PHYTOCHEMICAL SCREENING AND EVALUATION OF
APHRODISIAC AND ANTIHYPERLIPIDEMIC ACTIVITY OF
POLYHERBAL EXTRACT OF DAUCUS CARO...
Dr. V. RAVICHANDIRAN, M.Pharm., Ph.D.,
Director
School of Pharmaceutical Sciences.
Vels University, Pallavaram, Chennai-11...
Dr. J. Anbu, M. Pharm. Ph.D
Professor and Head,
Department of Pharmacology
School of Pharmaceutical Sciences.
Vels Univers...
DECLARATION

The work presented in thesis entitled “Phytochemical screening and evaluation of

aphrodisiac and antihyperli...
ACKNOWLEDGEMENT
First and foremost I express my deepest sense of gratitude and faithfulness to God‟s grace
which has enabl...
Phytochemical

screening

and

evaluation

of

aphrodisiac

and

antihyperlipidemic activity of polyherbal extract of Dauc...
CONTENTS

SL. NO.

PARTICULARS

PAGE NO.

1

INTRODUCTION

1-10

2

PLANT PROFILE

11-16

3

17
AIM and SCOPE OF WORK

4

...
INTRODUCTION

8
Phytochemical

screening

and

evaluation

of

aphrodisiac

and

antihyperlipidemic activity of polyherbal extract of Dauc...
Over the past decades, herbal medicine has become a thing of global significance with
medicinal and economic implications....
in treatment of erectile dysfunction but have some common side effects like headache,
flushing, blurred vision etc.

TABLE...
16.
17.
18.

19.

20.

21.

22.

Argyreia nervosa

Adhoguda

Convolvulaceae

Root

Jack tree

Moraceae

Fruit, Seed

Azadi...
34.

35.

Chenopodium album L.
Chlorophytum
tuberosum Baker.

36.
37.

38.

Cissus

White goosefoot

Chenopodiaceae

Seed
...
ANTIHYPERLIPIDEMIC STUDY:
Antihyperlipidemic drugs lower the levels of lipids and lipoproteins in blood in elevated
condit...
HYPERLIPIDEMIA CAN BE OF MANY CLASSES:
I.

Secondary hyperlipidemia:
This type of hyperlipidemia associated with diabetes,...
LIPOPROTEIN CLASSES:
Lipoproteins are macromolecules consisting of lipid substances (cholesterol and
triglycerides) noncov...
Table 3: Characteristics
Sl.
No.
1.

and Function of Plasma Lipoproteins

Lipoprotein Diameter(nm) Lipid contained
class
C...
Atorvastatin will be used as standard drug for the experiment. Statins are the most
efficacious and best tolerated class o...
PLANT PROFILE

19
PLANT PROFILE:
Daucus carota sativus
PLANT DESCRIPTION
Kingdom

Plantae – plant

Subkingdom

Trancheobionta – vascular pla...
and worm troubles 1, 2. Indian material media mention Daucus carota as a brain tonic 2. It has
an hypotensive, hypoglycaem...
Musa paradisiaca sapientum
Kingdom

Plantae

Division

Magnoliophyta

Class

Liliopsida

Order

Zingiberales

Family

Musa...
Picture: Musa paradisiaca sapientum plant
MEDICINAL USES:

The roots are anthelmintic, antiscorbutic, antibilious, alterat...
Cucumis melo
Kingdom

Plantae

Subkingdom

Tracheobionta – vascular plants

Superdivision

Spermatophyta – seed plant

Div...
PARTS USED:
Fruit, seed, peel, leaf, stem etc.

Picture: Cucurmis melo plant
MEDICINAL USES:
Cucumis melo (Cucurbitaceae) ...
AIM and SCOPE OF WORK

26
AIM and SCOPE OF WORK:
From literature review of the above mentioned plants, it can be concluded that Daucus
carota and Mu...
REVIEW OF LIERATURE

28
REVIEW OF LIERATURE
Daucus carota:
1. Kamlesh Singh et.al 2011, In vivo antioxidant and hepatoprotective
activity of metha...
3. Mithun Vishwanath K Patil et. al.2012, Pharmacological evaluation of
ethanolic extract of Daucus carota Linn root formu...
improvement in memory of young and aged rats in the behavioural models. DCE also
reversed the amnesia induced by scopolami...
In this study the ethanolic extract of Daucus carota l. seeds were evaluated for antimicrobial
activity against some patho...
13. A.H. Gilani et.al.2000, Hypotensive action of coumarin glycosides
from Daucus carota
In this study extracts of aerial ...
Musa paradisiaca:
17. A. S. Alabi et.al.2013 , Beneficial effects of low dose Musa paradisiacal
on the semen quality of ma...
20. Samuel Adetunji Onasanwo et.al.2013, Anti-ulcer and ulcer healing
potentials of Musa sapientum peel extract in the lab...
hyperglycaemic activity while the bulb of the plant can be used in ethano therapy of
diabetes mellitus37.

24. Santosh Kum...
Antihypertensive, atherosclerotic, Antioxidant, Diuretic, Wound healing, Anti-allergic,
Antimalarial, Anti-snake venom act...
34.

K. Ravishankar et.al.2012, Evaluation of diuretic effect of ethanolic

seed extracts of Macrotyloma uniflorum and Cuc...
In this study chloroform, methanolic and aqueous extracts of Cucumis melo fruit peel wera
evaluated against triton induced...
DESIGN OF INVESTIGATION

40
DESIGN OF INVESTIGATION
1. Plant collection and authentication
2. Extraction and Phytochemical screening
3. Selection of s...
METHODOLOGY:
1. EVALUATION OF APHRODISIAC POTENTIAL:
The sexually active Swiss albino mice (25-35g) are required for this ...
normal for food and water. Measurable hypercholesterolemia will be established
within 18 h54.
Adult healthy rat weighing 1...
REFERENCES

44
REFERENCES
1. Kritikar K. R., Basu B. D., “Indian Medicinal Plants,” 2nd ed., Periodical Export,
New Delhi, 1991.
2.

Nadk...
12. Hosamath PV. Evaluation of antimicrobial activity of Litsea Glutinosa. International
Journal of Pharmaceutical Applica...
23. Hiader Radhi Malih. Inhibitory effect of some plant extracts on growth of some
bacteria and fungi pathogens. Thi-Qar M...
33. Shanthy Sundaram, Shadma Anjum, Priyanka Dwivedi, Gyanendra Kumar Rai.
Antioxidant Activity and Protective effect of B...
42. P. Prabha, Thirunethiran Karpagam, A. Sohna Chandra Packiavathy. Indigenous
anti-ulcer

activity

of

Musa

sapientum
...
51. Kaur Manpreet, Arora R. Antioxidant activity of Cucumis melo var. Agrestis seeds
for their therapeutic potential. Inte...
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  1. 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. 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. 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. 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. 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
  6. 6. 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) 6
  7. 7. CONTENTS SL. NO. PARTICULARS PAGE NO. 1 INTRODUCTION 1-10 2 PLANT PROFILE 11-16 3 17 AIM and SCOPE OF WORK 4 18-27 REVIEW OF LIERATURE 5 DESIGN OF INVESTIGATION 6 28-30 30-36 REFERENCES 7
  8. 8. INTRODUCTION 8
  9. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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
  19. 19. PLANT PROFILE 19
  20. 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. 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. 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. 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. 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. 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
  26. 26. AIM and SCOPE OF WORK 26
  27. 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
  28. 28. REVIEW OF LIERATURE 28
  29. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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. 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
  40. 40. DESIGN OF INVESTIGATION 40
  41. 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. 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. 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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