Bp க்கு வெண்தாமரைசூரணம்

PRECLINICAL STUDY OF SIDDHA DRUG
VENTHAMARAIYATHI CHOORANAM FOR IT’S

THROMBOLYTIC, VASODILATOR, HYPOLIPIDEMIC &
CARDIO PROTECTIVE ACTIVITIES
Dissertation submitted to
THE TAMILNADU DR. MGR MEDICAL UNIVERSITY
CHENNAI-600032
In partial fulfilment of the requirements
for the award of the degree of
DOCTOR OF MEDICINE (SIDDHA)
BRANCH-II-GUNAPADAM
POST GRADUATE DEPARTMENT OF GUNAPADAM
THE GOVERNMENT SIDDHA MEDICAL COLLEGE
TIRUNELVELI-627002
OCTOBER 2016
GOVT. SIDDHA MEDICAL COLLEGE,
PALAYAMKOTTAI
DECLARATION BY THE CANDIDATE
I hereby declare that this dissertation entitled “Pre-clinical study of herbal formulation
Venthamaraiyathi chooranam for its Thrombolytic, Vasodilator ,Hypolipidemic, &
Cardioprotective activities” is a bonafide and genuine research work carried out by me
under the guidance of Dr.A.KINGSLY M.D(s), Reader, Head of the Department, Post
Graduate Department of Gunapadam, Govt. Siddha Medical College, Palayamkottai, Tirunelveli-
02 and the dissertation has formed the basis for the award of any Degree, Diploma, Fellowship or
other similar title.
Date: Signature of the Candidate
Place: Palayamkottai Dr.J.Indrakumar

PALAYAMKOTTAI
CERTIFICATE BY THE GUIDE
This is to certify that the dissertation entitled “Pre-clinical study of herbal formulation
Venthamaraiyathi chooranam for its Thrombolytic, Vasodilator ,Hypolipidemic, &
Cardioprotective activities” is submitted to The Tamilnadu Dr.M.G.R.Medical
University,Chennai-32 in partial fulfilment of the requirements for the award of degree of M.D
(Siddha) is the bonafide and genuine research work done by Dr. J.Indrakumar. Under my
supervision and guidance and the dissertation has formed the basis for the award of any Degree,
Diploma, Associateship, Fellowship or other similar title.
Date: Signature & Seal of the Guide

Place: Palayamkottai Dr.A.Kingsly M.D(s) Reader
PALAYAMKOTTAI
BONAFIDE CERTIFICATE
This is to certify that the dissertation entitled “Pre-clinical study of herbal formulation
Venthamaraiyathi chooranam for its Thrombolytic, Vasodilator, Hypolipidemic, &
Cardioprotective activities” is a bonafide work done by Dr.J.Indrakumar, a candidate of
Government Siddha Medical College, Palayamkottai, Tirunelveli-02 in partial fulfilment of the
University rules and regulations for award of M.D (siddha) - Gunapadam under my guidance and
supervision during the academic year of 2016.
Name & Signature of the Name & Signature of the

Head of department Principal
ACKNOWLEDGEMENT
However it would not have been possible without Siddhar’s blessings, who has
given me power and courage to accomplish this dissertation work with success.
I wish to express my gratitude and acknowledge to the Vice-chancellor,
The Tamilnadu Dr.M.G.R. Medical University, Chennai, for giving me the permission to
do this dissertation work and to approve my dissertation topic for research.
I wish my deep gratitude to Commissioner of Indian Medicine and Homeopathy, Chennai
for giving permission to undertake this research work.
I would like to express my deepest appreciation to all those who provided me the
possibility to complete this dissertation.
I sincerely thank to Dr. S. Soundararajan, M.D(s)., Former Principal, Government
Siddha Medical College, Palayamkottai who permitted and initiated this dissertation work and I also
thank to Dr.S.Victoria, M.D(s)., Principal, Government Siddha Medical College, Palayamkottai for
her continuous support for carring out this study.
I My sincere thanks to my guide Lecturer Dr.A.Kingsly M.D(s), Reader, Head
of the Department, Department of PG Gunapadam. Government Siddha medical college,
Palayamkottai, for his valuable guidance and hopeful support for completion of my whole
study.
I sincerely thank to Dr. M. Ravichandran, M.D(s).,Ph.D., Reader, Former Head of the
Department of PG Gunapadam, Government Siddha Medical College, Palayamkottai
I wish to express my deep gratitude and sincere thanks to
Dr Dr.G.Essaky Pandian M.D(S) Lecturer, PG Gunapadam Government siddha medical
college, Palayamkottai for providing much needed attentions helpful suggestions for
dissertation.
I would like to give special thanks to Dr. Anthony duraichi M.D(s), Lecturer
Government siddha medical college, Palayamkottai for his helpful suggestions and
encouragement for preclinical studies.
I would like to give special thanks to Dr.S.Justus Antony M.D(S) Lecturer,
Government siddha medical college, Palayamkottai for his helpful suggestions .
I acknowledge my special thanks to Dr.M.KalaivananM.Sc, Ph.D., Department
of Pharmacology, Post Graduate study centre, Government Siddha Medical College,
Palayamkottai, for his constant help and encouragement to complete the pharmacological
work successfully.
I also acknowledge my thanks to Mrs.N.Nagaprema, M.Sc.,M.Phil., Head of
the Department of Bio-chemistry, Government Siddha Medical College, Palayamkottai for
her kind help and suggestions on biochemical aspects of this dissertation.
I am happy to express my thanks to Dr. S. Sutha, M.Sc., Ph.D., Head of the
Department of Herbal Botany and Herbal Pharmacognosy, Govt. Siddha Medical College,
Palayamkottai for her kind help in botanical aspect of this study and valuable suggestions
regarding drug identification.
I acknowledge my thanks to Dr. R.Murugesan, Ph.D, Scientific Officer, Grade I
Staff, IIT, Chennai for Instrumental analysis of ICP-OES, SEM and FTIR.
I express my sincere thanks to Dr. N. Chidambaranathan, HOD, Dept. of
Pharmacology, K.M.University, Madurai for their excellent help in pharmacological study
and other guidance to do the research work.
I express my sincere thanks to Dr. Arihara Sivakumar, Associate Professor,
Dept. of Pharmacology, K.M.C.H. College of pharmacy, Coimbatore. For their excellent
help in pharmacological study and other guidance to do the research work
I also acknowledge my thanks to Aravindh Herbal Labs, Pvt ltd,
Rajapalayam for Physico chemical analysis .
I also acknowledge to the Mrs.Poongodi, Librarian, Government Siddha medical
college, Palayamkottai.
My special thanks goes to my Sister Dr.R.Gnanasundari M.D(s)., and his
family for valuable support and encouragement and very helpful work in completing my
dissertation.
I am also thankful to all my college staffs for their kind co-operation for my
study. I should express my greatfulness to all My Classmates and P.G. Gunapadam
Students for lending their helping hands whenever needed during the course of the study.
The great walls to build my dissertation are my lovable family members they
support me in all kinds to complete this work in a good way. I sincerely express my love to
My respectable father Mr.R.Jayam and my dearest mother Mrs.P.Meenatchi for their
sincere encouragement and inspiration throughout my research work and lifting me uphill
this phase of life. I owe everything to them. Besides this, several people have knowingly and
unknowingly helped me in the successful completion of this project.
Finally, I am very thankful to Mr. M.Maharaja, & Mrs. Rajeswari, Maharaja DTP
Services Tiruchendur road, Palayamkottai for their help in bringing out this work at fulltilt.
ABBREVATIONS
ALT - Alanine transaminase
ANOVA- - Analysis of variance
AST - Aspartate transaminase
CMC - Carboxyl Methyl Cellulose
CDC - Council of Disease control and prevention
CPCSEA - Committee for the purpose of control and supe
DC - Differential Count
EDTA - Ethylene Diamine Tetra Aceticacid
ESR - Erythrocyte Sedimentation Rate
FTIR - Fourier transform infrared spectroscopy
Hb - Haemoglobin
DC - Differential Count
EDTA - Ethylene Diamine Tetra Aceticacid
ESR - Erythrocyte Sedimentation Rate
FTIR - Fourier transform infrared spectroscopy
IAEC - Institutional Animal Ethical Committee.
ICP-OES - Inductively coupled plasma optical emission spectrometry
Ig E - Immunoglobulin E
LDH - Lactate Dehydrogenase
MCV - Mean Corpuscular Volume

VTC - Venthamaraiyathi Chooranam
OECD - Organisation for Economic Co-operation and Development
PCV - Packed Cell Volume.
PGE - Prostaglandin E
RBC - Red Blood Corpuscles
SEM - Scanning electron microscope
TLC - Thin Layer Chromatography
HPTLC - High performance thin layer chromatography

S.No TITLE Page No.
1. INTRODUCTION 1
2. AIM & OBJECTIVES OF THE STUDY 3
3. REVIEW OF LITERATURE 4
3.1 ELETTARIA CARDAMOMUM, Linn 4
3.1.1. Gunapadam Aspect 4
3.1.2. Botanical Aspect 7
3.1.3. Lateral Research 11
3.2. ZINGIBER OFFICINALE 12
3.3. PIPER LONGUM 23
3.4. CUMINUM CYMINUM 29
3.5. GLYCYRRHIA GLABRA 36

3.6. ANETHUM SOWA 42
3.7. NELUMBO NUCIFERA 48
3.8. Pharmaceutical Review 56
3.9. Disease Review 58
3.9.1. Siddha Aspect 58
3.9.2. Modern Aspect 63
4. MATERIALS AND METHODS 71
4.1 Preparation of the drug 71
4.2. Standardization of the drug 75
4.2.1. Physico Chemical Analysis 75
4.2.2. Chemical Analysis 78
4.2.3. Instrumental Analysis 80
4.3. Toxicological study 89

4.3.1. Acute Toxicity Study 89
4.3.2. Sub Acute Toxicity Study 94
4.4 Pharmacological study 110
4.4.1 Thrambolytic ACtivity 110
4.4.2 Vasodilator Activity 115
4.4.3 Hypolipidemic Activity 118
4.4.4 Cardioprodective Activity 121
4.5 Micro biological Analysis 123
5. RESULT & DISCUSSION 124
6. SUMMARY 162
7. CONCLUSION 165
8. FUTURE SCOPE 166
9. BIBILIOGRAPHY 167
10. ANNEXURE 171

TABLE CONTENTS
Table. Page.
Title of the Table
No No.
1. Acute toxicity study (Animals were marked on Body) Numbering and Identification 91
2. Cage label and body marking on the animal Numbering and Identification 91
3. Animal dose level 92
4. Sub acute toxicity (Animals were marked on Body) Numbering and Identification 95
5. Cage label and body marking on the animal Numbering and Identification 96
6. Animal dose level 97
7. Estimation of Urea Reagents 107
8. Estimation of Uric acid Reagents 108
9. Thrombus induction dose level 110
10. Organoleptic characters 124

11. Physicochemical analysis results 125
12. Chemical Analysis Results 129
13. ICP OES Results 133
14. FTIR Analysis Results 137

15. Effect of test drug on body weight physical parameters result 142
16. Effect of Test drug on total clotting tail measurement 143
17. Effect of Test drug on Bleeding Time and Clotting Time 144
18. Effect on Test drug APTT, PT, Fibrinogen Concentration and ECLT in Rats 146
19. Effect on Test drug in Lipid Profile 151
20. Effect of Test drug on AST, ALT, ALP, CPK and LDH Enzyme activities in 156
doxorubicin Treated Rats
21. Antimicrobial activities results 159
FIGURE CONTENTS
Figure
Title of the Figure Page. No.
No.
1. Ingredients of VTC ( a & b) 73, 74
2. Scanning Electron Microscope 80
3. FT-IR Apparatus 83
4. ICP-OES Apparatus 87
5. TLC Sheet 128
6. SEM Result 131
7. Graph of FTIR Analysis 136
8. Total Clotting Tail Length 143
9. Bleeding Time and Clotting Time Average 145
10. Effect of APTT, PT, Fibrinogen Concentration, ECLT In Rats 147
11. Vasodilator Result Graph (a, b & c) 148, 149
12. Antimicrobial Activity Results (a, b & c) 159, 160

( Kindly make sure that the minutes of the meeting duly signed by all the participation are
maintained by the college office.)
1. INTRODUCTION
Siddha system is considered to be one of the ancient systems of medicine in the

world. Siddha is a complete holistic medical system that has been practiced in India for
more than thousands of years.
dml<hiI!npqbqz<!dbqviI!npqui<
“dml<
kqml<
kqml<hm!olb<Riel<!OsvUl<!lim<miI!
iI!
! dml<jh!uti<g<Gl<!dhibl<!nxqf<Ok!
k!
! dml<jh!uti<k<Oke<!dbqIuti<k<!OkOe”/
OkOe /!
! ! ! ! ! .!kqVlf<kqvl<!
So Siddhars devised healing techniques and drugs to alletiate diseases, live long
and have a healthy soul that is healthy uyir.
! Siddha means "Perfection" or "Eternal bliss". Siddhi refers to the eight
supernatural powers that are attainable by man. Those who attained these supernatural
power or perfection or siddhi were called “Siddhars". Siddhars realised that if the body is
made strong and perfect, they could get rid of death and diseases.
Siddha System of Medicine is one of the classical system of medicine consisting
of Herbal , Mineral and Animal origin. This system approaches the diseases both
mentally and physically.
Its origin strictly belongs to southern part of India. This system has its own
basic principle i.e mukkutram ( Vatha, Pitta and Khapa) and theories of Five Elements
(Aimpootham). “Health sustainment or decline is defined by the Normal or abnormal
state of the Humors” .
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! ! !!!nx<
!!!!nx<xK!Ohix<xq!B{qe<”
- kqVg<Gxt<!
The doshas within any person keep changing constantly due to lifestyle, foods and
environment.The loss of balance among the humors causes energy disharmony and
physical and mental disequilibrium which may appear at any time and become the cause
of diseases. According to the Siddhar ‘,Aagasthir’ the diseases are widely classified into
4448 types. Kuruthi Azhal Noi is one the disease in the above classification.
SCIENTIFIC VALIDATION OF VENTHAMARAIYATHI CHOORANAM 1

Symptoms of Kurthi Azhal Noi are Weight gain, loss of energy, vomiting,
cough, indigestion , burning sensation of the body, bleeding in vomitus and phlegm,
Heaviness of Head, dryness of body, hesitation of food , excess secretion of saliva,
irritation of throat and shortness of breath
“hqk<kf<!kiOe!!GVkqbqx<!Oxie<Xl<”
So the disequilibrium of pitha reflects as a vascular diseases. The symptoms
quoted under “ Kuruthi Azhal Noi” in Noinadal and Noi muthal Nadal Thiratu more or
less correlate with Hypertension .
Hypertension
Hypertension is a chronic condition of concern due to its role in the causation of
coronary heart disease, stroke and other vascular complications. It is the commonest
cardiovascular disorder, posing a major public health challenge to population in all socio-
economic and epidemiological transition.
It is very common disease with annul incidence of more than 10 million cases for
per year in India .
The symptoms of Hypertension are pulsating headache, insomnia, dizziness, lack
of concentration, loss of memory, shortness of breath and occasional palpitation.
Normal Blood pressure - 120/80 mmHg
Pre Hypertension - 140/90 mmHg
Hypertension stage I - 140-159/90-99 mmHg
Stage II - Above 160 /100mmHg
With the increasing of side effects and dependance of Modern drugs. It has
become imperative to find out an Appropriate Siddha treatment for pithathikam that is
way a preclinical study has been conducted to assess the effect of the drug
Venthamaraiyathi Chooranam in essentional Hypertension.

2. AIM AND OBJECTIVES
AIM:
The aim of this study is to validate the Thrombolytic,Vasodilator,
Hypolipidemic and Cardioprotective activity of Venthamaraiyathi Chooranam and
thus ensuring a holistic approach by controlling the blood pressure levels, significantly
decreasing the development and progression of complication of Kuruthi Azhal Noi (HT).
OBJECTIVES:
The main objective of the present study is to highlight the safety and efficacy of
Venthamaraiyathi Chooranam in the treatment of Kuruthi Azhal Noi, the following
methodology was adopted to evaluate the drug and its standardization studies.
Collection of literature evidence regarding the trial medicine.
Identification of the drugs in the Venthamaraiyathi Chooranam .
Preparation of the trial medicine as per the text.
Physico-chemical analysis of Test drug.
Evaluation of the toxicity of Test drug.
Evaluation of Thrombolytic activity of the Test drug by in vivo Clot lysis in
rat models.
Evaluation of Hypolipidemic activity of Test drug by models of wistar
albino rats.
Evaluation of vasodilator activity of Test drug Student organ bath method.

Evaluation of Cardioprotective activity of Test drug by against Doxorubicin
induced myocardial toxicity in albino rats.

3. REVIEW OF LITERATURES
3.1. GUNAPADAM ASPECT
4/2/2/!Wzvqsq
4/2/2/!Wzvqsq!
Wzvqsq!
) Elettaria cardamomum, Maton)!
!
Other Name :
Anji, Thudi, Korangam
Types:
Peria Elam,
Chirttialam,
Kattuyelak-kay,
Malayelam
Habitat :
Cultivated for its fruits in many parts of Western of Southern India,
Part Used:
Dried ripe seeds, oil from fruits
Taste - Spicy
Character - Veppam (Hot potency)
Class - Spicy
Actions:
Stimulant
Carminative
Stomachic
General Characters: (Pothu gunam)

gUt<!kiZG!kr<gtqz<!
‘oki{<jm!uib<gUt<
Okie<Xl<!Ofibkq!sivl<he<!Olgk<kiz<!!
d{<jm!Ohiz<!wPr<!gm<c!!gqvqs<svl<!
q!uq]
dpjz!uif<kq!sqzf<kq!uq <Svl<!!
]RSvl<

m!oug<jg!usqkig!Ofib<
h{<jm!oug<jg!usqkig!Ofib<!gisLl<!!
hiPR<
hiPR<!Osil<h<!hq{quqf<K!fm<mLl<!!
ogz<zil<!!
n{<jm!bQjtue<<!hqk<kl<!!-jug<ogz<
!!!
Nz!lir<glp<!!Wz!!lVf<kOk”!! !
)Okjvbv<!G{uigml<*!
It cures Throat diseases, oral diseases, cough , dysuria, expectoration, and animal
bites and Pitha diseases
Therapeutic Uses:
Cardamom seeds which have a fragrant taste and aromatic odour . They are
generally used as a masticatory (as a spice and a flavor agent ). Cardamoms are
employed to a small extent in Europe for flavoring sweet meats. They are valuable in
many stomach complaints. An oil extract from the fruits is used in both Pharmacy and
Perfumery.
Cardamom may be safely used us a carminative in convalescence after diarrhea.
In the form of tincture are powder , cardamoms are used , both in eastern and western
systems of medicine , as a frequent adjunct to other stimulants, bitter and purgatives .
A decoction of cardamom together with their pericarp and jiggery added is a
popular home remedy to relieve giddiness caused by biliousness. A compound powder
containing equal parts of cardamom seeds, ginger, clove and caraway is a good stomachic
in half drachms doses in a tonic dyspepsia . Powder made of equal parts of parched
cardamom seeds, aniseeds and caraway seeds given in one teaspoonful doses good
digestive.
Other preparation of Elarasi
1. Elathi Kirutham
Dosage : 1-2 teaspoon (bd)
Indications : Cough , cold, Peptic Ulcer, Indigestion
( Kannusamy , Parambarai Vaithiyam, 2012, 9th edition , p 237.)

2. Vallarai Kirutham
Dosage : 2-3 teaspoon
Duration : 40 days ( bd)
Indicatons : Scabies , Venereal disease, Primary complex.
3. Maha Elathi chooranam

Dosage : Thirikadi piramanam
Indications : Fever, Vomiting ,Headache, Bone Tuberculosis,
Chest burning
4. Thalisapathiri Chooranam
Indication : Tuberculosis, Body Burning, Peptic Ulcer, Jaundice .
5. Seethobalathi Chooranam
Indiacation : Venereal disease, Diarrhoea, Tuberculosis
6.Asuvaganthi Legium
Dosage : 5 -10 gm Bd
Indication : Health Tonic for Tuberculosis Patients.
Strengthening. Infertility and fertility.
7. Ingi Legium
Dosage : 5-10 gm Bd
Indication : Cure for Pitha diseases, Control for
Diarrhoea, Indigestion

3.1.2. BOTANICAL ASPECT
Elettaria cardamomum
Scientific classification
Kingdom : Plantae
(unranked) : Angiosperms
(unranked) : Monocots
(unranked) : Commelinids
Order : Zingiberales
Family : Zingiberaceae
Genus : Elettaria
Species : E. cardamomum
Binomial name : Elettaria cardamomum(L.) Maton
Common name : Cardamom, Malabar cardamom ,
Ceylon cardamom
VERNACULAR NAMES
Hindi : Elaichi
Marathi : Elachi
Tamil : Elam Ancha
Malayalam : Elatarri
Telugu : Elaki
Kannada : Elakki
Urdu : Elaichi
Sanskrit : Trutih
Description
Overview:
A herbaceous perennial growing up to 6 m tall. Leaf-shoots arising from a stout
rhizome. Growing in a thick clump of up to 20 leafy shoots.

Leaves:
Dark green, long and sword-shaped. The underside is paler and may have a
covering of tiny hairs.
Flowers:
Borne on a flowering stalk that can reach over 1 m in length. The pale green
flowers contain both male and female parts. One of the petals is white and streaked with
violet.
Fruits:
Pale green to yellow and elongate oval-shape. Each fruit has three chambers filled
with small aromatic seeds, each about 3 mm long. The fruits and seeds dry to a straw-
brown colour and are widely used as flavouring.
Uses
Food and drink
The dried fruits and seeds of cardamom are used to add a unique taste to rice,
meat (including hamburgers, pizza sausages and Swedish meatballs), vegetables and
other savoury dishes. Whole and ground cardamom seeds are added to flavor coffee, tea,
liqueurs, ice cream, confectionery and baked goods (including Danish pastries).
Cardamom is highly valued in Kashmir as an essential ingredient of the fragrant
drink kahwa and sweet Kashmiri black tea.
Cardamom essential oil is extracted from the seeds. It is mainly used in the
flavouring of processed foods and drinks such as cordials, bitters and liqueurs and
occasionally in perfumery.
Cardamom oleoresin has similar applications to the essential oil. It is mainly used
to flavour meat products with a short shelf life, such as sausages. Because the oil has
antibacterial activity it has been added to foods as a preservative at low levels. It is used
in low quantities so it does not affect the flavour of the food.

Traditional medicine
In Ayurvedic medicine, cardamom is used to treat disorders of the stomach and
urinary system, asthma, bronchitis and heart problems. When mixed with neem and
camphor, cardamom is used as a nasal preparation to treat colds. An infusion of
cardamom can be used as a gargle to relieve sore throats, which has led to its use in
cough sweets.
Cardamom seeds have been used in a range of preparations. Roasted seeds were
boiled with betel nuts (fruits of the palm Areca catechu) to make a drink that is used to
treat indigestion and nausea. They are also added to tea to make a tonic to relieve the
symptoms of stress due to overwork or depression. Cardamom seeds are given to
patients with bad breath and a capsule of cardamom taken with honey is reputed to
improve eyesight.
The traditional uses of cardamom to treat skin conditions have attracted the
attention of those developing plant-based cosmetics, especially as it has been used
traditionally to treat areas of the body that have red-pigmentation. It is often incorporated
into soaps and hand creams.
Blood Pressure
As a diuretic and fiber rich spice, cardamom significantly lowers blood pressure.
Blood Clots
Cardamom prevents dangerous blood clots by preventing platelet aggregation and
the sticking to the artery walls.
Antioxidant
Many of the vitamins, phytonutrients, and essential oils in cardamom act as
antioxidants, cleaning up free radicals and resisting cellular aging.
Pathogens
The volatile essential oils in cardamom inhibit the growth of viruses, bacteria,
fungus, and mold.

Anti-inflammatory
Like ginger and turmeric, its relatives, cardamom has some anti-inflammatory
properties that limit pain and swelling, especially in mucus membranes, the mouth, and
throat.
Hiccups
Cardamom is an anti-spasmodic that can help get rid of hiccups. This also applies
to other involuntary muscle spasms, like stomach and intestinal cramps

3.1.3. LATERAL RESEARCH
Blood pressure lowering, fibrinolysis enhancing and antioxidant activities of
cardamom (Elettaria cardamomum).
Abstract
Elettaria cardamomum (L.) Maton. (Small cardamom) fruit powder was evaluated
for its antihypertensive potential and its effect on some of the cardiovascular risk factors
in individuals with stage 1 hypertension. Twenty, newly diagnosed individuals with
primary hypertension of stage 1 were administered 3 g of cardamom powder in two
divided doses for 12 weeks. Blood pressure was recorded initially and at 4 weeks interval
for 3 months. Blood samples were also collected initially and at 4 weeks interval for
estimation of lipid profile, fibrinogen and fibrinolysis. Total antioxidant status, however,
was assessed initially and at the end of the study. Administration of 3 g cardamom
powder significantly (p<0.001) decreased systolic, diastolic and mean blood pressure and
significantly (p<0.05) increased fibrinolytic activity at the end of 12th week. Total
antioxidant status was also significantly (p<0.05) increased by 90% at the end of 3
months. However, fibrinogen and lipid levels were not significantly altered. All study
subjects experienced a feeling of well being without any side-effects. Thus, the present
study demonstrates that small cardamom effectively reduces blood pressure, enhances
fibrinolysis and improves antioxidant status, without significantly altering blood lipids
and fibrinogen levels in stage 1 hypertensive individuals.

4/3/2/!Sg<
4/3/2/!Sg<G!
) Zingiber officinale)!
!
Other Name :
Artharagam, Arukkan, Athagam, Upakullam, Ularntha Inji, Sudupathiram,
Chukku, Chundi, Chondi, Choupannam, Chouvarnam, Navasuru, Naakaram,
Manoushatham, Vichvapechajam, Vidamoodiya amirtham, Verkompu. Dried Zingier is
called Chukku
Habitat :
Ginger is cultivated in many part of India. On large scale in the warm, moist
regions, chiefly in Madras , Cochin and Thavancore, and to a some what less extent in
Bengal and Punjab.
Part Used:
Tuber ( Dry )
Taste - Spicy
Character - Veppam (Hot potency)
Class - Spicy
Actions
Stimulant
Carminative
Stomachic

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uizgh!!
&zl<!-jvh<hqVlz<!&g<GfQv<!uizgh! !
Okimlkq!sivf<
Okimlkq!sivf<!okimv<uik!Ge<lfQv<k!
<
Okiml<N!ll<!Ohig<GR<!Sg<<G/”!

It cures Indigestion, Chest Burning, Sour Belching, Oral diseases , Respiratory
diseases, Diarrhea, Ulcer, Sinusitis, Abdominal unfortable, Ear pain, Face disease, Head
ache , Anemia, Cold fever.
Therapeutic Uses:
Ginger is prepared from the dried rhizomes. Ginger being aromatic and
pleasantly pungent, is commonly used us a spice and the preparation of condiments
curries ginger bread , and conserve and syrup are made from the fresh younger rhizomes.
Rhizomes are also pickled.
Dried ginger is of two types , peeled and unpeeled , the latter being mearly the
cleaned rhizomes dried in the sun in the cases of dry specimen the out layer should be
scraped off . when the fresh drug is used for extracting in the juice , the supernatant fluid
alone should be used and the sediment discarded (Chunnam). “Ginger was at one time
much employed for spicing beer and the modern equivalent, ginger beer, is highly
esteemed today as a beneficial cordial in a cold weather” (chopra).
Dry ginger is much used as a carminative adjunct along with black pepper and
long pepper under the name of trikatu. Ginger is extremely valuable in dyspepsia,
flatulence, colic, vomiting, spasms and other painful affection of the stomach and the
bowels an attended by fever; for cold , cough, asthma, dyspepsia and the indigestion is
highly recommended a preparation called “allaepauk” or ginger Jam or conserve; it
consist of ginger juice, water and sugar in sufficient quantites .
Other Preparation of Chukku

1. Mullangi Legium
Dosage : 5- 10 gm Bd
Indication : Loss of appetite , vomiting, Abdominal Pain, Peptic Ulcer
pain cure for the best this medicine . (Milk must taken
after medicine)

2. Kandakathiri Legium
Dosage : 5 -10 gm bd
Indication : Peptic Ulcer, Indigestion, Constipation, vomiting,
Throat irritation etc.
3. Sangu Mathirai
Dosage : 1 bd Hot water
Indication : Constipation, fever, cough, Tabe warm ,
4. Mantha Mathirai
Dosage : 65 gm tablet od for 1 week
Indication : Respiratory disease, Fever and Rhinitis, Body Pain.
5. Jeevaracha amirtha Chooranam

Dosage : Thirikadi Piramanam
Indication : Indigestion
6. Vaivilangathi Chooranam
Indication : Gastritis , Hiccup , Vomiting, Asthma and Pitha disease
7. Thair Chunndi Chukku Chooranam

Indication : Indigestion, loss of appetite, Flatulence,
Abdominal uncomfortable

Zingiber Officinale
Classification
According to Bentham and Hooker’s Classification (1862-83) Zingiber Officinale
Rose. in Trans. Linn. SOC is classified as follows.
Kingdom - Vegetable Kindom
Division - Spermatophyta
Sub-Division - Angiospermae
Class - Monocotyle donae
Series - Epigynae
Family - Scitominae
Sub –Family - Zingiberaceae
Genas - Zingiber
Species - Officinale
Vernacular Names:
Tamil - Dried – Shukku, Chukku –
English - Ginger
Sanskrit - Dried –Sunta- Nagara , Nagaram,
Hindi - Adrak, ada
Ger - Ingwer
Kannada - (Dried) – Vona – Shunti
Malayalam - Chukka
Telugu - Dried – Sonti
Bengali and - Dried – Sonth
Panjabi - Fresh – Adrak, Ada, Adi
Gujarathi - Adu

Distribution:
Widely cultivated in tropical Asia, Ginger is cultivated in many parts of India, and
on large scale in the warm, moist regions, chiefly in madras, cochin and Travancore, and
to a some what less extent in Bengal and the Punjab.
Botanical Description
It is an underground rhizome.
Habit:
A herbaceous, rhizomatous perennial, reaching up to 90cm in height under
cultivation.
Root :
Adventitious
Leaves:
Narrow, distichous, Sub-sessile, linear – lanceolate, 17.0 cm x 1.8cm, dark green,
evenly narrowed to form a slender tip with stem –clasping sheaths.
Inflorescence:
Spikes terminating Elongate Peduncles, sheathed by scarious bracks. Spikes
Oblong-cylindric, 1.5-3 inch long 4-1 inch wide, glabrous. Pedeincle slender, Sheathing
scales glabrous, about inch long. Lip 3 – 10bed, mid lobe oblong – ovate, lateral short,
ovate, obtuse
Floral Characters:
Flowers : Zygromarphic, asymmetrical, bisexual, epigynous. Flowers in spikes ,
Scape radical or terminating the leafty stem. bracts persistent, usually 1- flowered.
Greenish with a small dark purple or Purplish black tip in radical spikes 3.8 – 7.5 cm long
and 2.5cm diameter . on peduncles 15-30 cm. long stamen dark puple, as long as the lip,
rather shorter than the corolla.

Perianth : 6 in two whorls (3+3) distinguishable in to calyx and corolla.
Calyx : Cylindric, Shortly 3- lobes
Corolla : Tubular, Unequally three lobed. Lobes calceolate,
the upper concave.
Androecium
Stamens 6, in two whorls, only median posterior stamen of the inner whorl is
functional and epipetalous, the other two stamens of the innter whole are united to form a
2-lipped staminodium (or) labellum, the anterior stamen of the outer whorl is absent and
the remaining two modified into petaloid staminodes (or) absent.
Gynoecium:
Corples 3; syncarpous, ovary inferior, Axile Placentae, Ovules many, style and
stigma 1, the slender style held in a groove in the fertile stamen.
Fruit : An oblong, tardily dehiscing capsule.

Seeds : Large, globose, arillate.
Part used : Rhizone
Rhizome : Thick and Fleshy
Propagation : By rhizomes
Adulteration
Adulteration of ginger by exhausted or not well prepared on limed drug can be
determined by the above standards.
Sometimes in exhausted ginger, capsicum (or) Paradise grains are added to

increased pungency, Paradise grains are the seeds of aframomum melgueta
(Zingiberaceae) pungency of capsicum and paradise grains is not destroyed by boiling
with alkali while it is destroyed in case of ginger. Adulteration of ginger by capsicum and
paradise grains can be determined by this way.

Phytochemical Aspects:
Active Constituents:
Ginger Contains 1 to 2 % volatile oil, camphene, phellandrene, zingiberene,
Zingiberol, eucalyptol, Citral, bomecol, Linalol, Methyl heptenone, Monyl – Alcohol
esters of acetic, caprylic acid, yellow oil gingerol, Phenols, resin, Shogaol
5 to 8.7% pungent principle, resinous mass and starch. Volatile Oil is responsible
for the aromatic smell and consists of Zingiberene 6% sesquiterpene hydrocarbon
Zingiberol a sesquiterpene alcohol and bisabolene. Gingerol is a yellow pungent oily
Liquid and yields gingerone a ketone and aliphatic aldehyde shogaol is formed by loss of
water from gingerol. Shogaol and gingerone are less pungent. The pungency of gingerol
and ginger is destroyed when boiled with 5% potassium hydroxide or other alkalies.
Chemical Compositions:
The composition of ginger varies according to the type and the agroclimatic
conditions under it is grown.
Vitamins
Thiamine - 0.06mg / 100gm
Riboflavin - 0.03 mg/100gm
Niacin - 0.60 mg/ 100gm
Vitamin-c - 6.0 mg/ 100gm
Carotene - 40 mg/ 100gm
Ginger contains small quantities of glucose fructose and sucrose, rattinose is

probably present in traces.
The principal carbohydrate of the rhizome is starch. Besides starch the rhizomes
are reported to contain Pent sans. (7.6% on dry basis in one sample). Ginger contains
1.60-2.44% Nitrogen on dry basis.
Extraction of the freeze – dried and powdered rhizome by the conventional
Protein – solvents showed that albumin globuline, prolamine and glutelin formed
respectively – 35.6, 16.9, 11.0 and 17.9% of the total proteins. 18.6% of the total proteins
remained un extracted.

The free amino acids present in ginger include glutamic acid, aspartic acid,
serine, glycine threonine, alanine, glutamine arginine,aminobutyric acid, valine, leucines,
proline, phenylalanine, asparagines, lysine, cystine, histidine and pipecolic acid.
(Connell Mclachlan. J.Chromat)
Essential Oil
The Characteristic Pleasant and aromatic odour of ginger is due to an essential oil
which can be separated from the rhizome by stream distillation. The oil is generally
obtained from unscripted ginger. The oil containing cells are particularly numerous in the
epidermal tissue and scraped ginger is therefore poor in oil content. The essential oil
derived from the dried ginger known in trades as oil of ginger.
Ginger
Ginger was known in china as early as 400 B.C. It was also used as a spice by the
Greeks and Romans who considered it an Arabian product because it was received from
India by the way of the Red Sea. It was introduced in to Jamaica and other islands of the
west indies by the Spaniards and ginger was exported from the west indies to spain in
considerable quantity even during the year 1547 A.D.
Ginger has been under cultivation in India from times immemorial the plant does
not occur in a truly wild state, and rarely flowers, though the cultivated plants on the west
coast of India are said to bear flowers quite frequently in October.
Cultivation
For cultivation the rhizome is cut into pieces and each piece containing a bud is
planted into trenches in well-drained and loamy soil in March or April Sandy loan rich in
humens and warm humid conditions are ideal for its cultivation.
The plant requires about 80 inches rainfall per year and if rainfall is in adequate
water may be supplied by irrigation. Collection is done in December or January when the
plants wither after flowering period. Rhizomes are carefully dug out, aerial stems, fibrous
roots and buds are removed. They are washed to remove mould and clay attached to
them. Rhizome is peeled on that surface as well as between the fingers and thoroughly
washed in running water. Drug is then dried completely by keeping in the sun on mats

which are covered over. nights and in rainy and cloudy seasons. If moisture is present,
drug may become moudly. After drying it loses about 70% of its weight 1500 kg
rhizomes are required per hectare Around 12.500 kg of rhizome per hectare is obtained.
Curing of Ginger
After the flowers have disappeared and the stems have withered. ginger is ripe for
collection. The rhizome are dug up and prepared for the market in different ways.
Ginger is marketed both in the peeled and unpeeled condition, the forms as
scraped ginger and later us unscraped ginger. In Scraped ginger the epidermal layer of
the fresh rhizome is scraped off with the help of a sharpened bamboo- Splinter and then
washed in water and dried in the sun for F-10 days as the essential oil is contained in the
epidermal cells, excessive or careless scraping results in the loss of this oil and depreciate
the quality of the spice. the quality of dry ginger, cured at Dodahoo, Himachal Pradesh,
in the bright sun has been found to be better than that cured in a closed over with artificial
heat. Iron – Knives are not used for scraping as they leave stains on the product.
In the middle – East countries which buy a very large part of the Indian produce
there is a higher demand for white, polished rhizomes free from specks or spots. For this
purpose the raw rhizomes are soaked in water for a day and later in thick milk of lime
(1kg. slaked time/ 12 liters of water) This material is dried in the sun and then rubbed
with gunny bag pieces to remove the last remnants of he skin. the treatment imparts a
smooth finish to the final product.
In certain localities the lime treated rhizomes are treated further with sulphur
dioxide fumes at the rate 3.2 kg sulphur per tonne of rhizomes for 12 hours in specially
constricted chambers to bleach the product.
In Jamaica, a small quantity of lime-Juice is sometimes added to the water used
for washing the pealed rhizomes to produce whiter ginger.
Gradual drying is said to give a better product and so the rhizomes are dried in the
morning and then air – dried in shade during the rest of the day this is continued for 8-12
days. The dry rhizomes are graded on the basis of colour and size before packing the
product for market or shipment.

Medicinal Uses in Siddha
Uses
Used to treat Indigestion hyperacidity, cough, Asthma, dyspepsia, Polyuria,
ascites, diarrhea, oedema, stomach disorders, anemia, disease of kapham, cardiac
diseases.
In painful affections of the bowels stomach, etc., infusion of dry ginger is given
with the addition of a tablespoonful or two of castor oil the dose of the infusion. In
chronic rheumatism, infusion of south (1 in 24) taken warm just before going to bed, the
body being covered with blankets so as to produce copious perspiration, is often attended
with the best results. The same treatment has also been found beneficial in cold or cat
arrhol attacks and during the cold stage of intermittent fever.

3.2.3. LATERAL RESEARCHES
Zingiber officinale Mitigates Brain Damage and Improves Memory Impairment in
Focal Cerebral Ischemic Rat
Abstract
Cerebral ischemia is known to produce brain damage and related behavioral
deficits including memory. Recently, accumulating lines of evidence showed that
dietary enrichment with nutritional antioxidants could reduce brain damage and
improve cognitive function. In this study, possible protective effect of Zingiber
officinale, a medicinal plant reputed for neuroprotective effect against oxidative
stress-related brain damage, on brain damage and memory deficit induced by focal
cerebral ischemia was elucidated. Male adult Wistar rats were administrated an
alcoholic extract of ginger rhizome orally 14 days before and 21 days after the
permanent occlusion of right middle cerebral artery (MCAO). Cognitive function
assessment was performed at 7, 14, and 21 days after MCAO using the Morris
water maze test. The brain infarct volume and density of neurons in hippocampus
were also determined. Furthermore, the level of malondialdehyde (MDA), superoxide
dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in cerebral
cortex, striatum, and hippocampus was also quantified at the end of experiment.
The results showed that cognitive function and neurons density in hippocampus of
rats receiving ginger rhizome extract were improved while the brain infarct volume
was decreased. The cognitive enhancing effect and neuroprotective effect occurred
partly via the antioxidant activity of the extract. In conclusion, our study
demonstrated the beneficial effect of ginger rhizome to protect against focal
cerebral ischemia.

4/4/2/!kqh<
4/4/2/!kqh<hqzq!
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) Piper longum)
!
Other Name :
Aargathi, Unsaram, Ulavainasi, Kaaman, Kudaari, Kolakam, Koli, Kolaiyarukki,
Saram, Saadi, Thulavi, Mahathi, Sunai, Chounndi, Thanduli, Kanam, Kalini, Paanam,
Pippili, Vaitheki, Ampu, Aathi Marunthu
Habitat :
This plant is indigenous to northeastern and southern India and Ceylon and
cultivated in Eastern Bengal.
Types:
Arichi Thippili
Aanai Thippili
Part Used:
Vegetable and Rice
Taste - Sweet
Character - Thatpam (Coolant )
Class - Sweet
Actions:
Stimulant
Carminative

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It cures Kapha disease and General Tonic.
Therapeutic Uses:
Old long piper is more efficacious in medicine than fresh article ( U.C.Dutt).
powdered long piper administrated with honey will relieve cough, cold, asthma ,
hoarseness and hiccup. For catarrh and hoarseness a mixture of long piper root, long
piper, black piper and ginger in equal part is a useful combination . A compound powder
consisting of the same ingredient equal parts and called chuturushana chooranam is
useful in colic and flatulence besides cough and coryza . it was tested and found
successful. Dose is 10 to 60grm twice a day. A compound powder consisting of long
piper , ginger , black piper , cinnamon and caraway in equal part is a good expectorant .
Cow milk 350 ml was boiled and then 8 gm of Thippili powder added , mixed well
and taken in night time will cure cough, gastritis, dyspnea, and Thiridhosa.
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Thippili powder taken with honey will cure Tinea.
Other preparation Thipili
1. Thippili Chooranam
Indication : Indigestion, cough , Asthma, Tuberculosis.
2.Dharunasura Kiyalam
Dosage : 60 ml
Indication : Fever and associated disease
3. Kasakulanthaga Mathirai
Dosage : 1 bd
Indication : Respiratory disease

4. Balasanjeevi Mathiri
Dosage : 1 (30mg ) tablet bd
Indication : Cough, Fever and associated disease
5. Thippili Legium
Dosage : 5-10 gm
Indication : Abdominal Pain, Tuberculosis, cough, Asthma , Wheezing

Piper Longum, Linn
Scientific Classification:
Kingdom : Plant
Division : Angiosperms
Order : Piperales
Family : Piperaceae
Genus : Piper
Species : Longum
Vernacular Names:
English : Long pepper
Telugu : Pippilu
Malayamlam : Hippili
Sanskrit : Pippali
Duk : Pipliyan
Persian : Daraife-fil
Bengali : Pipul
Gujarati : Lindi pippal
Oriya : Pippalimula
Kanada : Hippali, modi
Punjabi : Maghan
Urdu : Filfil Daraz
Habit:
It is a creeping aromatic herb, slender climber.
Stems:
Creeping jointed
Leaves:
Lower leaves are broadly ovate, deeply cordate with big lobes at base, upto 10 cm
long, upper leaves are dark green, oblong-ovate, cordate.

Flowers:
Flowers are in spikes, bracts or male spikes narrow and female spikes circular.
Inflorescence:
Inflorescence is spike
Fruits:
Fruits are small, ovoid, shining, blackish green in colour, 2.5 to 4 cm long, light
green when immature.
Chemical Constituents:
The fruit of long pepper contains piperine (4-5%) and piplartine alkaloid, starch,
resin, gumand fat.
The dried fruits on steam distillation yield 0.7% of essential oil with spicy odour.
Besides that it contains sesamin and piplasterol Major chemical constituents are piperine,
piperlongumine (piplartine), piperlonguminine and also methyl – 3,4,5 –
trimethoxycinnamate.
Others:
Sesamin, a lignan, Dihydrostigmasterol and two low melting unstable compounds,
one of which appeared to be isobutylamide of an unsaturated acid, n-isobutyl-deca-trans-
2-trans-4 dien eamide, essential oil consisting of n-hexadecane n-heptadecane, n-
octadecane etc.
Therapeutic actions:
Carminative, stimulant, stomachic, flatulence
The dried unripe fruits are useful in cold, cough, chronic bronchitis and Diarrhoe
It is also used in liniments for rheumatic pains and paralysis. Fruits and roots are
used in Ayurvedic and Unani systems of drugs.
It is also used to treat insomnia, epilepsy, obstruction of bile duct and gall
bladder, dysentery and leprosy
Used for the diseases of respiratory track, cough and bronchitis as counter-irritant
Analgesic when applied locally for muscular pains and inflammation general
tonic and Haematinic.

Piperine from the Fruits of Piper longum with Inhibitory Effect on Monoamine
Oxidase and Antidepressant-Like Activity
Abstract :
A bioassay-guided isolation of the ethanol extract from the fruits of Piper longum
yielded a known piperidine alkaloid, piperine, as a monoamine oxidase (MAO) inhibitor.
Piperine showed an inhibitory effect against MAO-A (IC50 value: 20.9 µM) and MAO-B
(IC50 value: 7.0 µM). Kinetic analyses by a Lineweaver–Burk plot clearly indicated that
piperine competitively inhibited MAO-A and MAO-B with Ki values of 19.0±0.9 µM and
3.19±0.5 µM, respectively. The inhibition by piperine was found to be reversible by
dialysis of the incubation mixture. In addition, the immobility times in the tail suspension
test were significantly reduced by piperine, similar to that of the reference antidepressant
fluoxetine, without accompanying changes in ambulation when assessed in an open-field.
These results suggest that piperine possesses potent antidepressant-like properties that are
mediated in part through the inhibition of MAO activity, and therefore represent a
promising pharmacotherapeutic candidate as an antidepressant agent.

4/5/2/!sQ
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Cuminum cyminum!
Other Name :
Asai, Seeri, Upakumpapeesam, Narseeri, Thutthasampalam, Pirathi-viga,
Pithanasini, Posanakudori, Methiyam
Habitat :
Extensively cultivated as a cold season crop on the plains and as a summer crop in
the hills in northern India Himalayas and the Punjab, Baluchisthan, Kashmir, Kumaon,
Garhwal etc. also imported from Persia and Asia Minor.
Part Used:
Seeds
Taste - Spicy and Sweet
Character - Thatpam (Coolant )
Class - Sweet
Actions:
Stimulant
Carminative
Stomachic
Astringent

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It cures the Abdominal pain , Oral diseases, Hepatic, Tuberculosis, Calculi,
Bleeding dysentery and asthma.

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Cuminum seeds separately taken directly all diseases will cure. Just add to
Palm sugar then taken will cure the cough disease.
!
Therapeutic Uses:
Cumin seeds are largely used as a condiment or space in curries, pickles, etc., i.e,
in cookery they are medically useful in hoarseness of voice, dyspepsia and chronic
diarrhea, the dose is from 10 to 30 grains.
Seeds are also cooling in effect and therefore form an ingredient of most
prescriptions for gonorrhea chronic diarrhea and dyspepsia, externally they are applied
in the form of poultice to allay pain and irritation of worms in the abdomen; seeds
reduced to powder, mix with honey salt and clarified butter are applied to scorpion bite.
Seeds mixed with lime juice are administered in cases of bilious nausea in pregnant
female and cumin seeds taken internally shortly after child birth increases secretion of
milk.
A quantity of seeds lightly smeared with ghee, put in to pipe and smoke relief the
hiccup. a confection called Jirakady modaka is prepared thus : - take of the three
myrobalans tubers of cyperus rotundus , watery extract of gulancha , prepared talc,
flowers of mesua ferrea , leaf called Thejapatra.
Other preparation of Seerakam

1. Keelanelli Karkam
Dosage : 10-30 gm
Indication : Jaundice ,Liver diseases
2. Mullangi Chooranam
Indication : Diarrhoea, peptic ulcer, Abdominal pain and gastritis, indigestion

3. Bhaskaralavana chooranam
Dosage : ¼ rubai wt
Indication : Pitha disease, indigestion, flatulence , loss of appetite
4.Sharasvatha chooranam
Dosage : Arai kal Rubai wt.
Indication : Increase memory power
5. Nannari Mathirai
Dosage : 1 (130 mg ) tablet bd
Indication : gastritis , burning micturation, insominia
6. Seeraga Legium
Dosage : 5-10 gm Piramanam
Indication : Indigestion,loss of appetite, pitha and associated diseases
7. Ingi Legium
Dosage : 5-10 gm Bd
Indication : Cure for Pitha diseases, Control for Diarrhoea,
Indigestion

3.4.2. BOTANICAL ASPECTS
Cuminum cyminum, Linn
Scientific classification:
Kingdom : Plant
Class : Dicotyledonae
Subclass : Polypetalae
Family : Umbelliferae
Genus : Cuminum
Species : Cyminum
Botanical Name: Cuminum cyminum, Linn
Vernacular Names:
Tamil : Seeragam German : Kreuzkummel
Bengali : Jira Burma : ziya
English : Cumin Arabic : Kamuna
Hindi : Zira Gujarathi : Jiru
Marathi : Jiraghi Syria : Kemun
Telugu : Jiraka
Urdu : Jirah
Identification of the Family:

Leaves have sheathing bases. Stems hollow. Inflorescence umbels. The
involucres of bracts stand out prominently, G (2). Ovary inferior, two chambered with
one pendulous anatropous ovule in each loculus. Fruit cremocarp and odoriferous.
(Genera about 270: species about 2700)
Identification of the plant:
The slender annular herb about 1 feet height with a much branched angular or
striated stems bearing 2 or 3 particle linear leaves twice or thrice 3 partite, ultimate
segments filiform. Umbels compound, rays few, bracts and bracteoles, several, linear,
rigid, calyx-teeth small, subulate, unequal. Petals oblong or obovate, emarginate, white
often unequal fruits cylindrical.

Description:
Habitat:
It is cultivated throughout the temperate, sub-tropical regions like in India, Persia
and Afghanistan.
Habit:
A small slender,erect and annual herb about with the much branched angular and
striated stem.
Leaves:
2 or 3 ,bipinnate,dissected,ultimate segments linear,filiform,sheaths white
margined.
Inflorescence:
In compound umbels.
Flowers:
Bisexual, regular, actinomorphic epigynous, white in terminal leaf opposed, few
rayed.
Calyx:
Calyx teeth small on the inferior ovary
Corolla:
5 petals at various sizes, free, yellow in color
Androecium:
5 stamens, free
Gynoecium:
Bicarpellary, syncarpous pistil, inferior ovary, 2 chambers, one ovule in each
chamber.

Fruit:
The fruits are greyish about ¼ inch long, tapering towards both base and apex and
compressed laterally with ridges covered by pupillose hairs. The hairs may be absent in
some forms. .
Chemical Constituents:
The chief constituent is cumaldenyde C10 H12 O (P-iso-propyl benzaldehycte)
which forms nearly 20 to 40% of the oil. Besides the aldehyde, the oil contains P-
cymene, Pinene, dipentene, cumene, cuminic alchohol, B-phellandrene and –terpenol.
Seeds analysis carbohydrates 36.3%, moisture 11.9, protein 18.7 fibre 12, calcium
1.08, phosphorous 0.49%, Iron 31mg/100gms, Vitamin A-870 I.u/100g and vitamin C
3mg/100g.
Therapeutic uses:
Cumin seeds are aromatic and spicy, extensively used as condiment. In digenous
medicine, cumin seeds hare long been considered stimulant and carminative, stomachic,
astringent and useful in diarrhoea and dyspepsia.

Effects of the fruit essential oil of Cuminum cyminum Linn. (Apiaceae) on
pentylenetetrazol-induced epileptiform activity in F1 neurones of Helix aspersa
Abstract
The effect of the fruit essential oil of Cuminum cyminum Linn. (Apiaceae) (syn.
Cuminum odorum Salisb) on the epileptiform activity induced by pentylenetetrazol (PTZ)
was evaluated, using intracellular technique. The results demonstrated that extracellular
application of the essential oil of Cuminum cyminum (1% and 3%) dramatically
decreased the frequency of spontaneous activity induced by PTZ in a time and
concentration dependent manner. In addition it showed protection against
pentylenetetrazol-induced epileptic activity by increasing the duration, decreasing the
amplitude of after hyperpolarization potential (AHP) following the action potential, the
peak of action potential, and inhibition of the firing rate. These membrane effects suggest
cellular mechanisms by which the essential oil of Cuminum cyminum can inhibit the
PTZ-induced epileptic activity.
Keywords
• Cuminum cyminum;
• Anticonvulsant effect;
• Pentylenetetrazol;
• Current clamp

3.5.GUNAPADAM ASPECT!
4/6/2/!nkqlKvl<
4/6/2/!nkqlKvl<!
(Glycyrrhiya glabra)
Other Name :
Athingam, Atti, Mathookam, Kunriver
Habitat :
Arabia, Persian gulf , Afghanistan, Turkestan, Asia minor , Siberia etc., but the
root is cultivated in the Punjab, Sub Himalayan tracts from the Chenab east wards,
Sinthu, Peshawar valley , Burma and Andaman Island
Part Used:
Peeled Root
Taste : Sweet
Character : Thatpam (Coolant )
Class : Sweet
Actions:
Emollient
Demulcent
Mild Expectorent
Laxative
Tonic

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ilijz!
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It cures diseases like hiccup, vitiligo, eye disease, bone, thristiness, Burning
Micturation, toxicity and etc.
Therapeutic Uses:
Roots also used in scorpion sting . root in infusion decoction , extract or lozenge
is useful as a demultent in inflammatory affection or irritable condition of the bronchial
tube bowels and catarrh of the Genito Urinary passages as cough hoarseness sore throat,
asthma, dysuria etc., also used as a tonic and as slight laxative.
It is much used as adjunct in pharmaceutical preparation as compound decoction

and tincture of aloes , compound mixture and confection of Senna etc., also used for
flavouring infusion , lozenges , oil and ghritas. Liquid extract is especially in the
disguising the bitter or acid taste of many nauseous drugs. Particularly senna (leaf),
aloes, Chloride of Ammonium, Senega, Hyocyamus, turpentime etc and sweeten tobacco.
Other preparation of Athimathuram

1. Thapa Sura Kiyalam
Dosage : 10 ml Children, 30ml adult
Indication : Thrist, vomiting
2. Drashathi chooranam
Indication : Urinary Track Infection, Burning Micturation,
3. Athimathura Chooranam
Indication : Vomiting, Nausea, giddiness, bitterness in mouth,
pitha disease.
4.Irumal Mathirai
Dosage : 1 (500mg ) tablet 4 hours once
Indication : Cough
5. Nellikai Legium
Dosage : 5- 10 gm
Indication : Renal disease, Immune power, general tonic, vomiting,
jaundice, Anemia, Palpitation. Giddiness

Glycyrrhiya glabra
Scietific Classification
Kingdom : Plantae
(unranked) : Eudicots
(unranked) : Rosids
Order : Fabales
Family : Fabaceae
Subfamily : Faboideae
Tribe : Galegeae
Genus : Glycyrrhiza
Species : G. glabra
Binomial name : Glycyrrhiza glabra
Vernacular Name
Common name : Licorice, Liquorice, Sweetwood
Hindi : Jethi-madh, kubas-susa, mithilakdi
Kannada : Atimadhura, jestamaddu
Malayalam : Atimadhuram, erattimadhuram
Marathi : Jashtimadh
Sanskrit : Jalayashti, klitaka, madhu, madhu-yashtikam
Tamil : Adimaduram
Telugu : Athimathuram
Urdu : Mulhatti, mulathi
Habitat:
Dry, open scrubland, damp ditches or near streams; often in soils with high
nitrogen content.

Description
It is a herbaceous perennial, growing to 1 m in height, with pinnate leaves about
7–15 cm (2.8–5.9 in) long, with 9–17 leaflets. The flowersare 0.8–1.2 cm (1⁄3–1⁄2 in)
long, purple to pale whitish blue, produced in a loose inflorescence. The fruit is an oblong
pod, 2–3 cm (3⁄4–1 1⁄6 in) long, containing several seeds. The roots are stoloniferous.
Chemistry
The scent of liquorice root comes from a complex and variable combination of
compounds, of which anethole is up to 3% of total volatiles. Much of the sweetness in
liquorice comes from glycyrrhizin, which has a sweet taste, 30–50 times the sweetness of
sugar. The sweetness is very different from sugar, being less instant, tart, and lasting
longer.
The isoflavene glabrene and the isoflavane glabridin, found in the roots of
liquorice, are phytoestrogens.
Cultivation and uses

Liquorice, which grows best in well-drained soils in deep valleys with full sun, is
harvested in the autumn two to three years after planting.Countries producing liquorice
include India, Iran, Afghanistan, the People’s Republic of China, Pakistan, Iraq,
Azerbaijan, Uzbekistan, Turkmenistan, and Turkey.
The world's leading manufacturer of liquorice products is M&F Worldwide,
which manufactures more than 70% of the worldwide liquorice flavours sold to end
users.
Medicine
Glycyrrhizin has also demonstrated antiviral, antimicrobial, anti-inflammatory,
hepatoprotective, and blood pressure-increasing effects in vitro and in vivo, as is
supported by the finding that intravenous glycyrrhizin (as if it is given orally very little of
the original drug makes it into circulation) slows the progression of viral and autoimmune
hepatitis. In one clinical trial liquorice demonstrated promising activity, when applied
topically, against atopic dermatitis. Additionally, liquorice may be effective in treating
hyperlipidaemia (a high amount of fats in the blood).] Liquorice has also demonstrated

efficacy in treating inflammation-induced skin hyperpigmentation. Liquorice may also be
useful in preventing neurodegenerative disorders and dental caries.
The antiulcer, laxative, antidiabetic, anti-inflammatory, immunomodulatory,
antitumour and expectorant properties of liquorice have been investigated.
The compound glycyrrhizin (or glycyrrhizic acid), found in liquorice, has been
proposed as being useful for liver protection in tuberculosis therapy, but evidence does
not support this use, which may in fact be harmful.
Known hazards:
It has been reported that excessive liquorice consumption can lead to cardiac
dysfunction and severe hypertension.

In vitro and in vivo neuroprotective effect and mechanisms of glabridin, a major
active isoflavan from Glycyrrhiza glabra (licorice)
Abstract
Stroke is a life-threatening disease characterized by rapidly developing clinical
signs of focal or global disturbance of cerebral function due to cerebral ischemia. A
number of flavonoids have been shown to attenuate the cerebral injuries in stroked
animal models. Glabridin, a major flavonoid of Glycyrrhiza glabra (licorice), possesses
multiple pharmacological activities. This study aimed to investigate whether glabridin
modulated the cerebral injuries induced by middle cerebral artery occlusion (MCAO) in
rats and staurosporine-induced damage in cultured rat cortical neurons and the possible
mechanisms involved. Our study showed that glabridin at 25mg/kg by intraperitoneal
injection, but not at 5mg/kg, significantly decreased the focal infarct volume, cerebral
histological damage and apoptosis in MCAO rats compared to sham-operated rats.
Glabridin significantly attenuated the level of brain malonyldialdehyde (MDA) in MCAO
rats, while it elevated the level of two endogenous antioxidants in the brain, i.e.
superoxide dismutase (SOD) and reduced glutathione (GSH). Co-treatment with glabridin
significantly inhibited the staurosporine-induced cytotoxicity and apoptosis of cultured
rat cortical neurons in a concentration-dependent manner. Consistently, glabridin
significantly reduced the DNA laddering caused by staurosporine in a concentration-
dependent manner. Glabridin also suppressed the elevated Bax protein and caspase–3
proenzyme and decreased bcl–2 induced by staurosporine in cultured rat cortical neurons,
facilitating cell survival. Glabridin also inhibited superoxide production in cultured
cortical neurons exposed to staurosporine. These findings indicated that glabridin had a
neuroprotective effect via modulation of multiple pathways associated with apoptosis.
Further studies are warranted to further investigate the biochemical mechanisms for the
protective effect of glabridin on neurons and the evidence for clinical use of licorice
Keywords
• Glabridin
• Stroke

3.6.GUNAPADAM ASPECT!
4/7/2/!skGh<
4/7/2/!skGh<jh!!
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) Anethum graveolens)
!
Other Name :
Shoyikkeerai vithai, Mathurikai
Habitat :
Cultivated in Indian Gardens for Culinary Purpose “ As the fruit of Indian Variety
is much more narrowly winged then the variety met with in Europe, Its is considered by
same to belong to a distinct specie anitham sowa or peudecanum sowa
Part Used:
Leaf, Flower and Seed
Taste : Sweet and Spicy
Character : Veppam (Hot potency )
Class : Spicy
Actions:
Stimulant
Carminative
Deobstruent
Diuretic
Emmenagogue
Stomachic
Antispasmodic

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It cures bleeding disorder, headache, Earpain , Rhinitis, and etc., Strengthens the
liver, lung and stomach
Therapeutic Uses:
Essential oil contain in the fruit and distilled water of the fruit are much used in
flatulence , hicc up and colic and abdomen pain in children and adult. It may combined
with Na2co3 or little of lime water in hiccup and flatulence.
It is used to diminish the griping of purgatives , and the tormina of dysentery .
and infusion of the bruised fruits or seeds ( 1 in 30 ) is also very useful. Seeds bruied and
boiled in water and mixed with the roots are applied externally in rheumatic and others
swelling of the joints. Seeds are used as a worm remedy especially in horses.
Leaves are moistened with little oil and warmed and applied to the boils and
abscesses to hasten suppuration .
Other preparation of Sathakuppai

1. Panjakavviya Legium
Dosage : 5-10 gm bd
Indication : Menorrhoghea, Gonococcal arthritis ,
Peptic ulcer, gastritis,
2. Masha thylam
Indication : Tremor in Head and hands
3. Panjakavviya kirutham
Dosage : 5 -10 ml bd
Indication : Menorrhoghea, Gonococcal arthritis , Peptic ulcer,
gastritis, Anemia, Jaundice, Ascities
4.Karpurathi Chooranam
5. Poosanikai Nei
6. Soothaga vayu pattai legiyam

!
3.6.2. BOTANICAL ASPECT!
Anethum graveolens
Scientific classification
Kingdom : Plantae
(unranked) : Eudicots
(unranked) : Asterids
Order : Apiales
Family : Apiaceae
Genus : Anethum L.
Species : A. graveolens
Binomial name : Anethum graveolens L.
Vernacular Names
Sans : misroya
Eng : Dill
Hind : sowa
Punj : soya
Maha : shepu
Guj : surva-nu-bi
Telu : shataupivittulu
Tamil : satakuppi
Mal : chatukuppa
Can : sabbasige
Singh : sadakuppa
Malay : adaspudus
BOTANICAL DESCRIPTION
Anethum graveolens L. is the sole species of the genus Anethum, though
classified by some botanists in the related genus Peucedanum as Peucedanum graveolens
(L.) A variant called east Indian dill or Sowa(Anethum graveoeloens var sowa Roxb. ex,
Flem.) occurs in India and is cultivated for its foliage as a cold weather crop throughout
the Indian sub-continent, Malaysian archipelago and Japan.

Plant description
Anethum grows up to 90 cm tall, with slender stems and alternate leaves finally
divided three or four times into pinnate sections slightly broader than similar leaves of
fennel. The yellow flower develops into umbels.The seeds are not true seeds. They are
the halves of very small, dry fruits called schizocarps. Dill fruits are oval, compressed,
winged about one-tenth inch wide, with three longitudinal ridges on the back and three
dark lines or oil cells (vittae) between them and two on the flat surface. The taste of the
fruits somewhat resembles caraway. The seeds are smaller, flatter and lighter than
caraway and have a pleasant aromatic odor.
Cultivation
Dill prefers rich well-drained, loose soil and full sun. It tolerates a pH in the range
5.3 to 7.8. It requires warm to hot summers with huge sunshine levels; even partial shade
will reduce the yield substantially. The plant quickly runs into seeds in dry weather. It
often self sows when growing in a suitable position. Propagation is through seeds. Seeds
are viable for 3–10 years. The seed is harvested by cutting the flower heads off the stalks
when the seed is beginning to ripe.
Chemical content:
Volatile oil, dillanoside, xanthone glucoside, coumarins, scopoletin,
esculetin, bergapten, umbelliferone, umbelliprenine, daempferol, 3-gluucuronide,
vicenin 6, 8-di-C-glucosyl-5.7.3′-trihydroxyflavone, flavonoids, petroselinic acid
triglyceride, B-sitosterol glucoside, phenolic acids,caffeic, ferulic, and chlorogenic,
carvone, d-limonene, ax-phellandrene, dihydrocarvone, eugenol, B-phellandrene, ax-
pinene, anethole, dillapiole, myristicin, carveol, B-caryophellene, ax-phellandrene,
limonene, terpinene, ax-pinene, dillapiole, myristicin, coumarans.
Medicinal uses
Anethum is used as an ingredient in gripe water, given to relieve colic pain in
babies and flatulence in young children. The seed is aromatic, carminative, mildly
diuretic, galactogogue, stimulant and stomachic. The essential oil in the seed relieves
intestinal spasms and griping, helping to settle colic. The carminative volatile oil

improves appetite, relieves gas and aids digestion. Chewing the seeds improves bad
breath. Anethum stimulates milk flow in lactating mothers, and is often given to cattles
for this reason. It also cures urinary complaints, piles and mental disorders.
Sathakuppai use for % Daily Value*
Total Fat 15 g 23%
Saturated fat 0.7 g 3%
Polyunsaturated fat 1 g
Monounsaturated fat 9 g
Cholesterol 0 mg 0%
Sodium 20 mg 0%
Potassium 1,186 mg 33%
Total Carbohydrate 55 g 18%
Dietary fiber 21 g 84%
Protein 16 g 32%
Vitamin A 1% Vitamin C 35%
Calcium 151% Iron 90%
Vitamin D 0% Vitamin B-6 15%
Vitamin B-12 0% Magnesium 64%
*Per cent Daily Values are based on a 2,000 calorie diet. Your daily
values may be higher or lower depending on your calorie needs.

Evaluation of Safety and Protective Effect of Combined Extract of Cissampelos
pareira and Anethum graveolens (PM52) against Age-Related Cognitive
Impairment
Abstract
The present study aimed to determine acute toxicity, the protective effect, and
underlying mechanism of PM52, a combined extract of Cissampelos pareira and
Anethum graveolens, against age-related cognitive impairment in animal model of age-
related cognitive impairment. PM52 was determined as acute toxicity according to
OECD guideline. Male Wistar rats, weighing 180–220 g, were orally given PM52 at
doses of 2, 10, and 50 mg/kg at a period of 14 days before and 7 days after the
bilateral administration of AF64A via intracerebroventricular route. All animals were
assessed according to spatial memory, neuron density, MDA level, the activities of
SOD, CAT, GSH-Px, and AChEI effect in hippocampus. It was found that all doses of
PM52 could attenuate memory impairment and neurodegeneration in hippocampus.
The possible mechanisms might occur via the suppression of AChE and the decreased
oxidative stress in hippocampus. Therefore, our data suggest that PM52 may serve as
food supplement to protect against age-related cognitive impairment such as mild
cognitive impairment (MCI) and early phase of Alzheimer’s disease. However, further
researches are still essential.

4/8/2/!kiljv
4/8/2/!kiljv!
kiljv!
) Nelumbo nucifera )
!
Other Name :
Aravintham, Ellimanai, Sooriynatpu, Ponmanai, Vintham, Pundareegam,
Pathumam, Kamalam, Nalinam, Mulari, Mundagam, Malunthi, Sarogam, Koganagam,
Indai, Kanjam, Appusam, Amporagam, Chalasam, Vanasam, Varisam, Saraseerugam,
Pankerugam, Sarorugam, Pankasam
Habitat :
This large aquatic herbs with its elegant sweet scented flowers is generally met
with in tanks and ponds throughout India.
Part Used:
Flower, Seed, Tuber
Taste : Sweet and Astringent
Character : Thatpam (Coolant )
Class : Sweet
Actions:
Cooling
Astringent
Expectorant
Sedative
Demulcent
Tonic
Nutrient

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Therapeutic Uses:
Flower and filaments and juice of flower stalk are useful in diarrhea , cholera
and in liver complaints and also in fevers; it is recommended also as cardiac tonic .
compound decoction it is useful in bilious fevers . “the root, flower, stalk and leaves in
the form of infusion are used in fever as a refrigerant and diuretic” ( Chopra). Honey
formed in the flowers by the bees feeding upon the padma is called padma mathu or
maharantha. This is very useful in eye disease. Syrup of flower is used in cough to
check of Hemorrhoid from bleeding piles and Menorhogia and dysentery . Tubes of
white lotus boiled in gingili oil are rubbed on the head to cool head and eyes.
Express juice is also employed instead of speice of tuber. Root is mucilaginous and
even in piles.
Seeds are used as an application in leprosy other skin affection. Lotus flower
and fresh leaves ground with sandal wood and or embolic myrobalans also form a
cooling application to the forehead in the cephalalgia , to the skin in erysipelas and
cure other external inflammation.
Other Preparation of Venthamarai

1. Venthamarai Kudineer
Dosage : 15- 30 ml
Indication : Cardiac Tonic
2. Neithal Nei
3. Soodamani Mathirai
4. Aruganver Kizhangu

3.7.2. BOTANICAL ASPECTS
Nelumbo nucifera
Scientific classification:
Botanical Name(s) : Nymphaea Lotus, Nymphaea Nouchali
Family Name : Nymphaeaceae
Kingdom : Plantae
Division : Magnoliophyta
Class : Magnoliopsida
Order : Nymphaeales
Family : Nymphaeaceae
Genus : Nymphaea
Species : N. lotus
Vernacular Names:
English : Sacred lotus
Hindi : Kanwal, Kamal
Sanskrit : Ambuja
Tamil : Ambal, Thamarai, Padma, Pankaja, Kamala;
Bengal : Padma
Gujarat : Suriyakamal
Malayalam : Tamara
French : Nelumbo
German : Indische lotosblume
Persian : Nilufer.
Habitat:
Throughout warmer parts of India, in tanks, ponds and ditches
Description:
A species of water lily, white lotus is a perennial plant growing to a height of 45

cm. Also known as tiger lotus, it grows in clear, warm, still and slightly acidic waters.
The lily pads can be seen floating on water, while the blossoms rise above the water. The

flowers are white in color sometimes, with a pink tinge. The leaves vary from green to
red-brown, with a number of purple spots. Tiger lotus is native to the Nile and is grown
in various parts of East Africa and Southeast Asia. It is often used as an aquarium plant.
Plant Chemicals
Apomorphine, nuciferine, phytosterols, bioflavonoids, phosphoiesterase, glucose,
fructose, sucrose, mannitol, galacturonic acid, raffinose, amoni acids.
Uses & Benefits of White Lotus

White lotus was used in ancient Egypt, as a key to good health, sex and re-
birth.
The plant is an aphrodisiac for both men and women and a general remedy for
all kind of illnesses.
Continued use of tiger lotus enhances sexual vigor and general good health.
It is a tonic richer than ginseng, pain reliever richer than arnica, circulation
stimulant richer than ginkgo biloba and sexual stimulant richer than Viagra.
The flowers of white lotus are used for preparing tea that creates a warm,
euphoric glow.
The dried flowers are smoked by themselves or mixed with other herbs to add
flavor to smoking mixtures.
The effects of tiger lotus are enhanced when soaked in wine or other alcohol.
The plant is effectively used to increase memory and create a feeling of
euphoria and ecstasy, without the use of narcotics.
Its rhizomes are cooling, sweet, bitter and tonic and used in diarrhea,
dysentery, dyspepsia and general debility.
White lotus is used internally in treatment of gastrointestinal disorders and
jaundice.
The leaves are used in cutaneous, subcutaneous parasitic infection, eye
treatments and pregnancy.
The seeds are used in sauces, condiments, spices and flavorings.
(ref: Read more at http://www.iloveindia.com/indian-
herbs/nymphaea-lotus.html#BWgzZFJgGWzLfS2k.99)

Chemical constituents
Flower
Several flavonoids have been identified in the stamens of N.nucifera . These
include kaempferol and seven of its glycosides: kaempferol 3-O- β -D-galactopyranoside,
kaempferol 3-O-β-D-glucopyranoside , kaempferol 7-O-β -D- glucopyranoside ,
kaempferol 3-O-a-L-rhamnopyranosyl-( 1-6)-β -D-glucopyranoside , kaempferol 3-O-a-
L-rhamnopyranosyl-(1-2)- β -D-glucopyranoside, kaempferol 3-Oa-L-rhamnopyranosyl -
(1-2)- β- D-glucurono-pyranoside , kaempferol-3- O- β- D-glucurono-pyranoside ,
kaempferol 3-O-β-D-glucuronopyranosyl methylester , myricetin 3 0 ,5 0 -dimethylether
3-O- β -D-glucopyranoside , quercetin 3-O- β -D-glucopyranoside , nelumboroside A
and nelumboroside B.
It also contains two isorhamnetin glycosides: isorhamnetin 3-O-β-D-
glucopyranoside and isorhamnetin 3-O-a-L-rhamnopyranosyl- (1→6) -β -D-gluco-
pyranoside . Some non-flavonoid compounds, including adenine, myo-inositol, arbutin
and β-sitosterol glucopyranoside , have also been identified in stamen extract
PHARMACOLOGICAL ACTIVITIES
Flower
I. Hypoglycaemic activity
Sun-dried flower powder of N.nucifera, as well as the aqueous and alcoholic
extract of the flower, produced significant hypoglycaemia in fasting normal albino
rabbits. There was no significant difference in the activities of 1000 mg/kg of the test
drug (sun-dried powder of the flower) and equivalent amounts of the extracts. Glucose
tolerance studies with normal rabbits showed that oral doses of both extracts, equivalent
to 1000 mg/kg of the test drug, produced significant depression of the peak rise in fasting
blood sugar after glucose load; the effects of both extracts were 50% to that produced by
250 mg/kg of tolbutamide. A study of glucose tolerance curves shows that the duration of
hyperglycaemia was also notably reduced compared with the controls. In normal rabbits,
the extract at a dose of 1000 mg/kg significantly lowered hyperglycaemia induced by
subcutaneous injection of 0.5 mg/kg adrenaline hydrochloride.

II. Antioxidant activity
The potential of N. nucifera stamens to scavenge 1,1-diphenyl-2-picrylhydrazyl
(DPPH) free radicals and peroxynitrites (ONOO–), and the inhibition of total ROS
generation by kidney homogenates using 2v,7’-dichlorodihydrofluorescein diacetate
(DCHF-DA) was examined.The methanol extract showed strong antioxidant activity in
the ONOO– system and marginal activity in the DPPH and total ROS systems. Similarly,
seven known flavonoids were isolated from lotus stamens, most of which also showed
potent antioxidant activity. The glycosides nelumboroside A, nelumboroside B,
isorhamnetin glycoside and isorhamnetin rutinoside isolated from N. nucifera stamens
showed potent antioxidant activity in DPPH and ONOO– assays.[45] ‘Yunyupju’, a
liquor made from the blossoms and leaves of lotus, has been reported to have antioxidant
activity. The maximum scavenging activity on hydroxyl radicals (40%) could be
achieved when lotus liquor was more than 500 µg. Lotus liquor also has a potent
superoxide radical scavenging activity with value of 0.93 unit mg−1 as superoxide
dismutase equivalents with an IC50 value of 1.07 ± 0.04 mg.
III. Antipyretic activity

The ethanol extract of stalks of N. nucifera was evaluated for its antipyretic
potential on normal body temperature and yeast induced pyrexia in rats. In the model of
yeast provoked elevation of body temperature, the stalk extract showed significant
activity in both models at oral doses of 200 and 400 mg/kg. The stalk extracts showed
dose-dependent lowering of body temperature up to 4 h; the results were comparable to
those with paracetamol.
IV. Aldose reductase inhibitory activity

Two glycosides, namely kaempferol 3-O-a-L-rhamnopyranosyl-(1,6)-_-D-
glucopyranoside and isorhamnetin 3-O-a-Lrhamnopyranosyl-( 1!6)-_-D-glucopyranoside,
isolated from the methanol extract of stamens of N. nucifera exhibited a high degree of
inhibitory activity against rat lens aldose reductase in vitro, with IC50 values of 5.6 and
9.0 mM, respectively.

V. Antibacterial Activity
The hydroethanolic extract of flowers of N.nucifera Gaertn in vitro was reported
to possess antibacterial activity. The antibacterial activity was screened against different
bacterial strains like Escherichia coli Klebsiella pneumonia Pseudomonas aeruginosa
Bacillus Subtilis Staphylococcus aureus by detecting zone of inhibition and minimum
inhibitory concentration (MIC). The maximum zone of inhibition was exhibited by
N.nucifera flowers against Escherichia coli (14mm), Bacillus Subtilis (13mm) and
Staphylococcus aureus (11mm). The moderate zone of inhibition was found against
Klebsiella pneumonia (10mm) and Pseudomonas aeruginosa (8mm). Gram-negative
bacteria were more susceptible to the N.nucifera flower extracts than gram-positive
bacteria which contradict the previous reports that plant extracts are more active against
gram-positive bacteria than gram-negative bacteria. These results were compared with
the standard antibiotic chloramphenicol (30µg/ml)
VI. Antiplatellet activity

The antiplatelet activity of hydroethanolic extract of N.Nucifera flowers using
platelet-rich plasma in different concentrations (100 500µg/ ml) was reported. N.nucifera
flower extracts showed dose-dependent effective antiplatelet activity with maximum
activity at 500µg/ml concentration; prevention of platelet aggregation was 50% of that
achieved with standard aspirin.
( ref: Nishkrati R . Mehta , Nelumbo nucifera (Lotus) A review on Ethanobotany

Phytochemistry and Pharmacology ( IJPBR) 2013,1(4):152-167)

Effects of Nelumbo nucifera Rhizome Extract on Cell Proliferation and Neuroblast
Differentiation in the Hippocampal Dentate Gyrus in a Scopolamine-induced
Amnesia Animal Model
Abstract
A large aquatic herb, Nelumbo nucifera Gaertn, has psychopharmacological
effects similar to minor tranquillizers and antistress agents. This study examined the
effects of Nelumbo nucifera rhizome extracts (NRE) on cell proliferation and neuroblast
differentiation in the hippocampal dentate gyrus (DG) of a rat model of scopolamine-
induced amnesia. Immunohistochemical markers included Ki67, an endogenous marker
for active cell cycle, and doublecortin (DCX), a marker for immature neurons and
migratory neuroblasts. Scopolamine was administered for 28 days via an ALzet
minipump (44 mg/mL delivered at 2.5 µL/h). NRE was administered by gavage, 1 g/kg
per day for 28 days. The administration of scopolamine significantly reduced the number
of Ki67- and DCX-immunoreactive cells in the DG, whereas scopolamine did not induce
any significant changes in mature neurons in the DG. The administration of NRE
significantly ameliorated the scopolamine-induced reduction of Ki67- and DCX-
immunoreactive cells in the DG. In addition, the administration of NRE significantly
restored the scopolamine-induced reduction of brain-derived neurotrophic factor in DG
homogenates. These results suggest that NRE can ameliorate the scopolamine-induced
reductions of cell proliferation, neuroblast differentiation and BDNF levels. Copyright ©
2010 John Wiley & Sons, Ltd.

3.8. PHARMACEUTICAL REVIEW
CHOORANAM
Definition:
Chooranams are fine powders of drugs. The term Chooranam may be applied to
the powder of a single drug or a mixture of two or more drugs, which are powdered
separately prior to their being mixed to homogeneity.
Equipment Required:
1. A mortar and pestle
2. A fine sieve or fine cloth of close mesh (sieve no:100 mesh)
Note:
In large scale manufacture, in factories comminutors, pulverisers and ball mills
are employed for powdering. Sieving is performed by mechanical sifters which handle
large quantities of drugs in a short time.
Process of preparation:
The drugs enumerated in the recipe are clean and in well dried state.
The drugs which are to be used in the preparation should be taken from recently
collected material.
However, drugs like embelia fruits, senna, long pepper, jiggery and cow’s ghee
are prepared from fairy aged stock, provided they are not infested with pests, deteriorated
or spoiled or developed rancidity.
In general, the aromatic drugs are slightly fried, in order to enhance their aroma
and milling properties. Any extraneous material, organic or inorganic should be removed
from the drugs by close inspection.
The chooranam should be so find as to be called amorphous and should be never
damp. The fineness of the sieve should be 100 mesh or still finer.
Storage:
The prepared dry powder should be allowed to cool by spreading and mixing
prior to packing. They should be stored in rightly stoppered glass, polythene or tin
containers, or in polythene or cellophane bags and sealed. These bags should in turn be
enclosed in card board boxes.

The powder (Chooranam) is said to retain its potency for two months and then
gradually deteriorate. However if properly packed, and stored they keep good for a year.
The chooranam to facilitate easy handling and to assure exact dosage of administration
could be pressed into tablets with the addiction of a suitable binder. These tablets could
be packed in bottles or tubes made either of glass or plastics or packed in strips of metal
foil or plastic sheets. In industry, the tablets are made, counted and packed by electronic
devices.

3.9. DISEASE VIEW
3.9.1. SIDDHA ASPECT
KURUTHI AZHAL NOI ( HYPERTENSION)
Ratha Pitham
As the name suggests the disease is due to excessive production of blood and
heat in the body. In this disease, due to internal and external variations the heat is
aggravated; this causes blood to become impure; this impure blood does not flow in its
usually pathway but oozes out from the eye, nose, mouth , anus, vagina, urinary tract and
skin and makes it as dangerous disease.
Usually, doshas stinulate the diseases. However , while naming this disease the
dosha was not named first but blood has been named first. This is because it is only the
blood which ecposed the mailfunctioned heat and hence blood was named first and the
dosha ( Heat) was named next.
Aetiopathogenesis
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The following factors may be responsible for producing the disease; Excessive
wandering in the sun rays , eating of food item in excess which generate heat, such as
salt, bitter and sour substances, excessive indulgence insex; when yoga for controlling
five senses and for protecting the body is not properly practiced; excessive heat dosha
may dilute or concentrate the blood due to stimulation of vayu and may produce the
disease. Further , due to bad effect of heat, belly, left and right portion of the liver,
lungs, urinary bladder, large intestine, small intestine and heart will be damaged; the

impure blood come out from these organs and may produce the disease.
Types of disease:
The disease may be classified into eight types as follows:
1. Vali kuruthiazhal Noi ( Vatha blood heat disease)
2. Thee kuruthiazhal Noi ( Pitha blood heat disease)
3. Iya kuruthiazhal Noi ( Kapha blood heat disease)
4. Vali thee kuruthiazhal Noi (Vathapitha blood heat disease)
5. Vali Iya kuruthiazhal Noi (Vathakapha blood heat disease)
6. Iyavali kuruthiazhal Noi (Kaphavatha blood heat disease)
7. Iyaazhal kuruthiazhal Noi (Kaphapitha blood heat disease)
8. Mukkutra kuruthiazhal Noi (Tri-dosha blood heat disease)
9. Some other group of ancient physicians have classified this disease into
four types depending upon the direction of flow of blood as follows:
10. 1.Maelnoakku Kuruthi azhal Noi ( Upward directional blood flow)
11. Keelnoakku kuruthiazhal Noi ( Downward directional blood flow)
12. Irunoakku kuruthiazhal Noi ( Bi directional blood flow)
13. Allavilla noakku kuruthiazhal Noi (Unlimited directional blood flow)
14. It is considered that the above mentioned four types of classification was
made to identity the vayu which is responsible for the blood direction and
for controlling the same.
Prodromal sysmptom:
Before the disease is clinically apparent the following signs and symptoms may
occur. Heaviness of head, anorexia, severe cough, constriction of the skin due to body
dryness , excessive salivation , non vegetarian smell from the mouth or ointment smell,
severe nausea with vomiting, blood or bile strained vomiting, liking for sour substance
and development of vomiting after eating them, burning sensation of throat and ulcers
of the mouth. Later the following signs and symptoms may appear; Hoarseness of
voice, pinching cough, mild wheezing , ageusia, emaciation; eyes cheek and throat may
become yellow, red or green in colour . Finally the patient may lie down on the mat.

1. Vali kuruthiazhal Noi
In the disease the clinical features are similar to that of blood heat disease. In
addition the following effects of vatha dosha may also appear; Body pain; the blood may
attain black colour and and ooze out in diluted form, sometimes the blood may appear
with froath; the stool will be hardened and passed like the stool of goat; painful
defecation with blood in the stool.
2. Thee kuruthiazhal Noi

In this disease, the pitha dosha gets aggravated and hence the fire produces
various calamities. There will be excessive discharge of blood . As a consequence of
excessive activity of pitha the blood will be discoloured , become diluted and the colour
will appear like water in which dried leaves is soaked. Sometimes , bile will also be,
found mixed with the blood. Due to these the body will be pale and appear yellow in
colour. Eyes, tongue and skin etc. will appear green. Sometimes the blood may also
appear bright red in colour. Because of mixing of bile with the blood , the body becomes
greenish in colour and the patient will appear like jaundiced.
3. Iya kuruthiazhal Noi

In this disease the heart disease is formed because of Iyam . The blood which is
discharged will appear slightly pale mixed with excessive mucus . It will smell like meat
and becomes hardened . In addition , there will be uncontrolled cough, slight fever and
running nose.
4. Vali thee kuruthiazhal Noi

The signs and symptoms of this disease are similar to that of vali kuruthi azhal
noi mentioned above. In addition certain signs and symptoms caused by thee dosha may
also be formed.
5. Vali Iya kuruthiazhal Noi

The signs and symptoms of this disease are similar to that of vali kuruth azhal
disease. In addition , blood will be discharged with some or many of the clinical features
of Iya kuruthi azhal disease.

6. Iyavali kuruthiazhal Noi
In this disease, the signs and symptoms will be similar to that of excessive kapha
and kuruthi azhal disease. In addition, signs and symptoms of the vatha dosha may also
appear.
7. Iyaazhal kuruthiazhal Noi

In this disease, the signs and symptoms will be similar to that of kuruthiazhal and
excessive kapha diseases. In additional , some of the signs and symptoms of the zhal
dosha may be also found.
8. Mukkutra kuruthiazhal Noi

In this disease, all the three doshas become heperactive at the same time causing
serious harm to the body. The signs and symptoms caused because of these, will appear
similar to that of sanni disease. In addition, all the signs and symptoms of kuruthiazhal
will be also found. This is not an easily curable disease.
General signs and symptoms of the disease:

Because of the activity of the tridosha, there will be cough, vomiting and
wheezing. The cough and vomiting will be associated with excessive blood. Sometimes
bleeding may be found from the nose, eye, ear, mouth, urinary tract and skin. The body
strength will be reduced day by day. The skin may also shrink; it may become dry and
may bleed. In view of the blood loss, the body will become pale, breathlessness,
tiredness, polydipsia and hiccup may also appear. In additional , there will be swelling
of face , upper and lower limbs.
1.Maelnoakku Kuruthi azhal Noi ( Upward directional blood heat disease)

In this diseases, due to excessive heat, udhaana vavy is stimulated. This adversely
affects the blood resulting in vomiting , cough, sneezing, hiccup and flatulence. There
will be vigorour cough followed by vomiting. The vomiting may be stained with fresh
frothy blood. If the udhaana vayu also affects abdomen, in association with kapha
present there,bit makes the blood dry and causes pain in the abdomen; there may be

vomiting with drak coloured blood clots. If the disease attacks bronchi, left and right
portion of the liver, there will be pain in these sites and oozing of blood; as a results there
will be fatigue ,stickly sweating dryness of eye and paleness of body.
2. Keelnoakku kuruthiazhal Noi ( Downward directional blood heat disease)

In this disease, abaaba vayu becomes hyperactive due to excessive heat. As a
result the large intestine, urinary bladder, and ureter which are found below the level of
umbilicus are affected. The samaana vayu also becomes hyperactive and produces
inflammation of small intestine and kidney. In view of these , there will be excessive
diarrhea, frequent urination and heamorrhage. There will be burning pain micturation
and itching in the anus with burning pain .
3. Irunoakku kuruthiazhal Noi ( Bi directional blood heat disease)

In the disease, the abaana as well as udhaana vayu are stimulated at the same
times by the pitha . Hence, the clinical features of both upward and downward
directional blood heat diseases will be found. Due to excessive activity of the downward
directional blood heat disease , there will be excessive diarrhea, excessive urination with
heamorrhage. Due to excessive activity of upward directional blood heat disease, there
will be vomiting with blood. In view of the haemorrhage from upper and lower orifices,
the body health will be severely affected.
4. Allavilla noakku kuruthiazhal Noi (Unlimited directional blood heat disease )

In this disease, the following four directional factors are stimulated at a time by
the fire:
1) Abaanan ( downward directional)
2) Udhaanan ( upward directional)
3) Samaanan (mid directional)
4) Viyaanan (diffuse directional)
There will be haemorrhage from all the body orifices. Of all the four factors
mentioned above, the viyaana vayu is much affected and produces uncontrollable burning
sensation in the eye, ear and skin; the blood will ooze from the body and eye will
appear yellow in colour.

3.9.2. MODERN ASPECT
HYPERTENSION
Hypertension (HTN or HT), also known as high blood pressure (HBP), is

a long term medical condition in which the blood pressure in the arteries is persistently
elevated. High blood pressure usually does not cause symptoms. Long term high blood
pressure, however, is a major risk factor for coronary artery disease, stroke, heart
failure, peripheral vascular disease, vision loss, and chronic kidney disease.
High blood pressure is classified as either primary (essential) high blood
pressure or secondary high blood pressure. About 90–95% of cases are primary, defined
as high blood pressure due to nonspecific lifestyle and genetic factors. Lifestyle factors
that increase the risk include excess salt, excess body weight, smoking, and alcohol. The
remaining 5–10% of cases are categorized as secondary high blood pressure, defined as
high blood pressure due to an identifiable cause, such as chronic kidney disease,
narrowing of the kidney arteries, an endocrine disorder, or the use of birth control pills.
Blood pressure is expressed by two measurements, the systolic and diastolic pressures,
which are the maximum and minimum pressures, respectively. Normal blood pressure at
rest is within the range of 100–140 millimeters mercury (mmHg) systolic and 60–90
mmHg diastolic. High blood pressure is present if the resting blood pressure is
persistently at or above 140/90 mmHg for most adults. Different numbers apply to
children. Ambulatory blood pressure monitoring over a 24-hour period appears more
accurate than office best blood pressure measurement.
High blood pressure affects between 16 and 37% of the population globally. In
2010 hypertension was believed to have been a factor in 18% (9.4 million) deaths.
Signs and symptoms

Hypertension is rarely accompanied by any symptoms, and its identification is
usually through screening, or when seeking healthcare for an unrelated problem. Some
with high blood pressure report headaches (particularly at the back of the headand in the
morning), as well as lightheadedness, vertigo, tinnitus (buzzing or hissing in the ears),
altered vision or fainting episodes. These symptoms, however, might be related to

associated anxiety rather than the high blood pressure itself On physical examination,
hypertension may be associated with the presence of changes in the optic fundus seen
byophthalmoscopy The severity of the changes typical of hypertensive retinopathy is
graded from I–IV; grades I and II may be difficult to differentiate. The severity of the
retinopathy correlates roughly with the duration and/or the severity of the hypertension
Hypertensive crisis
Severely elevated blood pressure (equal to or greater than a systolic 180 or
diastolic of 110) is referred to as a hypertensive crisis. Hypertensive crisis is categorized
as either hypertensive urgency or hypertensive emergency, according to the absence or
presence of end organ damage, respectively.
In hypertensive urgency, there is no evidence of end organ damage resulting from
the elevated blood pressure. In these cases, oral medications are used to lower the BP
gradually over 24 to 48 hours
Pregnancy
Hypertension occurs in approximately 8–10% of pregnancies Two blood pressure
measurements six hours apart of greater than 140/90 mm Hg is considered diagnostic of
hypertension in pregnancy High blood pressure in pregnancy can be classified as pre-
existing hypertension, gestational hypertension or pre-eclampsia.
Children
Failure to thrive, seizures, irritability, lack of energy, and difficulty breathing can
be associated with hypertension in neonates and young infants. In older infants and
children, hypertension can cause headache, unexplained irritability, fatigue, failure to
thrive, blurred vision, nosebleeds, and facial paralysis.
Causes
Primary hypertension
Hypertension results from a complex interaction of genes and environmental
factors. Numerous common genetic variants with small effects on blood pressure have
been identified as well as some rare genetic variants with large effects on blood pressure.

Blood pressure rises with aging and the risk of becoming hypertensive in later life is
considerable. Several environmental factors influence blood pressure. High salt intake
raises the blood pressure in salt sensitive individuals; lack of exercise, obesity,
stress, and depression can play a role in individual cases. The possible role of other
factors such as caffeine consumption, and vitamin D deficiency are less clear. Insulin
resistance, which is common in obesity and is a component of syndrome X (or
the metabolic syndrome), is also thought to contribute to hypertension. Events in early
life, such as low birth weight, maternal smoking, and lack of breast feeding may be risk
factors for adult essential hypertension, although the mechanisms linking these exposures
to adult hypertension remain unclear. An increased rate of high blood urea has been found
in untreated people with hypertensive in comparison with people with normal blood
pressure, although it is uncertain whether the former plays a causal role or is subsidiary to
poor kidney function.
Secondary hypertension
Secondary hypertension results from an identifiable cause. Kidney disease is the
most common secondary cause of hypertension. Hypertension can also be caused by
endocrine conditions, such as Cushing's syndrome, hyperthyroidism, hypothyroidism,
acromegaly, Conn'syndrome or hyperaldosteronism, hyperparathyroidism andpheochrom
ocytoma Other causes of secondary hypertension include obesity, sleep
apnea, pregnancy, coarctation of the aorta, excessive liquorice consumption and certain
prescription medicines, herbal remedies and illegal drugs. Arsenic exposure through
drinking water has been shown to correlate with elevated blood pressure.
Pathophysiology
In most people with established essential hypertension, increased resistance to
blood flow (total peripheral resistance) accounts for the high pressure while cardiac
output remains normal. There is evidence that some younger people
with prehypertension or 'borderline hypertension' have high cardiac output, an elevated
heart rate and normal peripheral resistance, termed hyperkinetic borderline
hypertension. These individuals develop the typical features of established essential
hypertension in later life as their cardiac output falls and peripheral resistance rises with

age. Whether this pattern is typical of all people who ultimately develop hypertension is
disputed. The increased peripheral resistance in established hypertension is mainly
attributable to structural narrowing of small arteries and arterioles, although a reduction
in the number or density of capillaries may also contribute. Whether increased active
arteriolarvasoconstriction plays a role in established essential hypertension is
unclear. Hypertension is also associated with decreased peripheralvenous
compliance which may increase venous return, increase cardiac preload and, ultimately,
cause diastolic dysfunction.
Pulse pressure (the difference between systolic and diastolic blood pressure) is
frequently increased in older people with hypertension. This can mean that systolic
pressure is abnormally high, but diastolic pressure may be normal or low — a condition
termed isolated systolic hypertension. The high pulse pressure in elderly people with
hypertension or isolated systolic hypertension is explained by increased arterial stiffness,
which typically accompanies aging and may be exacerbated by high blood pressure.
EPIDEMIOLOGY
Hypertension in India
High blood pressure (BP) is a major public health problem in India and its
prevalence is rapidly increasing among both urban and rural populations. In fact,
hypertension is the most prevalent chronic disease in India.
The prevalence of hypertension ranges from 20-40% in urban adults and 12-17%
among rural adults. The number of people with hypertension is projected to
increase from 118 million in 2000 to 214 million in 2025, with nearly equal
numbers of men and women.
A survey of 26,000 adults in South India showed a hypertension prevalence of
20% (men 23% and women 17%) but 67% of those with hypertension were
unaware of their diagnosis. Majority of hypertensive subjects still remain
undetected and the control of hypertension is also inadequate. This calls for urgent
prevention and control measures for hypertension.
Recent (2012) studies show that for every known person with hypertension there
are two persons with either undiagnosed hypertension or prehypertension.

Reducing blood pressure can decrease cardiovascular risk and this can be
achieved by lifestyle measures in mild cases and should be the initial approach to
hypertension management in all cases. This includes dietary interventions weight
reduction, tobacco cessation, and physical activity.
A large-scale study undertaken in Tamil Nadu recently has established the
high prevalence of hypertension in rural India. The study reported 21.4 per
cent hypertension prevalence in about 10,500 people aged between 25 to 64 in
11 villages in the State with both sexes being affected equally. It was
published in the International Journal of Public Health. Urban areas had a
prevalence of hypertension of 22-30 per cent.
Investigation
ECG
X-Ray of the Chest
Echo Cardiography
Urine shows usually no change except RBC in Malignant hypertension
Blood Urea may be high in case of malignant hypertension
Serum Electrolytes
Ultra Sonogram is also helpful to find out renal disease
In other tests include IV urogram, Isotope renogram, CT Scan MRI etc.,
Complications:
Cardiac hypertensive Heart Disease
Left ventricular hypertophy develps in 10% to 30% of chronic cases.
Cerebral
Cerebrovascular complicationare more closely related to systeolic rather than
diastolic BP
1. Cerebral haemorrhate
2. Cerebral thrombosis
3. Lacunar infarcts
4. Hypertensive encephalopathy
5. TIA
6. Subarachnoid haemorrhage.
7. Dementia - both vascular and Alzheimer’s types

Retinal:
1. Dimness of vision
2. Thickening of arteries with narrowing of lumen, haemorrhage and
exudates, papiloedea, as metnioned before.
3. Detachment of retina, vitreous haemorrhage
Renal:
1. Renal arteriosclerosis (nephrosclerosis)
2. Uraemia
3. Renal infarct.
Aortic dissection
The marory cause of its is hypertension
Atherosclerotic complications
Many paitents of hypertension die out of these complications but the relationship
is much less close than other complications
Mode of Termination
1. Acute letft ventricular failure (60%)
2. Cerebral haemorrhage and allied episodes ( 35%)
3. Uraemia rare (5%)
Management
According to one review published in 2003, reduction of the blood pressure by
5 mmHg can decrease the risk of stroke by 34%, of ischaemic heart disease by 21%, and
reduce the likelihood of dementia, heart failure, and mortality from cardiovascular
disease.
Prevention
Much of the disease burden of high blood pressure is experienced by people who
are not labeled as hypertensive . Consequently, population strategies are required to
reduce the consequences of high blood pressure and reduce the need for antihypertensive
drug therapy. Lifestyle changes are recommended to lower blood pressure, before starting
drug therapy. The 2004 British Hypertension Society guidelines proposed the following
lifestyle changes consistent with those outlined by the US National High BP Education

Program in 2002 for the primary prevention of hypertension:
Maintain normal body weight for adults (e.g. Body mass index 20–25 kg/m2)
Reduce dietary sodium intake to <100 mmol/ day (<6 g of sodium chloride or
<2.4 g of sodium per day)
Engage in regular aerobic physical activity such as brisk walking (≥30 min per
day, most days of the week)
Limit alcohol consumption to no more than 3 units/day in men and no more than
2 units/day in women
Consume a diet rich in fruit and vegetables (e.g. At least five portions per day);
Effective lifestyle modification may lower blood pressure as much as an
individual antihypertensive drug. Combinations of two or more lifestyle modifications
can achieve even better results.
Lifestyle modifications
The first line of treatment for hypertension is lifestyle changes, including dietary
changes, physical exercise, and weight loss. Though these have all been recommended in
scientific advisories, a review by Cochrane found no evidence for effects of weight loss
diets on death or long-term complications and adverse events in persons with
hypertension. The review did find a decrease in blood pressure. Their potential
effectiveness is similar to and at times exceeds a single medication. If hypertension is
high enough to justify immediate use of medications, lifestyle changes are still
recommended in conjunction with medication.
Dietary changes shown to reduce blood pressure include diets with low
sodium, the DASH diet, vegetarian diets and high potassium diets.
Physical exercise regimens which are shown to reduce blood pressure
include isometric resistance exercise, aerobic exercise, resistance exercise, and device-
guided breathing. Stress reduction techniques such as biofeedback or transcendental
meditation may be considered as an add-on to other treatments to reduce hypertension,
but do not have evidence for preventing cardiovascular disease on their own.

Medications
Several classes of medications, collectively referred to as antihypertensive
medications, are available for treating hypertension.
First line medications for hypertension include thiazide-diuretics, calcium channel
blockers, angiotensin converting enzyme inhibitors and angiotensin receptor
blockers. These drugs may be used alone or in combination; the latter option may serve to
minimize counter-regulatory mechanisms that act to revert blood pressure values to pre-
treatment levels. The majority of people require more than one medication to control their
hypertension.

4. MATERIALS AND METHODS
4.1 Drug selection:
In this pre clinical study of Herbal Drug Venthamaraiyathi Chooranam is taken
as a trial drug for Thrombolytic,vasodilator, Hypolipidemic and Cardioprodective
activity from the Siddha literature “Pharmacopoeia of hospital of Indian medicine”
authored by Dr.v.Narayanaswami.
Ingredients
Elarasi(Elettaria cardamomum,.) - 2.5g(3%)
Chukku(Zingiber officinale, linn.) - 5g(6%)
Thippili(Piper longum, linn.) - 7.5g(9%)
Adhimadhuram(Glycyrrhiza glabra, linn.) - 10g(12%)
Sadhakuppai(Anethum sowa, linn.) - 12.5g(15%)
Seeragam(Cuminum cyminum, linn.) - 15g(18%)
Venthamarai Poo Ithalkal(Nelumbo nucifera, linn.) - 30g(36%)
Purification of Drugs
Elarasi
Fried at low flame and taken
Chukku
Cleaned with clean cloth and the outer skin is scrubbed off.
Sadhakuppai
Impurities are removed and dried in sunlight.
Thippili
Soaking it with lemon juice and. Fried at low flame and taken.
Adhimadhuram
Fried at low flame and taken
Seeragam
Impurities are removed and dried in sunlight.
Venthamarai Poo Ithalkal
Impuritis are removed and dried cool dark places

Process of preparation:
All the above mentioned ingredients are taken in the specified quantity and
roasted and powdered and filtered individually (fine process).Then all are thoroughly
mixed to make Ven thamaraiyathi chooranam.
Shelf life :
3 months
Dosage:
500mg to 1g two times a day
Vehicle:
Milk
Indication:
Kuruthi Alal Noi

Fig. 1 (a) : INGREDIENTS OF VENTHAMARAIYATHI CHOORANAM
Elettaria cardamomum - Seed Zingiber officinale, linn – Dried Rhizome
Piper longum, linn - Dry Fruit Cuminum cyminum, linn - Seeds
Glycyrrhiza glabra, linn - Root Anethum sowa, linn - Seed

Fig. 1 (b) : VENTHAMARAIYATHI CHOORANAM
NELUMBO NUCIFERA, LINN.- PETALS
VENTHAMARAIYATHI CHOORANAM

4.2 STANDARDIZATION OF THE DRUG:
The identification of herbal drugs & mineral drugs an authenticated by
P.G.Gunapadam Department & Herbal Botany Dept. GSMC, Palayamkottai.
Standardization of drugs helps in confirming its identify and determination of its quality
and effectiveness. Standardization of herbomineral drug is based on qualitative and
quantitative analysis through physico-chemical properties and instrumental studies.
Physico-chemical analysis and elemental analysis of this herbomineral formulation have
been done in Govt. Siddha medical college, Palayamkottai. IITM (FTIR in Dept of
Chemistry) and SEM in Dept of mechanics, Chennai.
Organoleptic character
The organoleptic characters of the sample drug were evaluated.1gm of the test
drug was taken and the colour, texture, particle size and other morphology were viewed
by naked eye under sunlight. Then the result is noted.
4.2.1 PHYSICO CHEMICAL ANALYSIS

Physicochemical studies of the trial drug have been done according to the WHO
guidelines.
Determination of Ash Values:

Total Ash:
3g is accurately weighed and incinerated in a crucible dish at a temperature not
exceed 450oC until free from carbon. It is then cooled and weighed. The % w/w of ash
with reference to the air-dried powder is calculated.
Water Soluble Ash:

The total ash is obtained as the above method for preparation of total ash. The ash
is boiled for 5minutes with 25ml water. The insoluble ashes is collected using filter paper
and washed with hot water and then transferred to the silica crucible then ignite for
15minutes at temperature not exceeding 450oC. The silica crucible and residue are
weighed until constant weight is attained for determination of weight of insoluble ash.
The weight of the water soluble ash is determined by substracting the weight of insoluble
ash from the weight of total ash.

Acid insoluble Ash:
The total ash is obtained as the above method for preparation of total ash. The ash
is boiled for 5minutes with 25ml 10% Hcl. The insoluble ashes is collected using filter
paper and washed with hot water and then transferred to the silica crucible then ignite for
15minutes at temperature not exceeding 450oC. The silica crucible and residue are
weighed until constant weight is attained.
Determination of Extractive Value:

Alcohol Soluble Extractive Value:
3g of test drug powder is weighed and macerated with 100ml of ethanol in a
closed container for 24 hours. The resulting solution is shaken continuously for 6 hours
and allowed to stand and soak for 18 hours. The solution is filtered and evaporated of the
filtrate in a flat bottomed shallow dish and dried at 105oC then cooled and weighed.
Water soluble Extractive value:

3g of test drug powder is weighed and macerated with chloroform and water,
respectively, at 80oC for 24 hrs. The resulting solution is shaken continuously for 6 hours
and allowed to stand and soak for 24 hrs then filtered. The solution from both chloroform
and water respectively is filtered and evaporated of the filtrate in a flat bottomed shallow
dish and dried at 105oC then cooled and weighed.
Loss on Drying:
The powdered drug is dried in the oven at 100- 105°C to constant weight. The
result was noted.
Thin layer chromatography (TLC)

Thin-layer chromatography(TLC) is a chromatographic technique that is useful
for separating organic compounds. Because of the simplicity and rapidity of TLC, it is
often used to monitor the progress of organic reactions and to check the purity of
products.TLC is a simple, quick and inexpensive procedure that gives how many
components are in a mixture.TLC is also used to support the identity of a compound in a

mixture when the Rf of a compound is compared with the Rf of a known
compound(preferably both run on the same TLC plate).Chromatography works on the
principle that different compounds will have different solubilities and adsorbtion to the
two phases between which they are to be partitioned. As the solvent rises by capillary
action up through the adsorbent, differential partitioning occurs between the components
of the mixture dissolved in the solvent stationary adsorbent phase. The more strongly a
given component of a mixture is adsorbed on to the stationary phase, the less time it will
spend in the mobile phase and the more slowly it will migrate up the plate.

4.2.2 CHEMICAL ANALYSIS:
Preliminary Basic and Acidic radical studies:
Preparation of the extract:
5gms of the test drug is weighed accurately and placed in a 250ml clean beaker.
Then 50ml of distilled water is added and dissolved well. Then it is boiled well for about
10 minutes. It is cooled and filtered in a 100ml volumetric flask and then it is made up to
100ml with distilled water. This preparation is used for the qualitative analysis of acidic/
basic radicals and biochemical constituents in it.
QUALITATIVE ANALYSIS FOR BASIC RADICALS:

Test for Calcium:
2ml of the above prepared extract is taken in a clean test tube. To this add 2ml of
4% Ammonium oxalate solution. Formation of white precipitate indicates the presence of
calcium.
Test for Iron (Ferric):
The extract is acidified with glacial acetic acid and potassium ferro cyanide.
Formation of blue colour indicates the presence of ferric iron.
Test for Iron (Ferrous):
The extract is treated with concentrated Nitric acid and ammonium thio-cyanate
solution. Formation of blood red colour indicates the presence of ferrous iron.
Test for Zinc:
The extract is treated with potassium ferro-cyanide. Formation of white
precipitate indicates the presence of zinc.
QUALITATIVE ANALYSIS FOR ACIDIC RADICALS:

Test for Sulphate:
2ml of extract is added to 5% barium chloride solution. Formation of white
precipitate indicates the presence of sulphate.
Test for Chloride:
The extract is treated with silver nitrate solution. Formation of white precipitate
indicates the presence of chloride.

Test for Phosphate:
The extract is treated with ammonium molybdate and concentrated nitric acid.
Formation of yellow precipitate indicates the presence of phosphate.
Test for Carbonate:
On treating the extract with concentrated hydrochloric acid giving brisk
effervescence indicates the presence of carbonate.
Test for starch:
The extract is added with weak iodine solution. Formation of blue colour
indicates the presence of starch.
Test for albumin:
The extract is treated with Esbach`s reagent. Formation of yellow precipitate
indicates the presence of albumin.
Test for tannic acid:
The extract is treated with ferric chloride. Formation of bluish black precipitate
indicates the presence of tannic acid.
Test for unsaturation:
The extract is treated with potassium permanganate solution. The dis-
colourization of potassium permanganate indicates the presence of unsaturated
compounds.
Test for the reducing sugar:
5ml of Benedict`s qualitative solution is taken in a test tube and allowed to boil
for 2 minutes and added 8-10 drops of the extract and again boil it for 2 minutes. Any
colour change indicates the presence of reducing sugar.
Test for amino acid:
One or two drops of the extract is placed on a filter paper and dried it well. After
drying, 1% Ninhydrin is sprayed over the same and dried it well. Formation of violet
colour indicates the presence of amino acid.

4.2.3. INSTRUMENTAL ANALYSIS
Fig. 2. Scanning Electron Microscope (SEM)
The microstructure of the powders was examined using a Hitachi S 3000H

scanning electron microscope (Fig ). The scanning Electron Microscope is one of the
most versatile instruments available for the examination and analysis of the micro
structural characteristics of solid objects. The primary reason for the SEM’s usefulness is
the high resolution which can be obtained when bulk objects are examined; values of the
order of 5nm (50degreeA) are usually quoted for commercial instruments. Advanced
research instruments have been described which have achieved resolutions of about
2.5nm (25 degree A). Any solid material can be studied. Sample size is limited to
specimens less than about 10µm in diameter. An electron beam passing through an
evacuated column is focused by electromagnetic lenses onto the specimen surface. The
beam is then rastered 0ver the specimen in synchronism with the beam of a cathode ray
tube display screen. In eleastically scattered secondary electrons are emitted from the
sample surface and collected by a scintillator, the signal from which is used to modulate
the brightness of the cathode ray tube. In this way the secondary electron emission from
the sample is used to form an image on the CRT display screen. (Goldstein, et. al., 1992)

Differences in secondary emission result from changes in surface topography. If
(elastically) back-scattered electrons are collected to form the image, contrast results
from compositional differences. Cameras are provided to record the images on the
display screen. Since an electron is a charged particle, it has a strong interaction with the
specimen (due to coulomb interaction). When an electron beam images on a specimen, it
is scattered by atomic layers near the surface of the specimen. As a result, the direction of
electron motion changes and its energy is partially lost. Once an incident electron
(primary electron) enters a substance, its direction of motion is influenced by various
obstructions (multiple scattering), and follows a complicated trajectory which is far from
a straight line. Also, when electrons with the same energy are incident on the specimen
surface, a portion of electrons is reflected in the opposite direction (back scattered) and
the remainder is absorbed by the specimen (exciting X- rays or other quanta in the
process). If the specimen issufficiently thin, the electron can pass all the way through the
specimen (transmitted electrons, scattered or non-scattered).
The depth at which various signals are generated due to electron beam – specimen
interaction indicates the diffusion area of the signals in the specimen in addition to the
local chemistry of the specimen. Secondary electrons mainly indicate information about
the surface of a specimen. Since secondary electrons do not diffuse much inside the
specimen, they are most suitable for observing the fine-structures of the specimen
surface. That is to say, sharp scanning images with high resolution can be expected from
secondary electrons, because of the smaller influence on resolution by their diffusion.
As the incident electron energy increases, the probability of incident electrons
colliding with elemental components of the specimen and releasing secondary electrons
also increases. In other words, as the incident energy increases, the emission of electrons
from the specimen also increases. However, as the energy increases beyond a certain
level, the incident electrons penetrate deeper into the specimen with the result that the
specimen derived electrons use up most of their energy to reach the specimen surface.
Consequently, the electron emission yield decreases. Therefore, the peak secondary
electron emission yield occurs at a specific entry level of the incident electrons.
In order to verify the existence of a substance and recognize its shape, the image
contrast must be well defined. In other words, even if a system boasts extremely high

resolution, if image contrast is poor, it would be extremely difficult to determine the
existence of a substance, let alone recognize its shape. Another important feature of the
SEM is the three-dimensional appearance of the specimen image, which is a direct result
of the large depth of field. The SEM is also capable of examining objects at very low
magnification. This feature is useful in viewing particle size and shape of any
composition at various stages of preparation in siddha system as well as other fields.
The large depth of field available in the SEM makes it possible to observe 3-
dimensional objects in stereo. Today, a majority of SEM facilities are equipped with X-
ray analytical capabilities. Thus topographic crystallographic and compositional
information can be obtained rapidly, efficiently and simultaneously from the same area.
The author was chosen this analysis for detecting Particle size of the classical
siddha herbal drug Venthamaraiyathi Chooranam . SEM results of Venthamaraiyathi
Chooranam was represented in results section.

FOURIER TRANSFORM - INFRA RED SPECTROSCOPY (FT-IR)
Fig. 3: FTIR Apparatus
FTIR-Mechanism

Introduction
Vibrational spectroscopy is an extremely useful tool in the elucidations of
molecular structure. The spectral bands can be assigned to different vibrational modes of
the molecule. The various functional groups present in the molecule can be assigned by a
comparison of the spectra with characteristic functional group frequencies. As the
positions of the bands are directly related to the strength of the chemical bond, a large
number of investigations including intermolecular interactions, phase transitions and
chemical kinetics can be carried out using this branch of spectroscopy. In IR
spectroscopy, the resonance absorption is made possible by the change in dipole moment
accompanying the vibrational transition. The Infrared spectrum originates from the
vibrational motion of the molecule. The vibrational frequencies are a kind of fingerprint
of the compounds. This property is used for characterization of organic, inorganic and
biological compounds. The band intensities are proportional to the concentration of the
compound and hence qualitative estimations are possible. The IR spectroscopy is carried
out by using Fourier transform technique.
Principle
Infra red spectroscopy involves study of the interaction of electromagnetic
radiation with matter. Due to this interaction, electromagnetic radiation characteristic of
the interacting system may be absorbed (or emitted). The experimental data consist of the
nature (frequency of wave length) and the amount (intensity) of the characteristic
radiation absorbed or emitted. These data are correlated with the molecular and electronic
structure of the substance and with intra- and inter molecular interactions.
Source : Nernst Glower
Beam splitter : It is made up of a transparent material. Thin films of

Silicondeposited on Potassium bromide (KBr)
Bromide (KBr) Detectors : Deutrated TriGlycine Sulphate (DTGS).
MIR Range : 4000 to 450 cm-1
Resolution : 4.0 cm-1

Sampling Techniques
There are a variety of techniques for sample preparation depending on the
physical form of the sample to be analyzed.
Solid : KBr or Nujol mull method.
Liquid : Csl / TIBr Cells
Gas : Gas cells
KBr Method
The sample is grounded using an agate mortar and pestle to give a very fine
powder.
The finely powder sample is then mixed with about 100mg dried KBr salt.
The mixture is then pressed under hydraulic press using a die to yield a
transparent disc and measure about 13mm diameter and 0.3mm in thickness.
Nujol Mull Method
The sample is ground using an agate mortar and pestle to give a very fine
powder.
A small amount is then mixed with nujol oil to give a paste and this paste is
then
applied between two sodium chloride plates.
The plates are then placed in the instrument sample holder ready for scanning.
Liquids
Viscous liquids can be smeared in the cell and directly measured.
For dilute solutions, liquid cells and variable path length cells are employed.
Measurements Techniques
The procedure for recording the %T or %A is as follows:
Air is first scanned for the reference and stored. The sample is then recorded and
finally the ratio of the sample and reference data is computed to give required %T
or %A at various frequencies.
Study of substances with strong absorbance bands and weak absorbance bands
aswell as possible.
Small amount of samples are sufficient
High resolution is obtained.

Procedure
Preparation of samples for infrared measurements and infrared spectra
Typically,1.5 mg of protein, dissolved in the buffer used for its purification, were
centrifuged in a 30 K Centric on micro concentrator (Amicon) at 3000_g at 4oC
until a volume of approximately 40 Al.
Then, 300 Al of 20 mM This buffer, prepared in H20 or 2H20, pH or p2H 7.2,
were added and the sample concentrated again. The p2H value corresponds to the
pH meter reading + 0.4. The concentration and dilution procedure was repeated
several times in order to completely replace the original buffer with the Tris
buffer.
The washings took 24 h, which is the time of contact of the protein with the 2H20
medium prior FT-IR analysis. In the last washing, the protein was concentrated to
fine a volume of approximately 40 Al and used for the infrared measurements.
The concentrated protein sample was placed in CaF2 windows and a 6 Am tin
spaceror a 25 Am Teflon spacer for the experiments in H20 or 2H20, respectively.
FT-IR spectra were recorded by means of a Perkin-Elmer -Spectrum-1 FT-IR
spectrometer using a deuterated triglycine sulfate detector.
At least 24 h before, and during data acquisition, the spectrometer was
continuously purged with dry air at a dew point of 40oC. Spectra of buffers and
samples were acquired at 2 cm_1 resolution under the same scanning and
temperature conditions. In the thermal denaturation experiments, the temperature
was raised in 5oC steps from 20 to 95oC.
Before spectrum acquisition, samples were maintained at the desired temperature
for the time necessary for the stabilization of temperature inside the cell (6 min).
Spectra were collected and processed using the SPECTRUM software from
Perkin- Elmer. Correct subtraction of H20 was judged to yield an approximately
flat baseline at 1900-1400 cm_ 1, and subtraction of 2H20 was adjusted to the
removal of the2H20 bending absorption close to 1220 cm_ 1

INDUCTIVELY COUPLED PLASMA OPTICAL EMISSION
SPECTROMETRY (ICP-OES),
Fig. 5: ICPOES Apparatus

Inductively coupled plasma optical emission spectrometry (ICP-OES)is an
analyticaltechnique used for the detection of trace metals. It is a type of emission
spectroscopy thatuses the inductively coupled plasma to produce excited atoms and ions
thatemit electromagnetic radiation at wavelengths characteristic of a particular element.
Theintensity of this emission is indicative of the concentration of the element within the
sample.
Mechanism
The ICP-OES is composed of two parts: the ICP and the optical spectrometer.
TheICP torch consists of 3 concentric quartz glass tubes. The output or “work” coil of the
radiofrequency (RF) generator surrounds part of this quartz torch. Argon gas is typically
usedto create the plasma.
When the torch is turned on, an intense electromagnetic field is created within
thecoil by the high power radio frequency signal flowing in the coil. This RF signal is
createdby the RF generator which is, effectively, a high power radio transmitter driving
the “workcoil” the same way a typical radio transmitter drives a transmitting antenna.

The argon gasflowing through the torch is ignited with a Tesla unit that creates a brief
discharge arcthrough the argon flow to initiate the ionization process. Once the plasma is
“ignited”, theTesla unit is turned off.
The argon gas is ionized in the intense electromagnetic field and flows in a
particularrotationally symmetrical pattern towards the magnetic field of the RF coil. A
stable, hightemperature plasma of about 7000 K is then generated as the result of the
inelastic collisionscreated between the neutral argon atoms and the charged particles. A
peristalticpump delivers an aqueous or organic sample into a nebulizer where it is
changed into mistand introduced directly inside the plasma flame. The sample
immediately collides with theelectrons and charged ions in the plasma and is itself broken
down into charged ions. Thevarious molecules break up into their respective atoms which
then lose electrons andrecombine repeatedly in the plasma, giving off radiation at the
characteristic wavelengths ofthe elements involved.
Within the optical chamber(s), after the light is separated into its
differentwavelengths (colors), the light intensity is measured with a photomultiplier tube
or tubesphysically positioned to “view” the specific wavelength(s) for each element line
involved,or, in more modern units, the separated colors fall upon an array of
semiconductor photodetectors such as charge coupled devices (CCDs). In units using
these detector arrays, theintensities of all wavelengths (within the system’s range) can be
measured simultaneously,allowing the instrument to analyze for every element to which
the unit is sensitive all atonce. Thus, samples can be analyzed very quickly.
The intensity of each line is then compared to previously measured intensities of
known concentrations of the elements, and their concentrations are then computed by
interpolation along the calibration lines. In addition, special software generally corrects
for interferences caused by the presence of different elements within a given sample
matrix.Examples of the application of ICP-OES include the determination of metals,
arsenicpresent in Traditional medicines, and trace elements bound to proteins. ICP-OES
is widelyused in minerals processing to provide the data on grades of various streams, for
theconstruction of mass balances.
The author used for elemental identification and quantitative compositional
information of the VENTHAMARAIYATHI CHOORANAM .

4.3. TOXICITY STUDY
A) ACUTE TOXICITY STUDY
FEMALE WISTER RATS TO EVALUATE TOXICITY PROFILE OF
VENTHAMARAIYATI CHOORANAM
OBJECTIVES
The aim of this Study is to evaluate the toxicity of the test substance
VENTHAMARAIYATI CHOORANAM, when administered orally to Female Wister
Rats with different doses, so as to provide a rational base for the evaluation of the
toxicological risk to man and indicate potential target organs.
Guidelines followed:
(a) OECD Guidelines No. 423,
Study Design and Controls:
Female Wister Rats in controlled age and body weight were selected.
Venthamaraiyathi Chooranam was administered at 5 mg/kg, 50 mg/kg, 300
mg/kg, 1000 mg/kg, and mg/kg body weight as (Water) as suspension along
with blank.
The results were recorded on day 0, with single oral dosing period of 14 days.
EXPERIMENTAL PROCEDURE
ANIMALS
Supply
A total of 15 Female Wister Rats with an approximate age of 6 weeks and purchased
from M/s.Venkateshwara Enterprises Pvt. Ltd, Bangalore. On their arrival a sample of
animals was chosen at random and weighed to ensure compliance with the age requested.
The mean weights of Female Wister Rats were 100-150 g respectively. The animals were
housed in metabolic cages (55 x 32.7 x 19 cm), with sawdust litter, in such a way that

each cage contained a maximum of 3 animals of the same sex.
All animals underwent a period of 20 days of observation and acclimatization
between the date of arrival and the start of treatment. During the course of this period, the
animals were inspected by a veterinary surgeon to ensure that they fulfilled the health
requirements necessary for initiation of the Study.
Housing
The Female Wister Rats were housed in metabolic cages (55 x 32.7 x 19 cm),
placed on racks. From the week before initiation of the treatment, each cage contained a
maximum of 6 mice of the same sex and treatment group.
Each cage was identified by a card, color coded according to the dose level. This
card stated the cage number, number and sex of the animals it contained, Study number,
test substance code, administration route, dose level and Study Director's name, date of
the arrival of the animals and initiation of treatment.
The temperature and relative humidity were continuously monitored. Lighting
was controlled to supply 12 hours of light (7:00 to 19:00 hours) and 12 hours of dark for
each 24-hour period.
The cages corresponding to each experimental group were distributed on racks in
such a manner that external factors, such as environmental conditions, were balanced as
far as possible.
DIET
All the rats had free access to a pelleted rat diet. The diet was analyzed by the
manufacturer to check its composition and to detect possible contaminants.
Water
The water was offered ad libitum in bottles.

ADMINISTRATION ROUTE AND PROCEDURE
The test substance was administered orally. The Female Wister Rats belonging to
the control group were treated with the vehicle (Water) at the same administration volume
as the rest of the treatment groups.
Numbering and Identification

The animals were marked on body with picric acid solution prepared in water.
The marking within the cage was as below.
Table-1 Numbering and Identification
Group No Animal Marking

1 Head
2 Body
3 Tail
The group no., cage no., sex of the animal and animal no. were identified as
indicated below using cage label and body marking on the animals
Table-2 Numbering and Identification
Cage No Group No Animal Marking Sex

1 I H,B,T Female
2 II H,B,T Female
3 III H,B,T Female
4 IV H,B,T Female
5 V H,B,T Female
Doses
The doses for the study were selected based on literature search and range finding
study. Following the period of fasting, the animals were weighed and then drug was
administered orally as single dose using a needle fitted onto a disposable syringe of
approximate size at the following different doses.

Table-3 Doses
GROUP DOSE
Group-I 5 mg/kg
Group-II 50 mg/kg
Group-III 300 mg/kg
Group-IV 1000 mg/kg
Group-V 2000 mg/kg
The test item was administered as single dose. After single dose administration
period, all animals were observed for day 14.
Dose Preparation
Venthamaraiyathi Chooranam was added in distilled water and completely

dissolved to form oral for administration. The dose was prepared of a required
concentration before dosing by dissolving, in distilled water. It was mixed well. The
preparation for different doses was vary in concentrations to allow a constant dosage
volume.
Administration
The test item was administered orally to each Female Wister rats as single dose
using a needle fitted onto a disposable syringe of appropriate size at the following
different doses. The concentration was adjusted according to its body weight. The volume
was not exceeding 10 ml/kg bodyweight. Variability in test volume was minimized by
adjusting the concentration to ensure a constant volume at all dose levels.
Observation period
All animals were observed for any abnormal clinical signs and behavioral
changes. The appearance, change and disappearance of these clinical signs, if any, were
recorded for approximately 1.0, 3.0 and 4.0 hours post-dose on day of dosing and once
daily thereafter for14 days. Animals in pain or showing severe signs of distress were
humanely killed. The cageside observation was included changes in skin, fur, eyes and

mucous membranes, occurrence of secretions and excretions. Autonomic activity like
lacrimation, piloerection, pupil size and unusual respiratory pattern, changes in gait,
posture, response to handling, presence of clonic or tonic movements, stereotypes like
excessive grooming and repetitive circling or bizarre behavior like self-mutilation,
walking backwards etc were observed. At the 14th day, sensory reactivity to stimuli of
different types (e.g. auditory, visual and proprioceptive stimuli) was conducted. Auditory
stimuli responses were measured by clicker sound from approximately 30 cm to the rats;
visual stimuli response were measured with the help of shining pen light in the eye of rats
and placing a blunt object near to the eye of rats. Response to proprioceptive stimuli was
measured by placing anterior/dorsal surface of animals paw to the table edge. The
responses of reactions for these three exercises were normal in animals belonging to both
the controls as well as drug treatment dose groups.
Mortality and Morbidity

All animals were observed daily once for mortality and morbidity at
approximately 1.0, 3.0 and 4.0 hours post dose on day of dosing and twice daily (morning
and afternoon) thereafter for 14 days.

B. SUB-ACUTE TOXICITY STUDY
WISTER RATS TO EVALUATE TOXICITY PROFILE OF

VENTHAMARAIYATHICHOORANAM
1. Objective
The objective of this ‘Sub-Acute Toxicity Study of Venthamaraiyathi
Chooranam ON Wister Rats’ was to assess the toxicological profile of the test item
when treated as a single dose. Animals should be observed for 28 days after the drug
administration. This study provides information on the possible health hazards likely to
arise from exposure over a relatively limited period of time.
2. Test Guideline Followed

OECD 407 Method - Sub-Acute Toxic Class Method (Repeated Dose 28-Day Oral
Toxicity Study in Rodents)
3. Test Item Detail

Name: Venthamaraiyathi Chooranam
4. Test System Detail

The study was conducted on 5 male 5 female Wister rats. These animals were
selected because of the recommended rodent species for oral studies as per followed
guideline and availability of Animals 8-12 weeks old male and female rats were selected
after physical and behavioral examination. The body weight range was fallen within ±
20% of the mean body weight at the time of Randomization and grouping. The rats were
housed in standard laboratory condition in Polypropylene cages, provided with food and
water adlibitum in the Animal at M/s. Sree Venkateshwara Enterprises Pvt. Ltd,
Bangalore. The experimental protocol was approved by Institutional Animal Ethical
Committee as per the guidance of Committee for the Purpose of Control and Supervision
of Experiments on Animals (CPCSEA), Ministry of Environment and Forest, government
of India.

5. Acclimatization
The animals were selected after veterinary examination by the veterinarian. All
the selected animals were kept under acclimatization for a week.
6. Randomization & grouping

One day before the initiation of treatment (days 0- last day of acclimatization), the
selected animals were randomly grouped into three different groups containing minimum
6 male animals per group.
7. Numbering and Identification

The animals were marked on body with picric acid solution prepared in water.
The marking within the cage was as below.
TABLE-4 NUMBERING AND IDENTIFICATION
Group No Animal Marking
H,B,T,HB,NM (MALE)
1. CONTROL
H,B,T,HB,NM (FEMALE)
2. LOW DOSE OF
H,B,T,HB,NM (MALE)
300MG/KG
3. MIDDLEDOSE OF
H,B,T,HB,NM (MALE)
600MG/KG
4. HIGH DOSE OF
H,B,T,HB,NM (MALE)
900MG/KG
The group no., cage no., sex of the animal and animal no. were identified as
indicated below using cage label and body marking on the animals:

Table – 5 Husbandry
Cage Group No Animal Marking Sex
No
H,B,T,HB,NM
Male
1. 1. CONTROL H,B,T,HB, NM
Female
2. LOW DOSE OF
H,B,T,HB,NM Male
2.
VENTHAMARAIYATHICHOORANAM 300mg/kg H,B,T,HB, NM Female
3. MIDDLEDOSE OF H,B,T,HB,NM
Male
3. H,B,T,HB, NM
VENTHAMARAIYATHICHOORANAM 600mg/kg Female
4. HIGH DOSE OF
H,B,T,HB,NM
4. Male
VENTHAMARAIYATHICHOORANAM 900mg/kg H,B,T,HB ,NM
Female
Husbandry
Housing
The Wister rats were housed in standard polypropylene cages with stainless steel
top grill. Paddy husk was used as bedding. The paddy husk was changed at least twice in
a week. From the week before initiation of the treatment, each cage contained a
maximum of 6 mice of the same sex and treatment group.
Environmental conditions
The animals were kept in a clean environment with 12 hour light and 12 hour dark
cycles. The air was conditioned at 22±30C and the relative humidity was maintained
between 30-70% with 100% exhaust facility. The cages corresponding to each
experimental group were distributed on racks in such a manner that external factors, such
as environmental conditions, were balanced as far as possible.

Feed & feeding schedule
‘Sai Durga Animal Feed, Bangalore. Feed was provided adlibitum throughout the
study period, except over night fasting (18-20 hours) prior to dose administration. After
the substance has been administered, food was with held for a further 3-4 hours.
Water
The water was offered adlibitum in bottles. There was periodically analyzed to
detect the presence of possible contaminants
Doses
The doses for the study were selected based on literature search and range finding
study. Following the period of fasting, the animals were weighed and then extract was
administered orally as single dose using a needle fitted on to a disposable syringe of
approximate size at the following different doses.
Table-6 Dose level
TEST GROUP DOSE TO ANIMALS NUMBER OFANIMALS

(mg/kg body-weight/day)
Group-1 1. CONTROL 10 (5MALE and5 FEMALE)
2. LOW DOSE OF
Group-II VENTHAMARAIYATHICHOORANAM 10(5MALE and 5 FEMALE)
300mg/kg
3. MIDDLEDOSE OF
Group-III VENTHAMARAIYATHICHOORANAM 10(5MALE and 5 FEMALE)
600mg/kg
4. HIGH DOSE OF
Group-IV VENTHAMARAIYATHICHOORANAM 10(5MALE and 5 FEMALE)
900mg/kg
period, all animals were observed for 28 days.

Dose Preparation
VENTHAMARAIYATHICHOORANAM was added in distilled water and completely

dissolved to for oral for administration. The dose was prepared of a required
concentration before dosing by dissolving Venthamaraiyathi Chooranam in distilled water. It
was mixed well. The preparation for different doses was vary in concentrations to allow a
constant dosage volume.
Administration
The test item was administered orally to each rat as single dose using a needle
fitted on to a disposable syringe of appropriate size at the following different doses. The
concentration was adjusted according to its body weight. The volume was not exceeding
10 ml/kg body weight. Variability in test volume was minimized by adjusting the
concentration to ensure a constant volume at all dose levels.
OBSERVATIONS
These observations were also performed on week-ends. The observations
included but were not limited to changes in skin and fur, in the eyes and mucous
membranes, in the respiratory, circulatory, central nervous and autonomous systems,
somatomotor activity and behavior.
Clinical signs of toxicity

All the rats were observed at least twice daily with the purpose of recording any
symptoms of ill- health or behavioral changes. Clinical signs of toxicity daily for 28
days.
Food intake
Prior to the beginning of treatment, and daily, the food intake of each cage was
recorded for period of 28 days and the mean weekly intake per rats was calculated.

Water intake
Water intake was checked by visual observation during the Study. In addition, the
water consumption in each cage was measured daily for a period of 28 days.
Bodyweight:
The body weight of each rat was recorded one week before the start of treatment,
and during the course of the treatment on the day of initial, 3rd, 7th, 10th, 14th, 17th,
20th, 24th and 28th days (day of sacrifice). The mean weights for the different groups
and sexes were calculated from the individual weights.
Blood Collection
Blood was collected through retro-orbital sinus from all the animals of different
groups on 28th day. The blood was collected in tubes containing Heparin/EDTA as an
anticoagulant. Animals were fasted over night prior to the blood collection.
LABORATORY STUDIES
During the 4th week of treatment, samples of blood were withdrawn from the
orbital sinus of 6 males from each group, under light ether anesthesia after fasting for 16
hours. The blood samples are used to evaluate Hematological parameters like RBC,
WBC, and PLATELETS etc….. The collected blood samples also centrifuged 10000 rpm
in 10 minutes to separate the serum. The separated serum used to evaluate biochemical
parameters like SGOT, SGPT, ALP and BILIRUBIN etc.,
Hematology
The following hematological parameters were analysed using Autoanalyser
Hb : Haemoglobin (g %)
PCV : Packed Cell Volume
WBC : White Blood Corpuscles (x103/cmm)
RBC : Red Blood Corpuscles (x106/cmm)
Blood Platelet count (x103/cmm)

Differential WBC count:
N : Neutrophils (%)
L : Lymphocytes (%)
M : Monocytes (%)
E : Eosinophils (%)
RDW : Red Cell Distribution Width.
MPV : Mean Platelet Volume
Clinical Biochemistry:
The following clinical Bio parameters were analysed using Auto analyser
Total serum protein (g/dl)
ALT/SGPT : Alanine amino transferase (U/L)
AST/SGOT : Aspartate amino transferase (U/L)
ALP : Alkaline serum phosphatase (U/L)
CHL : Cholesterol (mg/dL)
HDL : High density lipoprotein
TG : Triglyceride
TERMINAL STUDIES
Sacrifice and macroscopic examination
On completion of the 4 weeks of treatment, 18 Wister rats were sacrificed by
ether inhalation. A full autopsy was performed on all animals which included
examination of the external surface of the body, all orifices, cranial, thoracic and
abdominal cavities and their contents both in situ and after evisceration. As the number
of animals exceeded the number that could be sacrificed in one day, the autopsies were
carried out over three consecutive days at the end of the treatment period.
Organ weights:
After the macroscopic examination the following organs were weighed after
separating the superficial fat: Brain, Heart, Spleen Kidneys, Testes, Liver, Lungs,
pancreas and stomach.

HISTOPATHOLOGICAL STUDIES
Anatomy of the liver was studied immediately after sacrificing the animals. A
small portion was fixed in 10% neutral buffered formalin as described by Luna 14 . Thin
sections of 4-5 µm were taken, stained with Haematoxylin and Eosin and histology was
studied.
ESTIMATION OF HEMATOLOGICAL PARAMETERS:
Collection of blood for hematological studies

After the treatment period the animals were anaesthetized by ketamine
hydrochloride and the blood was collected from Retro-orbital sinus by using capillary
into a centrifugation tube which contains EDTA for haematological parameters The
haematological parameters like RBC, WBC and Hb percentage, Differential cell count,
MCV, MCHC, Hematocrit, MCH, platelet count were estimated by the following
procedures.
ENUMERATION OF RED BLOOD CELLS: 1 Ramnic 2007)
Reagents : RBC diluting fluid
Procedure:
Using a red blood cell pipette of haemocytometer, well mixed blood was drawn up
to 0.5 mark and RBC diluting fluid was taken up to mark II. The fluid blood mixture was
shaken and transferred onto the counting chamber. The cells were allowed to settle to the
bottom of the chamber for 2 min. See the fluid does not get dried. Using 45X or high
power objective the RBC’s were counted uniformly in the larger corner squares.
The cells were expressed as number of cells x1012/l

ENUMERATION OF WBC: 2 John 1972)
Reagents:
Turk’s fluid: Turk’s fluid was prepared by mixing 2ml of acetic acid with 100 ml
of distilled water. To this 10 drop of aqueous methylene blue 3 % w/v) was added. This
solution haemolysis the red cells due to acidity so that counting of white cells becomes
easy.
Procedure:
Using a white blood cell pipette of haemocytometer, well mixed blood was drawn
up to 0.5 mark and WBC diluting fluid was taken up to mark II. The fluid blood mixture
was shaken and transferred onto the counting chamber. The cells were allowed to settle to
the bottom of the chamber for 2 min. See the fluid does not get dried.
Using 10X or low power objective the WBC’s were counted uniformly in the
larger corner squares.
The cells were expressed as number of cells/10mm.
DIFFERENTIAL LEUCOCYTE COUNT: John 1972
Reagent:
Leishmann’s stain: 150mg of powdered leishmann’s stain was dissolved in

133ml of acetone free methanol.
Procedure:
A blood film stained with leishmann’s stain was examined under oil immersion
and the different types of WBCs were identified. The percentage distribution of these
cells was then determined. Smears were made from anticoagulant blood specimens and
stained with leishmann’s stain. The slides were preserved for counting the number of
lymphocytes and neutrophils, per 100 cells were noted.

From the different Leukocyte count and WBC count, absolute lymphocyte and
neutrophil count were calculated.
Number of neutrophils
_______________________________
Absolute neutrophil count = x TWBC
100
Number of lymphocytes
_______________________________
Absolute lymphocyte count = x TWBC
100
J. C. Dacie and S. M. Lewis, Practical haematology, London: Churchill Livingstone,

1984, pp. 5.
Measurement of biochemical parameters estimation

Haemoglobin (Hb), was estimated using whole blood. Remaining parameters
were measured in serum. All of the above biochemical parameters were estimated using
semi-autoanalyzer (Photometer 5010 V5+, Germany) with enzymatic kits procured from
Piramal Healthcare limited, Lab Diagnostic Division, Mumbai, India.
Determination of aspartate aminotransferase (AST)

Aspartate aminotransferase, also known as Glutamate Oxaloacetate Transaminase
(GOT) catalyses the transamination of L-aspartate and α keto glutarate to form
oxaloacetate and L- glutamate. Oxaloacetate formed is coupled with 2,4- Dinitrophenyl
hydrazine to form hydrazone, a brown coloured complex in alkaline medium which can
be measured colorimetrically.
Reagents
Buffered aspartate (pH 7.4); 2,4- DNPH reagent; 4N sodium hydroxide;
working pyruvate standard; solution I (prepared by diluting 1 ml of reagent 3 to 10 ml
with purified water).
Procedure
Rietman and Frankle method was adopted for the estimation of SGOT. (Reitmann
S, Frankel S, 1957. A colorimetric method for the determination of serum oxaloacetic and

glutamic pyruvate transminases. American Journal of Clinical Pathology.28: 56-63.The
reaction systems used for this study included blank, standard, test (for each serum
sample) and control (for each serum sample). 0.25 ml of buffered aspartate was added
into all the test tubes. Then 0.05 ml of serum was added to the test group tubes and 0.05
ml of working pyruvate standard into the standard tubes. After proper mixing, all the
tubes were kept for incubation at 37oC for 60 min, after which 0.25 ml each of 2,4-
DNPH reagent was added into all the tubes. Then, 0.05 ml of distilled water and 0.05 ml
of each serum sample was added to the blank and the serum control tubes respectively.
The mixture was allowed to stand at room temperature for 20 min. After incubation, 2.5
ml of solution I was added to all test tubes. Mixed properly and optical density was
measured in a spectrophotometer at 505 nm within 15 min.
The enzyme activity was calculated as:-

AST (GOT) activity in IU/L) = [(Absorbance of test - Absorbance of control)/
(Absorbance of standard - Absorbance of blank)] x concentration of the standard
Determination of alanine aminotransferase (ALT)

Alanine aminotransferase, also known as Glutathione Peroxidase (GPT) catalyses
the transamination of L-alanine and α keto glutarate to form pyruvate and L- Glutamate.
Pyruvate so formed is coupled with 2,4 – Dinitrophenyl hydrazine to form a
corresponding hydrazone, a brown coloured complex in alkaline medium which can be
measured colorimetrically.
Reagents
Buffered alanine (pH 7.4), 2,4–DNPH, 4N sodium hydroxide, working pyruvate
standard, solution I (prepared by diluting 1 ml of reagent 3 to 10 ml with purified water).
Procedure
Rietman and Frankle method was dopted for the estimation of SGPT. The reaction
systems used for this study included blank, standard, test (for each serum sample) and
control (for each serum sample). 0.25 ml of buffered alanine was added into all the test
tubes. This was followed by the addition of 0.05 ml of serum into the test group tubes and

0.05 ml of working pyruvate standard into the standard tubes. After proper mixing, all the
tubes were kept for incubation at 37oC for 60 minutes, after which 0.25 ml each of 2,4-
DNPH reagent was added into all the tubes. Then, 0.05 ml of distilled water and 0.05 ml
of each serum sample was added to the blank and the serum control tubes respectively.
The mixture was allowed to stand at room temperature for 20 min. After incubation, 2.5
ml of solution I was added to all test tubes. Mixed properly and optical density was read
against purified water in a spectrophotometer at 505 nm within 15 min.
The enzyme activity was calculated as:- ALT (GPT) activity in IU/L) =
[(Absorbance of test - Absorbance of control)/ (Absorbance of standard - Absorbance of
blank)] x concentration of the standard.
Determination of alkaline phosphatase (ALP)
Alkaline phoshatase from serum converts phenyl phosphate to inorganic phosphate and
phenol at pH 10.0. Phenol so formed reacts in alkaline medium with 4-aminoantipyrine in
presence of the oxidising agent potassium ferricyanide and forms an orange-red coloured
complex, which can be measured spectrometrically. The color intensity is proportional to
the enzyme activity.
Reagents:
Buffered substrate
Chromogen Reagent
Phenol Standard, 10 mg%
Procedure:
ALP was determined using the method of Kind (Kind PRM, King EJ, 1972. In-
vitro determination of serum alkaline phosphatase. Journal of Clinical Pathology 7: 321-
22\). The working solution was prepared by reconstituting one vial of buffered substrate
with 2.2 ml of water. 0.5 ml of working buffered substrate and 1.5 ml of purified water
was dispensed to blank, standard, control and test. Mixed well and incubated at 370C for
3 min. 0.05 ml each of serum and phenol standard were added to test and standard test
tubes respectively. Mixed well and incubated for 15 min at 370C. Thereafter, 1 ml of

chromogen reagent was added to all the test tubes. Then, added 0.05 ml of serum to
control. Mixed well after addition of each reagent and the O.D of blank, standard, control
and test were read against purified water at 510 nm.
Serum alkaline phosphatase activity in KA units was calculated as follows
[(O.D. Test-O.D. Control) / (O.D. Standard- O.D. Blank)] x 10
Determination of bilirubin
In toxic liver, bilirubin levels are elevated. Hyperbilirubinemia can result from
impaired hepatic uptake of unconjugated bilirubin, such a situation can occur in
generalized liver cell injury, certain drugs (e.g Rifampin and probenecid) interfere with
the rat uptake of bilirubin by the liver cell and may produce a mild unconjugated
hyperbilirubinemia. Bilirubin level rises in diseases of hepatocytes, obstruction to
bilirubin excretion into duodenum, in haemolysis and defects of hepatic uptake and
conjugation of Bilirubin pigment such as Gilbert’s disease.
Elevation of total serum bilirubin may occur due to:

Excessive haemolysis or destruction of the red blood cells.Eg:Haemolytic disease
of the new born.
Liver diseases.Eg.Hepatitis and cirrhosis.
Obstruction of the biliary tract.Eg.Gall stones.
The method is based on the reaction of Sulfonilic acid with sodium nitrite to form
azobilirubin which has maximum absorbance at 546nm in the aqueous solution. The
intensity of the color Produced is directly proportional to the amount of direct or total
bilurubin concentration present in the sample.
Reagents
1. Diazo A-(Reagent-R1) :Ready to use
2. Diazo B-(Reagent-R2):Ready to use
3. Bilirubin Activater :Ready to use

Procedure
Kind & King’s method was followed for the estimation of Bilirubin. Five hundred
µl of working reagent was added to 50 µl of rat serum & incubated for 5 min at 37°C.
Absorbance was measured AT 546 NM in semi auto analyzer against the standard.
The Bilirubin content was calculated using the following equation: Total bilirubin
(mg/dt) = Abs of the sample blank x 15.
Estimation of Urea
Urea is the nitrogen-containing end product of protein catabolism. States
associated with elevated levels of urea in blood are referred to as hyper uremia or
azotemia.
Method
Estimation of urea was done by Urease-GLDH: enzymatic UV test.
Principle
Urease
Urea + 2H2O 2NH4 + 2HCO3
2- Oxoglutarate +NH4+ +NADH GLDH
L- Glutamate +NAD+ + H2O
Table 7. Reagents
TRIS pH 7.8 120 mmol/l

2-Oxoglutarate 7 mmol/l
R1 ADP 0.6 mmol/l
Urease ≥ 6 KU/l
GLDH ≥ 1 KU/l
R2 NADH 0.25 mmol
R3 Standard 40 mg/dl

Procedure
a. Take 1000 µl of reagent-1 and 250 µl of reagent-2 in 5 ml test tube.
b. To this, add 10 µl of serum.
c. Mix well and immediately read the test sample at 340 nm Hg 334 nm Hg 365 nm
optical path 1 cm against reagent blank (2-point kinetic).
d. And note down the value.
Normal range: 10 – 50 mg/d
ESTIMATION OF URIC ACID

Uric acid and its salts are end products of the purine metabolism. In gout the
most common complication of hyperuricemia, ie. Increased serum levels of uric acid
lead to formation of monosodium urate crystal around the joints.
Method
Enzymatic photometric test using TOOS (N ethyl- N (hydroxyl -3- sulfopropyl)-
m- toluidin)
Principle
Uric acid + H2O + O2 uricase Allantoin +CO2 +H2O2
POD
TOOS + 4 aminoantipyrine + 2H2O2 Indamine + 3H2O
Table 8.reagents
R1 Phosphate buffer pH 7.0 100mmol/l
TOOS 1mmol/l
Ascorbate oxidase ≥1 KU/l
R2 Phosphate buffer pH 7.0 100mmol/l
4- amino antipyrine 0.3mmol/l
K4 (Fe( CN)6) 10µmol/l
Peroxidase ≥1KU/l
Uricase ≥50U/l

Procedure
a. Take 800µl of reagents -1 in a2ml centrifuge tube.
b. To this add 20µl of serum.
c. Mix well and incubate at 30°c for 5 minutes.
d. Then add 200µl of reagent2
e. Mix well incubate for 5min at 37°c
f. Measure the not down the values.
Normal range: 1.9-8.2mg/dl
ESTIMATION OF CREATININE:
Estimation of Creatinine by Jaffe Method (modified)
Principle:
Creatinine forms a coloured complex with picrate in alkaline medium. The rate of
formation of the complex is measured.
Reagents:
Reagent 1 Standard Creatinine (2mg/100ml)
Reagent 2 Picric acid solution
Reagent 3 Sodium hydroxide solution
Procedure:
Take 500 µl of reagent -2 and 500 µl of reagent -3 in a 5ml test tube. To this add
100 µl of serum. Mix well and immediately read the test sample at Hg 492 nm 1cm light
path and note down the values.
Normal range is 0.6 -1.1 mg/dl.

4.4. PHARMACOLOGY STUDY
4.4.1.EVALUATION OF IN VIVO CLOT LYSIS IN RAT MODELS
In vivo studies were carried out by the oral administration of aril and rind of
VENTHAMARAIYATHI CHOORANAM to examine the clot lysing ability,
antioxidative and cytotoxic effect.
Thrombus induction
Experimental rats were anaesthetized with i.p injection of ketamine hydrochloride

(100 mg/kg). To attain effective clot formation κ – carrageenan (1 mg/kg) dissolved in
saline was injected into the rat tail vein at a site 12 cm from the tip of the tail with a
ligation. After a period of 10 minutes, the ligature was removed. The length of the infarct
was monitored for thrombus formation. Once thrombus was formed, the animals were
treated with respective extracts and monitored for the reduction in the length of the
thrombus in rat tail for 30 days.
Table – 9 Dose level

DOSE TO ANIMALS
TEST GROUP
(mg/kg body-weight/day)
GROUP-1 CONTROL
GROUP-II ONLY CARRAGEENAN
GROUP-III CARRAGEENAN+STD
CARRAGEENAN+ LOW DOSE OF

GROUP-IV VENTHAMARAIYATHICHOORANAM 300MG/KG
CARRAGEENAN+ MIDDLEDOSE OF
GROUP-V VENTHAMARAIYATHICHOORANAM 600MG/KG
CARRAGEENAN+ HIGH DOSE OF

GROUP-VI
VENTHAMARAIYATHICHOORANAM 900MG/KG
period, all animals were observed for 30 days.

Estimation of D-dimer
(Marder and Francis, 1983)
Fibrin degradation products (FDP), which are highly heterogeneous soluble
fragments, are a result of 2simultaneous phenomena:
• Fibrinogen is coagulated by thrombin and factor XIIIa toform stabilized fibrin,
• The fibrin clot is dissolved by plasmin into solublefragments released into the
blood. The terminal productof fibrinolysis is D-dimer.
Principle
The assay principle combines a two-step enzymeimmunoassay sandwich method
with a final fluorescentdetection (ELFA). The Solid Phase Receptacle (SPhR) serves as
the solid phase with an anti-FDP monoclonal antibody adsorbed on its surface.
Procedure
The sample was taken and transferred into the wellcontaining an alkaline
phosphatase – labeled anti-FDP monoclonal antibody. The sample mixture was cycled in
and out of the SPhR several times to increase the reaction speed. The antigen binds to
antibodies coated on the SPhR and to the conjugate forming a “sandwich”. The remaining
free antigen sites were saturated by cycling the conjugate in the fifth well of the strip in
and out of the SPhR. Unbound components wereeliminated during the washing steps.
Two detection steps were then performed successively. Duringeach step, the
substrate (4-Methyl-umbelliferyl phosphate) wascycled in and out of the SPhR. The
conjugate enzymecatalyzes the hydrolysis of this substrate into a fluorescentproduct
(4-Methyl-umbelliferone), the fluorescence of whichis measured at 450 nm. The intensity
of the fluorescence isproportional to the concentration of antigen present in thesample.
Estimation of tissue plasminogen activator

(Kit method - Roche)
Tissue-type plasminogen activator (tPA) is one of two major physiologic
activators of plasminogen in plasma. The activation of plasminogen by tPA is dependent
on the presence of a fibrin cofactor. The binding of both tPA and plasminogen to fibrin is
mediated in part through lysine binding sites within the kringle structures of both enzyme

and substrate, but also through the finger domain of tPA. Activation of plasminogen by
tPA occurs by cleavage after residue Arg560 to produce the two-chain active serine
protease plasmin.
Principle
Affinity-purified antibody to tPA is coated onto the wells of a microtitre plate. Any
remaining binding sites on the plastic wells are blocked with an excess of bovine serum
albumin. The plates are washed and plasma or other fluids containing tPA are applied.
The coated antibody will capture the tPA in the sample. After washing the plate to remove
unbound material, a peroxidase conjugated second antibody to tPA is added to the plate to
bind to the captured tPA. After washing the plate to remove unbound conjugated
antibody, the peroxidase activity is expressed by incubation with O-enylenediamine
(OPD). After a fixed development time the reaction is quenched with the addition of
H2SO4 and the colour produced is quantified using a microplate reader. The colour
generated is proportional to the concentration of tPA present in the sample.
Reagents
1. Capture antibody (TPA–EIA-C)
2. Detecting antibody (TPA-EIA-D)
3. Coating Buffer: 50 mM Carbonate: 1.59g of Na2CO3 and 2.93g of NaHCO3 up to
1 litre. Adjust pH to 9.6.
4. PBS (base for wash buffer and blocking buffer): 8.0g NaCl, 1.15g Na2HPO4, 0.2g
KH2PO4 and 0.2g KCl, up to 1 litre. Adjust pH to 7.4, if necessary.
5. Blocking Buffer: PBS-BSA (1%, w/v) Dissolve 2.5 g of Bovine Serum Albumin
(Sigma-RIA grade) in 200 ml of PBS. Adjust pH to 7.4, if required, then make up
to 250 ml with PBS.
6. Sample Diluent (HBS-BSA-T20): 5.95g HEPES (free acid), 1.46 g NaCl, 2.5 g
Bovine Serum Albumin dissolved in 200 ml H2O. Add 0.25 ml of Tween- 20,
check and adjust pH to 7.2 with NaOH, then make up to a final volume of 250 ml
with H2O.

7. Substrate Buffer (Citrate-Phosphate buffer - pH 5.0) 2.6g Citric acid and 6.9g
Na2HPO4 up to a final volume of 500 ml with purified H2O.
8. OPD Substrate (o-Phenylenediamine.2HCl) TOXIC!: Make up immediately
before use. Dissolve 5mg OPD in 12 ml substrate buffer then add 12 µl 30%
H2O2.
9. Stopping Solution: 2.5 M H2SO4 (Where stock sulphuric acid is 18 M, add 13.9
ml to 86 ml H2O).
Procedure
1. Coating of plates: Dilute the capture antibody 1/100 in coating buffer (in a
polypropylene tube) and immediately add 100 µl per well in the plate. Incubate
for 2 hours at ambient temperature or overnight at 2-80C.
2. Blocking: Empty contents of plate and add 150 µl of blocking buffer to every well
and incubate for 60 minutes at 220C. Wash plate X 3 with wash buffer.
3. Preparation of tPA Reference Standards: Reconstitute vials of tPA standard and
tPA/PAI-1 deficient plasma according to manufacturer’s instructions. After
reconstitution, dilute the tPA standard into tPA/PAI-1 deficient plasma to achieve
six reference standard plasmas with final tPA concentrations of 50, 25, 12.5, 6.25,
3.13 and 1.56 ng/ml respectively.
4. Samples: Reference plasmas prepared in step 3 and test plasmas are diluted 1/4
and 1/8 in HBS-BSA-T20 sample diluent. Samples should be run in duplicate.
Apply 100 µl/well and incubate plate at 220C for 90 minutes. Wash plate X 3 with
wash buffer.
5. Detecting Antibody: Dilute the detecting antibody 1/100 in HBS-BSA-T20
sample diluents and apply 100 µl to each well. Incubate plate at 220C for 90
minutes. Wash plate X 3 with wash buffer.
6. OPD Substrate: Apply 100 µl of freshly prepared OPD substrate to every well.
Allow colour to develop for 5-10 minutes then stop colour reaction with the
addition of 50µl/well of 2.5 M H2SO4. The plate can be read at a wavelength of
490 nm.

Determination of Bleeding Time
(Brown, 1993)
Apparatus
Blood lancet, spirit, pieces of filter paper, stop watch
Procedure
With usual aseptic precautions, a puncture wound was inflicted. It is to be more
deep than usual and should be done in a standard manner. The time of puncture and first
appearance of the blood was noted. The blood was gently blotted with a filter paper. Care
should be taken not to press or wipe the wound. It was repeated with a fresh piece of
filter paper for every 10 seconds till no blood appears on the paper. The number of filter
paper that shows the blot of blood on it was counted. Number of blot x 10 seconds will be
bleeding time. Similar experiments were carried out on a healthy animal and the result
obtained was considered as a control value. Normal bleeding time is 1 – 5 minutes.
Blood clotting time

The ability of N. laevis extract to inhibit in vitro coagulation of blood was
quantitatively assayed following a standard procedure adopted by Koffuor and Amoateng
(2011). Briefly, 1.0 ml of whole blood drawn from the marginal ear vein of rabbits was
added separately to 0.2 ml of 5 % and 10 % w/v of NLMLE in test tubes placed in a
water bath at 37 °C. The time taken for the blood to clot in the separate concentration of
the extract was recorded. A repeat of each test was performed to obtain five determinants
in coagulation times exerted with differences in concentration of the extract. Controls
were coagulation time for blood in 0.2 ml distilled water or blood alone as baseline
clotting times.
Determination of euglobulin clot lysis time

(Nordbyet al., 1980)
The euglobulin clot lysis time is a test that reflects the overall fibrinolytic activity
of plasma.

Principle and Method:
Venous blood is collected into chilled tubes containing trisodium citrate as an
anticoagulant and placed on ice. The sample is then centrifuged at 4°C and the plasma
sample is collected, diluted with acetic acid and incubated on ice for 15 minutes. A
precipitate forms [the euglobulin fraction of plasma] which contains plasminogen,
plasminogen activators [primarily t-PA] and fibrinogen. The supernatant is collected by
centrifugation in refrigerated centrifuge at 4°C. The supernatant is discarded and the
precipitate is dissolved in buffer. This is then clotted with thrombin and the time to clot
lysis is determined by inspection every 15 minutes. A control plasma sample collected at
the same time must be run in parallel.
4.4.2. VASODILATOR ACTIVITY OF STUDENT ORGAN BATH METHODS
Materials and Methods

Chemicals
Phenylephrine chloride (PE), Nω-nitro-L-arginine (L-NA), atropine sulfate,
acetylcholine chloride (Ach), rutin hydrate, oleuropein, kaempferol disaccharides, and
quercetin were purchased from Sigma-Aldrich Chemical, USA. All other chemicals used
were of analytical grade.
Animals
Wistar rats (Rattusnorvegicus) and ICR mice (Musmusculus) used were obtained
from the National Laboratory Animal Center,Baide Metha Pharmacy College,
Thuraipakkam, Chennai. All animals were acclimatized for 1 week before starting of the
experiments in controlled environmental conditions (25 ± 1°C) with a 12 h light/dark
cycle and allowed access to standard food and tap water ad libitum. Animal welfare was
under control by IACUC of the Baide Metha Pharmacy College.
Plant Material and Extraction

The flowers Venthamarai were collected from thovalai , kanniyakumari dist in
October 2015. The specimen voucher was TISTR no. 160309, and the samples were
deposited at TISTR. Approximately 3.1 grams of the flowers were dried at 50°C for 42 h,
powdered, and macerated in 18 mL of 95% ethanol at room temperature overnight. After

elution, the flower residues were repeatedly macerated with equal volume of ethanol
overnight and eluted again. The ethanol elutes were combined, filtered through
Whatmann filter paper no. 42, and evaporated under reduced pressure at 50°C. The
semisolid light yellow materials were stored in desiccators until used. Percentage yield of
the extract was 17.68% yield (w/w). The extract was dissolved in 0.05%
dimethxylsulfoxide (DMSO) for in vitro experiments and in 1% Tween or 1% gum
tragacanth for in vivo studies.
Vasodilatation Effect Test
Each rat was killed by cervical dislocation. Its thorax was opened, and the aortic
vessel was removed from fat and connective tissues and kept in a Petri dish containing
Krebs' solution. Two adjacent aortic rings of 3-4 mm in length were cut. In only one ring,
endothelium was removed mechanically by gently rubbing the intimal surface of the
vessel using the method as previously described. The organ bath contained the Krebs-
Henseleit solution (NaCl 119, KCl 4.7, CaCl2 2.5, MgSO4·7H2O 1.0, KH2PO4 1.2,
NaHCO3 25.0, and glucose 11.1 mM). This solution was maintained at 37°C and
continuously bubbled with 95% O2 and 5% CO2. The two rings were individually
suspended horizontally between two stainless steel hooks in a 20 mL organ bath
containing Kreb's solution. One of the hooks was fixed to the bottom, and the other was
connected to a force displacement transducer that connected to a MP100 (BIOPAC
System, Inc., Model MP100 WSW) for the isometric tension record. The stabilization
period was of 45 min under a resting tension of 1.0 g and the solution was changed every
15 min to prevent the accumulation of metabolites. After equilibration, the rings were
preconstricted with 1 × 10−6 M phenylephrine (PE) until the responses curve reached
plateaus (5–8 min), and dilator responses to 1 × 10−5 M acetylcholine (Ach) were
detected. The absence of the relaxation to acetylcholine was taken as evidence that the
vessel segment was functionally denuded of endothelium, and at least 70% vasodilatation
to acetylcholine for theendothelium-intact thoracic aorta rings was observed.
Effect of the VenthamaraiChoornam Extract on PE-Induced Tonus in the
Endothelium-Intact or Endothelium-Denuded Thoracic Aorta Rings
After 45 minutes re-equilibration, the rings with and without endothelium intact
were pre-contracted with 1 × 10−6 M PE for 20 minutes. After several washings and re-

equilibration for 30 minute, five different concentrations of the VTC extracts (50, 100,
200, 300, and 400 µg/mL) were added 5 min and the rings were incubated with PE for
another 20 min. The contractions were measured by comparing the developed tension
before and after the addition of the extract and expressed as percentage of contraction
from induced tonus.
Effect of the Extract on PE-Induced Tonus in the Endothelium-Intact Thoracic
Aorta Rings with Atropine and Nω-Nitro-L-arginine
After 45 min re-equilibration, the endothelium-intact aortic rings were pre-
constricted with 1 × 10−6 M PE for 20 min. After several washings and re-equilibration
for 30 minutes, the rings were exposed to atropine (1 × 10−6 M) or Nω-nitro-L-arginine
(L-NA), a nitric oxide synthase inhibitor (1 × 10−4 M), for 5 minutes. Then, the same
serial concentrations of the extracts were added, and the percentage of the contractions
before and after the addition of the extracts was determined as described

4.4.3. HYPOLIPIDEMIC ACTIVITY OF SIDDHA FORMULATION
VENTHAMARAIYATHI CHOORANAM IN HYPERLIPIDEMIC
MODELS OF WISTAR ALBINO RATS
INTRODUCTION:
Many people with diabetes have conditions called “risk factor” that contribute to
atherosclerosis and its complications. These include high blood pressure, excess weight
and high blood glucose levels. Dyslipidemia further raises risk of atherosclerosis in
people with diabetes. Dyslipidemia affects people with type 2 diabetes more often than
those with type 1 diabetes. The most common Dyslipidemia in diabetes is the
combination of high triglycerides and low HDL levels. People with diabetes may also
have elevated LDL cholesterol.
Among the drugs available to treat Dyslipidemia, statins are often the first choice
for lowering total and LDL cholesterol levels, other drugs that lowers cholesterol include
cholesterol-adsorption blockers, bile acids, sequestrants, and nicotinic acids. These may
be used in combination, if a single drug is not effective in reaching target levels. Fibrates
and extended release niacin may be used to lower triglycerides (or) raise HDL cholesterol
levels.
Hyperglycemia and Dyslipidemia are significant and independent risk factors for
the vascular complications and suggested to cause cardio vascular pathological changes
in diabetic states through the following molecular mechanism, formation and
accumulation of advanced glycation products, increased oxidative stress, activation of
proteinkinase C pathway , increased activity of hexosamine pathway and vascular
inflammation and the impairment of insulin action in the vascular tissues. As siddha
formulation venthaa marayathy chooranam have been traditionally claimed for the
treatment of HYPOLIPIDEMIC, hence in the present study an attempt has been made to
screen the siddha formulation Venthamaraiyathi Chooranam for the Hypolipidemic
activity..
The present investigation is undertaken to study the effect of siddha formulation
Venthamaraiyathi Chooranam on changes in Total cholesterol, Triglycerides, HDL,
LDL, VLDL, AI, and LDL/HDL.

Materials and methods
Animals
Wister albino rats were obtained from central animal house, K.M.College of
pharmacy, Madurai. The animals were given standard rodent diet and water ad libitum
throughout the study. The rats used in the present study were maintained in accordance
with guidelines of the national institute of nutrition, Indian council for medical
research, Hyderabad, India and study approved by Institutional animal ethical
committee.
Materials:
• Siddha formulation venthaa marayathy chooranam
• Cholesterol extra pure for feeding purpose was obtained from S D fine-chem.
limited, Mumbai, India. Coconut oil was used as a vehicle for cholesterol
feeding.
• Atorvastatin was obtained from Micro labs, Bangalore, India.
Experimental procedure:
All the animals were weighed and divided into five groups each of six animals.
Group I : Normal control.
Group II : Cholesterol control. Fed cholesterol at a dose of
400mg/kg body weight for 30 days.
Group III : fed cholesterol as in group II and Atorvastatin
1mg/kg body weight from days 15 to day 30. (3)
Group IV : fed cholesterol as in group II and siddha formulation
Venthamaraiyathi Chooranam at a dose of 200mg/kg body
weight from days 15 to day 30.
Group V : Fed cholesterol as in group II and siddha formulation
Venthamaraiyathi Chooranam at a dose of 400 mg/kg body
weight from days 15 to day 30.
At the end of 30 days all the rats were sacrificed, blood was collected, allowed
to clot and serum was obtained by centrifugation. The serum samples were used for
various biochemical procedures.

Biochemical analysis:
The serum was analyzed for total cholesterol, triglycerides, high density
lipoprotein (HDL), low density lipoprotein (LDL), very low density lipoprotein
(VLDL), by using standard protocol methods. (Auto Analyzer)
Atherogenic index (AI) and LDL-C/HDL-C ratio

• The AI was calculated by the following formula
• AI = (total cholesterol – HDL-C)/HDL-C
• LDL-C/HDL-C ratio was calculated as the ratio of plasma LDL-C to HDL-C
Levels
Statistical analysis
• All the values were expressed as mean ± SEM.
• Data was analyzed by one way analysis of variance (ANOVA) followed by
Newmankeuls multiple test.
• P values <0.05 were considered as statistically significant.

4.4.4. CARDIOPROTECTIVE EFFECT OF VENTHAA MARAYATHY
CHOORANAM AGAINST DOXORUBICIN INDUCED MYOCARDIAL
TOXICITY IN ALBINO RATS
INTRODUCTION
Doxorubicin (DOX), an anthracycline antibiotic, is an excellent drug for the
treatment of a wide variety of human solid tumors and leukemias. However, its clinical
uses are limited by seriously high incidence of cardiotoxicity. An initial acute effect
includes hypotension and transient electrocardiographic abnormalities. Meanwhile, the
chronic effects may occur several weeks or months after cumulative DOX administration.
Cardioxmyophathy is dose dependent which accounts for high mortality . The possible
mechanisms proposed for cardiotoxic effects of DOX include free radical induced
myocardial injury, lipid peroxidation, mitochondrial damage, decreased activity of Na+–
K+ adenosine triphosphate, vasoactive amine release and cellular toxicity . Increased
oxidative stress and release of free radicals, including super oxide anion (O2-) and other
reactive oxygen intermediates as well as endogenous antioxidant deficits have been
suggested to play a major role in Dox-induced cardiomyopathy and heart failure .
Additionally, DOX has a very high affinity by cardiolipin, a phospholipid that is present
in mitochondrial membranes of heart, resulting in the accumulation of DOX inside
cardiac cell Moreover, in recent years, it has been observed that there is a growing
interest in the usage of natural antioxidants as a protective strategy against
[10]
cardiovascular-related problems in experiments such as ischemia reperfusion and
Dox-induced cardiotoxicity .The aim of this study was to evaluate the carioprotective
properties of Venthamaraiyathi Chooranam against Dox-induced cardiotoxicity in rats.
Chemicals and drugs
Dox was a gift from Get Well Pharmaceuticals, India. Other chemicals were
obtained from Sigma and analyzing kits were obtained from ERBA. All the chemicals
were of the analytical grade.
Animals
Albino wister rat (150-225 gm) were produced from animal experimental
laboratory, and used throughout the study. They were housed in micro nylon boxes in a
control environment (temp 25±2°C) and 12 hrs dark /light cycle with standard laboratory

diet and water ad libitum. The study was conducted after obtaining institutional animal
ethical committee clearance. As per the standard practice, the rats were segregated based
on their gender and quarantined for 14 days before the commencement of the experiment.
They were fed on healthy diet and maintained in hygienic environment in our animal
house.
Treatment protocol
After one week of acclimatization, the animals were randomly divided into 5
groups of 6 animals and treated as follows- Group 1 served as Normal control and
received normal saline 5 ml/kg body weight (i.p.). Group 2 animals were treated with
Dox (2.5 mg/kg body weight, i.p.) in 6 equal injections alternatively for two weeks to
make a total cumulative dose of 15 mg/kg body weight. Group 3 animals were treated
with Dox (2.5 mg/kg body weight, i.p.) and Vitmanin E. Group 4 animals received
Venthamaraiyathi Chooranam with low dose and Dox (200 mg/kg body weight). Group
5 animals received VENTHAMARAIYATHI chooranam extract with high dose and Dox
(400 mg/kg body weight).
Evaluation of myocardial infarction

Biochemical analysis
Dissection and Homogenization
After the experimental period the rats were sacrificed by euthanasia method.
Blood was collected and serum supernatant was for the assay of marker enzymes
Alkaline Phosphatase(ALP), lactate dehydrogenase (LDH), Aspartate Transaminase
(AST), Alanine Transaminase (ALT) and Creatine Phosphokinase (CPK).
Statistical analysis
The results are expressed as mean ± S.E.M. The results were analyzed using one-
way ANOVA followed by Newman Keul’s multiple rang. Data was computed for
statistical analysis by using GraphPad InStat version 3.00 of GraphPad Software, Inc.
(San Diego, CA).

4.5. ANTI - MICROBIAL ACTIVITIES BY WELL DIFFUSION METHOD
Aim:
The antimicrobial activity of Venthamaraiyathi Chooranam was adapted through
Well diffusion method(Agar diffusion testing).
PRINCIPLE
The antimicrobials present in the plant extract are allowed to diffuse out into the
medium and interact in a plate freshly seeded with the test organisms. The resulting zones
of inhibition will be uniformly circular as there will be a confluent lawn of growth. The
diameter of zone of inhibition can be measured in millimeters.
Components of Muller Hinton agar medium:

Beef extract - 2gm/lit
Acid Hydrolysate of Casein - 17.5 gm/lit
Starch - 1.5 gm/lit
Agar - 17gm/lit
Distilled water - 1000 ml
PH - 7.3 0.1 at 250C
PROCEDURE (Murray et al., 1995)

Petriplates containing 20ml Muller Hinton medium were seeded with 24hr
culture of bacterial strains. Wells were cut and 20 µl of the plant extracts (aqueous) were
added. The plates were then incubated at 37°C for 24 hours. The antibacterial activity
was assayed by measuring the diameter of the inhibition zone formed around the well
(NCCLS, 1993). Streptomycin was used as a positive control. standard for an In-vitro
antimicrobial activity of Venthamaraiyathi Chooranam was screened against bacteria
strains such as Klebsiella pneumonia, Streptococcus mutans, Staphylococcus aureus,
Escherichia coli, Pseudomonas.aerugina, Enterococcus faecalis.
Murray PR, Baron EJ, Pfaller MA, Tenover FC, Yolken HR Manual of Clinical
Microbiology, 6th Ed. ASM Press, Washington DC, 1995; 15-18.

5. RESULTS AND DISCUSSION
STANDARDIZATION OF THE TEST DRUG

Standardisation of the drug is more essential to derive the efficacy, potency of the
drug by analysing it by various studies. Following are the results of physicochemical and.
Physical characterisation and estimation of basic and acidic radicals have been done and
tabulated.
Toxicological results of the drug and pharmacological activity of the drug were
derived. Its result has been tabulated and interpretation was made below. Thus it is to give
a complete justification to bring the effectiveness of the trial drug Venthamaraiyathi
Chooranam .
Table : 10 Organoleptic Characters
S.No. Parameter Results
1. a. Colour Yellowish L.Green
2. b. Odour Pleasant Odour
3. c. Sense of Touch Nice
4. d. Appearance Powder
5. e. Taste L.Bitter, L.Sweet
Interpretation:
The organoleptic characters of the drug Venthamaraiyathi Chooranam showed
that the colour of the chooranam is Yellowish L. Green in colour since prepared from
dry herbs and minerals, L. Bitter, L. Sweet in taste which might be responsible for the
activity mentioned earlier and on sight they are fine powder.
The fineness of the chooranam represents easy absorption and better

availability of the drugs.
The size of the particle is reduced through various stages like pounding,
sieving, filtering through white cloth (vasthirakaayam).
Only if the size of the particle is reduced to micro and nano particles, the drug
is easily assimilable in the digestive system.
The above processes reduced the size of the particle so that the chooranam
passes through the sieve.

TABLE : 11 PHYSICO CHEMICAL ANALYSIS
S.NO Parameter Results
1 Loss on Drying 6.3%
2 Total Ash 6.1%
3 Acid Insoluble ash 2.3%
4 Water soluble Extractive 26.7%
5 Alcohol Soluble Extractive 21.7%
6 Total Viable aerobic count 1.3 x 104 col/g
7 Total Enterobacteriaceae Nil
8 Total fungal count
9 Test for specific pathogens 3 x 10 2 col/g
10 Salmonella sp Nil
11 Staphylococcus aureus Nil
12 E.coli Nil
13 Pseudomonas aeruginosa Nil
Interpretation of Physico chemical analysis

Ash Values
The ash remaining following ignition of medicinal plant materials is determined
by three different methods which measure total ash, acid insoluble ash and water
insoluble ash.
The total ash method is designed to measure the total amount of inorganic
material remaining after ignition.
It measures the total inorganic content (ammonium, silica, potassium, calcium,
chloride, iron, etc.) present in the drug.
This includes both “physiological ash”, which is derived from the plant tissue
itself and “non-physiological ash” which is the residue of the extraneous matter like sand
and soil adhering to the plant surface.

Total Ash
The ash value of the drug was determined as 23.5% which is high when compared
to other herbo mineral drugs but the increased ash content may be due to the presence of
inorganic materials. Thus Ash value is a validity parameter describe and to assess the
degree of purity of a given drug
Acid insoluble ash:

The acid insoluble ash value of the drug denotes the amount of siliceous matter
present in the plant. The quality of the drug is better if the acid insoluble value is low. It is
2.3% for VTC
Water soluble ash

Water-soluble ash is the part of the total ash content, which is soluble in water. It
is 26.7%for VTC Water-soluble extractive value plays an important role in drug delivary
system.
Alcohol-soluble ash:
Alcohol-soluble ash is the part of the total ash content, which is soluble in
alcohol. It is 21.7% for VTC
These are indicating the approximate measure of chemical constituents of crude
drug.
The percentage of soluble matters present in the drug is determined by the values
of water extractive and ethanol extractive.
Based on the extractive value suitable solvent can be selected. It also gives the
percentage of drug which will correlate with the metabolism reactions.
Water-soluble extractive value plays an important role in evaluation of crude
drugs
The alcohol-soluble extractive value was also indicative for the same purpose as
the water-soluble extractive value

Loss on drying:
The total of volatile content and moisture present in the drug was established in
loss on drying.
Moisture content of the drug reveals the stability and its shelf-life.
High moisture content can adversely affect the active ingredient of the drug.
Thus low moisture content could get maximum stability and better shelf life.
Since the drug has low loss on drying, the moisture content is less which is
suitable for medicine preparation
Microbial Limit Tests

The total bacterial count and the total fungal count of the drug were found to be
within the WHO prescribed limits which indicate that the drug is free from microbial
condamination. The other pathogens like Escherichia coli, Salmonella sps,
Pseudomonoas aeruginosa and Staphylococcus aureus were found to be completely
absent in the drugs.

Thin Layer Chromotography :
Under UV 254nm and 366nm
Fig : 5 TLC sheet

Interpretion:
Under UV 254 nm and 366 nm test related to alkaloids, it shows major spots short
at Rf 0.74(violet), 0.50(violet), 0.49(violet), 0.36(violet), 0.29(violet), 021(green) and
long at Rf 0.74(blue), 0.50(blue), 0.49(blue) ,0.36(blue) ,0.29(blue), 0.21(green) . show 2
compounds are found.

CHEMICAL ANALYSIS
Table:12 Result of Acidic and Basic Radicals
S.No Test Inference
1 Test for calcium +ve
2 Test for sulphate +ve
3 Test for chloride +ve
4 Test for carbonate -ve
5 Test for starch +ve
6 Test of iron ferrous +ve
7 Test for phosphate +ve
8 Test for Albumin -ve
9 Test for Tannic acid -ve
10 Test for unsaturation +ve
11 Test for reducing sugar +ve
12 Test for amino acid +ve
13 Test for zinc -ve
Interpretation :
The chemical analysis of venthamarayathi chooranam contains the following
chemical constituents ferrous Iron, calcium, Sulphate, chloride,Starch, iron ferrous,
phosphate, Unsaturated, reducing sugar ,amino acid compounds.
Calcium
Presence of calcium improves the physical strength of skeletal tissue. Calcium
ions are necessary for muscle contraction. Calcium ions are necessary for the normal
transmission of nerve impulse
Unsaturated fats
Monounsaturated and polyunsaturated fats can replace saturated fat in the diet,
trans unsaturated fats should not. Replacing saturated fats with unsaturated fats helps to
lower levels of total cholesterol and LDL cholesterol in the blood.

Chloride
Chloride is an anion in the human body needed for metabolism (the process of
turning food into energy) It also helps keep the body's acid-base balance. The amount of
serum chloride is carefully controlled by the kidneys.
Chloride ions have important physiological roles. For instance, in the central
nervous system, the inhibitory action of glycine and some of the action of GABA relies
on the entry of Cl− into specific neurons
Starch
Digestive enzymes have problems digesting crystalline structures. Raw starch
will digest poorly in the duodenum and small intestine, while bacterial degradation will
take place mainly in the colon.
Iron
iron metabolism is the set of chemical reactions maintaining human
homeostasis of iron at both the systemic and cellular level. The control of this necessary
but potentially toxic metal is an important part of many aspects of human health and
disease. Hematologists have been especially interested in systemic
iron metabolism because iron is essential for red blood cells, where most of the human
body's iron is contained.
Phosphate
Parathyroid hormone(PTH), and calcitriol also regulate phosphate in the body. PTH
helps lower blood phosphate levels. It does this by reducing the reabsorption of
phosphates dissolved in urine in the kidneys, causing more excretion of phosphates.
Calcitriol raises the level of phosphate in the blood by promoting its absorption by the
intestine.
Amino acids
Amino acids derivatives include 5-HTP (5-hydroxytryptophan) used for
experimental treatment of depression L-DOPA (L-dihydroxyphenylalanine) for
Parkinson's treatment
VTC drug that inhibits ornithine decarboxylase and used in the treatment of sleeping
sicknes

SCANING ELECTRON MICROSCOPE (SEM) OFVENTHAMARAYATHI
CHOORANAM
Fig. 6
SEM Picture 28, 000 Magnification of VT
SEM Picture 5,000 Magnification of VT

SEM images showing the shape and size of the particle size of the drug VTC
Interpretation
The morphology of the VT C sample can be determined by Environmental SEM
(FEI Quanta). A representative portion of each sample must be sprinkled onto a double
side carbon tape and mounted on aluminium stubs, in order to get a higher quality
secondary electron image for SEM examination. We have observed from SEM
photographs that particles are spherical in shapes and sizes are in the range from 0.1
micron to 0.5 micron. Although the particle sizes of different batches showed similarity, it
seems that these particles are aggregates of much smaller particles. When dispersed in an
aqueous medium, these preparations form a negatively charged hydrophobic particle
suspension. This hydrophobicity gives these particles a tendency to aggregate together to
form larger particles. Chooranam exhibited larger sizes and agglomeration of the
particles. Therefore, the comparatively larger size may be due to the agglomeration of the
particles by repeated purification process involved in preparation.

ICP - OES RESULT:
Venthamarayathi Chooranam weight = 0.32515gm
Table No. 13
Elements symbol Wave length (nk) Concentration (Mg/ l)
Al 396.152 BDL
AS 188.979 BDL
Ca 315.807 12.170 mg/L
Cu 327.393 BDL
Fe 238.204 01.306 mg/L
Hg 253.652 BDL
K 766.491 33.801mg/L
Mg 285.213 01.304 mg/L
Na 589.592 24.310 mg/L
Ni 231.604 BDL
Pb 220.353 BDL
P 213.617 88.341 mg/L
S 180.731 01.324 mg/L
Zn 206.200 00.258 mg/L
Interpretation:
The result reveals the below detection limit (BDL) of Aluminimum (Al), Arsenic
(As), Copper (Cu), Mercury (Hg), Nickel (Ni), Lead (Pb). This reveals the safety of the
drug.
The result indicates the presence of Calcium (Ca), Iron, Pottasium (K),
Magnesium (Mg), Sodium (Na), Phosphorus (P), Sulphur (S), Zinc (Zn).
Calcium:
Calicum was associated with the lower risk of Hypertension.
By inhibiting calcium influx into cytosol, histamine release from mast cell,
acetylcholine release from cholinergic nerve endings
It also may increase the vasodilator effect of beta2-agonist by increasing the
receptor affinity
Iron:
The heme containing enzyme such as catalase and peroxidase protect cell against
portentially damaging highly reactive species.
Iron is essential for may numbers of biological functions such as growth,
reproduction, healing and immune function.

Iron is associated with effective immune competence of the body.
Iron plays an important role within the cells that produce the energy.
Potassium
Potassium dilates the arteries and relaxes the smooth muscles and increases the
blood flow (F.J. Haddy, 2012).
An adequate amount of potassium on consumption, reacts as vasodilator, brings
blood flow freely in body ,
Avoid the formation of clot and reduce the cause of stroke
It controls electrical activity of heart
Magnesium
Magnesium is a cofactor that regulates diverse biochemical reactions in the body,
including protein synthesis, muscle, nerve function, blood glucose control and
blood pressure regulation (Rude.R.K, 2012).
It is an important mineral to several body functions
It enhances the function of the endothelium, which is the inner most layer of
blood vessels. & mucus membrane
Zinc
Zinc is a micro nutrient and is required for wound healing.
Zinc has effects against viruses. {Rhino virus}.
Zing may be regarded as an antioxidant, protects the body against free radical
damage and cell damage.
Zinc is important for a healthy immune system
It enhances absorbtion of iron.
It can produce healthy veins and arteries that enhance the blood circulation
Sodium:
Sodium regulates the body’s acid base balance
Sodium is required for the maintenance of osmotic pressure and fluid balance
It is necessary for the normal muscle irritability and cell permeability

Phosphorus:
It is requried for the formation of phospholipids & Nucleic acids .
It palays a central role for the formation and utilization of high energy phosphate
compounds
Maintenance of PH in the blood as well as in the cells.
Sulphate
Sulphate may prevent the occurrence of any infection
Sulphate is potent anti oxidant activity in human body

Fourier Transform Infrared Spectroscopy Analysis
100.0
95
2131
90 VENTH CHO0OR
472
85 460
762
80
75
70
65
711
60
55
50
%T
544
45
40 873
35 940
917
30
25
2854
20
15
1425
10 2925 1632 1031
5 3381
0.0
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 450.0
cm-1
Fig : 7 Graph of FTIR analysis

Table : 14 Results of FTIR analysis of VTC
Frequency, cm-1 Bond Functional group

3381 (m) N – H stretch Primary, secondary, amines, amides
2925 (m) C – H stretch Alkenes

2854 (m) C – H stretch Alkenes
2131 (w) C (triple bond) C- stretch Alkynes
1632 (m) N – H bend Primary amines
1425 (m) C –C stretch (in – ring) Aromatics
1031 (m) C – N stretch Aliphatic amines
900 (s) C – H “oop” Aromatics
762 (m) O – H bend Carboxylic acids
711 (m) O – H bend Carboxylic acids
675 (s) C – H “oop” Aromatics
544 (m) C – CI stretch Alkyl halides
M = medium, w = weak, s=strong,
Interpretation:
The FTIR has proven to be a valuable tool for the charecterisation and
identification of compounds or functional groups present in the VTC
In FTIR the wavenumbers between 4000cm-1-400cm-1 is known as functional
group area. <400cm-1 wave number is known as finger print area. The corresponding
absorbtion frequency by FTIR shows the presence of, Alkanes,
Amines,Aromatics,,Aliphatic amines , Alkyl halides, Carboxylic acids. The C-H stretch
may be due to the presence of moisture content. Infrared bonds for inorganic materials
appear in the lower wave numbers than those observed for organic materials.
In cofirms that Venthamarayathi chooranam contains Alkanes,
Amines,Aromatics,,Aliphatic amines , Alkyl halides, Carboxylic acids

Amines groups:
Amines groups act as neurotransmitters.
It is involved in protein synthesis.
This group of substances has antihistaminic and analgesic activity.
Amines are a class of compounds derived from ammonia by replacement of
one or more effective antagonists of SSTR5 (Stomatostatin receptor 5)and are
used for treatment, control and prevention of disorders such as ,lipid disorders
and obesity
Alkanes groups:
Alkanes have little biological activity.
It is predominate in plants. They protect against bacteria and fungi
Alkynes:
Alkyne is an unsaturated hydrocarbon containing one carbon –carbon triple
bond,Some alkyne compounds which are physiologically acceptable salts are
used as MCH antagonist .Thus they influence eating behavior , reduce body
weight and addressing obesity which often leads to diseases such as diabetes
,dyslipidemias, high BP and CHD.
Alkyl Halides:
These are group of compounds derived from alkanes containing one or more
halogens. Some are used as anesthetics and antiseptic agents. Some of them
are used in medicine for the elimination of hook worms

TOXICITY STUDY RESULTS
ACUTE TOXICITY STUDY IN FEMALE WISTER RATS TO EVALUATE
TOXICITY PROFILE OF VENTHAMARAIYATI CHOORANAM
Result:
From acute toxicity study it was observed that the administration of
Venthamaraiyathi Chooranam at a dose of 2000mg/kg, to a rats. From acute toxicity
study it was observed that the administration of Venthamaraiyathi Chooranam at a dose
of 2000 mg/kg to the rats do not produce drug-related toxicity and mortality. So No-
Observed-Adverse-Effect- Level (NOAEL) of Venthamaraiyathi Chooranam is 2000
mg/kg.
Discussion
Venthamaraiyathi Chooranam was administered single time at the dose of
5mg/kg, 50mg/kg , 300mg/kg, 1000mg/kg and 2000mg/kg to rats and observed for
consecutive 14 days after administration. Doses were selected based on the pilot study
and literature review. All animals were observed daily once for any abnormal clinical
signs. Weekly body weight and food consumption were recorded. No mortality was
observed during the entire period of the study. Data obtained in this study indicated no
significance physical and behavioural signs of any toxicity due to administration of
Venthamaraiyathi Chooranam at the doses of 5mg/kg, 50mg/kg , 300mg/kg, 1000mg/kg
and 2000mg/kg to rats.
At the 14th day, all animals were observed for functional and behavioral
examination. In functional and behavioral examination, home cage activity, hand held
activity were observed. Home cage activities like Body position, Respiration, Clonic
involuntary movement, Tonic involuntary movement, Palpebral closure, Approach
response, Touch response, Pinna reflex, Sound responses, Tail pinch response were
observed. Handheld activities like Reactivity, Handling, Palpebral closure, Lacrimation,
Salivation, Piloercetion, Papillary reflex, abdominaltone, Limb tone were observed.
Functional and behavioral examination was normal in all treated groups. Food
consumption of all treated animals was found normal as compared to normal group.
Body weight at weekly interval was measured to find out the effect of
Venthamaraiyathi Chooranam on the growth rate. Body weight change in drug treated
animals was found normal.

SUB-ACUTE TOXICITY STUDY
WISTER RATS TO EVALUATE TOXICITY PROFILE OF
RESULT:
Clinical Signs:
All animals in this study were free of toxic clinical signs throughout the dosing
period of 28 days.
Mortality:
All animals in control and in all the treated dose groups survived throughout the
dosing period of 28 days.
Body weight:
Results of body weight determination of animals Table-1 from control and
different dose groups exhibited comparable body weight gain throughout the dosing
period of 28 days.
Food consumption:
During dosing and the post-dosing recovery period, the quantity of food
consumed by animals from different dose groups was found to be comparable with that
by control animals.
Organ Weight:
Group Mean Relative Organ Weights (% of body weight) are recorded in Table
No.4 Comparison of organ weights of treated animals with respective control animals on
day 29 was found to be comparable similarly.
Hematological investigations:
The results of hematological investigations (Table 4) conducted on day 29
revealed following significant changes in the values of different parameters investigated
when compared with those of respective controls; however, the increase or decrease in
the values obtained was within normal biological and laboratory limits or the effect was
not dose dependent.

Biochemical Investigations:
Results of Biochemical investigations conducted on days 29 and recorded in Table
2 revealed the following significant changes in the values of hepatic serum enzymes
studied. When compared with those of respective control. However, the increase or
decrease in the values obtained was within normal biological and laboratory limits.
Histopathology:
In histopathological examination, revealed normal architecture in comparison
with control and treated animal.
Discussion:
All the animals from control and all the treated dose groups up to 900 mg/kg
survived throughout the dosing period of 28 days.
No signs of toxicity were observed in animals from different dose groups
during the dosing period of 28 days.
Animals from all the treated dose groups exhibited comparable body weight
gain with that of controls throughout the dosing period of 28 days.
Food consumption of control and treated animals was found to be comparable
throughout the dosing period of 28 days
Haematological analysis conducted at the end of the dosing period on day 29,
revealed no abnormalities attributable to the treatment.
Biochemical analysis conducted at the end of the dosing period on day 29 no
abnormalities attributable to the treatment.
Organ weight data of animals sacrificed at the end of the dosing period was
found to be comparable with that of respective controls.
Histopathological examination revealed normal architecture in comparison
with control and treated animal.

PHARMACOLOGICAL STUDIES RESULT
1. THROMBOLYTIC ACTIVITY RESULT

EFFECT OF VENTHAMARAIYATHI CHOORANAM ON BODY WEIGHT
(PHYSICAL PARAMETER)
Table : 15 Results
ONLY CARRAGEE CARRAGEE CARRAGEE
GROU CONTR CARRAGEE
CARRAGEE NAN + VMC NAN + VMC NAN + VMC
P OL NAN +STD
NAN -300mg/kg -600mg/kg -900mg/kg
BODY
129±2.1
WEIG 133.8±1.167 126.5±1.821 134.2±0.9804 126.5±1.408 130.5±1.765
76
HT
Values are expressed as the mean ± S.D; Statistical significance (p)calculated by one way
ANOVA followed by dunnett’scP< 0.001, b
P< 0.01,aP < 0.05 calculated by
comparing treated group with CONTROL group
INBODY WEIGHT
150
Weight (mg/g)
100
50
0
kg
kg
kg
L
kg
N
O
g/
g/
g/
g/
TR
5m
0m
0m
0m
EE
N
O
30
60
90
G
IN
C
IR
C
R
VM
VM
VM
R
SP
A
A
C

EFFECT OF VENTHAMARAIYATHICHOORANAM ON
Table : 16
GRO CONTRO CARRAGEE
CARRAGEE NAN + VMC NAN + VMC NAN + VMC
UP L NAN +STD
Total
Tail
14.1±0.430 13.24±0.4214 13.96±0.6281 13.7±0.46368 13.84±0.5662 13.34±0.3668
Lengt
116 26 72 1 16 79
h
Total
Clotti
ng
12.68±0.3786 7.98±0.23537 5.64±0.44339
Tail 0±0 9±0.223607 6.76±0.2502
82 2 6
Lengt
h
Values are expressed as the mean ± S.D; Statistical significance (p)calculated by one way
ANOVA followed by dunnett’scP< 0.001, b
P< 0.01,aP < 0.05 calculated by
Fig. 8. Total Clotting Tail Length

EFFECT OF VENTHAMARAIYATHICHOORANAM ON BLEEDING TIME and
BLOOD CLOTTING TIME.
Table : 17 Stastical significance

GRO CARRAGEE
CONTROL CARRAGEE NAN + VMC NAN + VMC NAN + VMC
UP NAN +STD
Bleedi
1241.33±20.0 437.667±76.8 738.333±19.0 971.333±4.86 1077.33±18.0
ng 923±20.4385
577 117 8 256 89
time
blood
clottin 33.1667±0.90 24.1667±1.16 14.5±0.88506 25.5±0.88506 31.1667±0.90
6±0.68313
g time 9823 667 1 1 9823
(sec.)
Values are expressed as the mean ± S.D; Statistical significance (p)calculated by

one way ANOVA followed by dunnett’scP< 0.001, bP< 0.01,aP < 0.05 calculated by

sec/min tim/sec
CO
C
0
10
20
30
40
NT O
0
500
1000
1500
CA
R C N
RR O A TR
L R O
AG R L
A
A EE G
SP NA A EE
IR N
SP N
IR A
IN
IN N
5m
VM g/ 5m
C kg VM g/
30 C kg
Fig. 9
0m 30
VM 0m
g/ VM
C kg
C
g/
k
60 60 g
Bleeding time
0m 0m
VM g/ VM
C
SCIENTIFIC VALIDATION OF VENTHAMARAIYATHI CHOORANAM

g/
90
kg
C k g
90
blood clotting time (sec.)
0m 0m
g/ g/
kg k g
145
Table : 18 Effect on Partial Thromboplastin Time (APTT), Prothrombin Time (PT),
Fibrinogen Concentration AndEuglobulin Clot Lysis Time (ECLT) In Rats.
GROUP APTT(S) PT(S) Fibrinogen ECLT (S)

Concentration
Control 266±2.19089
18.26±0.475 34.93±1.536 238±1.93218
Only Carrageenan
18.36±0.423 36±0.730 486±7.89937*** 440±15.249***
Carrageenan
+STD 20.46±0.735 25.33±1.115 218±0.730297*** 206.67±2.764***
Carrageenan
+Vmc 300mg/Kg 23.06±1.187*** 31±0.632 307±6.46014*** 246±5.11208***
Carrageenan
+Vmc 600mg/Kg 17.73±0.830 45.33±3.451 292.667±19.25 270.667±3.67575
Carrageenan 278±5.26625*
+Vmc 900mg/Kg 20.33±0.735 50±6.366** 224.667±5.18116**

Prothrombin Time (PT) Activated Partial Thromboplastin Time
60 30
40 20
sec/min
sec/min
20 10
0 0
L
kg
kg
kg
kg
N
kg
kg
kg
N
L
kg
RO
O
NA
g/
g/
g/
g/
g/
g/
g/
g/
TR
5m
0m
0m
0m
NT
5m
0m
0m
0m
EE
EE
N
CO
30
60
90
AG
O
30
60
90
N
IN
C
RI
A
C
RR
IR
C
R
PI
VM
VM
VM
VM
VM
VM
R
SP
CA
AS
A
A
C
Fibrinogen Concentration Euglobulin Clot Lysis Time (ECLT)

600 500
400
400
sec/min
300
g/dl
200
200
100
0 0
L
kg
kg
kg
kg
kg
kg
kg
kg
N
N
RO
O
NA
A
g/
g/
g/
g/
g/
g/
g/
g/
TR
5m
0m
0m
0m
5m
0m
0m
0m
NT
EE
EE
N
30
60
90
G
O
30
60
90
G
N
IN
RA
C
RI
R
C
C
C
R
PI
VM
VM
VM
I
VM
VM
VM
R
R
SP
CA
AS
A
A
C
Fig. 10
Interpretation:
The test drug “Venthamaraiyathi Chooranam ” has got Significant thrombolytic
activity.

2. VASODILATOR ACTIVITY RESULT
Vasodilatation Effect Test

PE at 1 µM produced a steady-state contraction in the aortic rings with or without
endothelium. As shown in Figure 1, the extract (50–400 µg/mL) caused vasodilation of
the endothelium-intact thoracic aorta ring pre-constricted with phenylephrine in a dose-
response manner. However, this effect disappeared after preincubation of the aortic rings
with atropine (1 × 10−6 M), L-NA (1 × 10−4 M) or by removal of the vascular
endothelium.
Figure: 11 (a)

Figure: 11 (b)
Figure: 11 (c)

Dose-response curves for the vasodilation effects of the
ethanolicVenthamaraiyadhiChoornam extract on the preconstricted thoracic aorta rings.
Results are presented as means ± SD (n = 6), (Student’s t-test, *P < 0.05). (a)
Endothelium ..
Discussion
Venthamarai Flower is one of the most well-famous fragrant plants worldwide
and has been prescribed in folk medicines in many countries according to its
multipurpose actions. Venthamaraiyathi Choornam possessed vasodilation activity. As
shown in Figure 11(a), the vasodilation effect of the Venthamaraiyathi Choornam extract
was mediated by endothelial cells in the aortic vessel. The Choornam extract at a dose of
400 µg/mL reduced the contraction to lower than 43% of the maximal contraction. This
result suggests that the vasorelaxation effect of the ethanolic Venthamaraiyathi
Choornam extract is endothelium dependent. It has been reported that the vasorelaxant
property of most plant extracts was from flavonoids.
Thus, the vasodilation activity should be attributed to the high content of
flavonoid mixtures found in the Venthamaraiyathi Choornam extract.Preincubating the
thoracic aortic rings with atropine (10−6 M), a muscarinic receptor antagonist completely
blocked the relaxant activity of the extract (Figure 11(b)). The pharmacologically relaxant
effect of the Choornam extract was further determined. Pretreatment of endothelium-
intact aorta with L-NA (1 × 10−4 M) also reduced the vasodilation effect of the extract to
an extent equivalent to the effect in endothelium-denuded aorta (Figure 11(c)).
Nowadays, the mechanism of nitric oxide (NO) and the function of endothelial cells in
the relaxation of arteries are well described. NO is a potent vasodilator synthesized in the
endothelium by NO synthase and causes vascular relaxation. The result from the present
study suggests that the Venthamaraiyathi Choornam extract may exert its endothelium-
dependent relaxation activity by stimulating the nitric oxide release from the vascular
endothelium via muscarinic receptors.

3. HYPOLIPIDEMIC ACTIVITY RESULT
Table : 19 - Effect on siddha formulation ventha maraiyathy chooranam

in Lipid Profile
Total Tri
HDL LDL VLDL
GROUPS cholesterol glycerides AI LDL/HDL
(mg/dl) (mg/dl) (mg/dl)
(Mg/dl) (mg/dl)
Normal 50.70 ± 58.10± 27.40 ± 14.40 ± 31.22 ± 0.87 ±

0.54 ±
Control 1.80 0.90 1.20 0.75 1.15 0.50
Cholesterol 115.40 ± 162.65 ± 11.92 ± 30.95 ± 13.10 ± 8.51 ±

2.60 **(a)
Control 1.58**(a) 1.72**(a) 0.70**(a) 1.36**(a) 0.70**(a) 1.33**(a)
Standard 76.60± 83.15± 21.70 ± 20.15 ± 24.90 ± 2.34 ±

0.92 **(b)
Control 1.45**(b) 1.80**(b) 0.40**(b) 0.80**(b) 0.80**(b) 2.33**(b)
Treatment
91.80 ± 115.40 ± 17.50 ± 25.30 ± 17.85 ± 4.39 ±
control 1.46**(b)
1.30**(b) 1.92**(b) 0.50**(b) 0.60**(b) 0.42**(b) 1.48**(b)
200mg/kg
Treatment
84.45 ± 96.5 ± 20.30 ± 22.15 ± 21.30 ± 3.13 ±
control 1.08 **(b)
0.95**(b) 1.10**(b) 1.32**(b) 0.70**(b) 0.52**(b) 0.24**(b)
400mg/kg
Values are expressed as Mean ± SD.

Values were find out by using ONE WAY ANOVA followed by Newman Keul’s
multiple range tests.
** (a) values were significantly different from normal control at P< 0.01.
** (b) Values were significantly different from hyperlidemic control at P< 0.01.
Table shows the levels of Cholesterol, Triglycerides, HDL, LDL and VLDL of
control and experiment rats respectively. Serum of hyperlipidemic rats showed
significantly increased levels of Cholesterol, Triglycerides, LDL - C and low HDL – C,
when compared with normal rats. In rats treated with both doses of siddha formulation
Venthamaraiyathi Chooranam and Atorvastatin there was significant decrease in the
content cholesterol, TGs, LDL – C, and VLDL and increases HDL – C, when compared
with cholesterol control rats.

Atherogenic index (AI) and LDL – C / HDL – C ratio:
Table shows the changes of Atherogenic Index and LDL – C / HDL – C ratio in
control and treated rats. It appears clear form these results that the cholesterol induction
significantly affects the cardio vascular risk markers.
Indeed, AI was statistically increased in cholesterol control group 90% compared
with the values found in their normal control group.
Besides there were significant further increase of LDL – C / HDL – C ratio in
cholesterol control group compared to normal control group.
Promising results in lowering of AI by siddha formulation venthaa marayathy
chooranam was found in Table I. The siddha formulation Venthamaraiyathi Chooranam
showed an improvement of the cardio vascular risk level by decrease of AI in the treated
group by more than 73% and 63% (p < 0.01), when compared to the cholesterol control
group.
The ratio of LDL – C to HDL – C is also a protective indicator of cardio vascular
disease incidence. The cholesterol induction produced a significant increase of this
marker. In contrast, elevated ratio in treated group and Atorvastatin group returned to
basal value when the data were compared in the same period to the data found for
cholesterol rats.
DISCUSSION
The reduction of plasma total cholesterol was associated with a decrease
in its LDL fraction which is a major, potentially modifiable risk factor of cardio vascular
disease and the target of drug. Many suggest that the cholesterol lowering activity of this
product appears to be due to the enhancement of LDL – C catabolism through hepatic
receptors .(4)In addition siddha formulation ventha maraiyathy chooranam showed
protective action which is reported to have a preventive function against atherogenesis
since an independent inverse relationship between blood HDL – C levels and cardio
vascular risk incidence is reported.(5) The lipoprotein called “good cholesterol”
facilitates the mobilization of triglycerides and cholesterol from plasma to liver where it
is catabolised and eliminated in the form of bile acids. The possible mechanism of this
activity may result from the enhancement of lecithin cholesteryl acyl transferase (LCAT)

and inhibition of Hepatic Triglyceride Lipase (HTL) on HDL which may lead to a rapid
catabolism of blood lipids through enterohepatic tissues.(6)
It is also recently reported that triglycerides plays a key role in the regulation of
lipoprotein interaction to maintain normal lipid metabolism. Indeed, the elevated plasma
TG levels were associated with an increased incidence of coronary artery disease.
Moreover these higher plasma TG levels have been attributed mainly to increase
population of small, dense LDL deposits which are very atherogenic and enhanced
cholesteryl ester mass transfer from apolipoprotein B containing lipoproteins (VLDL and
LDL).(7) TG has also been proposed to be major determinant of cholesteryl
esterification, its transfer and HDL remodeling in human plasma.(8)
Siddha formulation Venthamaraiyathi Chooranam significantly suppress the
elevated blood concentration of TGs. This result suggests that the product is able to
restore, at least partially, the catabolism of TG. The underlying the mechanism of this
activity is not elucidated by the present study. However, as hypothesized by many works
with other plants, the restoration of catabolic mechanism of TGs would be due to an
increased stimulation of the lipolytic activity of Plasma Lipoprotein Lipase (LPL).(9)
Administration of siddha formulation Venthamaraiyathi Chooranam provides a
beneficial action on rat lipid metabolism with regard to the reduction of AI. Infact, the AI
was decreased in all treated groups. Similar results were reported by others when
studying the hypolipidemic effects of natural products .(10) This ameliorative action was
due to the plasma lipid lowering activity of different constituents of the plant.
It is also desirable to have higher plasma HDL and lower LDL – C to prevent
atherogenesis, since there is a positive correlation between an increased LDL – C / HDL
– C ratio and the development of atherosclerosis. Again, the administration of siddha
formulation Venthamaraiyathi Chooranam significantly suppress the higher values of
LDL – C / HDL – C ratio showing the beneficial effect of this preparation in preventing
atherosclerosis incidence.

This result is considered as an important for the treatment of hyperlipidemia
induced atherosclerosis and apparently validates the folk medicinal use of hyperlipidemic
patients in India.
Interpretation:
Hyperlipidemia is considered to be major risk factor for the premature
atherosclerosis and essentially the cholesterol in atherosclerotic plaque is derived from
that of circulatory cholesterol. The antihyperlipidemic effect on siddha formulation
Venthamaraiyathi Chooranam in particular could be considered as a possible therapeutic
value.

3. CARDIOPROTECTIVE ACTIVITY RESULT:
EFFECT OF VENTHAA MARAYATHY CHOORANAM AGAINST
DOXORUBICIN INDUCED MYOCARDIAL TOXICITY IN ALBINO RATS
Effect of Venthamaraiyathi Chooranam on Dox-induced cardiac toxicity was

established by measuring cardiac biomarker enzymes and endogenous antioxidants and
heart tissue histopathology.
General observation and mortality

The general appearance of all groups of animals was recorded throughout the
study. In Dox-treated group, the animal fur became scruffy and developed a pink tinge.
These rats also had red exudates around the eyes and nose, soft watery feces and
enlargement of abdomen. These observations were significantly less in Venthamaraiyathi
Chooranam treated group.
Serum enzyme biomarkers

Animals treated with Dox showed significant increase in the levels of CPK and
LDH compared to normal Tab.no 1. Venthamaraiyathi Chooranam + Dox treated group
shown significantly lower levels of CPK and LDH as compared to Dox treated group.The
myocardial damage in the various treated groups was determined by estimating the
activities of AST, ALT and ALP Tab no.1. These biochemical markers were significantly
increased in the Dox group compared to control (P<0.01). Venthamaraiyathi Chooranam
pretreatment group showed significant reduction in AST, ALT and ALP levels as
compared to Dox-treated group.

Table 20: Effect of Venthamaraiyathi Chooranam on AST, ALT, ALP, CPK and LDH
enzyme activities in doxorubicin-treated rats
Group AST ALT ALP CPK LDH
Group 1 66.60±1.40 30.60±1.10 122.82±3 153.4±3. 220.20±8.52
(Normal control) .12 60
Group 2 194.60±7.80*a 60.84±2.50 244.20±6 320.20±6 408.52±1
(Toxic control) *a .36*a .60*a 2.30*a
Group 3 8.30±2.32*b 38.20±1.80 16.30±4. 220.30±4 282.40±9.20
(Standard *b 34*b .25*b *b
control)
Group4 128.90±5.28*b 49.35±2.15 192.65±4 270.35±4 325.20±10.6
Treatment low *b .50*b .65*b 2*b
dose
Group 5 108.40±4.92*b 42.64±1.98 175.32±3 243.25±3 298.40±9.45

Treatment high *b .60*b .60*b *b
dose
Values are expressed as mean ± SD,

a* - values were significantly different from Normal control (GI) at P<0.01,
b* - values were significantly different from Toxic control (GI) at P<0.01.
DISCUSSION
The study reveals the cardioprotective effect of Venthamaraiyathi Chooranam
against Dox-induced cardiotoxicity in rats. Following evidence can be emphasized from
the present study. Venthamaraiyathi Chooranam has been traditionally used in medicine
and culinary practices in India, possesses cardioprotective, hepatoprotective and lipid
lowering properties. The present study is aimed to investigate the cardioprotective effects
of oral administration of Venthamaraiyathi chooranam against Dox -induced
cardiotoxicity.
In the Dox-treated group, the animal fur became scruffy and developed a pink
tinge which in the later days of observation period was followed by red exudates around
the eyes and nose. Necrosis was also observed at the site of Dox injection. These changes
were less pronounced in case of Venthamaraiyathi Chooranam pretreated group animals,

which accounts for the effective cell protecting property of Venthamaraiyathi
Chooranam with anti-inflammatory, antioxidant and antifibrotic effect.
The study revealed severe biochemical changes as well as oxidative damage in
the cardiac tissue after the chronic treatment with Dox. Transaminases such as ALT and
AST are liberated into the serum after extensive tissue injury. Because the heart muscle is
rich in both (especiallyAST), it suggests that their increased level is an indicator of
myocardial damage.
The Dox-treated group showed marked elevation in serum levels of AST and
ALT as compared to vehicle-treated group. The mild elevations of AST have been
associated with liver injury or myocardial infractions. Greater the injury size, higher the
activity of AST. This result implies that the Dox when taken for long period of time
could cause both liver and heart injury. A typical myocardial injury gives an AST/ALT
ratio greater than 1. However, AST/ALT ratio less than 1 are found due to release of ALT
from the affected liver [12]. Since the result showed AST / ALT ratio to be greater than 1
with higher doses over a long period of time, Dox is likely to lead myocardial damage.
In Venthamaraiyathi Chooranam +Dox treated group, AST and ALT levels
significantly decreased as compared to Dox treated group; therefore, present results
suggest that treatment of Venthamaraiyathi Chooranam may inhibit myocardial damage.
These findings confirm that Venthamaraiyathi Chooranam is responsible for
maintenance of normal structural and architectural integrity of cardiac myocytes, which
can be accounted for membrane stabilizing property of Venthamaraiyathi Chooranam , as
evident from the near normal serum enzymatic activities of AST and ALT.
The serum ALP, LDH and CPK enzyme activities are important measures of both
early and late phases of cardiac injury. It is reported that serum LDH and CPK were
increased after Dox administration in mouse. The present results are in good agreement
with our earlier findings [13] ALP activity on endothelial cell surface is responsible, in
part, for the conversion of adenosine nucleotides to adenosine, a potent vasodilator and
anti-inflammatory mediator that can protect tissues from the ischemic damage that results
from injury. This may account for the elevation of ALP in the Dox group, where tissue
injury and inflammation are prominent. On the other hand, CPK and LDH are not
specific for myocardial injury individually; however, evaluation of these enzymes

together may be an indication of myocardial injury. In the preventive group, i.e.
Venthamaraiyathi Chooranam +Dox, the ALP, LDH and CPK enzyme levels were
decreased to a level near to that of control group, suggesting that Venthamaraiyathi
Chooranam may protect the myocardial tissue against Dox toxicity.
Interpretation
In conclusion, the present study shows that the cardiotoxicity induced by Dox is
related with oxidative stress. Anti-proliferative, anti-initiation and free radical scavenging
properties of Venthamaraiyathi Chooranam may boost myocardial integrity and attenuate
the cardiac toxicity. Venthamaraiyathi Chooranam has shown to be cardioprotective,
which may be attributed to its potent antioxidant properties.

ANTI –MICROBIAL ACTIVITY RESULT:
Table : 21 Anti - Microbial Activities By Well Diffusion Method
Zone inhibition
Organism
s.no Susceptibility
(Culture)
Streptomycin Medicine
zone size size
1. E.coli Sensitive 20mm 18mm
2. Klebsiella pnemoniae Sensitive 24mm 11mm
3. Staphylococcus aureus Resistant 24mm _
4. Streptococcus mutant Sensitive 18mm 22mm
5. Enterococcus faecalis Moderate resistant 24mm 25mm

Pseudomonas aeruginosa 14mm _
6. Resistant
EFFECT OF VTC ON ANTI MICROBIAL ACTIVITY
Fig. 12 (a)

Fig. 12 (b)
Fig. 12 (c)

INTERPRETATION:
It was observed that anti microbial studies of Venthamaraiyathi Chooranam
showed that it is sensitive against Escherichia coli, Klebsiella pnemoniae, Streptococcus
mutant and moderately sensitive against Enterococcus faecalis when compared to the
standard drug (Streptomycin) which was evident from the zone inhibition.The herbel
drug VTC showed inhibition of the growth of the micro organism at 100mg/ml
concentration for the organism.Our result confirmed the traditional use of VTC has Anti
microbial activity.

6. SUMMARY
The test drug “Venthamaraiyathi Chooranam ” was selected from the

siddha literature “Pharmacopoeia of hospital of Indian medicine” authored by
Dr.V.Narayanaswami for its Thrombolytic, vasodilator, Hypolipidemic and
Cardioprotective activities. The dissertation started with an introduction
explaining about the siddha concept, prevalence of hypertension and role of the
test drug in treating it.
The test drug was prepared properly by the given procedure. All the ingredients
were identified and authenticated by the botanist and experts of
Gunapadam,Govt. Siddha medical college, Palayamkottai.
Review of literatue in various categories was carrid out siddha aspect botanical
aspect abd pharmaceutical review disclosed about thae drug and disease
pharmacological review was done to establish the methodologies
According to siddha aspect hypertension (Kruthi Azhal Noi ) is caused by

derangement of vatha pitha humors. Spicy taste of test drug diminish the vatha
humor the spicy taste where posses the seetha veriyam very much helpful to
demines the aggrevated vatha pitha humors.
Various analysis such as physicochemical , microbiological , phytochemical,

biochemical analysis, instrumental analysis were made . From the above analysis
we came who known the presence of active ingredients responsible for its activity.
Phytochemical analysis showed the presence of potassium, calcium, chloride,
iron,.
Biochemical analyisis showed the the presence of calcium,sulphate,

chloride,starch, iron ferrous, phosphate, unsaturation , reducing suger, amino
acids .thus from these results we come to known the effectiveness of the drugs is
due to the presence of these constituents.

The instrumental analysis FTIR showed the peak values presence of Alkanes,
Amines,Aromatics,,Aliphatic amines , Alkyl halides, Carboxylic acids
ICP-OES analysis of these drug shows heavy metals Like Arsenic, cadmium,
Nickel, copper and lead are found in below detecting level. The toxic metals are
found in BDL. It reveals the drug in safer for long term use. Calicum was
associated with the lower risk of Hypertension. Sodium regulates the body’s acid
base balance. Iron is essential for may numbers of biological functions such as
growth, reproduction, healing and immune function. The phosphorus is involved
in tissue repair Magnesium is regulates the blood pressure. Sulphate is potent anti
oxidant activity in human body. Zinc has got potent anti-microbial activity.
Acute and sub-acute toxicity were carried out in wistar albino rats according to
OECD guidelines(423). This drug has no acute toxicity as there was no mortality
seen sub-acute toxicity is carried by repeated dose of test drug for 28 days.
Mortality, the functional observations, haematological and bio-chemical
investigations were done. There were no significant changes in the bio-chemical
and haematological profile. So the toxicological study of these test drug,
venthamaraiyathi chooranam establish the safety of the drug for long time
administration.
In pharmacological studies, the thrambolytic activity is carried out in wistar

albino rats by using enzymeimmunoassay sandwich method. The test drug
venthamaraiyathi chooranam has significant thrambolytic activity.
The vasodilator activity of test drug venthamaraiyathi chooranam carried out

wistar albino rats by using students organ bath. The present study suggests that
the Venthamaraiyathi Choornam extract may exert its endothelium-dependent
relaxation activity by stimulating the nitric oxide release from the vascular
endothelium via muscarinic receptors.
The hypolipidemic activity of test drug venthamaraiyathi chooranam carried out

wistar albino rats models. Hyperlipidemia is considered to be major risk factor for

the premature atherosclerosis and essentially the cholesterol in atherosclerotic
plaque is derived from that of circulatory cholesterol. The antihyperlipidemic
effect on siddha formulation Venthamaraiyathi Chooranam in particular could be
considered as a possible therapeutic value.
Cardioprotective activity present study shows that the cardiotoxicity induced by

Dox is related with oxidative stress. Anti-proliferative, anti-initiation and free
radical scavenging properties of Venthamaraiyathi Chooranam may boost
myocardial integrity and attenuate the cardiac toxicity. Venthamaraiyathi
Chooranam has shown to be cardioprotective, which may be attributed to its
potent antioxidant properties
Anti-microbial study of the test drug carried out by well diffusion method. It is
observed that Venthamaraiyathi Chooranam is sensitive to Escherichia coli and
Streptococcus mutant & Klebsiella pnemoniae. These VTC has significant anti-
bacterial activity.
Result and discussion gives the necessary and essential justification to prove the
potency of test drug Venthamaraiyathi Chooranam with the scientific validation.
Thus the herbel formulation is validated for its safety efficiency in the
management of diarrhoea and it would be the way for a drug of choice.

7. CONCLUSION
It is concluded that the test drug Venthamaraiyathi Chooranam has significant

activity to control blood clot possess thrombolytic activity and control to cholesterol level
by possessing hypolipidemic action and also stimulating the nitric oxide release from the
vascular endothelium via muscarinic receptors some significant vasodilator activity and
boost myocardial integrity by possessing cardioprotective activity. Apart from this
Venthamaraiyathi Chooranam has anti-bacterial sensitivity against various pathogens.
Finally the drug Venthamaraiyathi Chooranam is scientifically validated by modern
techniques.

8.FUTURE SCOPE
Pre clinical evaluation of the drug Venthamaraiyathi Chooranam has been done
by physico chemical, bio chemical, Instrumental, pharmacological, toxicological, anti
microbial procedures. In future, the drug has to be validated by clinical trials and should
be used for patients.
The active principle which is responsible for the activity has to be find out,
through modern scientific analysis. Having made up of nano particles, Venthamaraiyathi
Chooranam holds extra promise for the treatment of hypertension.

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PRECLINICAL STUDY OF SIDDHA DRUG

VENTHAMARAIYATHI CHOORANAM FOR IT’S

THE TAMILNADU DR. MGR MEDICAL UNIVERSITY

In partial fulfilment of the requirements

for the award of the degree of

DOCTOR OF MEDICINE (SIDDHA)

POST GRADUATE DEPARTMENT OF GUNAPADAM

THE GOVERNMENT SIDDHA MEDICAL COLLEGE

DECLARATION BY THE CANDIDATE

Date: Signature of the Candidate

Place: Palayamkottai Dr.J.Indrakumar

CERTIFICATE BY THE GUIDE

Date: Signature & Seal of the Guide

Name & Signature of the Name & Signature of the

ALT - Alanine transaminase

ANOVA- - Analysis of variance

AST - Aspartate transaminase

CMC - Carboxyl Methyl Cellulose

CDC - Council of Disease control and prevention

CPCSEA - Committee for the purpose of control and supe

EDTA - Ethylene Diamine Tetra Aceticacid

ESR - Erythrocyte Sedimentation Rate

FTIR - Fourier transform infrared spectroscopy

EDTA - Ethylene Diamine Tetra Aceticacid

ESR - Erythrocyte Sedimentation Rate

FTIR - Fourier transform infrared spectroscopy

IAEC - Institutional Animal Ethical Committee.

ICP-OES - Inductively coupled plasma optical emission spectrometry

LDH - Lactate Dehydrogenase

MCV - Mean Corpuscular Volume

OECD - Organisation for Economic Co-operation and Development

PCV - Packed Cell Volume.

RBC - Red Blood Corpuscles

SEM - Scanning electron microscope

TLC - Thin Layer Chromatography

HPTLC - High performance thin layer chromatography

2. AIM & OBJECTIVES OF THE STUDY 3

3.1 ELETTARIA CARDAMOMUM, Linn 4

3.1.1. Gunapadam Aspect 4

3.1.2. Botanical Aspect 7

3.1.3. Lateral Research 11

3.2. ZINGIBER OFFICINALE 12

3.2.1. Gunapadam Aspect 12

3.2.2. Botanical Aspect 15

3.2.3. Lateral Research 22

3.3. PIPER LONGUM 23

3.3.1. Gunapadam Aspect 23

3.3.2. Botanical Aspect 26

3.3.3. Lateral Research 28

3.4. CUMINUM CYMINUM 29

3.4.1. Gunapadam Aspect 29

3.4.2. Botanical Aspect 32

3.4.3. Lateral Research 35

3.5. GLYCYRRHIA GLABRA 36

3.5.2. Botanical Aspect 38

3.5.3. Lateral Research 41

3.6. ANETHUM SOWA 42

3.6.1. Gunapadam Aspect 42

3.6.2. Botanical Aspect 44