EXTRACTION OF CARDAMOM BY DIFFERENT EXTRACTION METHODS AND ISOLATION OF MOLECULE AND IN-SILICO ANALYSIS OF THE MOLECULES IN LUNG CANCER

1. EXTRACTION OF CARDAMOM BY DIFFERENT EXTRACTION
METHODS AND
ISOLATION OF MOLECULE AND IN-SILICO ANALYSIS OF THE
MOLECULES IN LUNG CANCER
A Thesis
Submitted to
Centurion University of Technology and Management
In partial fulfilment of the requirements for the award of the degree of
MASTER OF SCIENCE (CHEMISTRY)
IN
School of Applied science
By
SHAKTI SWARUP MOHAPATRA
(shaktisworup3@gmail.com)
Department of Chemistry
School of applied science
Centurion University of Technology and Management, Bhubaneswar
Dist: Khordha-752050, Odisha, India
www.cutm.ac.in
May, 2021

2. DECLARATION
I declare that the thesis entitled “Extraction of cardamom by different
extraction methods and isolation of molecule and in-silico analysis of the
molecules in viral disease(Lung cancer).” is my own work conducted under the
supervision of, (Dr) preetha Bhadra, Asst. Professor, head of the Department
of Biotechnology and Dr. Bibhudutta Mishra, Asst. Professor, Department of
Zoology, School of Applied Science, Centurion University of Technology and
Management, Parlakhemundi , Odisha and approved by Research Advisory
Committee.
I, further declare that this thesis does not contain any part of any work which has
been submitted for the award of any degree either in the University or in any other
University or elsewhere without proper citation.
Place: Bhubaneswar Signature of the Candidate
Date: (Shakti Swarup Mohapatra)

3. CERTIFICATE - I
This is to certify that the thesis entitled “Extraction of cardamom by different
extraction methods and isolation of molecule and in-silico analysis of the
molecules in viral disease(Lung cancer).” submitted to School of Applied
Science is a piece of research work done by SHAKTI SWARUP
MOHAPATRA with Registration No. 190705100017 under my guidance and
supervision for the partial fulfilment of the degree of Master of Science in
Chemistry.
To the best of my knowledge and belief the thesis
1. Embodies the work of candidate himself.
2. Has duly been completed.
3. Fulfils the requirement of the ordinance relating to the M.Sc. Chemistry in Post
Graduate degree of the University.
Place: Bhubaneswar Mrs. PreethaBhadra
Date: __________ May 2021 Associate Professor
Department of Biotechnology
School of Applied Science
Centurion University of Technology & Management

4. iii
CERTIFICATE II
This is to certify that the thesis entitled “Extraction of cardamom by different extraction
methods and isolation of molecule and in-silico analysis of the molecules in viral disease(Lung
Cancer)” has been prepared and submitted by Shakti Swarup Mohapatra with Registration No.
190705100017 in partial fulfilment of the requirements for the award of degree of Master of
Science (Chemistry) in discipline of school of applied science has been examined and approved
by the Evaluation Committee after viva-voce.
Research Advisory Committee:
Member Mrs. Preetha Bhadra
Assistant Professor
Department of Biotechnology
School of Applied Science
Centurion University of Technology and Management Paralakhemundi
Member Dr. Yashaswi Nayak
Associate Professor & HOD
Department of Zoology
School of Applied Science
Centurion University of Technology and Management Bhubaneswar
Member Dr. Bibhudutta Mishra
Associate Professor
Department of Zoology
School of Applied Science
Centurion University of Technology and Management Bhubaneswar
Date of final viva-voce:
Head
Department of Chemistry,
School of applied science, Centurion
University of Technology and Management,
Bhubaneswar
Dean
School of applied science, Centurion
University of Technology and
Management, Bhubaneswar

5. iv
Acknowledgement
With great pleasure, I accept this opportunity to express my heartfelt gratitude
and respectful respects to Mrs. Preetha Bhadra, Associate educator, Department
of Biotechnology, School of Applied Science, for his vivacious interest,
commendable advice, rousing direction, unmistakable fascination for planning,
constructive ideas, and untiring dedication.
Dr. Atia Arzoo, HOD of Department of Chemistry, School of Applied Science,
and Dr. Bibhudutta Mishra, Assistant Professor and member of my warning advisory
group, for their moving direction, helpful analysis, and significant ideas during the
course work and beyond, I lack the words to express my profound gratitude, regard,
and sincere gratitude.
Dr. Yashaswi Nayak, Dean (Academic), School of Applied Science, Centurion
University of Technology and Management, for his guidance and convenience
during my examination work.
I'd like to express my deep gratitude to Dr. Pradipta Banerjee, Associate
Professor in the Department of Biotechnology, for his kind cooperation and
invaluable assistance throughout my examination.
I'd like to thank my closest companions, Subhalaxmi Das, Biswajit Mohapatra,
Lokanath, Chinmaya Behera, Nabalipi, Aparna , pritiparna , Ashis Majhi and
Satyajit Rout for their lovely company throughout my visit, and in particular for their
enthusiasm, devotion, begrudging co-activity, motivation, and friendliness, as well
as the several divisions who assisted me greatly with my examination work.
Mr. Santhosh Rout, Chief Librarian, and all members of the School of Applied
Science library staff for their persistent cooperation and availability of necessary
materials throughout my examination. Any exclusion from these brief statements
does not reflect a lack of gratitude. Last but not least, I now endeavour to convey my
genuine gratitude to the Almighty, the Creator and Preserver, with a high sense of
thanksgiving, in honour of God and in memory of so many and splendid favours and
endowments.
Place: Bhubaneswar (Shakti Swarup Mohapatra)
Date

6. v
TABLE OF CONTENTS
Chapter
No.
Title Page No.
I INTRODUCTION 1-4
II REVIEW OF LITERATURE 5-9
III MATERIALS AND METHODS 10-25
IV IN SILICO ANALYSIS OF PHYTOCHEMICALS 26-32
V RESULTS AND DISCUSSION 32-33
VI SUMMARY AND CONCLUSIONS 33-35
VII
REFERENCES 35-38
VIII
APPENDICES

7. vi
LIST OF TABLES
Table No. Particulars Page No.
3.1 Results of Super Critical fluid Extraction 13
3.2 Results of total carbohydrate by Anthrone method 25
4.1.1.a Results of total carbohydrate by Anthrone method 25
4.1.1.b Results and Discussions Molecular Docking 33
LIST OF FIGURES
Fig No. Title Page No.
1 Image of Cardamom 1
3.1 Map of Gajapati District showing Paralakhemundi 10
3.3.1 Pilot panel 11
3.3.2 Raw material of Cardamon 11
3.3.3 Cardamon in Blender 11
3.4.1 NaOH and dilute HCl 14
3.4.2 sample showing colorless 15
3.5.1 Test tube with Glucose monosaccharide 16
3.5.2 Barfoed’s reagent 17
3.5.3 Solutions mixing with Barfoed’s reagent 17
3.6.2 Paper chromatography chamber 19
3.7.1 TLC paper after KMnO4 spray 20
4.1 3D Images of ligands 30
4.2 3D images of protein LC4lvt 32

8. vii
ABSTRACT
Shakti Swarup Mohapatra Reg. No:190705100017
Year of Admission: 2019 Degree: M.Sc. (Chemistry)
Department of Chemistry
This article discussed about history of cardamom and its type, it’s chemical composition,
essential oil and oleoresin present in it . The different types of major vitamins found in cardamom
are niacin, pyridoxine, riboflavin, thiamin, vitamin C. Past researches and work regarding
antioxidant activity, antimicrobial activity, anti-inflammatory activity and anti insecticide activity
is also mentioned. This article gives a brief idea about different health benefits of cardamom. Three
types of Lung cancer and effectiveness of cardamom on Lung cancer is discussed briefly. Past
work shows that phytochemicals 1,8 Cineole and alpha terpineol exhibit anti Lung cancer
properties. Vitamin C, riboflavin, niacin, pyridoxine is important for a healthy Lung and can
prevent Lung cancer to some extent. It has been described that Elettaria cardamom plant extract
can be used to cure Lung cancer.
Phytochemicals are also known as phytonutrients are the plant chemical having some
biological importance. Researchers believed that in future they will able to classify about 40000
phytochemicals. Phytochemicals are not essential to body as vitamin and minerals but they can be
consumed to prevent disease. Phytochemicals rich food mainly includes fruits, vegetables, whole
grains, nuts, tea, spices. Phytochemicals have antioxidant, anti-inflammatory, anti-proliferative
and show anti-cancer properties. Cardamom is one of the known perennial spice plant which dried
fruit placed in top rank when the ranking of spices is done on the basis of price. Spices in foods
mainly act as a flavoring agent ,as a medicine sometimes food preservative since ancient times.
The essential oil and oleoresin obtained from seeds and seed pods by super critical carbon dioxide
fluid extraction which contains different phytochemicals. Lung cancer is caused by various
factors.
Key Words: Elettaria cardamom, BIOVIA, Phytochemical, Metabolites, Discovery studio,
kaempferol

9. 1
Chapter - I
INTRODUCTION
Elettaria cardamom(L) Cardamom is an evergreen herb also referred as queen of species
mainly used in adding flavor to the foods and sweets. Cardamom is perennial plant has a long
erect stem (pseudo stem) which is formed by leaf sheaths layered one over the other. A fully
matured plant attained 2–5-meter height. Rhizome is the real stem present below the soil. The
fully grown leaf length is about 30-50 cm having diameter 10-15 cm.
Cardamom Maton is popularly known as Queen of species due to its pleasant spicy taste,
commonly used as flavoring agent for preparation of delicious foods, Cardamom herb belongs
to Zingiberaceae family foundly native in western ghats of southern India and take third
position of world-wide spices. Cardamom is the third most expensive spice obtained from the
seeds of perennial herb. In India three states are famous for cultivation of cardamom are Kerala,
Karnataka, tamandu.
Systematic classification of cardamom
Kingdom_ plantae
Order-zingiberales
Family-zingiberaceae
Genus-Elettaria
Species-cardamomum
1.1PROPERTIES OF CARDAMOM
In Ayurveda cardamom essential oils is used as tonic for better gut health, aroma, nausea,
vomiting and morning sickness in pregnant lady. The major medicinal properties include the
following:
1. Antioxidant effect
2. Anti-microbial activity
3. Anti-inflammatory effect
4. Anti insecticide effect
5. Cytotoxic activity:_
I. Antioxidant effect
Oxidative stress is a factor which is responsible for causing of several chronic and
degenerative diseases such as diabetics, cancer, immune
dysfunction.(Haliwell,2000:Metodiewa and Koska,2000). Cardamom seeds are rich in
antioxidant substances that balancing free radicals by prohibiting oxidation of other
constituents. Cardamom seeds have an average level of antioxidant properties due to the

10. 2
presence of some phenol compound as quercetin ,kaempferol and luteolins. Natural
antioxidants are better than synthetic forms (saeed et al .,2014) Essential oil of cardamom act
as antioxidant like phytonutrients and vitamins, also help in scavenging free radicals , so
cellular aging is reduced.(Nanasombat and wimuttigosol,2011; Saeed et al.,2014).
II. Anti-microbial activity
Cardamom essential oil shown anti-bacterial and anti-microbial activity against
s.aureus,E.coli,s.typhi and c.albicans(Abdullah et al.,2018) Bacillus pulmilus and Listeria
monocytogenes(Bano et al.,2016).so cardamom essential oil plays an important role in
promoting safe and novel antibiotics. In modern medicine essential oil has potential broad
spectrum anti-bacterial and anti-viral activities that could be used to prevent from pathogens
and food-spoilage organisms (Mejdi et al.,2015)
The majority of studies focused on the anti-bacterial activity of cardamom extracts.
III. Anti-inflammatory effect
Anti-inflammatory activity was observed by introducing 0.1 milliliter of one percent
carrageenan in rat right hind paw by injection. Iflammation of the paw was measured by
plethysmometer. It was found that cardamom oil when given orally at a dose of 0.280ml/kg was
more effective than a dose of 0.175ml/kg and the inflammation was reduced significantly.
(Sermugapandian et al., 2018).
A dose of 175 and 280 microlitre per kg of cardamom oil extract from seed and a dose of 30mg/kg
of indomethacin was given against oedema induced by carrageenan in male albino rat which
exhibit anti-inflammatory function. (Al-Zuhair et al., 1996).
IV. Anti insecticide effect
Both flavors and fragrance of cardamom is due to the presence of volatile oil of cardamom
extract have a harmful effect on pest, insect. (Shayya et al., 1991)
V. Cytotoxic activity:_
Cardamom capsule extracts increased the cytotoxic effect of natural killer cell by indicated
their anti-cancer activity.(Majdalawieh and carr,2010)
Raksamiharja et al ,2012) stated that cardamom essential oil increased the amount of
lymphocyte, CD4+ and CD8+ in doxorubicin in a dependent manner by treating in rats.
(Elguindy et al.,2016) additionally examined that cardamom oil or geraniol decreases the
level of TNF, IL1 in diethyl nitrosamine actuate rats. These scientists also examined that
cardamom essential oil was an immune stimulant agent for chemotherapy.
1.2 OTHER BENEFITS OF CARDAMOM
I. Promotes Heart Health
The antioxidant properties present in cardamom can enhance heart health as it brings down
blood pressure levels. Cardamom contains fiber, which help in lowering cholesterol. Black
cardamom is more effective to improve heart health than green cardamom.

11. 3
II. Cancer Prevention
Cardamom act as potential natural agent in anti-cancer treatment. It delays, inhibits, and reverse
the formation of cancer to some extent. Due to its anti-inflammatory property, it decreases the
cancer cell growth.
III. Diabetes Treatment
As cardamom is rich in manganese which can lower the risk of diabetes. Manganese helps in
regulating blood sugar levels. It is also concluded from human studies that people with diabetes
have less manganese.
IV. Flavoring in food_:
Cardamom is mostly used in Indian kitchen as a flavoring for savoury and sweet dishes.
For its antibacterial properties of cardamom oil, it is used as a food preservative. Dried
fruits and seeds of cardamom are used for preparing rice, meat, vegetable dishes for
spicy flavor. Cardamom seeds are used for aroma and flavor in coffee ,tea and bakery
items. It is mostly worthy in Kashmir as a required ingredient for its fragrant and sweet
flavor.
V. Oral Health
Cardamom contains antimicrobial properties which inhibit the growth of mouth bacteria. It also
fights with the bad smell of mouth. It is the main composition in some toothpaste due to its
antibacterial properties.
The plant has pointed leaves and bears white flowers followed by a pod which contain 15 to 18
number of seeds. The two main genera of cardamom include Elettaria and Amomum. Elettaria
is regarded as true cardamom which is light green in colour and small in size. Amomum is black
to dark brown in colour and bigger in size in comparison to Elettaria. In the pharmaceutical
industry, cardamom extract dissolve in alcohol is mainly used as flavor and aroma, in cosmetic
industry cardamom is used in exotic perfumes (Lawrence, 1979).
Now it is cultivated in other countries like srilanka, Thailand, Vietnam, Guatemala,
Tanzania. Total production of cardamom in worldwide is evaluated as 30,000MT. recently
Guatemala is producing a great amount of cardamom approximately about 11000-12000MT.
above all ,the best quality of cardamom is cultivated in India. Maximum consumption of
cardamom in worldwide is found in western Asian countries as Saudi Arab, India these
countries share 60% world consumption. Cardamom plant grows in thick clump of up to 20
leafy shoots and can reach a height in between 2 to 6m.A perennial reed like herb with long
,silky and blade shaped leaves . Its stem bears small, yellow flowers with purple tips. the fruit
is about half inch long and turn into grey when ripe. The leaves are dark green, and swored
shaped.
There are two types of cardamom.one is small green cardamom or true dried cardamom
and are sweetly fragrant with a slightly purgent flavor. Black or large cardamoms are coarser

12. 4
in flavor and scent and used in pickles, meat dishes. . cardamom seeds are used for traditional
medicine applications and also for pharmaceutical applications. The Medicinal applications
include for control of asthma, teeth and gum infections, nausea, diarrhea as well as cardiac,
digestive and kidney disorders.
The essential oil and other bioactive compound derived in cardamom seeds contribute
to their characteristics flavor, aroma and utilized as a functional food, pharmaceutical,
medicinal and nutraceutical field.(Hamzaa et al.,2012)In food and dairy industries cardamom
essential oil has commercially used as flavoring agent in breads, cakes, sweets, curry powder
and pickles. It can use in cosmetics, lotions, perfumes due to its fragrance.(madhusoodanan et
al.,2002)
In cardamom such bioactive compounds are majorly present as alpha-pinene, sabinene,
1,8-cineole, linalool, alpha-terpineol, alpha-terpinyl acetate. Minor constituents of cardamom
essential oil are limonene,4-terpineol, alpha-pinene, beta-pinene, myrcene, octanal, (E)-
nerolidol, cis sabinene hydrate, geranylacetate, cis-sabinene hydrate acetate, beta-
caryophyllene and trans-linalooloxide in 2.9%, 1.4%, 1.1%, 0.8%, 0.2%,0.7%, 0.6%, 0.3%,
0.2%,0.2%,0.1% respectively.( Mejdi et al.,2015)
All bioactive compound has therapeutic benefits including antioxidant, anticancer,
antimicrobial, antifungal, antiviral activities.(Nirmala,2000; Marongiu et al.,2004: Hamzza et
al.,2012: Winarsi et al.,2014) Recent studies claimed that flavonoids,terpenoids, a thocyanins,
alkaloids constituents from cardamom were used for controlling cardiovascular,
pulmonary,kidney and lung associated disorders (Vaidya and Rathod,2014)
Some studies showed that the cardamom essential oil has anticarcinogenic effects by inhibiting
damage to adult DNA by aflatoxin B1 in a microsomal enzyme intermediated reaction.(Hashim
et al.,1994). The phytochemicals of cardamom essential oil as 1,8-cineole and limonene have a
protective property against cancer development.
Cardamom is responsible for stimulate and secretion of bile acid in stomach further
aiding in digestion. The spice prevents other gastrointestinal aliments like acid reflux,diarrhea
etc.(Amma et al.,2010) cardamom can work in treating bad breath. When we take a mixture of
species, including the seeds of anise, cardamom and fennel-bad breath cannot be a problem
anymore. In histoplasmosis a condition in which one of the symptoms is lack appetite
cardamom can also treats well. Green cardamom reliefs from sore throat in cases of cold and
cough. When in gargling Cardamom seeds can add in boiled water and it reduced the
inflammation of throat
Family of rhino viruses are fastidious and can be spread in living organisms. Second most
common cause of cold and fever are the corona virus. Corona viruses are identified about 20%
of colds (Tyrrell,1988). The genetic material of corona virus is single stranded RNA that can
cause infections similar to rhino viruses. Mainly rhino virus is responsible for one half of cold
like illness within the genus enterovirus and family picornaviridae. Of the 160 serotypes
generally HRV are divided into three different species HRV-A,B and C. The incubation period
from the time of infection until the onset of symptoms were 24 to 72 hours and that infections
would hold for 3_7 days. The virus was not only present in nasal discharge of the patient but
also in nasal washings obtain during the incubation time. Transmission of infection spread by
hand to hand( direct or indirect) to the nose or eyes of a normal person.(Sung et al.,1987). The
main aim of this study is to identify the accurate phytochemical of Elettaria cardamom having
medicinal properties that will help to cure Lung cancer.

13. 5
REVIEW OF LITERATURE
In this chapter, a review of past research works in the field has been compiled to enable
better understanding of the research in various regions, method of analysis on the research
subject. The chapter is presented under the following headings. 2.1 Concept and Importance
of roles of cardamom on Lung Cancer
2.2 Overviews of Cardamom Elettaria
2.3 Phytochemicals present in Cardamom
2.4 Pharmacological properties and health benefits of Cardamom
2.5 Lung Cancer and its effects
2.6 Causative agent and its overviews
2.7 Role of cardamom on Lung Cancer
2.8 Insilco analysis and molecular docking
2.1 Overviews of Cardamom Elettaria
Vijayan et al.,(2018) reported that in multinational spices market small cardamom occupied a
quality position for its use in food, cosmetics, medicinal fields. Mostly Karnataka, Kerala,
tamandu is known for best production of evergreen herbs. For better cultivation of cardamom
monsoons is best that is, more than 450 kg hectare raining and altitude having 600-1200 meters
above MSL. Using proper management of cultivation produced best quality of cardamom and
commercially supplied to other areas.
Govindarajan et al.,(1982) were reported that in multinational spice market small cardamom
has occupied a good position for its use in food and dairy industries, cosmetics and medicinal
fields. Mostly in Karnataka, Kerala, tamandu is known for cultivation of the evergreen herbs.
For better cultivation of cardamom herb, altitude having from 600-1200 meters above MSL,
weather having above 450 kg per hectare raining. Using proper management of cultivation get
produced best quality of cardamom herb and commercially supplied to other areas.
Jadav and Mehta(2019) were documented that cardamom is native to India, also found in other
countries. Cardamom is third most expensive spices and known as queen of spices. Cardamom
essential oil mainly composed of alpha-terpinyl acetate and 1,8-cineole. It has lots of medicinal
properties such as antiseptic, carminative, digestive,diuretic,stimulant,stomachic,

14. 6
antimicrobial and anti-inflammatory activities. Mostly cardamom is used as flavouring agent
in food and dairy industries due to its pleasant aroma and spice in taste.
2.2 Phytochemicals present on Cardamom
Charles(2012) reported that the chapter describes the botanical description on cardamom plant
parts, types of seeds and their uses. Cardamom is evergreen perennial herb with attractive white
flowers, the stems develop into small green capsules .In India cardamom is mostly cultivated
in Karnataka, Kerala, tamandu. Cardamom is found in Indians kitchens due to its heavy use for
pleasant aroma and spicy in taste. Cardamom contains many bioactive compounds such as
minerals, carbohydrate, vitamins, proteins, fat, fatty acids. Mostly the essential oil composed
of alpha-terpinyl acetate, 1,8-cineole, limonene, linalyl acetate, and linalool. Cardamom fruit
have many essential uses with antioxidant properties.
Rajpura and samratha (2018) were demonstrated that the evergreen cardamom is mostly known
for its aromatic favour, spicy and sweet in taste, also for health benefits. Cardamom is native
to India but also it is cultivated in other countries like srilanka, Tanzania, Guatemala, there are
two types of cardamom, one is black or large cardamom, other one is green or small cardamom.
Cardamom is third most expensive spice and known as queen of spices in the world. Cardamom
essenrial oil contain many bioactive compounds as myrcene, 1,8-cineole, sabinene, alpha-
pinene, beta-pinene, linalyl acetate, limonene. The recent studies briefly describe about
bioactive compounds, commercial uses, pharmacological activities, and health benefits.
Ahmed et al.,(2019) were examined the phytochemicals present in cardamom and explore the
composition of essential oil and their anti-bacterial and anti-microbial activities. The
composition of cardamom essential oil was alpha-ionone, eucalyptol, santolina alcohol, 1,6-
octadiene-3-ol, 3.7-dimethyl , 2,6-octadiene-1-ol,3-ol,3,7-dimethyl-
(Z),cinnamaldehyde,(E),Terpinene-4-ol.1,6,10-dodecatrien-3ol,3,7,11-trimethyl having
percentage (29.295%),(18.85%), (9.23%), (8.19%),(7.71%), (4.20%),(4.06%), (3.82%).
2.4 Pharmacological properties and health benefits of cardamom
Ashokkumar et al.,(2020) were documented that small cardamom(Elettaria
cardamomum) belongs to family Zingiberaceae has a lots of useful properties. In ancient times
as well as in current times cardamom seeds are used for medicinal applications like like control
of asthma, kidney and cardiac disorders, prevents from teeth and gum infection. Cardamom
seeds have used in other beneficial fields like in dairy and food industry it can add in spices for
preparation delicious foods, it can add in flavouring agent in sweets. cakes, breads, cookies.
Cardamom essential oil has contains minerals, proteins, lipids, carbohydrates, fatty acids
having different health benefits including anti-microbial, antiviral, anticancer activities.
Meenakshi and Muralitharan(2015) were demonstrated that the paper explains about
scientific classifying, about parts of plants( seeds,leaveas,fruits,flowers). Cardamom seed are
extracted by stream distillation method and yields are 1-5%. The cardamom essential oils are
composed of limonene, alpha-terpineol,linalool,linalyl acetate,citronellol,nerol, geraniol,

15. 7
methyl eugenol. Cardamom has anti-microbial and anti-bacterial activities, also it can useful
for digestive and stomachic and cancer prevention. Cardamom oil has other benefits as it is
effective for insect bites, ingestion of light poison, oral infections and it can be used in
cosmetics due to its cooling properties. Cardamom extracts are more essential against
streptococcus mutans and candida albicans. Its medicinal properties are briefly mentioned in
this chapter.
Korikanthimathm et al.,(2000) were documented that cardamom(Elettaria
cardamomum) belongs to zingiberacae family is native to south India. Cardamom is widely
used as a flavoring agent in food preparations as well as in sweets, cakes and spicy pickles. In
this article geo ecological requirements, economic importance, cultivation areas and their
output, traditional use, Morden use in medicinal fields.
2.5 Lung Cancer and its effect
Lungs are a pair of spongy air filled organ of lower respiratory tract within the thoracic
cavity on either side of heart within ribs.it has 1.3 kg. The main reason of lungs cancer is
smoking, and consumes alcohol. When lungs cell gets mutate is grow uncontrollably form a
tumor.
The tissue of the lung can be affected by various respiratory infections including
pneumonia and lung cancer. Lung illnesses are the absolute most regular clinical conditions on
the planet. A huge number of individuals have lung disease in the US alone . Smoking,
contaminations and qualities cause most lung illnesses. Smoking tobacco is by for the main
source of cellular breakdown in the lungs. About 80% of cellular breakdown in the lungs
passing’s are brought about by smoking and numerous others are brought about by openness to
used smoke. A variety of things caused cellular breakdown in the lungs like tobacco smoke
vapes Maryjane, radon, asbestos, molecule contamination.
This kind of most lung cancers happens especially in present day or former smokers,
however it's also the maximum not unusual place kind of lung most cancers visible in non-
smokers. It is extra not unusual place in girls than in men, and it's miles much more likely to
arise in more youthful human beings than different sorts of lung most cancers.
Considering both men and women Lung cancer is the world's second most spreader
cancer. Smoking is the major fact that leads to lung cancer and leads to 75-80% of deaths due
to lung cancer. Small cell carcinomas along with the non-small cell carcinomas are the two
major classification part of lung cancer. Normal cell proliferation and homeostasis are
governed by regulatory circuits which have defects during lung cancer. multistep fashion arises
in phenotype when lung can get transformation occurs from normal to malignant. By clonal
expansion the chain of genetic along with epigenetic in giving into invasive cancer.
2.5. Causative agents and its overview
Lung cancer is a leading cause of malignancy-related death in Japan, and its incidence is still
rising steeply. Various factors, including cigarette smoking, asbestos, and diet, have been
reported to correlate with the development of lung cancer. Of these factors, cigarette smoking
is believed as the major carcinogen for lung cancer. Recent studies indicate that cigarette smoke

16. 8
carcinogens cause genetic damages at both oncogenes(K-ras) and tumor suppressor genes(p53)
of lung cancer, and hence initiate and promote the development of lung cancer.
Mining and usage of asbestos in industry or manufacture (asbestos cement products, thermal
and electrical insulation in construction and shipyard work, brakes, textile industry) . It seems
that asbestos fibers size (long and thin) is a strong predictor of lung cancer mortality. Even
though there is still a controversy in the literature, probably chrysotile is considered less
carcinogenetic than amphibole forms of asbestos.
Usage of arsenic and arsenic compounds (antifungal outdoor wood preservatives, agricultural
industry of pesticides, herbicides and insecticides, manufacture of non-ferrous alloys, glass-
manufacturing, electronics industry).
Exposure to beryllium and beryllium oxide (nuclear technology, X-ray and radiation
technology, dental applications and as beryllium-copper alloys in the electronics, aerospace
technology, automotive).
Second-hand tobacco smoke (passive-smoking) represents an occupational exposure for
workers in bars, restaurants, public buildings and educational institutions especially in countries
without smoke free legislations in public places.
Workers in aluminium production who are primarily exposed to polycyclic aromatic
hydrocarbons and also to sulfur dioxide and fluorides, various aluminium compounds,
chromium and nickel. The risk for lung cancer seems to be increased but studies are still
controversial.
of these agents is probably due to increased rigidity of the capsid protein, thereby inhibiting
disassembly.
2.5. Role of cardamom on Lung Cancer
Cancer is that the most generally familiar reason for human mortality. Plant-derived natural
merchandise presently comprises a big amount of commercially available antineoplastic
drugs. Chemoprevention with food phytochemicals is meanwhile thought-about united of the
foremost important ways for cancer control. Acting on multiple targets within the cellular
signal pathways and therefore the regulation of important molecules in carcinogenesis by food
phytochemicals and natural harvests was operative in Counteracting, preventing, delaying, or
plummeting the risk of cancer. As urged by experimental proof and phytochemical
composition, cardamom will have potential health edges against many sorts of cancer.
(Hussain T et, al, 2008)
Anticarcinogenic effects of the volatile oil of cardamom are unconcealed over in vitro studies.
because it reserved the harm to adult desoxyribonucleic acid by bioweapon B1 during a
granule protein intermediated reaction. this might be attributed to bioactive elements in a
necessary oil that has doable anti-cancer functions. Bhattacharjee et al. explicit that the
phytochemicals elements of cardamom oil such as terpene and 1, 8-cineole had shown a
defensive result against cancer evolution. (Saeed Met, al,2018)
The initiation of the detoxification enzymes may be a basic feature within the influences of
cardamom in antagonizing numerous chemical-induced carcinogenesis. The detoxifying
protein scheme plays a vital role in decisive the ultimate destiny of
carcinogens/procarcinogens and its resulting impact on carcinogenesis. Regulation of
different detoxifying enzymes is mediate by the transcription issue Nrf2 (nuclear factor blood
cell 2–related issue 2) binding to the inhibitor rejoinder element (ARE)/electrophilic response

17. 9
part (EpRE) placed within the organizer region of connected genes to trigger the expression
of inhibitor and cellprotective genes. bodily process of cardamom has been shown to rescue
the nuclear expression of Nrf2 within the mouse Blood of DMBAtreated mice, that suggests
that the cardamom treatment would possibly trigger activation of Nrf2 despite the harmful
effects of DMBA, to take care of cellular physiological condition throughout aerobic stress.
additionally, to Nrf2 activation, oral administration of cardamom to 7,12-
dimethylbenz[a]anthracene (DMBA)-treated mice up-regulated the clinical test detoxification
enzymes, comparable to glutathione-Stransferase, antioxidant glutathione reductase,
superoxide dismutase, and enzyme in addition as abridged glutathione via activation of
nuclear issue erythroid-2-related factor two transcription factor in mice and blocked NF-kB
activation and down-regulated cyclo-oxygenase- two expression. (Goyal SN et, al, 2015)
It has been revealed that cardamom extract or geraniol as a part of cardamom oil was
extremely effective in thwarting diethylnitrosamine (DENA)-induced hepatocellular
carcinoma through obstruction aerophilic stress, decreasing professional inflammatory
cytokines like TNF-α and IL-1β and NF-κB, and conjointly decreasing amino acid enzyme
(ODC). The possible for cardamom extract chemoprevention is also attributed to the
synergistic influence of the Phytomolecules within the cutting. several anticarcinogenic
compounds that might function potential NF-kB inhibitors are now being industrialized for
anti-cancer treatment. (Surh YJ et, al, 2008)
2.6 In silico analysis and molecular docking
Agoni et al(2020) documented that the rhino virus inhibition through capsid and
described role of benzothiophene. Their efforts have been shown that benzothiophene
derivative inhibits HRV virus replication. The derivative bound to HRV-B14, shown similar
activity as capsid inhibitor i.e., pleconaril. In this in silico report residues and interaction maps
are analyzed that binding of benzothiophene in the HRV-B14 active site deforms it at first
and condensed the interaction profile. This was distinguished by hydrogen bonds, reduce van
der waals interactions and increased residue flexibility.
Pagadala et al.,(2017) experimented that molecular docking is a process by which binding of
target protein into ligands. In drug discovery for prevention of disease molecular docking is
used as a tool. Within the docking process the risk factors as mutant, irritant, tumorigenic,
solubility and affinities of ligands are check.
cardamo
m
Anti-
michrobi
a
Anti-
Cancer
Gastroprote
ctive
Anti-
inflamentor
y
Cardio
Vascular
protectiv
e

18. 10
Chapter - III
MATERIALS AND METHODS
3.1 About the chapter
In this chapter, the procedure followed in different extraction methods, separation of
molecules using different tests, collection of those data, analysis of bioactive compounds is
described. The details of the methodology followed for the present investigation are presented
under this chapter.
3.2 Working Area
The study has been done on different extraction procedures such as super critical fluid
extraction, solvent extraction, and some of the experiments are done in laboratories using
chemicals and equipment. But supercritical fluid extraction process is an industrial process, so it
has been done in GTF(Gram Tarang Food) at Paralakhemundi, dist-Gajapati. Using trial machines
essential oils are extracted from raw materials.
The findings of this study have been discussed after analysis of the collected data through
such extraction processes. The procedures and using materials had been summarized in the study.
The results and discussions have been concluded in the last.
Fig 3.1: Map of Gajapati District showing Paralakhemundi

19. 11
Methods-
• Extraction methods
• Insilico analysis
3.3 Super Critical fluid Extraction
Principle-:
Super critical fluid extraction is a process of
separating one component from another using super
critical fluid as the C𝑂2 act as liquid extracting
solvent. Substance with a pressure and temperature
above its critical point where distinct gas and liquid
phase do not exist. Mostly C 𝑂2 are used as
supercritical fluid.
Materials required-:
I. Pilot control panel
II. Two condenser
III. Chiller
IV. C𝑂2 pump
V. C𝑂2 cylinder
VI. Geyser
VII. Bypass pipe
VIII. BPRV( back pressure regulator valve)
IX. Extractor
X. Separator
XI. Raw material( cardamom 4 kg)
XII. Heat exchanger
XIII. Collector
XIV. C𝑂2 storage tank
Procedure:-
Freshly we have to prepared cardamom powder by grinding of cardamom seeds, then filled in the
extractor. liquid carbon dioxide from storage tank with a siphon attachment was passes through a
cooling bath and was then compressed to the operating pressure by an HPLC pump.
Fig-3.3.1:Pilot panel

20. 12
Under high pressure and temperature C𝑂2 gets supercritical. Compressed carbon dioxide flowed
into the extraction vessel of 50mm length containing cardamom samples. The extraction column
containing the raw material to be extracted was in a thermostatically controlled water bath, the
temp. Inside the extractor being controlled by a digital controller within a accuracy of (0.1k).
Fig-3.3.2 : Raw material(cardamom)
Fig-3.3.2 : cardamom in Blender Fig-3.3.3 : Cardamom essential oil

21. 13
Sample Wt. After
mixing
(in gm)
Pressure
(in bar)
Temp
(in ℃)
Time Amount of
extraction
(in gm)
1 907.5 gm 100 35 11.12-11.42 58
150 40 11.42-12.12 7.5
200 45 12.12-12.42 0.1
Total 65.5
2 995 gm 100 35 2.15-2.45 80
150 40 2.45-3.15 9
250 45 3.15-3.45 3
Total 92
3 800 gm 100 45 4.20-4.50 34.2
150 45 4.50-5.00 4.5
250 45 5.00-5.20 4
Total 42.7
4 955 gm 100 35 11.45-12.15 57.5
150 40 12.15-12.45 10
200 40 12.45-1.15 4
Total 71.5
CONCLUSION:-
Lowest temperature is required for cardamom extraction. In the above tables the pressure,
temperature are mentioned . At temperature 35℃ and 100bar pressure highest amount i.e. 58 gm
of cardamom essential oil is extracted.

22. 14
QUALITATIVE TEST
3.4 Flavonoid test:-
Principle: -
Flavonoids help regulate cellular activity and fight off free radicals that cause oxidative
stress in the body. It helps the body function more efficiently while protecting it against
everyday toxins and stressors. Flavonoids are also powerful antioxidant agents.
Materials required: -
• Test tubes
• Samples
• NaOH solution
• 𝐻2O solution
Procedure: -
The stock solution overnight EtOH and 𝐻2O as well as heated EtOH and 𝐻2O was taken in
different test tube and added simultaneously few drops of NaOH and HCl solution. An
intense yellow colour was appeared in the test tubes.
Fig-3.4.1 : NaOH and dilute HCl

23. 15
Objective: -
When addition of dilute NaOH and HCL solution, if it is colorless, it indicates that presence
of flavonoid.
Fig – 3.4.2: sample showing colorless
Results: -
In heated EtOH and H2O as well as in overnight EtOH solution was colorless. Flavonoids are
present in this solution.

24. 16
3.5 Barfoed’s test: -
Principles: -
Barfoed’s test is used for distinguishing monosaccharides usually react in about 1-2
minute while the reducing disaccharides take much longer time between 7-12
minutes to react with the reagent. Brick red color is obtained in this test which is due
to formation of cuprous oxide.
Reagents for Barfoed’s test: -
• Glucose (dilute solution) 2ml
• Cardamom extracts 2ml
• Barfoed’s reagent: cupric acetate in 1% acetic acid
• Water bath
• Dry test tubes
• Pipettes
Procedure: -
Take 2ml of cardamom samples in dry test tube. Take 2ml distilled water diluted in glucose in
another test tube.
Fig- 3.5.1: Test tube with Glucose monosaccharide

25. 17
Add 2ml of Barford’s reagent to all test tubes. Keep boiling in water bath at 70C. temperature. wait
for the development of brick red precipitate. Note the time taken to change the color.
Fig-3.5.2: Barfoed’s reagent Fig-3.5.3: Solutions mixing
with Barfoed’s reagent
Fig-3.5.4: Water Bath
Results: -
Glucose solution is being red color due to presence of monosaccharide. But no color change in our
cardamom samples, because it contains disaccharide.
3.6 Paper chromatography: -

26. 18
Principle: -
In paper chromatography separation of component is distributed between phases of liquid. One
phase of liquid is water that is held the pores of the filter paper and the other liquid is the mobile
phase that travels along with the filter paper. Separation of the mixture is the result that is obtained
from the differences in the affinities towards the mobile and water phase while travelling under
capillary action between the pores of the filter paper.
Materials required: -
• Chromatography chamber
• Whatman’s paper
• Buffer
• Pencil
• Paper clip
• Samples
• Glucose and fructose (standard solution)
• Coloring reagent
Procedure: -
Take whatman’s paper in 9.8cm and level in 2cm. then mark G,F,S,F with pencil. then Prepare a
buffer solution containing N-butanol, acetic acid and water in 4:1:5 ratio. Prepare the coloring
reagent containing 𝐾2C𝑂3(10gm), KMn𝑂4(1.5gm), NaOH(1gm). In the paper drop by drop add
standard solution by micropipette then dry.
Fig- 3.6.1: drying by drier

27. 19
After that the buffer is placed in the chamber then prepared paper fit in the chromatography
chamber with clip then salute tape it because for not evaporate.
Fig-3.6.2: Paper chromatography chamber
The solute arises in the paper, we will wait until the solvent buffer reach in certain level. After
approx. 2 hours mark the line by pencil and dry it. Then spray the coloring reagent in the paper
and yellow color appears in some areas.
Fig-3.6.3: showing monosaccharide in G and F
Results: -
The yellow color appears in G and F pores. But in samples yellow color is not found. That
concluded that the sample does not contain monosaccharide. It contained disaccharide.

28. 20
3.7 Thin layer Chromatography
Principle of TLC
Thin layer chromatography uses a thin glass plate coated with either aluminum oxide or silica gel
as the solid phase. The mobile phase is a solvent chosen according to the properties of the
components in the mixture. The principle of TLC is the distribution of a compound between a solid
fixed phase (the thin layer) applied to a glass or plastic plate and a liquid mobile phase (eluting
solvent) that is moving over the solid phase. A small amount of a compound or mixture is applied
to a starting point just above the bottom of TLC plate. The plate is then developed in the developing
chamber that has a shallow pool of solvent just below the level at which the sample was applied.
The solvent is drawn up through the particles on the plate through the capillary action, and as the
solvent moves over the mixture each compound will either remain with the solid phase or dissolve
in the solvent and move up the plate.
Plate preparation:
TLC plates are usually commercially available, with standard particle size ranges to improve
reproducibility. They are prepared by mixing the adsorbent, such as silica gel, with a small amount
of inert binder like calcium sulphate (gypsum) and water. This mixture is spread as thick slurry on
an unreactive carrier sheet, usually glass, thick aluminum foil, or plastic. The resultant plate is
dried and activated by heating in an oven for thirty minutes at 110 °C. The thickness of the
adsorbent layer is typically around 0.1- 0.25 mm for analytical purposes and around 0.5- 2.0 mm
for preparative TLC.
Capillary spotters:
Place a melting point capillary and in the dark blue part of the Bunsen burner flame. Hold it there
until it softens and starts to sag. Quickly remove the capillary from the flame and pull on both ends
to about 2-3 times its original length. If you pull the capillary inside the flame, you will have a
"piece of art", but not a good spotter. Allow the capillary to cool down, and then break it in the
middle. Make sure to break off the closed end on one of them.
Spotting the plate:
The thin end of the spotter is placed in the dilute solution; the solution will rise up in the capillary
(capillary forces). Touch the plate briefly at the start line. Allow the solvent to evaporate and spot
at the same place again. This way you will get a concentrated and small spot. Try to avoid spotting

29. 21
too much material, because this will deteriorate the quality of the separation considerably
(‘tailing’). The spots should be far enough away from the edges and from each other as well. If
possible, you should spot the compound or mixture together with the starting materials and
possible intermediates on the plate.
Fi- 3.7.1: TLC paper after KMnO4 spray
Fig 3.7.2: TLC plate in UV visible machine

30. 22
QUANTITATIVE TEST
3.8 Estimation of total carbohydrate by Anthrone method:
Anthrone method principle: -
Carbohydrates are dehydrated by conc. 𝐻2𝑆𝑂4 to form furfural. Active form of the reagent is
anthranol, the enol tautomer of anthrone, which reacts by condensing with the carbohydrate
furfural derivative to give a green colour in dilute and blue colour in concentrated solutions, which
is determined calorimetrically. The blue-green solution shows absorption maximum 620 nm.
Materials required:-
I. Equipment’s:-
• UV spectrophotometer
• Water bath
• Centrifuge
II. Chemicals/ reagents:-
• Anthrone reagent
• Glucose
• 𝑁𝑎2𝐶𝑂3
• 𝐻2𝑆𝑂4
III. Glass wares and others:-
• Test tube, test tube stand, pipettes, beaker, ice, tissue paper, aluminium foil, wash bottle
Reagents:-
•Anthrone reagent- 200mg Anthrone add in chilled 𝐻2𝑆𝑂4. Use freshly prepared
reagent for the assay.
•Glucose stock solution-200 mg glucose per mL of distilled water.
•Concentrated 𝐻2𝑆𝑂4 and 5ml water was taken and 𝐻2𝑆𝑂4 must be cold.
Procedure:-
First 0.1gm of raw material extract was taken in a beaker and then add 5ml of 2.5N HCl. It would
kept in water bath for 15-20 mins. Then it would cooled down and then 𝑁𝑎2𝐶𝑂3was added until
effervescence stops. Sample contain 𝑁𝑎2𝐶𝑂3 is to be diluted upto mark. Then 200mg anthrone
reagent was added to the 𝐻2𝑆𝑂4.

31. 23
Fig-3.8.1 : Test tubes in cool water
Then 10ml sample in test tube put in centrifuge.( 5 min, 2500 rpm). In a test tube 200ml
sample(after centrifuge) and 800ml of distilled water was added. Then it was taken in water bath
for 5mins. Water bath should be preheated at 100C, after sample put temperature should be 60-
70C.
Fig-3.8.2 : Sample containing HCl

32. 24
Fig-3.8.3 : Anthrone solution
After that cardamom samples and reference(Anthrone soln.+ chilled H2SO4) taken in water bath.
Then Put the test tubes with reference in ice water until it cool down. Then dilute it 3 microliter
distilled water, 200 ml discard and 200 ml sample added. Put in UV-vis spectroscopy and set
wave length 630 with basic mode.
Fig-3.8.4 : centrifuge

33. 25
Fig-3.8.4 : water bath
Results-
Concentration O.D at 630 nm
0 0
200 0.228
600 0.556
800 0.813
1000 0.839
Sample O.D. at 630 nm Concentration
(µg/ml)
Mahua 0.265 258.6
Amla 0.746 793
Ginger 0.475 491.9
Cardamon 0.356 359.7
Garlic 0.135 114.1

34. 26
4. In silico analysis of Phytochemicals from Elettaria cardamomum against
Lung Cancer.
4.1 About the chapter
In this chapter, the procedure followed in-silico analysis, super critical CO2 extraction,
solvent extraction, qualitative test and quantitative test. The details of the methodology followed
for the present investigation are presented under this chapter.
4.2 Material and methods-
Software:-
For analysis of phytochemical screening it was need to predict the molecular interaction
strength by using Biovia Discovery studio software(Dassault systems of France). Machine learning
techniques of BIOVIA software utilizes to predict the level of molecular interaction in between
the phytochemical as ligand and protein as a receptor.
Research Design
The study was based mainly extraction of Cardamom. Accordingly, after a thorough and
meaningful formulation of the problems, specific objectives were decided. In the light of these
objectives, an effort was made to make a thorough review of the relevant literature relating to the
previous researches in this field. Objectives were given in the introduction chapter.
The findings of this study have been discussed after analysis of the collected data through
in-silico analysis, extraction of Cardamom, quantitative and qualitative analysis of extracted
sample of ginger were subsequently summarized. The findings had been summarized after
throwing light on all the major aspects of the study. The conclusion and the suggestions were made
to make the curing of Lung cancer using Cardamom.
4.3 Definition of terms
Phytochemical:
"Phytochemicals (from Greek Phyto, meaning "plant") are chemicals produced by plants
through primary or secondary metabolism. They generally have biological activity in the plant host
and play a role in plant growth or defenses against competitors, pathogens, or predators".
Cancer:
Cancer is a group of diseases involving abnormal cell growth with the potential to invade
or spread to other parts of the body. These contrast with benign tumors, which do not spread.
Other names: Malignant tumor
Symptoms: Lump, abnormal bleeding, prolonging
Treatment: Radiation therapy, surgery, chemotherapy

35. 27
4.4 Methodology-
4.4.1 Selection of plant metabolites-:
Small cardamom (Elettaria cardamomum(L) Maton is perennial herb, native to India. Cardamoms
are the capsules of dried fruits in different genera belongs to the family zingiberaceae. The
cardamom essential oil has antiviral properties against viral disease like Lung Cancer. It may also
be added to drinking water to eliminate the germs contained there. Elaichi is rich in antioxidants,
black cardamom helps in combating the symptoms of cold and cough. The essential oil derived
from its seeds act as an antiseptic which reliefs from sore throat. Different research and studies
give idea about cardamom essential oil contains sabinene, geraniol, myrcene, linalyl acetate, alpha
pinene, alpha-terpineol, beta-terpineol, terpinyl acetate, linalool, limonene, eugenol, nerol, cineol.
It has known that to treat Lung Cancer with use of cardamom seed as a natural remedy. This study
focused on identification of particular metabolites responsible for controlling Lung Cancer
infections.
A collection of 10 phytocompounds from Elettaria cardamomum were taken as ligands for
docking analysis. The three-dimensional coordinates for ligands were obtained from PubChem
database (https://pubchem.ncbi.nlm.nih.gov). These ligands were then cleaned up, calculated 3D
coordinates and generated ligand conformations by applying ‘prepare ligand protocol’ of
Discovery Studio 4.0.
After preparation, the compounds were filtered based on the molecular properties for
predicting their solubility and permeability in drug discovery. The best known of the physical
property filters is “Lipinski’s rule-of five and Veber rules”, which focuses on bioavailability. The
rule states that the compounds have molecular mass less than 500 Daltons, not more than 5
hydrogen bond donors, not more than 10 hydrogen bond acceptors and an octanol-water partition
coefficient XlogP not greater than 5. The filtered compounds were then downloaded in ‘.csv’
format for toxicity assessment.
4.4.2 Drug likeness and toxicity prediction of the ligands
Drug-likeness is a qualitative concept used in drug design to evaluate how the substance acts
like drug with respect to factors like bioavailability. The molecular properties which influence
Mutagenic, Tumorigenic, Irritant and Reproductive are recognized as a long side therapeutic
potency as key determinants of whether a molecule can be successfully developed as a drug. These
parameters are responsible for about 60 percent failures of all drugs in the clinical phases and so
the prediction of OSIRIS property explorer(https://www.organic-chemistry.org) explore the drug-
likeness of the select ligands.

36. 28
Besides taking the Lipinski’s rule of five into account, Osiris predicts the drug likeness and
toxicity in a fragment-based approach. The input for Osiris is the ISOSMILES notation of the
ligands and the output is a tabulation of parameters that indicate the likelihood of a molecule to be
drug like or not.
For this prediction, Lipinski’s rule of five is a key component taken into account. According
to Lipinski’s rule, a ligand molecule should have the following properties to become a good drug
candidate. The molecular weight of that ligand should be below 500 Daltons, the cLogP value
should be less than 5, solubility value should be greater than -4, TPSA value should be less than
or equal to 140, drug likeness values must be in positive range and drug score should be greater
than 5.
Those ligands of the phytocompounds which have passed the toxicity parameters of the
Lipinski' rule was then downloaded from the Corina Online web server(https://www.mn-am.com)
in ‘.pdb’ format. Then the .pdb files was converted to. pdbqt files using the software Open
Babel(http://openbabel.org) which was then used for Virtual screening studies of those five
phytocompounds against Lung cancer .
Fig-4.1.1(a): 3-ethyl-7-methyloct-6-en-3-ol

37. 29
Fig- 4.1.1.(b) : 2-(5-ethyl-5-methyloxolan-2-yl)propan-2-ol
Fig- 4.1.1.(c) : 2-phenylchromene-3,4-dione

38. 30
Fig- 4.1.1.(d) : 5-propan-2-ylbicyclo[3.1.0]hex-3-en-2-one
Fig- 4.1.1.(e) : 5,7-dihydroxy-8-(3-methylbut-2-enyl)-2-phenyl-2,3-dihydrochromen-4-one

39. 31
4.1.2 LC4lvt protein :-
Bcl_2-Navitoclax (ABT-263) Complex
Classification: APOPTOSIS REGULATOR/INHIBITOR
Organism(s): Homo sapiens
Expression System: Escherichia coli
Mutation(s): Yes
The structure of LC4lvt is , an antiviral drug that act as a viral capsid inhibitor. Pleconaril is a
small molecule inhibitor with the activity against rhino viruses. It binds to a hydrophobic pocket
in the major capsid protein and prevents uncoating of the viral genome. LC4lvt protein in viral
capsid has been downloaded from RCSB site, and this proteins are involved for causing Lung
Cancer.
Fig- 4.1.2 : Protein Lc4lvt
4.1.2 Molecular docking:-
Protein preparation:-
The protein prepared for molecular docking using discovery studio. The PDB(protein database)
code of the LC4lvt protein in capsid was identified as a PDB format from the RCSB website. All
the water molecules are deleted. Add polar hydrogens select the ligand and using this ligand site,
define the binding site. the active site residues that are selected , the binding site spear should be
created.
Ligand preparation:-
Ligands are downloaded from the website PUBCHEM. ligands are downloaded as
SDF file in xl sheet from the data bases. In Osiris explorer property calculate the

40. 32
mutagenic, irritant ,tumorigenic, cLogp, solubility, molweight, drug likeness, drug-
score values. then to convert this smiles format into PDB format using Corina online.
the PDB files are renamed in ligand cid no. To convert PDB file to pdbqt file using
3rd
software open babel . Open the ligand in auto Dock as in PDB format.
Protein-ligand interaction:-
The protein molecule was treated as a receptor molecule and plant metabolites was
treated as the ligands. The docked protein and ligand are prepared for docking
process.
In molecular docking process the protein-ligand interaction has been done by using
BIOVIA Discovery Studio. In this process first the pdbqt file has converted to PDB
file using open babel. Out of 20 poses of ligand a single pose file should be created.
Then single pose PDB file in ligands and docked protein open in discovery studio.
Then successfully protein-ligand interaction has been shown. The interactions are
visualized in 2D diagram to see the interaction showing active site .

41. 33
CHAPTER-IV
Results and Discussions:-
Docked ligands are successfully binds in the active sites of LC4lvt protein. Protein-
ligand interaction signifies the bonds are interacts between the residues. Molecular
docking is based on the grid and it is developed for the perfections. in (table-1) shows
that different bonds as van darwaals, conventional hydrogen bond, pi-sulfur bonds,
pi-pi stacked, pi-Alkyl and amide-pi stacked bonds interacts with the residues and
close neighbouring residues. Osiris properties in all ligands has been shown in
(table-2). Calculate the risk factors like solubility, irritant, mutagenic, tumorigenic,
druglikeness, drugscore, reproductive effect and molweight values and affinity of
the ligands. Interaction map of 3-ethyl-7-methyloct-6-en-3-ol , 2-(5-ethyl-5-
methyloxolan-2-yl)propan-2-ol,2-phenylchromene-3,4-dione,5-propan-2-
ylbicyclo[3.1.0]hex-3-en-2-one,5,7-dihydroxy-8-(3-methylbut-2-enyl)-2-phenyl-
2,3-dihydrochromen-4-one as shown in the Fig-4.1.1.(a), Fig- 4.1.1.(b), Fig-
4.1.1.(c), Fig- 4.1.1.(d), Fig- 4.1.1.(e) above respectively.

42. 34
Ligand cid
no
Residues interacting
with ligands
Close neighbouring
residues
Affinity
124049 MET A:112, ALA
A:146, TYR A:105,
LEU A:134,PHE
A:101, ARG A:143
VAL A:153,PHE A:109,
Asp A:108, GLU A:149,
PHE A:150, VAL A:130,
GLU A:133, GLY A:142
265703 LEU A:198, ALA
A:97, ASP A:100,
VAL A:145
TRP A:141, PHE A:195,
TYR A:199, PHE A:101,
TYR A:105, ARG A:104
526764 VAL A:130, MET
A:112, PHE A:150,
LEU A:134, ALA
A:146
GLU A:133, VAL A:153,
PHE A:109, ASP A:108,
GLU A:149, PHE A:101
527426 ALA A:97, VAL
A:145
ASP A:100, PHE A:101,
GLY A:142, TYR A:199,
TRP A:141, PHE A:195,
LEU A:198
5317140 PHE A:101, LEU
A134, ALA A:146,
ASP A:108, VAL A:130,
MET A:112, PHE A:109,
VAL A:153, GLU A:149,
PHP A:150

43. 35
cid no- 124049 cid no.- 265703
cid no.- 526764
cid no.- 527426 cid no.- 5317140

44. 36
Conclusion-:
It was early studied known that Elettaria cardamomum plant has medicinal
action against viral disease . Elaichi is rich in antioxidants, black cardamom helps in
combating the symptoms of cold and cough, while the oil derived from its seeds acts
as an antiseptic which is help to cure against lungs cancer. Using autoDock vina
software molecular docking action was performed to identify the phytochemicals (3-
ethyl-7-methyloct-6-en-3-ol,2-(5-ethyl-5-methyloxolan-2-yl)propan-2-ol,2-
phenylchromene-3,4-dione,5-propan-2-ylbicyclo[3.1.0]hex-3-en-2-one,5,7-
dihydroxy-8-(3-methylbut-2-enyl)-2-phenyl-2,3-dihydrochromen-4-one) which can
have a dominant role with the protein LC4lvt . This work gives information about
the presence of this metabolites are most functional constituents in cardamom plant
that reduces the risk of Lung Cancer.

45. 37
Chapter – V
References
Mays Hadi Jebur, Ilham, A. Bnuyan, Amean, A. Yasri and Nada Khazal Kadhim
Hindi (2014). Antimicrobial Effect of Seed Extracts, leaves and Crude oil of
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