Phytochemical screening and evaluation of aphrodisiac and antihyperlipidemic activity of polyherbal extract of daucus carota sativus
1. PHYTOCHEMICAL SCREENING AND EVALUATION OF
APHRODISIAC AND ANTIHYPERLIPIDEMIC ACTIVITY OF
POLYHERBAL EXTRACT OF DAUCUS CAROTA SATIVUS (SEED),
MUSA PARADISIACA SAPIENTUM (STEM) AND CUCUMIS MELO
(PEEL)
A Phase-1 Dissertation Submitted to
VELS UNIVERSITY
CHENNAI – 600117
In partial fulfilment of the requirements
For the award of the Degree of
MASTER OF PHARMACY
IN
PHARMACOLOGY
SUBMITTED BY
Register No. 12403102
Under the Guidance of
Dr. J. Anbu, M. Pharm. Ph.D.
NOVEMBER-2013
DEPARTMENT OF PHARMACEUTICAL ANALYSIS
SCHOOL OF PHARMACEUTICAL SCIENCES, VISTAS
VELS UNIVERSITY, CHENNAI-600117
2. Dr. V. RAVICHANDIRAN, M.Pharm., Ph.D.,
Director
School of Pharmaceutical Sciences.
Vels University, Pallavaram, Chennai-117
CERTIFICATE
This is to certify that the dissertation entitled “Phytochemical screening and
evaluation of aphrodisiac and antihyperlipidemic activity of polyherbal
extract of Daucus carota sativus (seed), Musa paradisiaca sapientum
(stem) and Cucumis melo (peel)” submitted by Reg No. 12403102 to the Vels
University in partial fulfilment for the award of the Degree of Master of Pharmacy is an
independent bonafide work of the candidate carried out under the guidance of Dr. J.
Anbu, M. Pharm. Ph.D. School of pharmaceutical sciences during the academic year
2013 – 14.
.
Place: CHENNAI
SIGNATURE
Date:
3. Dr. J. Anbu, M. Pharm. Ph.D
Professor and Head,
Department of Pharmacology
School of Pharmaceutical Sciences.
Vels University, Pallavaram, Chennai-117
CERTIFICATE
This is to certify that the research work entitled “Phytochemical screening and
evaluation of aphrodisiac and antihyperlipidemic activity of polyherbal
extract of Daucus carota sativus (seed), Musa paradisiaca sapientum (stem) and
Cucumis melo (peel)” submitted by Reg. No. 12403102 to the Vels University in partial
fulfilment for the award of degree of master of pharmacy is an independent bonafide work of the
candidate carried out in the department of Pharmacology during the academic year 2013 – 14.
Place: CHENNAI
Date:
SIGNATURE
4. DECLARATION
The work presented in thesis entitled “Phytochemical screening and evaluation of
aphrodisiac and antihyperlipidemic activity of polyherbal extract of
Daucus carota sativus (seed), Musa paradisiaca sapientum (stem) and
Cucumis melo (peel)‟‟ Under the direct supervision and guidance of Dr. J. Anbu, M.
Pharm. Ph.D. Professor and Head of Pharmacology Vels University, Chennai – 600117.
This work is original and has not been submitted in any part or full for the award of any
degree or diploma of any other university.
Place: CHENNAI
(Reg. No. 12403102)
Date:
4
5. ACKNOWLEDGEMENT
First and foremost I express my deepest sense of gratitude and faithfulness to God‟s grace
which has enabled to finish my project work successfully. I am glad to have the blessings of
God in the implementation of our thought of doing this project.
I sincerely and extremely thankful to my esteemed my guide and Head of Department Dr. J.
Anbu, M. Pharm. Ph.D., and
Department of Pharmacology, for their excellent
suggestions, constant inspiration and personnel encouragement throughout my project work.
I would like to extend my sincere gratitude to Dr.V. Ravichandran, M.Pharm, Ph.D.,
Director, School of Pharmaceutical Sciences, Vels University.
I take pleasure to express my sincere thanks to Mrs. S. Nithya. M.pharm, assistant
professor, Department of Pharmaceutical Analysis for her valuable suggestions and
encouragement.
I take opportunity to express my thanks to all the Department members of Pharmacology,
for guidance, support and encouragement rendered for doing my project work.
Above all I pay my sincere thanks to my parents and family members for their blessings and
wishes which enabled me to go through all the difficulties. I will always remember the
support and encouragement given by my classmates and friends.
Reg. No.12403102
5
9. Phytochemical
screening
and
evaluation
of
aphrodisiac
and
antihyperlipidemic activity of polyherbal extract of Daucus carota sativus
(seed), Musa paradisiaca sapientum (stem) and Cucumis melo (peel)
INTRODUCTION
Plants have been one of the important sources of medicines since the beginning of human
civilization. Plants are a source of many biologically active products and now days they are
of great interest to the pharmaceutical industry. Medicinal plants contain natural chemicals,
which are acceptable to human and animal systems. All these chemicals cannot be
synthesized in laboratories. There is a growing demand for plant based medicines, health
products, pharmaceuticals, food supplements, cosmetics, etc. There exists a plethora of
knowledge and information and benefits of herbal drugs in our ancient literature of
Ayurveda and Unani medicine. One of the earliest treatises of Indian medicine, the Charaka
samhita (1000B.C) mentions the use of over 2000 herbs for medicinal purpose.
Daucus carota Linn. commonly known as “carrot" belongs to the family Apiaceae
(Umbelliferae) and is cultivated almost all over the world as a useful vegetable. Carrot is
widely consumed as an aphrodisiac and nervine tonic and its scraped root is used as a local
stimulant for indolent ulcers. Different parts of this plant are used in Indian systems of
medicine for the treatment of a broad spectrum of aliments including kidney dysfunction,
asthma, dropsy, inflammation, leprosy and worm troubles
hypoglycaemic and hepatoprotective role
3, 4, 5
1, 2
. It has an hypotensive,
. Musa paradisiaca sapientum (Musaceae) or
banana is widely used in ancient medicinal system. Musa paradisiaca sapientum fruits are
aphrodisiac, anthelmintic, antiscorbutic, antibilious, alterative, depurative and tonic. They
are useful in venereal diseases, helminthiasis, scabies, leprosy, toothache, skin diseases,
debility, anaemia, cachexia and intrinsic haemorrhage.
Cucumis melo (Cucurbitaceae) is commonly known as wild melon. Cucumis melo peels are
antihyperlipidemic in nature. It is heavy to digest, nutritious, aphrodisiac, relieves burning
sensation, relieves tiredness, diuretic, useful in blood disorders etc. In Ayurveda it is used to
cure Vata disease (Neurological diseases) and Pitta diseases (Bile related disease).
9
10. Over the past decades, herbal medicine has become a thing of global significance with
medicinal and economic implications. Wide spread use of herbs throughout the globe has
raised serious concerns over its quality, safety, and efficacy.
The main aim of our investigation is to potentiate the aphrodisiac and antihyperlipidemic
effect of the selected plants namely Daucus carota, Musa paradisiaca sapientum and
Cucumis melo by making a polyherbal formulation.
APHRODISIAC:
Aphrodisiac is the word derived from Aphrodite, the Greek goddess of sexual, love and beauty.
An aphrodisiac is defined as an agent (food or drug) that arouses sexual desire. Sexual
dysfunction is an inability to achieve a normal sexual intercourse including premature
ejaculation, retrograded, retarded or inhibited ejaculation, erectile dysfunction, arousal
difficulties (reduced libido), compulsive sexual behaviour, orgasmic disorder and failure of
detumescence. It is increasing worldwide due to etiological factors and aging. Several types of
treatment are claimed in the modern medicine but due to their serious side effects and higher
cost, search of natural supplement from medicinal plants as an aphrodisiac substance is
significantly increased6.
Sexual dysfunction can be a result of a physical or psychological problem.
• PHYSICAL CAUSES: Many physical and/or medical conditions can cause problems
with sexual function. These conditions include diabetes, heart and vascular (blood vessel)
diseases like hyperlipidemia which can lead to atherosclerosis and blockade of the heart
vessels, neurological disorders and hormonal imbalances, chronic diseases such as
kidney or liver failure, alcoholism and drug abuse. In addition, the side effects of certain
medications, including some antidepressant drugs, can affect sexual desire and function.
• PSYCHOLOGICAL CAUSES: These include work related stress and anxiety,
concern about sexual performance, marital or relationship problems, depression, feelings of
guilt, and the effects of a past sexual trauma etc.
Phosphodiesterases are the class of enzymes that exists in nearly all tissues regulating the
second messengers cAMP and cGMP involved in many diverse physiological functions.
Phosphodiesterase inhibitors are used for management of clinical disorders such as
dementia, depression, cardiovascular diseases, diabetes, chronic obstructive pulmonary
diseases, colitis, toxicities, pulmonary hypertension and erectile dysfunction
7,
8
.
Phosphodiesterase 5 inhibitors (PDEІ) i. e., sildenafil, vardenafil, tadalafil etc. are now used
10
11. in treatment of erectile dysfunction but have some common side effects like headache,
flushing, blurred vision etc.
TABLE 1- PLANTS CONTAINING APHRODISIAC POTENTIALS9
Sl.
Common Name
Family
Part Used
Bhindi
Malvaceae
Root
Abelmoschus moschatus
Musk mallow
Malvaceae
Seed
3.
Abrus precatorius L.
Crab's Eye
Paplionaceae
4.
Abutilon indicum (Linn.) Thuthi
Malvaceae
Seed, root
5.
Acacia catechu Wild.
Mimosaceae
Heartwood
Amaranthaceae
Root
Araceae
Rhizome
No.
1.
2.
7.
Name of Plant
Abelmoschus esculantus
(L)
Achyranthes
Catechu
aspera Apamarg,
Linn.
Latjeera
8.
Acorus calamus Linn.
Sweet flag
9.
Actiniopteris
radiata Morshikha
Seed
Actinopteridaceae Plant
Sw.
Whole
Allium sativum L.
Garlic
Liliaceae
Bulb
Allium cepa L.
Piaz
Liliaceae
Bulb
Aloe vera
Dhritkumari
Liliaceae
13.
Amaranthus spinosus L.
Chaulai
Amaranthaceae
14.
Asparagus
Asparagus
Liliaceae
Root
Peanut
Fabaceae
Seeds
10.
11.
12.
racemosus
Willd.
15.
Arachis hypogaea Linn.
Gel extract from
Leave
Leaves, Whole
Plant
11
12. 16.
17.
18.
19.
20.
21.
22.
Argyreia nervosa
Adhoguda
Convolvulaceae
Root
Jack tree
Moraceae
Fruit, Seed
Azadirachita indica
Neem
Meliaceae
Root
Aristolochia indica L.
Iswaramul
aristolochiaceae
Whole plant
Bacopa monnieri L.
Brahmi
Scrophulariaceae
Whole plant
Caesalpiniaceae
Seed
Caesalpiniaceae
Bark
Artocarpus
heterophyllus
Camel‟s
Bauhinia vahlii
Bauhinia
Foot
Climber
variegate
Linn.
Bauhinia
23.
Blepharis edulis Linn.
. Utangan/ Shikhi Acanthaceae
Seeds
24.
Boerhavia diffusa L.
Punarnava
Nyctaginaceae
Root
25.
Bombax ceiba Linn.
Silk-Cotton Tree
Bombacaceae
Bark
26.
Butea frondosa Roxb.
Papilionaceae
Whole plant
Arhar
Fabaceae
Root
Carica papaya L.
Papita
Caricaceae
Fruit
Cannabis indica L.
Indian hemp
Cannabinaceae
Leaf
Cannabis sativa
Bhang
Cannabinaceae
Leaf
Capsicum annuum L.
Capsicum
Solanaceae
Seed
Kasondhi
Fabaceae
Leaf
Vandangul
celastreaseae
Seed
27.
28.
29.
30.
31.
32.
Cajanus
cajan
(L.)
Millsp.
Cassia
occidentalis
Linn.
33.
Celastrus
wild.
paniculatus
Flame-of-theforest
12
13. 34.
35.
Chenopodium album L.
Chlorophytum
tuberosum Baker.
36.
37.
38.
Cissus
White goosefoot
Chenopodiaceae
Seed
Safed musli
Liliaceae
Whole plant
quadrangularis Edible stemmed
Linn.
Vine
Cocculs cardifolia Linn.
Guduchi
Commiphora mukul
Vitaceae
Menispermaceae
Hook. ex Stocks,
Indian bdellium Burseraceae
Root
Stem, leaf, Root
Root, leaf
Tree
39.
Desmodium gangeticum
Linn.
Fabaceae
Desmodium
(Papilionaceae)
Root
Coriander
Apiaceae
Leaf
Ebenaceae
Flower
40.
Coriandrum Sativum
41.
Diospyros melanoxylon East
Indian
Roxb.
ebony
Dolichos lablab Linn.
Flat bean, sem
Fabaceae
Seeds
Daucus carota L.
Carrot
Umbelliferae
Root
44.
Dalbergia sissoo
Shisham
Fabaceae
Wood
45.
Emblica officinalis
Gaerth Emblic
euphorbiaceae
Fruit
42.
43.
13
14. ANTIHYPERLIPIDEMIC STUDY:
Antihyperlipidemic drugs lower the levels of lipids and lipoproteins in blood in elevated
condition. The antihyperlipidemic drugs have attracted considerable attention because of
their potential to prevent cardiovascular diseases by retarding the accelerated atherosclerosis
in hyperlipidemic individual. The major cause of death in the Western world today is
vascular disease, of which the most prevalent form is atherosclerotic disease. The current
prediction is that within the year 2020 cardiovascular diseases, notably atherosclerosis will
become the leading global cause of total disease burden, defined as the years subtracted
from healthy life by disability or premature death an important factor in causing
atherosclerosis is hyperlipidemia
10
. Although many causative factors of this heart disease
are recognized (e.g., smoking, stress, diet), atherosclerotic disease can be treated through
medication or surgery. Hyperlipidemia is the most prevalent indicator for susceptibility to
atherosclerotic heart disease; it is a term used to describe elevated plasma levels of lipids
that are usually in the form of lipoproteins. Atherosclerosis may be defined as the
degenerative changes in the intima of medium and large arteries. This degeneration includes
the accumulation of lipids, complex carbohydrates, blood and blood products and is
accompanied by the formation of fibrous tissue and calcium deposition on the intima of
blood vessels. These deposits result in a narrowing of the lumen (inside diameter) of the
artery and a decrease in blood supply to the area served by the artery. When these fatty
deposits occur in the coronary arteries, the patient experiences coronary artery disease.
Lowering blood cholesterol levels can arrest or reverse atherosclerosis in the vessels and can
significantly decrease the incidence of heart disease.
Figure: Difference between healthy blood vessel and the hyperlipidemic blood vessel.
14
15. HYPERLIPIDEMIA CAN BE OF MANY CLASSES:
I.
Secondary hyperlipidemia:
This type of hyperlipidemia associated with diabetes, myxo-edema,
nephritic syndrome, chronic alcoholism etc.
II.
Primary hyperlipidemia:
A single gene defect:
This type of hyperlipidemia occurs due to familial causes and called as
„monogenic‟ or genetic hyperlipidemia.
Multiple genetic, dietary and physical activity related causes:
This type is called as „polygenic‟ or multifactorial.
Table 2: TYPES OF PRIMARY HYPERLIPOPROTEINEMIAS
Types
Disorder
Cause
Occurrence
Elevated
Plasma
plasma
lipids
lipoprotein
І
Familial lipoprotein
CH
TG
G
Very rare
Chylomicron
↑↑
↑↑↑
G
Less common
LDL
↑↑
N
MF
Commonest
LDL
↑
N
G
Rare
IDL, Chy.
↑
↑
lipase deficiency
ІІa
Familial
hypercholesterolemia
ІІb
Polygenic
hypercholesterolemia
ІІІ
Familial
dysbetalipoproteinemia
Rem.
ІV
Hypertriglyceridemia
MG, G
Common
VLDL
N
↑↑
V
Familial combined
G
Less common
VLDL, LDL
↑
↑
hyperlipidemia
CH- Chylomicron, TG- Triglycerides, G- Genetic, MF- Multifactorial, Chy. Rem.Chylomicron remnants, VLDL- Very- low- density lipoprotein, LDL- Low- density
lipoprotein, IDL- Intermediate density lipoprotein
15
16. LIPOPROTEIN CLASSES:
Lipoproteins are macromolecules consisting of lipid substances (cholesterol and
triglycerides) noncovalently bound with protein and carbohydrate. These combinations
solubilise the lipids and prevent from forming insoluble aggregates in the plasma. They have
a spherical shape and consist of a non polar core surrounded by a monolayer of
phospholipids whose polar groups are oriented towards the lipid phase of plasma. Included
in the phospholipid monolayer are a small no of cholesterol molecules and proteins termed
appoproteins. The appoproteins appear to be able to solubilise lipids for transport in an
aqueous surrounding such as plasma.
The various lipoproteins found in plasma can be separated by ultracentrifugal techniques
into chylomicrons, very- low- density lipoprotein (VLDL), intermediate- density lipoprotein
(IDL), low density lipoprotein (LDL), and high density lipoprotein (HDL). These correlate
with the electrophoretic separations of lipoproteins as follows: chylomicrons, pre-βlipoproteins (VLDL), broad β-lipoprotein (IDL), β-lipoprotein (LDL) and α- lipoprotein
(HDL).
Chylomicrons contain 90% triglycerides by weight and originate from exogenous fat from
the diet. They are the least dense of the lipoproteins and migrate the least under the
influence of electric current. Chylomicrons are normally absent from plasma after 12 to 24
hours of fasting.
Very- low- density lipoprotein (VLDL) is composed about 60% triglycerides, 12%
cholesterol and 18% phospholipids. It originates in the liver from free fatty acids. Although
VLDL can be isolated from plasma, it is catabolised rapidly into IDL, which is degraded
further into LDL.
Low density lipoprotein (LDL) consists of 50% cholesterol and 10% triglycerides. This is
the major cholesterol carrying protein. In normal persons, this lipoprotein accounts for about
65% of plasma cholesterol and is of major concern in hyperlipidemic disease states. The
LDL is formed from the intravascular catabolism of VLDL. The HDL is composed of 25%
cholesterol and 50% protein and accounts of about 17% of total cholesterol in plasma.
16
17. Table 3: Characteristics
Sl.
No.
1.
and Function of Plasma Lipoproteins
Lipoprotein Diameter(nm) Lipid contained
class
Chylomicron
Source of
Function
lipid
100-150
TG >> CHE
Diet
Dietary TG
transport
2.
Chylomicron
30-50
CHE >> TG
Diet, chy.
remnant
3.
VLDL
Dietary CH
transport
40-80
TG >> CHE
Liver
Endogenous TG
transport
4.
IDL
30-35
CHE > TG
VLDL
Transport CHE &
TG to liver, source
of LDL
5.
LDL
20-25
CHE
IDL
Transport CH to
tissue and liver
HDL
5-10
Phospholipid,
Tissues,
Removal of CH
CHE
6.
cell
from tissues
membrane
LIPOPROTEIN METABOLISM:
Lipids can be obtained either from de novo biosynthesis in the tissues or from the diet. The
rate of entering cholesterol and triglycerides in circulation depend on the supply of the lipid
and proteins necessary to form the lipoprotein complexes. Reduction of plasma lipid by
controlling the diet can delay the development of atherosclerosis and use of drugs can
decrease the assimilation of lipids into the body and can decrease the mortality of cardiac
disease.
Drugs used to treat hyperlipidemia are HMG-CoA reductase inhibitor or statin
derivatives (lovastatin, simvastatin, pravastatin, atorvastin, rosuvastatin etc.); bile acid
sequestrants (cholestyramine, cholestipol); lipoprotein lipase activators (clofibrate,
gemifibrozil, bezafibrate, fenofibrate); inhibitors of lipolysis and triglyceride synthesis
(nicotinic acid) and other compounds(ezetimib, gugulipid, β- sitosterol)
17
18. Atorvastatin will be used as standard drug for the experiment. Statins are the most
efficacious and best tolerated class of hypolipidemic drugs. They completely inhibit
conversion of 3-Hydroxy- 3-Methyl glutaryl coenzyme A to mevalonate by the enzyme
HMG-CoA reductase. It is the rate limiting step in cholesterol synthesis. Therapeutic dose
of statin derivates reduce CH synthesis by 20-50%. Atorvastatin is the potent and appears to
have the highest LDL-CH lowering efficacy at maximal daily dose of 80mg. Plasma t1/2 of
atorvastatin is 18-24 hours. It has additional antioxidant property.
18
20. PLANT PROFILE:
Daucus carota sativus
PLANT DESCRIPTION
Kingdom
Plantae – plant
Subkingdom
Trancheobionta – vascular plants
Superdivision
Spermatophyta - flowering plant
Division
Mangoliposida – dicotyledone
Subclass
Rosidae
Order
Apliales
Family
Apiaceae – carrot family
Genus
Daucus L.
Species
Daucus carota L.
Synonyms
Queen anne‟s lace, Bird‟s Nest
Hindi
Gazar
DISTRIBUTION:
Daucus carota Linn., commonly known as “Carrot” belongs to the family (Apiaceae,
Umbelliferae) and is cultivated almost all over the world as a useful vegetable. Native to
Europe and the Mediterranean region; extensively cultivated in Punjab, Haryana, Uttar
Pradesh and Madhya Pradesh for its fleshy tap roots which are eaten raw or cooked. Wild
Carrot: Nativeto Europe, Africa and Asia. It grows at 3,000 – 3,600 m in the Himalayas.
PARTS USED
Root, seed, flower etc.
MEDICINAL USES:
Daucus carota is commonly known as “Carrot” belongs to the family Apiaceae
(Umbelliferae) and is cultivated almost all over the world as a useful vegetable. Different
parts of this plant are used in Indian systems of medicine for the treatment of a broad
spectrum of aliments including kidney dysfunction, asthma, dropsy, inflammation, leprosy
20
21. and worm troubles 1, 2. Indian material media mention Daucus carota as a brain tonic 2. It has
an hypotensive, hypoglycaemic and hepatoprotective role3, 4, 5. Carrot is widely consumed as
an aphrodisiac and nervine tonic and its scraped root is used as a local stimulant for indolent
ulcers. The British Herbal Pharmacopoeia recommends Daucus carota Linn. (wild carrot)
for its diuretic activity. The ethanolic extract of seeds exhibited diuretic effect in dogs.
Aqueous extract of carrots showed hepatoprotective activity against CCl4 induced hepatic
damage in mice liver.
Roasted roots - prescribed in palpitation, burning micturation, cough and bronchitis. Carrot
increases the quantity of urine and helps the elimination of uric acid; also lowers blood
sugar. Juice a rich source of carotene.
Seeds - diuretic, emmenagogue, spasmolytic (prescribed in anuria and sexual debility).
Wild carrot - diuretic and antilithic (used for kidney stones, cystitis and in gout).
Seeds – emmenagogue and also used for hot flushes of the menopause.
ACTIVE CONSTITUENTS:
Daucus carota commonly known as “Carrot” is one of the most important vegetables and
belongs to the family of Apiaceae. Its main active constituents are volatile oils, steroids,
tannins, flavonoids and carotene have been isolated.
Picture: Daucus carota seeds
21
22. Musa paradisiaca sapientum
Kingdom
Plantae
Division
Magnoliophyta
Class
Liliopsida
Order
Zingiberales
Family
Musaceae
Genus
Musa
Species
Musa paradisiaca, Musa sapientum
Synonyms
Kadali, kala etc.
BOTANICAL DESCRIPTION:
Musa paradisiaca sapientun is a stout, stoloniferous, perennial herb, 2-8 m tall. Leaves
oblong, 1-3 x 0.2-0.3 m, suddenly truncate at both ends, acuminate or emarginate; petioles
0.5- 1 m on long sheaths forming pseudostems. Flowers unisexual, in a cymose
inflorescence subtended by a large bract and all partial inflorescences arranged spirally on a
long, drooping, stout axis. Bracts large, broadly ovate, 20-40 x 15-30 cm, brownish red,
truncate at base; lower bracts subtending female and distal ones male flowers. Fruits are
oblong to fusiform, generally 15-25 cm long and fleshy.
DISTRIBUTION:
It is extensively cultivated in India. It‟s a familiar topical fruit. From its native Southwestern
Pacific home, the banana plant spread to India by about 600 BC and later on it spread all
over the tropical world. It is possibly the world's oldest cultivated crop. It even spread into
the Islands of the Pacific and to the West Coast of Africa as early as 200-300 BC13.
PARTS USED:
Root, leaf, fruit, stem, flower
22
23. Picture: Musa paradisiaca sapientum plant
MEDICINAL USES:
The roots are anthelmintic, antiscorbutic, antibilious, alterative, depurative and tonic. They
are useful in venereal diseases, helminthiasis, scabies, leprosy, toothache, skin diseases,
debility, anaemia, cachexia and intrinsic haemorrhage.
The tender leaves are useful in scabies, inflammations, eye diseases, blisters and burns.
The fruits are astringent, emollient, cooling, anthelmintic, aphrodisiac, antiscorbutic,
laxative, demulcent and nutrient. They are useful in dipsia, haemoptysis, diabetes, gastritis,
dyspepsia with flatulence and acidity, chronic dysentery, diarrhoea, scurvy, helminthiasis,
scabies, pruritus, bronchitis, pharyngeal disorders, nephropathy, strangury, menorrhagia,
metrorrhagia and general debility.
The ashes obtained by burning the plant are antiscorbutic, anthelmintic, stomachic and
useful in hyperacidity, heartburn, colic and verminosis.
The flowers are astringent and good for dysentery, bronchial asthma, dysmenorrhoea,
menorrhagia, diabetes, ascites and dropsy.
The inflorescence axis (stem) is very specific for renal and vesical calculi. The stems are
also having haemostatic, hypoglycaemic effect.
23
24. Cucumis melo
Kingdom
Plantae
Subkingdom
Tracheobionta – vascular plants
Superdivision
Spermatophyta – seed plant
Division
Magnoliophyta – flowering plant
Class
Magnoliopsida – dicotyledons
Subclass
Dilleniidae
Order
Violales
Family
Curcubitaceae – cucumber family
Genus
Cucumis L.
Species
Melo L.
Synonyms
Kharbujam(hindi), mask melon(english)
BOTANICAL DESCRIPTION:
Picture: Cucurmis melo fruit
Cucumis melo Linn belonging to Cucurbitaceae family, commonly known as Muskmelon or
sweet melon (in English), Kharbuja (in Hindi), Ervaru (in Sanskrit). Leaves about 7.5 cm
diameter, orbicular-reniform in outline, 5-angled or lobed, scabrous on both surfaces and
also often with soft hairs; lobes not deep nor acute; petiole 5 cm; petals 1.6 cm. Female
peduncle sometimes 5 cm. Fruit spherical ovoid elongated or contorted, glabrous or
somewhat hairy, not spinous nor tuberculate.
24
25. PARTS USED:
Fruit, seed, peel, leaf, stem etc.
Picture: Cucurmis melo plant
MEDICINAL USES:
Cucumis melo (Cucurbitaceae) is commonly known as wild melon, cantaloupe, small gourd;
wild musk melon is an annual. The fruits can be used as a cooling light cleanser or
moisturizer for the skin and has stomachic properties. They are also used as a first aid
treatment for burns and abrasions. Seeds are antitussive, digestive, antioxidant, diuretic,
antiulcer, febrifuge and vermifuge. The extract of seed oil was reported for Antifungal
activity14. Further, the phytochemical studies of Cucumis melo seeds revealed the presence
of flavonoids15.
. Cucumis melo peels are antihyperlipidemic in nature. It is very sweet in taste, cool in
potency and delicious to eat. It is heavy to digest, nutritious, aphrodisiac, relieves burning
sensation, relieves tiredness, diuretic, useful in blood disorders etc. In Ayurveda it is used to
cure Vata disease (Neurological diseases) and Pitta diseases (Bile related disease).
25
27. AIM and SCOPE OF WORK:
From literature review of the above mentioned plants, it can be concluded that Daucus
carota and Musa paradisiaca sapientum both are having aphrodisiac potential and all of them
are having a very good antihyperlipidemic activity. So, our aim is
To potentiate the aphrodisiac and antihyperlipidemic effect of the selected
plants namely Daucus carota, Musa paradisiaca sapientum and Cucumis melo by
making a polyherbal formulation.
To study various parameters involved in this study.
The scope of the study can be further explored for its probable mechanism of
action and active principle mean for such biological activity.
To study the effect of the polyherbal extract in a dose dependent manner.
27
29. REVIEW OF LIERATURE
Daucus carota:
1. Kamlesh Singh et.al 2011, In vivo antioxidant and hepatoprotective
activity of methanolic extracts of Daucus carota seeds in experimental
animals.
In this study the In vivo antioxidant and hepatoprotective activity of methanolic extract of
Daucus carota seeds was evaluated in experimental animals.
Methanolic extracts of
Daucus carota seeds was used for hepatoprotection assessment. Oxidative stress were
induced subcutaneously in rats by thioacetamide, in test groups,standard and toxic control
group. Two test groups received Daucus carota seeds extract at different doses. Silymarin
was used as standard. Control group received only vehicle. On the 8th day animals were
sacrificed and liver enzyme like serum glutamic pyruvic transaminase (SGPT), serum
glutamic-oxaloacetic transaminase (SGOT) and alkaline phosphatase (ALP) were estimated
in blood serum and antioxidant enzyme like superoxide dismutase (SOD), catalase (CAT),
glutathione reductase (GRD), glutathione peroxidase (GPX), glutathione-S-transferase
(GST) and lipid peroxidation (LPO) were estimated in liver homogenate. The methanolic
extract of Daucus carota seed extract showed a significant dose dependent reduction of
oxidative stress and hepatoprotective effect14.
2. Kamlesh Singh et.al. 2010 , Hypolipidemic activity of ethanolic extract
of daucus carota seeds in normal rats.
In this study Hypolipidemic activity of ethanolic extract of Daucus carota seeds in Normal
rats was evaluated. Rats were grouped into 4 groups. The control group was treated only
with vehicle. The standard group treated with lovastatin for a period of one week. Test
groups were treated with different doses of Daucus carota seed extract for same period.
After one week blood samples were collected by retro orbital route and lipid profiles were
estimated in serum. The test extract showed a dose dependent reduction of oxidative stress
and lipid levels in experimental animals15.
29
30. 3. Mithun Vishwanath K Patil et. al.2012, Pharmacological evaluation of
ethanolic extract of Daucus carota Linn root formulated cream on wound
healing using excision and incision wound model
In this study ethanolic extract of Daucus carota L. root was used to evaluate wound healing
activity on excision wound model and incision wound model. Soft paraffin based creams
containing different concentration of ethanolic extract of Daucus carota L. (EEDC) root was
formulated and evaluated for pharmaceutical parameters such as rheological properties, pH,
skin irritation and external characters. The test extract showed a dose dependent quicker
process in wound healing activity may be due to the antioxidant and antimicrobial potential
of the text extract16.
4. Mani Vasudevan et.al.2006, Antinociceptive and Anti-inflamatory
Properties of Daucus carota Seeds Extract
In this study ethanolic extract of Daucus carota seeds (DCE) was investigated for antiinflammatory and analgesic activity at different doses. For evaluation of inflammation
carrageenan, histamine and serotonin-induced paw edema served as acute models and
formalin induced arthritis served as a chronic model in rats. The acetic acid induced
writhing response and formalin-induced paw licking time in the early and late phases of
mice were used to assess analgesic activity. the higher dose of DCE were inhibiting
carrageenan, histamine and serotonin-induced paw edema as well as formaldehyde-induced
arthritis successfully. In all doses DCE significantly attenuated the writhing responses
induced by intraperitonial injection of acetic acid and late phase of pain response induced by
an subplantar injection of formalin in mice17.
5. Vasudevan mani et.al.2010 , Anti-Dementia Potential of Daucus carota
Seed Extract in Rats
In this study ethanolic extracts of Daucus carota seeds were investigated for the antidementia potential in rats. The ethanolic extract of Daucus carota (DCE) was administered
orally thrice daily for seven successive days to different groups of young and aged rats. To
evaluate behavioural parameters for testing memory Elevated plus-maze, Hebb-Williams
maze and hexagonal swimming pool etc were used. Diazepam, scopolamine and ageinginduced amnesia served as the interoceptive behavioural models. DCE showed significant
30
31. improvement in memory of young and aged rats in the behavioural models. DCE also
reversed the amnesia induced by scopolamine and diazepam. These parameters suggest that
D. carota seeds appear to be a promising candidate for improving memory and it has a
potential in the management of Alzheimer patients18.
6. Baljinder Singh et al.2010 , pharmacological potential of plant used as
aphrodisiacs
According to this review article D.carota linn. Roots are having aphrodisiac activity19,20.
7. Muhammad Afzal et.al.2013 , Comparison of protective and curative
potential of Daucus carota root extract on renal ischemia reperfusion
injury in rats
In this study the protective and curative potential of Daucus carota root extract was
evaluated in rats. Daucus carota root was extracted in different solvents like pet ether,
methanol etc and it was given to the rats 14 days prior and after induction of ischemia. After
treatment it showed a significant reduction in serum creatinine , uric acid and urea than the
disease control. These parameters suggests that Daucus carota root extract significantly
increase the kidney function in rats21.
8. A.H. Gilani et.al.2000 , Hypotensive action of coumarin glycosides from
Daucus carota
In this study the hypotensive activities are evaluated on normotensive anaesthetised rats.
The cumerin glycosides obtained from the aerial parts of Daucus carota shows dose
depended arterial hypotensive activity. In vitro studies showed that both compounds caused
a dose - dependent inhibitory effect on spontaneously beating guinea pig atria as well as on
the K+-induced contractions of rabbit aorta at similar concentrations. These results indicate
that the glycosides can act through blockade of calcium channels. This K+ blocking effect
may be responsible for the blood pressure lowering effect of the compounds observed in the
in vivo studies22.
9. Hiader Radhi Malih 2008, Inhibitory effect of some plant extracts on
growth of some bacteria and fungi pathogens
31
32. In this study the ethanolic extract of Daucus carota l. seeds were evaluated for antimicrobial
activity against some pathogenic fungi, Gram-positive and Gram-negative bacteria.
Ethanolic Daucus carota showed inhibitory activity against Alternaria alternata growth
and the ethanolic extract was also showed a ability to control clinically isolated Gramnegative, Gram-positive and pathogens23.
10.
Alka Sahrawat et.al. 2013, Antibacterial screening of Sapindus
mukorossi Gaertn. fruits and Daucus carota L. root extracts against
pathogenic bacteria
In this study taproots of Daucus carota L. were extracted using different solvents like
ethanol, methanol, bezene etc. and evaluated for antimicrobial activity. Daucus carota
showed antibacterial activity against drug resistant bacterial strains such as Shigella flexneri
and Klebsiella pneumonia24.
11.
Vikrant Arya et.al. 2011, a review on plants having anti-arthritic
potential
According to this review article ethanolic extract of Daucus carota contains antiinflammatory activity in formaldehyde induced paw arthritis in rats25.
12. Muhammad Afzal et.al.2013, Comparison of protective and curative
potential of Daucus carota root extract on renal ischemia reperfusion
injury in rats
In this study petroleum ether extract, fractional methanolic extract and direct methanolic
extract of root are evaluated for protective and curative potential on renal ischemia
reperfusion injury in rats. All the extracts showed a significant reduction in the levels of
serum creatinine, uric acid and urea as compared to the disease control and the kidney
function was significantly improved in case of post conditioning with Daucus carota root
extract26.
32
33. 13. A.H. Gilani et.al.2000, Hypotensive action of coumarin glycosides
from Daucus carota
In this study extracts of aerial parts of Daucua carota was used to isolate two cumarin
glycosides coded DC-2 and DC-3. These isolated compounds are evaluated for hypotensive
activity both in-vitro and in-vivo. These compounds showed a dose dependent blood
pressure lowering effect in normotensive anaesthetised rats. The hypotensive effect may be
due to the calcium channel blocking activity of the isolated glycosidal compounds27.
14. Ana Cristina Tavares et.al.2008, Essential oil of Daucus carota subsp.
halophilus: Composition, antifungal activity and cytotoxicity
In this study the essential oils of Daucus carota was evaluated for antimicrobial and
antifungal activity. It showed a significant antifungal activity in mouse skin dendritic cells.
The essential oils showed a dose dependent antifungal effect mainly due to the presence of
elimicin. It showed a very low human cell detrimental activity28.
15.
Anupam Bishayee et.al.1995, Hepatoprotective activity of carrot
(Daucus carota L.) against carbon tetrachloride intoxication in mouse
liver
In this study the extract of carrot was evaluated for its hepatoprotective activity against
carbon tetrachloride intoxication using mouse liver. Daucus carota extract significantly
reduced the elevated lavels of hepatic enzymes due to the administration of carbon
tetrachloride. It showed a significant protective effect in carbon tetrachloride induced
hepatic injury29.
16.
P. K. Majumder et.al.1998, Effect of the seed extract of carrot
(Daucus carota Linn.) on the growth of Ehrlich ascites tumour in mice
In this study petroleum ether extract of Daucus carota seeds were evaluated for antitumor
activity. The petroleum ether extract of Daucus carota seeds showed a significant reduction
in the growth of Ehrlich ascites tumour in mice30.
33
34. Musa paradisiaca:
17. A. S. Alabi et.al.2013 , Beneficial effects of low dose Musa paradisiacal
on the semen quality of male WISTER rats
in this study elaluation of semen quality of adult male rats are done by using Musa
paradisiacal green mature fruits. Flour of fruit was given orally with distilled water . a
significant increase was observed in lower dose group but the higher dose group shows
marked decrease in sperm cell concentration and percentage of normal spermatozoa. so, it
can be concluded that M.paradisiaca has a beneficial effect on enhancing male reproductive
functions31.
18. Atul kumar Gangwar et.al.2013, To evaluate the analgesic activity of
leaves of Musa sapientum linn.
In this study the analgesic activity of M.sapientum linn. Was evaluated using hot plate
method. Aqueous and ethanolic extract was prepared for evaluation. The prepared drugs are
administered intraperitonially and the reaction time was compared against the control and
standard group, high doses of both extract showed a significant analgesic activity32.
19.
Shanthy Sundaram et.al.2011, Antioxidant Activity and Protective
effect of Banana
Peel against Oxidative Hemolysis of Human
Erythrocyte at Different Stages of Ripening
In this study, evaluation of protective effect of peel extracts of unripe, ripe, and leaky ripe
banana fruit on hydrogen peroxide-induced hemolysis and their antioxidant capacity were
investigated. Musa paradisica peel at different stages of ripening (unripe, ripe, leaky ripe)
were treated with acetone mixture, which were partitioned in order of polarity with water,
ethyl acetate, chloroform (CHCl3), and hexane sequentially. The antioxidant activity was
evaluated by the red cell hemolysis assay, free radical scavenging (1,1-diphenyl-2picrylhydrazyl free radical elimination) and superoxide dismutase activities. The Folin–
Ciocalteu's reagent assay was used to estimate the phenolic content of extracts. This showed
that the unripe banana peel has higher antioxdent potency than ripe and leaky type33.
34
35. 20. Samuel Adetunji Onasanwo et.al.2013, Anti-ulcer and ulcer healing
potentials of Musa sapientum peel extract in the laboratory rodents.
In this studythe antiulcer and ulcer healing properties of Musa sapientum methanolic peel
extract are evaluated in laboratory rats. Models involved in this study are alcohol-induced,
aspirin-induced, and pyloric ligation-induced models, and for its ulcer healing activity study
acetic acid-induced ulcer models in rats are employed.the extract showed dose dependent
anti ulcer and ulcer healing activity. The experiment revealed that the anti-ulcer effect of the
extract may be due to its anti-secretory and cyto-protective activity. The healing of the ulcer
base might not be unconnected with basic fibroblast growth factors responsible for epithelial
regeneration34.
21. M.H. Borges et.al.2005, Neutralizing properties of Musa paradisiaca L.
(Musaceae) juice on phospholipase A2, myotoxic, hemorrhagic and lethal
activities of crotalidae venoms.
In this study nutralizing property of Musa paradisiaca juice on snake venoms.interaction
between snake venom proteins with Musa paradisiaca extract was examined. Crotalidae
venom induced Phosphodiesterase A2, myotoxic and hemorrhagic activities including
lethality on the mice were significantly inhibited by the extract. This effect may be due to
the presence of polyphenols and tannins as they are responsible for non-specific inactivation
of proteins. Musa paradisiaca showed significant protection against the toxic effect of snake
venom in-vitro,but it has no effect when the experiments were done in vivo35.
22. A. Weremfo et.al2011, Haemostatic Effect of the Stem Juice of Musa
paradisiaca L. (Musaceae) in Guinea Pigs.
In this study the effect of Musa paradisiacal stem extract was evaluated for possible
haemostatic activity in guinea pig using bleeding and clotting times. It was observed that
both clotting time and bleeding time were reduced significantly36.
23. Ekpo Benjamin et.al 2011, Evaluation of Hypoglycemic activity of
Musa Paradisiaca L.(Musaceae) in rat.
In this study the bulb and stem part of Musa paradisiacal was evaluated against blood
concentration in normoglycemic and alloxan induced diabetic rat. The stem shows
35
36. hyperglycaemic activity while the bulb of the plant can be used in ethano therapy of
diabetes mellitus37.
24. Santosh Kumar Singh et.al 2007, Assessment of glycemic potential of
musa paradisiaca stem juice.
In this study Musa paradisica stem juice was evaluated on blood glucose level of normal and
hypoglycaemic rats. The stem juice shows a significant increase in blood glucose level in
both the cases38.
25. R. Gopakumara pillai et.al 1995, The core of the pseudostem of musa in
the treatment of Urinary stones
In this study the core of the psudostem juice of Musa paradisiaca and Musa sapientum were
evaluated against urolithiasis in patients. The juice shows effect in curing urolithiasis,
especially of the calcium oxalate variety39.
26. Paul C. Onyenekwe et.al. 2013. Phytochemical Screening and Effect of
Musa paradisiaca Stem Extrude on Rat Haematological Parameters
In this study investigation of phytochemical composition and effect of Musa paradisiacal
stem extrude was calculated on haematological parameters in Albino Wister rats. The
extrude shows haematopoietic and immunomodulatory effect on animal model40.
27. M. Sarowar Hossain et.al. 2013. Antidiarrheal, Antioxidant and
Antimicrobial Activities of the Musa sapientum Seed.
In this study the Antidiarrheal, Antioxidant and Antimicrobial potential of Musa sapientum
seeds were evaluated using animal model. The result suggest that the seed extract is having
antidiarrheal potential along with antioxidant and antibacterial potentiality41.
28. P. Prabha et.al. 2013. Indigenous anti-ulcer activity of Musa sapientum
on peptic ulcer.
In this study the antiulcer potential of Musa sapientum was evaluated using indomethacin
induced ulcerogenic rats. The flavonidal content present in the drug showed protective
effect agaist used animal models42.
29. Mohammad Zafar Imam et.al. 2011. Musa paradisiaca L. and Musa
sapientum L. : A Phytochemical and Pharmacological Review
In this article the authors mentioned traditional uses, taxonomical classification,
phytochemical constituents present in the both species of banana. They are having
Antidiarrhoeal, Antiulcerative, Antimicrobial, Hypoglycemic, Hypocholesterolaemic,
36
37. Antihypertensive, atherosclerotic, Antioxidant, Diuretic, Wound healing, Anti-allergic,
Antimalarial, Anti-snake venom activities and mutagenic potential43.
30. Repon Kumer Saha et.al.2013. Medicinal activities of the leaves of
Musa sapientum var. sylvesteris in vitro
In this study the medicinal values of methanolic extract of the leaves of Musa sapientum.
The extract showed hemaglutination inhibitory activity in hydrogen peroxide induced
hemolysis inhibition activity of human red blood cells44.
31. Imam Md. Zafar et.al.2011. Antimicrobial and cytotoxic properties of
different exracts of Musa sapientum L. Subsp. sylvestris
In this study the different extracts of Musa sapientum L. was used to evaluate antimicrobial
and cytotoxic potentials are evaluated in eggs of brine shrimp. The extract was shown a
good antimicrobial activity against a wide range of bacteria as well as against pathogenic
fungi45.
32. K.R. Tandel et.al.2012. Evaluation of gastric antiulcerogenic action of
vegetable plantain banana (musa sapientum var. Paradisiaca) in aspirin
plus pylorus ligated albinorats.
In this study unripe dried power of banana was evaluated against aspirin plus pylorus ligated
albino rats. Orally induced banana powder showed a significant effect in reduction of ulcer
in aspirin with pyloric ligated rats46.
Cucumis melo
33. K. Ravishankar et.al.2012, In vitro antioxidant activity of ethanolic
seed extracts of Macrotyloma uniflorum and Cucumis melo for
therapeutic potential
In this study the ethanolic seed extract of Macrotyloma uniflorum and Cucumis melo are
evaluated for their antioxidant potential and for their therapeutic potential. In-vitroantioidant
study was carried out by nitric- oxide free radical scavenging assay. Presence of
phytochemicals like alkaloids, tannins, flavonoids, glycosides in both the extracts might be
responsible for their antioxidant activity47.
37
38. 34.
K. Ravishankar et.al.2012, Evaluation of diuretic effect of ethanolic
seed extracts of Macrotyloma uniflorum and Cucumis melo in rats
In this study the ethanolic seed extract of Macrotyloma uniflorum and Cucumis melo are
evaluated for their diuretic activity. The ethanolic extract of both the seed extracts showed a
significant diuretic activity and they show signergistic activity too48.
35. N. S. Gill et.al.2011, Evaluation of antioxidant and antiulcer activity of
Traditionally consumed Cucumis melo seeds
In this study the antioxidant and antiulcer potential of methanolic extract of Cucumis melo
seeds are evaluated against free radical scavenging assay by DPPH and pyloric ligation
method respectively. The methanolic extract of Cucumis melo showed a high antiulcer and
antioxidant potential49.
36. Mohd. Imtiyaz Ahmad et.al. 2012, Pharmacognostical studies and
establishment of quality parameters of Cucumis melo l. Cv. Namdhari
In this research article the pharmacognostical studies like morphological study, powder
microscopy, Physicochemical Standardization, Powdered drug reaction with different
reagents, Fluorescence Analysis, Phytochemical screening, Loss on drying, Bitterness value,
Foaming index etc are done50.
37. Kaur Manpreet et.al.2011, Antioxidant activity of Cucumis melo var.
Agrestis seeds for their therapeutic potential
In this study the antioxidant potentiallity of methanolic extract of Cucumis melo var.
Agrestis was evalated for its therapeutic potential. Presence of phytochemicals like
triterpenoids, alkaloids, tannins, flavonoids, coumarin glycosides, carbohydrates might be
responsible for the antioxidant activity51.
38. Jayant S. Bidkar et.al.2012, Anti-hyperlipidemic activity of Cucumis
melo fruit peel different extract in triton x-100 induced hyperlipidemia in
rats.
38
39. In this study chloroform, methanolic and aqueous extracts of Cucumis melo fruit peel wera
evaluated against triton induced hyperlipidemic rats. The methanolic extract of Cucumis
melo fruit peel showed a definit antihyperlipidemic activity in triton induced hyperlipidemic
rats52.
39
41. DESIGN OF INVESTIGATION
1. Plant collection and authentication
2. Extraction and Phytochemical screening
3. Selection of solvent
4. Bulk extraction
5. Institutional Animal Ethical Committee approval
6. Animal selection and procurement
Healthy adult mice weighing 25-35 g will be selected for aphrodisiac study and healthy
adult rats weighing 100-150 g will be selected for antihyperlipidemic studies.
7. Toxicity study – According to OECD-423
8. Grouping and selection of dose
For both the experiments the rats will be grouped into 5 groups
Group І
control(receive only CMC)
Group ІІ
test І (low test drug group)
Group ІІІ
test ІІ (high test drug group)
Group ІV
standard (receive standard drug)
Group V
receive the solvent, selected for bulk extraction
9. Selection of dose will depend on the toxicity study.
41
42. METHODOLOGY:
1. EVALUATION OF APHRODISIAC POTENTIAL:
The sexually active Swiss albino mice (25-35g) are required for this purpose. They will
be grouped separately and divided into 5 groups; each group consists of 6 animals.
Group І will receive CMC, group ІІ and group ІІІ will receive the test drug at different
doses, Group ІV will receive standard drug (sildenafil citrate) and the group V will
receive the selected vehicle for extraction of the drug, orally using the oral needle.
Sexual behaviour will observe in a dim light at day time in specially designed cages
having glass on all sides. The female mice should be in its oestrous phase when they
will introduce with male animals. The first 15 min will considered as acclimatization
period. The activities of male mice in each group will record individually for 60 min
after 30 min of drug administration on 1st, 7th and 14th day of drug treatment52, 53.
PARAMETERS TO BE NOTED:
i.
Mounting frequency,
ii.
Mounting latency
iii.
Intromission frequency,
iv.
Intromission latency
v.
Sperm count,
vi.
Sperm motility.
vii.
Level of serum testosterone, estrogen and progesterone.
2. EVALUATION OF ANTIHYPERLIPIDEMIC ACTIVITY:
(a) DIET INDUCED HYPERLIPIDEMIA: High fat /cholesterol supplements
to normal diet will be administered separately by intragastric tube once daily or
incorporated into diet pellets for 1-2 weeks.(like 400 mg/kg cholesterol in 5ml
coconut oil)54.
(b) TRITON INDUCED HYPERLIPIDEMIA: In rats, hypercholesterolemia
will be induced by administering Triton (isooctyl-polyoxy-ethylenephenol) at a dose
of 400mg/kg in saline suspension by intraperitoneal route. They will be allowed
42
43. normal for food and water. Measurable hypercholesterolemia will be established
within 18 h54.
Adult healthy rat weighing 100-150g will be required for this purpose. They will be grouped
separately and divided into 5 groups; each group consists of 6 animals. Group І will receive
CMC, group ІІ and group ІІІ will receive the test polyherbal drug at different doses, Group
ІV will receive standard drug (atorvastatin) and the group V will receive the selected vehicle
for extraction of the drug, orally using the oral needle. The rats will receive the polyherbal
drug for 15 days.
COLLECTION OF BLOOD SAMPLES
After treatment blood will be collected by retro orbital sinus puncture, under mild ether
anaesthesia in centrifugation tubes. Serum will be obtained by centrifugation. The
temperature and centrifugation speed should be maintained at 4ºC and 3000 rpm for 5 min
respectively.
PARAMETERS TO BE NOTED:
Serum lipid profiles
i.
serum Total Cholesterol,
ii.
Triglycerides,
iii.
LDL and
iv.
HDL
STATISTICAL ANALYSIS
All the data will be analysed statistically by one way ANOVA and students„t‟ test.
43
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