNOSTIC PROPERTIES OF CALOTROPIS
GIGANTEA, AND COMPARATIVE STUDY OF ANTIHELMENTIC
PROPERTY WITH A STANDARD DRUG
A PROJECT REPORT SUBMITTED
IN PARTIAL FULFILLMENT OF THE REQUIREMENT
FOR THE AWARD OF THE DEGREE OF
MASTER OF PHARMACY
2022-23
Under the guidance of
Dr. Prithwiraj Mohapatra
Professor
Department of Pharmacognosy,
Jeypore College of Pharmacy,
Rondapalli, Jeypore (K), Odisha-764002.
Name:Mr. Satyabrata Prusti
M.Pharm Regd. No:2108268017
Department of Pharmacognosy
Jeypore College of Pharmacy
Rondapalli, Jeypore-764002
SUBMITTED BY:

2. CONTENT
• INTRODUCTION
• LITERATURE
• PLANT PROFILE
• METHODOLOGY
• RESULT
• CONCLUSION

3. INTRODUCTION
The historical significance of medicinal plants
stretches back to the very origins of human
civilization. For millennia, plants have been an
integral part of human life, providing sustenance,
shelter, and healing properties. As early humans
began to explore their natural environment, they
discovered various plants that offered relief from
ailments, injuries, and diseases. The knowledge of
these medicinal properties was passed down through
generations, forming the foundation of traditional
medicine practices that continue to be relevant today

4. • In ancient Egypt, papyrus texts like the Ebers
Papyrus and the Edwin Smith Papyrus provide
detailed information about medicinal plants
and their therapeutic uses. The Egyptians
utilized a vast array of plant-based remedies to
treat various conditions, from skin ailments to
gastrointestinal issues.

5. • The knowledge of medicinal plants also spread
to other regions of the world. In India, the
traditional system of medicine known as
Ayurveda developed around 3,000 years ago.
Ayurveda, derived from the ancient Sanskrit
words "Ayur" (life) and "Veda" (knowledge),
focuses on holistic healing and the use of
herbal remedies to maintain harmony between
the body, mind, and spirit.

6. • The ancient Greeks, too, contributed
significantly to the understanding of medicinal
plants. The works of Hippocrates, known as
the "Father of Medicine," emphasized the
importance of using natural remedies derived
from plants and herbs.

7. • Throughout the medieval period, herbal
medicine continued to be a dominant form of
healthcare across different cultures. The
Arabic world played a crucial role in preserving
and expanding the knowledge of medicinal
plants, translating Greek and Roman texts and
adding their own discoveries to the field.

8. • The development of modern medicine in the
19th and 20th centuries brought about
significant changes in healthcare practices.
With the rise of synthetic drugs and
advancements in pharmaceuticals, the use of
medicinal plants declined in some parts of the
world. However, many traditional medicine
systems, especially in Asia and Africa,
continued to rely on herbal remedies for
primary healthcare.

9. • In recent decades, there has been a renewed interest in
medicinal plants and natural remedies. The search for
new drug compounds and the recognition of the
limitations of synthetic drugs have prompted scientists
and researchers to explore the rich biodiversity of
plants once again.
• Ethnobotany, a field of study that examines the
traditional knowledge of plants by indigenous cultures,
has gained importance. Ethnobotanists work closely
with local communities to understand their traditional
medicine practices and identify potential medicinal
plants for further research.

10. • Furthermore, the practice of using herbal
supplements and alternative medicine has
gained popularity in many parts of the world.
People are turning to natural remedies and
plant-based therapies for various health
conditions, seeking a more holistic approach to
healthcare.

11. • 1.2. Importance of Medicinal Plants in Healthcare
• The importance of medicinal plants in healthcare is immeasurable, as they have been a fundamental
source of healing and treatment for various ailments throughout human history. These plants possess
bioactive compounds with therapeutic properties that can address a wide range of health conditions,
making them invaluable assets in the medical field. The significance of medicinal plants in healthcare
can be understood through the following points:
• Natural Remedies: Medicinal plants offer natural remedies for numerous health issues, which is
particularly crucial in a world where synthetic drugs may come with side effects and risks. Natural
remedies derived from medicinal plants often have fewer adverse effects and can provide a gentler
approach to healing.
• Traditional Knowledge: Traditional medicine systems, deeply rooted in the use of medicinal plants,
have been passed down through generations. This accumulated knowledge encompasses the
identification, preparation, and application of various plant-based treatments. Preserving and integrating
this traditional knowledge into modern healthcare can expand treatment options and foster cultural
heritage.
• Accessible and Affordable Healthcare: Medicinal plants are often more accessible and affordable,
especially for rural and economically disadvantaged communities. Many people worldwide rely on
herbal remedies as their primary source of healthcare due to the cost-effectiveness and availability of
these plants.
• Broad Spectrum of Use: Medicinal plants offer a diverse range of applications. They can address
various health conditions, from common ailments like colds and digestive issues to more complex
diseases such as diabetes, hypertension, and even certain cancers.
• Source of Drug Discovery: Many modern pharmaceutical drugs have their origins in medicinal plants.
Natural compounds derived from these plants have served as the basis for the development of synthetic
drugs. Studying medicinal plants continues to provide valuable insights into potential drug discovery and
novel treatments.
• Complementary Medicine: In some cases, medicinal plants can complement conventional medical
treatments, enhancing overall healthcare outcomes. Integrating herbal remedies into treatment plans can
lead to a more holistic and comprehensive approach to patient care.

12. • Plant Conservation: The importance of medicinal plants extends beyond human
health. Conserving these plants and their habitats is vital for maintaining
biodiversity and supporting ecosystems. As medicinal plants are often harvested
for their healing properties, their conservation becomes essential to sustain their
availability.
• Antimicrobial Properties: Some medicinal plants possess potent antimicrobial
properties, which have been utilized for centuries to combat infections. In an era of
rising antibiotic resistance, exploring and utilizing plant-based antimicrobial
compounds could offer alternative treatment options.
• Health Promotion and Prevention: Medicinal plants can also be used for health
promotion and prevention of diseases. Certain plants contain antioxidants and
phytochemicals that contribute to overall well-being and help prevent chronic
conditions.
• Cultural and Ethnobotanical Significance: Medicinal plants hold cultural and
ethnobotanical significance in various societies. The use of specific plants for
healing is often intertwined with cultural beliefs and practices, strengthening
community ties and identity.

13. LITERTURE
• Barkha Darra Wadhwani, 2021: Calotropis procera, locally
known as Aak or Madar in Hindi and milkweed in English,
belongs to the Apocynaceae family and subfamily
Asclepiadoideae. It is a wild-growing tropical plant with a
significant cultural significance as its flowers are offered in
worship to Lord Shiva in Hinduism. The plant is used in
traditional folk medicine to treat various ailments, including
snake bites, body pain, asthma, epilepsy, cancer, sexual
disorders, and skin diseases. The review emphasized the
phytochemistry, pharmacology, toxicology, and therapeutic
potential of Calotropis procera, highlighting the need for
more comprehensive research in these areas.

14. • Adeel Liaquat Bhatti, 2023: Co3O4 nanostructures were prepared
using a green method by utilizing the milky sap of Calotropis
procera (CP) through a low-temperature aqueous chemical growth
method. The Co3O4 nanostructures were evaluated for oxygen
evolution reactions (OERs) and supercapacitor applications. The
study found that the prepared Co3O4 nanostructures exhibited
promising OER performance with low overpotential and Tafel slope,
along with good durability. The nanostructures also showed high
specific capacitance and power density, making them suitable for
energy conversion and storage devices. The unique properties of
Co3O4 nanostructures prepared using CP's milky sap were
attributed to surface oxygen vacancies, a relatively high amount of
Co2+, and a reduced optical band gap, induced by the presence of
reducing, capping, and stabilizing agents from the milky sap

15. • Ashish Singhai, 2021: Pavonia odorata
(Malvaceae), commonly known as
sugandhabala, was evaluated for anthelmintic
activity. Alcoholic and aqueous extracts were
tested in dose-dependent manners, measuring
paralysis time and death time. The alcoholic
extract showed significant anthelmintic
activity compared to the standard drug and was
more potent than the aqueous extract.

16. Plant Profile:
• Calotropis gigantea R. Br. (Linn) is a lactiferous
shrub commonly found in tropical and sub-tropical
regions worldwide. The plant possesses immense bio-
potential uses, including antimicrobial, anti-
inflammatory, anti-cancer, anti-leprosy, wound
healing, and skin disease treatment properties. This
study focuses on exploring the antimicrobial, anti-
inflammatory, and anti-tumor activities of Calotropis
gigantea R. Br. (Linn).
• Figure 1: Calotropis gigantea R. Br. (Linn)

17. • 1.1 Scientific Classification
– Kingdom: Plantae
– Phylum: Tracheophyta
– Class: Magnoliopsida
– Order: Gentianales
– Family: Asclepiadaceae
– Subfamily: Asclepiadaceae
– Genus: Calotropis
– Species: Gigantea
– Scientific Name: Calotropis gigantea R. Br. (Linn)
• 1.2 Origin and Distribution Calotropis gintea R. Br. (Linn) is native to the Indian subcontinent and
can be found across tropical regions of Asia, Africa, Gulf, and South America. It thrives in tropical,
sub-tropical, and arid climates, displaying drought resistance and some level of salt tolerance. The
plant can grow wild at altitudes of up to 900 meters above sea level, and it prefers sandy soils with a
mean annual rainfall of 300-400 mm.
• In summary, Calotropis gigantea R. Br. (Linn) is a versatile shrub with various medicinal properties,
making it a subject of interest for investigating its antimicrobial, anti-inflammatory, and anti-tumor
activities. The plant's wide distribution across tropical regions highlights its adaptability and
potential for various medicinal applications.

18. • 1.3.1 Plant Morphology
• Calotropis gigantea R. Br. (Linn) is a remarkable
shrub that can reach heights of up to 6 meters.
The plant's roots are extended, simple, and
branched, with a woody base covered in corky
bark. Its branches have a succulent appearance
and are densely covered in tomentose hairs,
giving them a velvety texture. One of the notable
features of this plant is the presence of a thick
white latex (milky sap) that circulates throughout
all parts of the plant.

19. • The flowers of Calotropis gigantea are both fascinating and
visually striking. They are bracteate, complete, and
bisexual, featuring five petals and five sepals. The flowers
are actinomorphic, meaning they have radial symmetry, and
are pentamerous, having five floral parts in each whorl.
They are hypogynous, which means that the flower parts are
attached to the receptacle below the ovary. The flowers are
also pedicellate, with pedicels measuring 1 to 3 cm in
length. They are arranged in dense inflorescences and occur
in multi-flowered, umbellate clusters with peduncled
cymes. These flower clusters arise from the nodes of the
plant and can appear either axillary (growing from leaf
axils) or terminal (at the tips of branches

20. • The fruit of Calotropis gigantea is simple, fleshy,
and inflated, taking on a sub-globose to obliquely
ovoid shape. The fruit can grow to be up to 10 cm
or more in diameter. Initially, the fruit is green
and gradually ripens to a yellowish-brown color.
As it matures, the fruit splits apart to reveal
multiple small, flat, and obovate seeds, each
approximately 6x5 mm in size. These seeds have
a silky white pappus attached to them, which
assists in their dispersal by wind.

21. • Calotropis gigantea R. Br. (Linn) holds significant medicinal
importance in traditional practices, where various parts of the plant
are utilized to address a range of health issues. The plant's extracts
have been employed as remedies for bronchitis, asthma, leprosy,
eczema, vertigo, toothache, and common cold, showcasing its
diverse therapeutic potential. One of its prominent applications lies
in wound healing, where the extracts are believed to promote the
regeneration of damaged tissues and facilitate the recovery process.
Additionally, traditional healers view the whole plant extracts as an
effective health tonic, implying its potential in enhancing overall
well-being and vitality. The presence of antimicrobial and anti-
inflammatory properties in various parts of the plant further
contributes to its medicinal value, making Calotropis gigantea a
valuable resource in traditional medicine systems

22. • . Here are some of the notable medicinal benefits of Calotropis gigantea:
• Wound Healing: The latex and extracts from Calotropis gigantea have been
used traditionally for wound healing. They are believed to possess
properties that promote tissue regeneration, reduce inflammation, and
accelerate the healing process.
• Anti-inflammatory Activity: The plant exhibits anti-inflammatory
properties that can help alleviate inflammation and related conditions. This
makes it potentially useful in managing inflammatory skin conditions and
other inflammatory disorders.
• Antimicrobial Effects: Calotropis gigantea has shown antimicrobial activity
against various bacteria and fungi. The plant's extracts have been explored
for their potential in combating microbial infections.
• Anticancer Potential: Studies have demonstrated the presence of bioactive
compounds in Calotropis gigantea that exhibit cytotoxic effects on cancer
cells. These compounds may hold promise in cancer therapy and research.

23. • Antioxidant Properties: The plant contains antioxidants that can neutralize free
radicals, which are harmful molecules linked to various diseases and aging.
Antioxidants help protect cells from oxidative stress and contribute to overall
health.
• Anti-parasitic Activity: Certain components of Calotropis gigantea have
demonstrated anthelmintic activity, meaning they can combat parasitic worm
infections.
• Respiratory Ailments: The plant has been used traditionally to manage respiratory
issues such as bronchitis and asthma, possibly due to its bronchodilatory and anti-
inflammatory effects.
• Leprosy Management: In traditional medicine, Calotropis gigantea has been
employed to treat leprosy and its associated symptoms.
• Anthelmintic Property
• Effective Against Parasitic Worms: Calotropis gigantea possesses anthelmintic
properties, making it effective in combating parasitic worm infections.
• Treatment of Helminth Infections: Studies have shown that various parts of
Calotropis gigantea exhibit anthelmintic activity, making it a potential treatment
option for helminth infections in both humans and animals.

24. • 1.6 Phytochemicals
• Plants are often referred to as nature's "chemical
factories" due to their ability to produce a wide
variety of organic compounds, known as
phytochemicals. These bioactive compounds
serve as natural defenses, protecting plants from
pathogens and predators. While phytochemicals
are non-nutritive, they have gained recognition
for their potential in managing various human
health conditions, including cancer.

25. • The medicinal value of a plant lies in the unique combination of
phytochemicals found in its various parts. Each plant contains
specific phytochemicals that impart distinct medicinal properties.
These compounds produce definite physiological actions within the
human body, making them valuable for therapeutic use. Some
common physiological effects of phytochemicals include
antioxidant, anti-inflammatory, and cytotoxic effects.
• Secondary metabolites, such as alkaloids, steroids, flavonoids,
terpenoids, and tannins, are examples of phytochemicals found in
smaller quantities in higher plants. These compounds contribute to
the plant's medicinal properties and have been studied for their
potential health benefits. Harnessing the power of phytochemicals
from medicinal plants offers promising opportunities for natural
medicine and disease management.

26. • Methodology
• CollectionofMaterial
•
• Fresh leaves were collected from the Calotropis gigantea plant
growing in neaby forest of Koraput, Odisha. The plant specimens
were authenticated (voucher specimen and the leaves were washed
in running water and air-dried. The fresh leaf was then studied for
pharmacognostic evaluation, including examination of
morphological and microscopic characteristics and some
preliminary phytochemical evaluation.
• The plant parts were dried separately under the shade, powdered,
stored in airtight containers and used for the studies.

27. •
• Organoleptic evaluation
•
• Organoleptic evaluation is a technique of qualitative study of organs of sense of the
plant. It provides some specific characteristics of the material which establishes the
identity and degree of purity of the plant. In the present study, the colour of the
outer and inner surface of the fresh and dried stem was observed. The features like
shape, odour, taste, texture and surface characteristics of the stem and the powder
were performed.
•
• Anatomical studies of Calotropis gigantea
•
• In this study, anatomical studies were conducted on the leaf, stem, and root of in
vitro germinated thirty days old plants. Healthy and normal organs were carefully
selected for the analysis. Samples of different organs were cut and removed from
the plants and then fixed in FAA (Farmalin: 5 ml, Acetic acid: 5 ml, 70% Ethyl
alcohol: 90 ml).

28. • Once the infiltration process was complete, the
specimens were cast into paraffin blocks, allowing the
tissue samples to be embedded within the paraffin wax.
This embedding process facilitates the sectioning of the
tissues for microscopic examination.
• Overall, the described procedure aims to preserve the
cellular structure of the plant organs and facilitate
microscopic analysis, enabling researchers to study the
anatomical features and characteristics of the leaf, stem,
and root of the in vitro germinated plants. The use of
paraffin wax embedding ensures that the tissues are
well-preserved and can be easily sectioned for further
observation and analysis.

29. • Overall, the combination of various staining
techniques, including toluidine blue, saffranin,
fast-green, and IKI, facilitated a comprehensive
analysis of the anatomical features and cellular
characteristics of the plant specimens. The
staining methods utilized in this study contributed
to a better understanding of the cellular
composition, structures, and functional properties
of the leaf, stem, and root tissues in the in vitro
germinated plants.

30. • Phytochemicalstudy
•
• The plant parts, including the leaf, stem, and root, were subjected to
extraction using various solvents based on their polarity. The
solvents employed for extraction included hexane, chloroform,
methanol, ethanol, and water. The cold percolation method was
utilized to obtain the extracts from the powdered plant material.
• To begin the extraction process, a predetermined amount of
powdered plant material was weighed and soaked in hexane for 72
hours. After the specified duration, the extract was collected by
filtering the contents. This procedure was repeated once more, and
the collected extracts were combined and concentrated on a water
bath, maintaining the temperature below the boiling point of the
solvent used. Subsequently, the concentrated extracts were placed in
a desiccator to facilitate further solvent evaporation.

31. • Qualitative chemical analysis of methanolic extract
• Qualitative chemical tests play a crucial role in identifying the presence of specific
chemical compounds in plant extracts, such as alkaloids and glycosides. These tests
are essential for pharmacognosists, phytochemists, and herbalists to analyze the
chemical composition of natural products and medicinal plants. In this context, the
following tests for alkaloids and glycosides are commonly used:
• Test for Alkaloids:
– Mayer's Test: Mayer's reagent is a solution of mercuric chloride in potassium iodide. When a
small amount of the solvent-free extract is mixed with a few drops of dilute hydrochloric acid
(HCl) and filtered, the resulting filtrate is subjected to Mayer's test. Addition of Mayer's reagent
to the filtrate forms a white or pale yellow precipitate if alkaloids are present in the extract. The
formation of the precipitate is due to the interaction between alkaloids and the reagent.
– Wagner's Test: Wagner's reagent consists of iodine dissolved in potassium iodide solution. In
this test, a few drops of Wagner's reagent are added to a separate portion of the extract. The
presence of alkaloids in the extract results in the formation of a reddish-brown precipitate upon
mixing with Wagner's reagent.
– Hager's Test: Hager's reagent is a saturated aqueous solution of picric acid. When 1 to 2 ml of
Hager's reagent is added to a few ml of the filtrate obtained after mixing the extract with dilute
HCl, a prominent yellow precipitate forms if alkaloids are present. This reaction is specific to
alkaloids and aids in their identification.

32. – Dragondroff's Test: Dragondroff's reagent is a mixture of
potassium mercuric iodide and potassium iodide in water. Upon
adding a few drops of Dragondroff's reagent to the extract, an
orange-yellow precipitate forms if alkaloids are present. This test
is particularly useful for detecting alkaloids that are not detected
by other tests.
• Test for Glycosides: Glycosides are a class of compounds
containing a sugar moiety linked to an aglycone (non-sugar)
component. To detect glycosides in the extract, the
following tests are commonly employed:
– Legal's Test: Legal's test involves the use of glacial acetic acid
and ferric chloride solution. After hydrolyzing the extract with
dilute HCl for a few hours, the hydrolyzed sample is treated with
Legal's reagent. Formation of a violet ring at the junction of the
two layers indicates the presence of cardiac glycosides.

33. Determination of Ash Value of Powdered Leaves of Calotropis gigantea
Procedure:
Weighing the Sample: Take a known amount of powdered leaves of Calotropis
gigantea, typically around 2 to 5 grams, and accurately weigh it using an analytical
balance.
Ashing the Sample: Place the weighed sample in a pre-weighed crucible and heat it
in a muffle furnace or a high-temperature oven at a specific temperature, usually
between 500°C to 600°C. Continue the heating process until all the organic material
in the sample is completely burnt, leaving behind the inorganic ash.
Cooling and Weighing the Ash: After ashing, remove the crucible from the furnace
and allow it to cool in a desiccator. Once it reaches room temperature, weigh the
crucible containing the ash to determine the weight of the inorganic residue.
Calculating the Ash Value: The ash value is calculated as the percentage of the
inorganic residue obtained from the original weight of the sample. Use the following
formula:
Ash value (%) = (Weight of ash / Weight of sample) x 100

35. Type of Ash Ash Value (%)
Total Ash 18.7
Acid-insoluble Ash 1.4
Water-soluble Ash 2.2
Ash Content of Calotropis gigantea

36. Standard (Albendazole)20 mg/ml
Readin
g
Number of
Pheretima
posthuma
Concentratio
n (mg/ml)
Paralysis
Time
(minutes)
Death Time
(minutes)
1 5 100 2.17 6.3
2 5 100 2.1 6.15
3 5 100 2.05 6.2
4 5 100 2.12 6.25
5 5 100 2.08 6.3
Readin
g
Number of
Pheretima
posthuma
Concentratio
n (mg/ml)
Paralysis
Time
(minutes)
Death Time
(minutes)
1 5 50 5.35 7.6
2 5 50 5.1 7.9
3 5 50 5.2 7.85
4 5 50 5.25 7.7
5 5 50 5.05 7.95
Methanolic extract 50 mg/ml

37. Reading
Number of
Pheretima
posthuma
Concentration
(mg/ml)
Paralysis
Time
(minutes)
Death Time
(minutes)
1 5 100 2.17 6.3
2 5 100 2.1 6.15
3 5 100 2.05 6.2
4 5 100 2.12 6.25
5 5 100 2.08 6.3
Reading
Number of
Pheretima
posthuma
Concentration
(mg/ml)
Paralysis
Time
(minutes)
Death Time
(minutes)
1 5 50 5.3 7.6
2 5 50 5.1 7.8
3 5 50 5.25 7.65
4 5 50 5 7.75
5 5 50 5.2 7.75
Methanolic extract 100 mg/ml
Aqueous extract 50 mg/ml

38. Readin
g
Number of
Pheretima
posthuma
Concentratio
n (mg/ml)
Paralysis
Time
(minutes)
Death Time
(minutes)
1 5 100 2.7 6.25
2 5 100 2.55 6.45
3 5 100 2.65 6.35
4 5 100 2.5 6.25
5 5 100 2.7 6.3
Extract
Concentration
(mg/ml)
Paralysis Time (P)
(minutes)
Death Time (D)
(minutes)
Control (CMC) - - -
Standard 20 3.28 ± 0.621 7.50 ± 0.215
Methanolic 50 5.14 ± 0.271 7.79 ± 0.271
100 2.123± 0.162 6.25 ± 0.261
Aqueous 50 5.14 ± 0.562 7.70 ± 0.261
100 2.61 ± 0.103 6.30 ± 0.312
Aqueous extract 100 mg/ml
Anthelmintic Potency Of
Methanolic And Aqueous Extract
Of Calotropis Gigantea

39. CONCLUSION
• The experimental findings from the study clearly indicate that
Calotropis gigantea exhibits potent anthelmintic activity against
Indian earthworms. The anthelmintic activity was assessed based on
the time taken for paralysis and death of the earthworms. At a
concentration of 100 mg/dL, the plant extract displayed highly
significant anthelmintic effects, causing paralysis and death of the
earthworms in a relatively short period.
• Comparing the different concentrations of methanolic extracts of
Calotropis gigantea with the standard drug Albendazole and the
control Carboxy Methyl Cellulose (CMC) provides valuable insights
into the plant's potential as an anthelmintic agent. The fact that the
plant extract demonstrated significant anthelmintic activity,
comparable to Albendazole, a commonly used synthetic anthelmintic
drug, highlights its potential therapeutic value.

40. THANK YOU