“PUNGAM VER CHOORANAM ”

PART I

ANTI-ULCER ACTIVITY OF

“PUNGAM VER CHOORANAM ”

(

Pongamia pinnata.linn

)

&

PART II

ANTI-UROLITHIATIC ACTIVITY OF

“VEDIYUPPU CHEYANEER”

The dissertation Submitted by S.RAJAPREHIDHA Under the Guidance of Dr.V.VELPANDIAN, M.D(s)

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

GOVT. SIDDHA MEDICAL COLLEGE, CHENNAI-106

DECLARATION BY THE CANDIDATE

I hereby declare that this dissertation entitled “Anti ulcer activity of Pungan Ver Choornam Pongamia pinnata(linn)” and “Anti urolithiatic activity of Vediyuppu Cheyaneer” is a bonafide and genuine research work carried out by me under the guidance of Dr.V.Velpandian M.D(S)., Lecturer, Post Graduate Department of Gunapadam, Govt.Siddha Medical College, Arumbakkam, Chennai-106 and the dissertation has not formed the basis for the award of any Degree, Diploma, Fellowship or other similar title.

Date: Signature of the Candidate

Place: Chennai Dr .S.Rajaprehidha

GOVT. SIDDHA MEDICAL COLLEGE, CHENNAI-106

CERTIFICATE BY THE GUIDE

This is to certify that the dissertation entitled “Anti ulcer activity of pungam ver choornam (Pongamia pinnata.linn) and Anti urolithiatic activity of Vediyuppu Cheyaneer ” is submitted to the Tamilnadu Dr.M.G.R.Medical University in partial fulfillment of the requirements for the award of degree of M.D (Siddha) is the bonafide and genuine research work done by Dr.S.Rajaprehidha,Under my supervision and guidance and the dissertation has not formed the basis for the award of any Degree, Diploma, Associateship, Fellowship or other similar title.

Date: Seal and Signature of the Guide

Place: Chennai

GOVT. SIDDHA MEDICAL COLLEGE, CHENNAI-106

ENDORSEMENT BY THE HOD, PRINCIPAL/HEAD OFTHE INSTITUTION

This is to certify that the dissertation entitled“Anti ulcer activity of Pungam Ver Cchooranam (Pongamia pinnata linn)” and “Anti urolithiatic activity of Vediyuppu Cheyaneer” is a bonafide work carried out by Dr.S.Rajaprehidha under the guidance of Dr.V.Velpandian M.D(S)., Lecturer, Post graduate department of Gunapadam, Govt.Siddha Medical College, Chennai - 106.

Seal and Signature of the HOD Seal and Signature of the Principal

Date: Date:

Place: Chennai Place: Chennai

ACKNOWLEDGEMENTS

Thanks to Almighty and may His peace and blessings be upon all his prophets for granting me the chance and the ability to successfully complete this dissertation.

I would like to express my deepest appreciation to my guide Dr.V.Velpandian.,M.D(s), Lecturer, Govt. Siddha Medical College, Chennai-106.

He continuously and convincingly conveyed a spirit of adventure in regard to research, without his guidance and help this dissertation work have not been possible.

I wish to express my sincere thanks to our honourable Principal Professor Dr.A.M.Abdul Kadher.,M.D(s), Government Siddha Medical College, Chennai-106, for permitting me to carry out this dissertation work.

I let my heartful thanks to our respectable Principal(i/c) Dr.V.Banumathy.,M.D(s)., for her valuable suggestions and encouragement throughout the cource of study.

I render my humble and sincere thanks to our respectable Professor Dr.K.Kumar.M.D(s), Head of the department, Gunapadam branch, Govt Siddha Medical College, Chennai-106 for his valuable guidance throughout this dissertation work.

I express my cordial thanks to Dr.S.Ayyasamy Ph.D., Assistant Lecturer, Govt. Siddha Medical College, Chennai-106., for his sincere guidance.

My deepest thanks to Dr.V.Pichayakumar.,M.D(s), Assistant Lecturer, Govt.

Siddha Medical College, Chennai-106., for extending his support in completing this project.

I render my special thanks to our former H.O.D Professor Dr.B.Malarvizhi.,M.D(s) for her valuable support and encouragement regarding in completing this project work

I register my special thanks to Dr.R.Sivagami.,M.D(pharmacology), Assistant Professor, Dept. of pharmacology,Kilpauk Medical College, Chennai for her

I register my special thanks to Mrs.Girija Srinivasan, Assistant Professor, Medicinal Botany, GSMC, Chennai-106., for her valuable support in doing phytochemical analysis for this dissertation work.

My special thanks to Prof.Selvaraj.,Ph.D., H.O.D, Department of Biochemistry, GSMC, Chennai-106., for letting me to carry out biochemical analysis

I wish to thank Dr.P.Vidhya M.B.BS, D.M.R.D., Radiologist, Aringar Anna Government Hospital of Indian Medicine and Homoeopathy, Chennai-106., for helping me in doing clinical studies regarding this project.

I would like to convey my regards to Dr.K.Ramalakshmi., Superintendent, Aringar Anna Government Hospital of Indian Medicine, Arumbakkam, Chennai-106, for their support in doing clinical studies.

I cordially register my thanks to Mr.V.Dhandayudhapani., Librarian, Ambedkar Library for his support in referring books in regard to this project work.

I am grateful to Dr.Sasikala Ethirajulu., R.O, CRIS, Chennai-106 and Dr.T.G.Menon for helping me in doing pharmacognostic studies.

My regards to Dr.Shegila, R.O, Chemistry department, CRIS, Chennai-106., for helping me to carry out physiochemical analysis

My deep sense of gratitude to Dr.J.Anbu, Ph.D., Vels University, Pallavaram, Chennai., who had laid a great support in completing the pharmacological studies successfully.

I express my reverential gratitude to Dr.V.Murugesan, Director, SAIF, IIT, Chennai., for his extended help in doing quantitative and qualitative analysis.

I immensely thank Mr.Sugendran of Government veritinary college, Vapery, Chennai for his valuable help in doing histopathological studies.

My regards to Mr.Syed amirjohn., BSc, DMRD., Chennai Labs, Aminjakarai, Chennai., for his help in doing haematological studies.

and assistance for giving wonderful moral support to continue my post graduation and for his encouragement given by him throughout my career.

I am very much indeed thankful to my brother S.M.Vijayaragavendar B.E., and my cousin T.Aruna Devi B.E., for their marvellous support in completing this dissertation work.

My special thanks to my parents and all my family members for their blessings and kind co-operation.

I honour my collagues for being with me and supporting me in developing this project.

My humble thanks to all my patients who had laid a great support in completing my clinical study.

Finally I wish to thank the authorities of Dr. M.G.R Medical University, Chennai for permitting me to conduct this study.

CONTENTS PART-I

S.No ANTI ULCER ACTIVITY OF PUNGAM VER CHOORNAM Page. No

1 INTRODUCTION 1

2 AIM AND OBJECTIVES 3

3 REVIEW OF LITERATURES 4

3. 1 BOTANICAL ASPECT 4

3. 2 GUNAPADAM ASPECT 8

3. 3 MODERN ASPECT 11

3.4 SIDDHA ASPECT OF THE DISEASE 12

3.5 MODERN ASPECT OF THE DISEASE 16

3.6 LATERAL RESEARCH WORK 21

4 MATERIALS AND METHODS 23

4.1 PREPARATION OF THE TRIAL DRUG 24

4.2 STANDARDIZATION OF THE DRUG 24

4.2.1 PHYSIO-CHEMICAL ANALYSIS 24

4.2.2 BIO-CHEMICAL ANALYSIS 28

4.2.3 TOXLOGICAL STUDY 36

4.2.4 PHARMACOLOGICAL ACTIVITY 37

4.3 CLINICAL STUDY 39

5 RESULTS AND DISCUSSION 45

6 CONCLUSION 55

7 SUMMARY 56

CONTENTS PART-II

S.No ANTI UROLITHIATIC ACTIVITY VEDIYUPPU CHEYANEER Page. No

1 INTRODUCTION 57

2 AIM AND OBJECTIVES 59

3 REVIEW OF LITERATURES 60

3. 1 MODERN ASPECT 60

3. 2 GUNAPADAM ASPECT 73

3.3 SIDDHA ASPECT OF THE DISEASE 98

3.4 MODERN ASPECT OF THE DISEASE 102

3.5 LATERAL RESEARCH WORKS 112

4 MATERIALS AND METHODS 117

4.1 PREPARATION OF THE TRIAL DRUG 117

4.2 STANDARDITIZATION OF THE DRUG 120

4.2.1 PHYSIO-CHEMICAL ANALYSIS 120

4.2.2 BIO-CHEMICAL ANALYSIS 121

4.2.3 TOXLOGICAL STUDY 125

4.2.4 PHARMACOLOGICAL ACTIVITY 126

4.3 CLINICAL STUDY 131

5 RESULTS AND DISCUSSION 138

6 CONCLUSION 161

7 SUMMARY 162

8 BIBLIOGRAPHY 163

ABBREVIATIONS

alb - Albumin

As - Arsenic

AT - After treatment

BDL - Below Detective Level

BT - Before treatment

Ca - Calcium

CaOx - Calcium Oxalate

Cd - Cadmium

Ck - Cork

CL - Cholesterol

CMC - Carboxy Methyl Cellulose

CNS - Central Nervous System

CPCSEA - Committee for the purpose of Control and Supervision of

CT - Computed topography

DC - Differential count

dep - deposits

DF - Deviation factor

DNA - De-oxy ribose nulclic acid

E - Erythrocytes

ESR - Erythrocyte sedimentation rate Experiments on animals

F - Fibres

Fe - Iron

Hg - Mercury

IAEC - Institutional Animal Ethical Committee

ICP-OES - Inductively Coupled Plasma Optical Emission Spectroscope

K - Potassium

KUB - Kidney Ureter Bladder

L - Lymphocytes

LD - Lethal dose

Mg - Magnesium

Mr - Medullary ray

MRI - Magnetic Resonance Imaging

Na - Sodium

NSAID - Non Steroidal Anti Inflammatory Drugs

OECD - Organization for Economic Co-operation and Development

P - Parenchyma

P - Phosphorous

P - Polymorphs

Pb - Plumbum

PUD - Peptic ulcer disease

PVC - Pungam ver choornam

S - Sulphur

Sc - Stone cell

Sco - Secondary cortex

SD - Standard deviation

SEM - Standard error of mean

Sph - Secondary phloem

Sxy - Secondary xylem

TC - Total count

TLC - Thin Layer Chromatography

V - Vessel

VCN - Vediyuppu Cheyaneer

TABLES

PART-I ANTIULCER ACTIVITY OF PUNGAM VER CHOORNAM

TABLE

NO SUBJECT P.NO

4.1 TLC OF PUNGAM VER CHOORNAM 25

4.2.1.1 PHYTOCHEMICAL ANALYSIS OF PUNGAM VER 33 4.2.2.1 BIO CHEMICAL ANALYSIS OF PUNGAM VER 29

4.3.1 AGE WISE DISTRIBUTION 42

4.3.2 SEX DISTRIBUTION 42

4.3.3 OCCUPATIONAL STATUS 42

4.3.4 PERSONAL HABITS 43

4.3.5 SOCIO ECONOMIC STATUS 43

4.3.6 GRADATION RESULT 43

4.3.7 IMPROVEMENT IN SIGNS AND SYMPTOMS 44

4.2.3.1 ACUTE TOXICITY 48

4.2.4.1 ANTI ULCER ACTIVITY 49

5.1 STATISTICAL ANALYSIS 53

TABLES

PART II- ANTI UROLITHIATIC ACTIVITY OF VEDIYUPPU CHEYANEER TABLE

NO SUBJECT P.NO

4.2.1.1 PHYSIOCHEMICAL ANALYSIS OF VEDIYUPPU CHEYANEER

120

4.2.1.2 PHYSIO CHEMICAL PROPERTIES 120

4.2.1.3 COLOUR, NATURE & YIELD OF VEDIYUPPU CHEYANEER

120

4.2.2.1 BIOCHEMICAL ANALYSIS OF VEDIYUPPU CHEYANEER

121

4.3.1 AGE WISE DISTRIBUTION 134

4.3.2 SEX WISE DISTRIBUTION 135

4.3.3 OCCUPATIONAL STATUS 135

4.3.4 PERSONAL HABITS 136

4.3.5 SOCIO ECONOMIC STATUS 136

4.3.6 GRADATION RESULT 136

4.3.7 IMPROVEMENT IN SIGNS AND SYMPTOMS 136

4.2.2 ICP-OES 141

4.2.3.1 ACUTE TOXICITY STUDY 143

4.2.3.2 DIURETIC ACTIVITY 144

TABLE NO

SUBJECT P.NO

4.2.4.1 ANTI UROLITHIATIC ACTIVITY 145

4.2.3.3 BODY WEIGHT OF ALBINO RATS 150

4.2.3.4 FOOD INTAKE OF ALBINO RATS 151

4.2.3.5 WATER INTAKE OF ALBINO RATS 151

4.2.3.6 HEMATOLOGICAL PARAMETERS 152

4.2.3.7 BIOCHEMICAL PARAMETERS 153

4.2.3.8 RENAL FUNCTION TEST 154

4.2.3.9 LIPID PROFILE 154

4.2.3.10 URINE ANALYSIS 155

5.1 STATISTICAL ANALYSIS 159

FIGURES

PART I -ANTI ULCER ACTIVITY OF PUNGAM VER CHOORNAM.

Fig 4.1 - Pongamia tree

Fig 4.2 - Pungam ver

Fig 4.3 - Pungam ver choornam

Fig 4.2.4.1 - Anti ulcer activity (control) Fig 4.2.4.2 - Anti ulcer activity (standard) Fig 4.2.4.3 - Anti ulcer activity (test)

PART II- ANTI UROLITHIATIC ACTIVITY OF VEDIYUPPU CHEYANEER.

Fig 3.1 - Impure Vediyuppu

Fig 3.2 - purified Vediyuppu

Fig 3.3 - Impure Venkaram

Fig 3.4 - Purified Venkaram

Fig 3.5 - Impure Navacharam

Fig3.6 - Purified Navacharam

Fig 3.7 - Impure Thurusu

Fig 3.8 - Purified Thurusu

Fig 3.9 - Impure karpooram

Fig 3.10 - Purified karpooram

Fig 3.11 - Palampuli

Fig 3.12 - Lemon

Fig 3.13 - Vediyuppu Cheyaneer

Fig 4.2.3.1 - Histopathology of Vediyuppu Cheyaneer.

1. INTRODUCTION

Mankind since time immemorial knows the healing power of nature. Nature has its harmony with constructive principles on physical, mental, moral, and spiritual planes and it has health promotive, disease preventive and curative as well as restorative powers.

No one knows for sure when humans began using herbs for medicinal purpose. The usage of plants for medicinal purposes dates back long before recorded history. Herbal medicine has been documented for almost 4000 years ago. These medicines have survived real world testing and thousands of years of human testing. Herbal medicine sometimes referred to herbalism is the use of herbs for their therapeutic or medicinal use.

Herbal medicine is the oldest form of health care known to man. Primitive man observed and appreciated the great diversity of plants available to him. Much of the medicinal use of plants seems to have been developed through observations of wild animals and by trial and error.

As time went on each tribe added the medicinal power of herbs in their area to the knowledge base. They methodically collected information on herbs and developed well defined pharmacopoeias. In India usage of plants for its medicinal value dates back several thousand years to the Rig-Veda (the collection of Hindu sacred verses).

There are countless remedies in nature which are magnalia die (i.e.), the mysteries of curing and healing hidden from eyes but open to spiritual perception of the wise. Our siddha system of medicine was propounded by such great saints known as siddhars supposed to have lived at a very early period. They were men of highly cultured intellectuals and spiritual faculties combined with supernatural powers

According to siddha medicine diet and lifestyle play a major role not only in health but also in curing diseases. Lifestyle changes have a major impact on health. Economic growth, modernization, socialization, urbanization has changed the life style of humans. The transition from a traditional to modern life style with a shift in eating habits, mental stress, and adoption of a sedentary lifestyle has lead to the increasing prevalence of new diseases in the human community.

Changes in dietary habits and lifestyle pattern leads to emerging of new diseases, of one which is the peptic ulcer disease. Gunman noi which is referred in noi nadal can be easily compared with peptic ulcer disease from its etiology, signs and symptoms.

Peptic ulcer disease is a serious gastrointestinal disorder that requires a well targeted therapeutic strategy. A number of drugs including proton pump inhibitors, H2 receptor antagonist are available for the treatment of peptic ulcer, but clinical evaluation of these drugs has shown incidence of relapse, side effects and drug interactions

According to latest WHO data published in April 2011 peptic ulcer disease deaths in India reached 108, 392, or 1.20% of total deaths. The age adjusted death rate is 12.37%. India ranks fifth in the world in peptic ulcer disease .

Keeping in view the distressing problem of this disease a search of novel molecules has been extended to herbal drugs that offer better protection and decreased relapse. Drugs of plant origin are gaining popularity and are being investigated for this disease.

In siddha system of medicine, the herbal drug”

PUNGAM VER CHOORNAM

” is selectee for the study in treating “

GUNMAM

”. However there are no records of systematic pharmacological studies. The present study was planned to assess the efficacy of this drug.

2. AIM AND OBJECTIVES Aim

Gastric and duodenal ulcers are illness that affects a considerable number of people in the world. Some of the causes of these disorders are smoking, stress, nutritional deficiencies and ingestion of non-steroidal anti-inflammatory drugs. The pathogenesis of gastro duodenal ulcers is influenced by various aggressive and defensive factors, such as acid-pepsin secretion, mucosal barrier, mucus secretion, blood flow, cellular regeneration and endogenous protective agents.

Hence in the present study an attempt was made to investigate the anti-ulcerogenic effects of Pungam Ver Choornam both pre-clinically and clinically

Objectives

 To accomplish the literature review of the drug.

 Collection, identification, authentication, and preparation of the drug.

 To evaluate the pharmacognostic features of this drug by microscopic studies.

 To evaluate phytochemical properties of the drug.

 Determination of LD50, ED50 and therapeutic index.

 To evaluate toxicological studies in the trial drug

 To evaluate the pharmacological activities of the drug

 To study the efficacy and to evaluate the safety of the drug clinically.

3. REVIEW OF LITERATURE 3.1 BOTANICAL ASPECT Pongamia pinnata(linn)

Synonyms

Pongamia pinnata(L) pierre Galedupa indica lam Dalbergia arborea wild Derris indica(Lam) Bennet Pongamia glabera

Taxonomical hierarchy.

Kingdom - Plantae

Division - Magnoliophyta

Class - Magnolipsida

Order - Fabales

Family - Leguminoseae

Genus - Pongamia

Species - pinnata

Vernacular names:

English - Indian beech

Tamil - Pungamaram

Hindi - Karanj, Kiramal Sanskrit - Karanja, Natkamala

Telegu - Kanugu chetu

Kannada - Honge mara

Bombay - Karanj

Maharastra - Kidamar

Punjab - Sukhchain

Bengal - Dahar, Karanja, Natakaranga

Oriya - Koranjo

Assam - Karchaw

Habit:

This tree is common all over India and met with from central Himalayas to southern India and Cyelon.

Varities

:

There are six types of verities found in India, they are Dahar karanja

Nata karanja Kanta karanja Makra karanja Bigh karanja Amba karanja

Part used:

Seeds, Stem, Leaves, Fruit, Root, and Oil from seeds.

Distribution:

The natural distribution of pungam is along coasts and river banks in India and Burma.

Native to the Asian subcontinent, this species has been introduced to humid tropical low lands in the Philippines.

Ecology:

Native to humid and subtropical environments, pungam thrives in area having an annual rainfall ranging from 500-2500mm in its natural habitat, the maximum temperature ranges from 27-38 degrees and the minimum to 16 degrees. Mature trees can withstand waterlogging and slight frost. This species grows to elevations of 1200m, but in the Himalayan foot hills is not found above. Pungam can grow on moist soil types. It is highly tolerant to salinity, highest growth rates are observed on well drained soils with assured moisture. Natural reproduction is profuse by seed and common by root suckers.

Vegetative characters

:

Pungu is a medium sized tree generally attains a height of about 8m-18m, a trunk diameter of more than 50m. The trunk is generally short with thick branches spreading into a dense hemispherical crown of dark green leaves. The bark is thin grey to grayish brown and yellow in the inside. The taproot is thick and long lateral roots are numerous and well developed.

Leaves:

The leaves are alternate compound pinnate leaves consist of 5-7 leaflets which are arranged in 2-3 pairs and a single terminal leaflet. Leaflets are 5-10cm long, 4-6cm wide and pointed at the tip.

The leaves are lopped for fodder and are said to act as a galactogogue. They are rich in nitrogen and are used as green manure for rice and sugarcane fields.

Flowers:

Flowers borne on racemes, are pink, light purple or white, fragrant in axillary racemes.

The flowers also furnish good manure for pot plants.

Stem bark

:

They are grayish green or brown in colour, smooth and are covered with tubercles. The stem bark is fibrous and is used for cordage.

Seeds

:

Pods are elliptical,3-6cm long 2-3cm wide, thick walled and usually contain single seed, seeds are 10-20cm long big oblong and light brown in colour, reniform broad wrinkled with reddish brown leathery testa.

The seeds are mainly valued for oil obtained from them which are mainly industrial and medicinal uses.

Roots:

The root is very thick and long and deep rooted it has numerous lateral branches. They are light brown in colour.

Medicinal uses of roots:

 The juice of the roots is used for cleaning foul ulcers and closing fistulous sores and for cleaning teeth and strengthening gums.

 The juice is also used in the treatment of gonorrhea.

 A paste of the roots is used as a local application in scrofulous enlargements

3.2 GUNAPADAM ASPECT

ò§F–Pungu

Pongamia pinnata (Linn) Pierre

ntW bga®

ò‹F, óªÂ, fuŠrf«, fuŠr«

ÏJ rhjhuzkhŒ ϪÂah KGtJ« gæuhf¡ Toa ku« Ïš fh£Lò§F v‹bwhU tifÍ©L. Ïu©o‰F« Fz« x‹nw.

ga‹gL« cW¥ò - Ïiy, ó, éij, nt®, v©bzŒ

Rit - if¥ò, Jt®¥ò

j‹ik - bt¥g«

ÃçÎ - fh®¥ò

brŒif - Jt®¥Ã, cl‰nj‰¿, ö¡Fâ¥òGtf‰¿

ò§f« nt® Fz«

thj¡fL¥ò kfh_®¢ir jhgRu«

thjF‹k«u¤j¤jhš tªÂLnehŒ–xJ»‹w g©òiuÍ« tšèlK« nghF« Âu©lU©nl g©òWò§f« nt®¡F¥ gh®

- mf°Âa® Fzthfl«.

ÏJ, thjF‹k«, thj¤Fil¢rš, _®¢ir, FUÂædhš Ãw¡»‹w nehŒfŸ, ò©, òiunahlš gh«ò eŠR Ïitfis Ú¡F«.

tH¡F

 ò§f« ntiu¥ òuâ Ú¡», bkšèajhf¢ Óé ghš ÃêªJ, mj‰F ne® nj§fha¥

ghš nr®¤J¡ fhŒ¢Á it¤J¡ bfh©L, xU bkšèa Jâæš eid¤J, Ãsit, Mwhj ò©fŸ, nkf¥ ò©fŸ M»aitfS¡F ngh£Ltu Ó¡»u¤Âš M¿¥

nghF«.

 ò§f« nt® r§f«nt®, Á‰whkz¡F nt® tif¡F 35 » és¡bf©bzŒ 1,400 äè ó©L¢ rhW 350 äè fLFnuh»â 9 »uh«, Kl¡f¤jh‹ 17 », Ïš Kjš _‹W nt®fisÍ« miu¤J és¡bf©izæš fyªJ, Ë k‰w ru¡FfisÍ«

rh‰iwÍ« nr®¤J fy¡»¢ Nçaòl« it¤J, j¡f msthf cŸS¡F¡ bfhL¤J tu, Niy, ò© fu¥gh‹ Kjèad ÔU«.

F‹k nehŒ¡F ò§f« nt® nrU« kUªJfŸ

1.tUdh »ahH«

nrU« ru¡FfŸ: ò§f« nt®, éšt«, ehÍUé, Á¤Âu_y«, mUbešè, bgçabešè, KU§ifg£il KŸS¡f¤jç, f©l§f¤jç, btŸis mHtz«, g¢ir mHtz«, fU¥ò mHtz«, bgU§FU«ignt®, fL¡fhŒó, Óikãynt«ò, f‰flfÁ§», nfhitky®, j©Ù®é£lh‹ »H§F.

Ïitfis nr®¤J »ahHä£L bfhL¡f nt©L«.

ÔU« nehŒ–F‹k«, kªjNiy, fgnuhf«.

2. rjhtçah fšf«

nrU« ru¡FfŸ: j©Ù®é£lh‹»H§F, ò§f« nt®, kukŠrš, f©Lghu§», Â¥Ãè, vŸ. Ïitfis nr®¤jiu¤J f‰fkh¡»¡ bfh©L Ãunah»¡fΫ

ÔUk nehŒ : F‹k«, F¿¥ghf Ïu¤j F‹k« ÔU«.

3. gu§»ah Nuz«

nrU« ru¡FfŸ: ò§f«nt®g£il, f©Lghu§», Â¥Ãè, Â¥Ãè_y«, njtbjU

nk‰Tw¥g£l ru¡Ffis rkvil vL¤J Nuzkh¡»¡ bfhŸs nt©L«.

JizkUªJ : vŸS »ahH«

ÔU« nehŒ : F‹k«

4. óÂfh Nuz«

nrU« ru¡FŸ : ò§Fnt®, br›éa«, Á¤Âu_y«, R¡F, Â¥Ãè, äsF, c¥ò nk‰f©l ru¡Ffis Nuâ¤J cgnah»¡fΫ

Jiz kUªJ : jæ®

ÔU« nehŒ : F‹k«, cÂu«, Å¡f«, gh©L.

5. Âuhakhz »Uj«

nrU« ru¡FŸ : bfh¤J¥ò§f‹ nt® - 4 gy¤ij40 gy« #y¤Âš ngh£L

#y¤Âš 1 ghf« ÛW«goahf »ahHŠ R©l¡fhŒ¢Á tof£o, mš fL¡fhŒÃŠR fLF nuhfâ, nfhiu»H§F, óidfhŠbrhç, Âuh£ir, ÑHhbešè, Ӫšbfho, ÏŠÁur«, beŒ, ghš Ïitfis tif¡F 8 gyŠ nr®¤J »Ujg¡Ftkhf fhŒ¢rΫ.

msΖ1/4 gy« rh¥ÃlΫ. / 15-30 ml

ÔU« nehŒfŸ–äjF‹k«, u¤jF‹k«, äjRu« FzkhF«.

6. ótur§fhŒ v©bzŒ.

7. ò‹D¡F méœj«.

3.3 MODERN ASPECTS Chemical constituents of roots:

The root bark contains two closely related derivatives.

Kanugin andDemethoxy kanugin Tetra-o-methyltisetin

Pongachromene

are also isolated from roots

The root bark from Australia has been reported to contain karanjin and pongapin and smaller amounts of two leniar furanoflavones, pinnatin and gamatin which are isomeric to karanjin and pongapin respectively.

Some of the anti cancer compounds are elucidated from the root extract, they are Paclitaxel

Vinblastin

Vincristine(sulphate) Teniposide

Fluoxetine

Oetoposide derivatives.

Actions:

Bitter Astringent Refrigerant Appetizer Insecticide

3.4 SIDDHA ASPECT OF THE DISEASE F‹k«

ntW bga® - Fšk«

Ïašò:Ϫnehæš c©Q« czÎ, brçahJ c©l Á¿J neu¤Â‰bfšyh«

tæ‰WŸjh§f Koahj vç¢riyÍ«, tèiaÍ« c©lh¡», c©l czit thªÂ vH¢

brŒJ c£br‹w czit gad‰wjh¡F«. xnu fhy¤Âš cliyÍ« kdijͧ

F‹w¢brŒJ cæiuÍ« khŒ¡f¢ ÁªÂ¡f brŒÍ« nehŒ Mjyhš Ïjid F‹kbkd¥

bgaç£ld®.

nehŒ tU« tê :

“brŒahd F‹k¤Â‹ njh‰wª j‹id¢

br¥Ãlnt Jt®¥ghd bghÁ¥Ã dhY«

kŒahd k§ifÍl‹ kUt yhY«

tifyhF§ »H§Ftif aUªj yhY«

cŒahd äsF tif Íiu¥ÃdhY«

cWgÁia al¡»L kªj¤jhY«

jŒahd r©lhs nfhg¤jhY«

rè¥ghY« F‹k« tªjilÍ« ghnu”

-ô» Áªjhkâ

 mÂf NLŸs bghU£fis òÁ¤jš

 mÂf Jt®¥ghd bghU£fis c©zš

 bg©nghf« mÂfkhf bfhŸsš

 »H§F tiffis mÂfkhf òÁ¤jš

 mÂfkhd fhu czÎfis bfhŸsš

 gÁia ml¡Fjš, g£oå ÏU¤jš, kd¢rè¥ò

 mÂf« nfhg§ bfhŸsš M»at‰whš F‹knehŒ Ãw¡F« v‹W ô» TW»wh®

nkY«,

 k©, cä, fš, öR ÏitfyªJ bghU£fis c©gjhY«

 RidÚ®, x£lk‰wÚ®, R©zh«ò fyªj Ú® Ïitfis mUªjyhY«

 nj§fhŒ ghš M»a brç¡f¡TlhŒ bghU£fis äFÂahf bfhŸtjhY«,

 tê jt¿ _¢ir mil¤J nahf ãiyæš ÏU¥ngh®¡F ϪnehŒ c©lhF«.

nehŒ v©:

“brŒant v©F‹k¢ braiy¡ nfshŒ brayhd thÍF‹kthj F‹k«

vŒant äjF‹k bkçF‹kkhF«

Vyhd tèF‹kŠ r¤Â F‹k«

ijant råF‹kŠ nr£g F‹k«

Rhfkh§ F‹k§ by£lh«”

F¿¥g¿ª bjh›bth‹whŒ¡ TWnthnk”

-ô» Áªjhkâ.

F‹k nehŒ ô» Kåtç‹ T‰Wgo v£L tif¥gL«.

1. thÍF‹k«

2. thjF‹k«

3. äjF‹k«

4. fg F‹k«

5. vçF‹k«

6. tèF‹k«

7. r¤ÂF‹k«

8. r‹åF‹k«

ÂUf©l Kåt® T‰¿‹ go F‹k¤ij v£L tifahf Ãç¤JŸsh®. mit 1. thjF‹k«

2. äjF‹k«

3. fgF‹k«

4. thjäjF‹k«

5. thj fg F‹k«

6. äj Ány¤Jk F‹k«

7. Âçnjhõ F‹k«

8. Ïu¤j F‹k«

nkY« Áy® Ïjid rhkhåa thjF‹k«, rhkhåa äj F‹k«, rhkhåa fg F‹w« vd _‹whfΫ Ãç¥g®.

nehæ‹ K‰F¿Fz§fŸ

 gÁæ‹ik

 gÁc©lhæD« czé‹ nkš éU¥gä‹ik

 thŒFk£lš

 thŒÚ® Cwš

 c©l czÎ vÂbuL¤jš

 tæW òu©L nehjš

 thªÂahjš

 òë¤nj¥g«

 tæW Ïiujš

Ïitna Ï‹ nehæ‹ K‰F¿Fz§fŸ.

bghJ¡F¿Fz§fŸ:

 ϪnehŒ bgU«gh‹ikÍÍ« ÏUg¤ijªJ taJ Kjš eh‰g¤ijªJ taJila M© k¡fS¡F tUtjhF«. ÁWgh‹ik bg©fS¡F« tUtJ©L.

 ÂObud gÁæ‹ik

 thŒ Fk£lš

 äJ äjhf thªÂ vL¤jš

 czÎ brçahkš V¥gälš

 òëna¥g«

 tæW gGthf ÏU¥gJ nghèU¤jš

 jh§f Koahj tæ‰W tè

K¡F‰w ntWghL

“bjh¥thjgªj kyhJ F‹k« tuhJ” -nju‹

 jfhj brašfshY«, czé‹ khWgh£lhY« tëF‰w« nflilªJ, Ñœneh¡F¡

fhiyÍ« ghÂ¥gila brŒJ Ïjdhš kygªj« c©lh¡»Í«, nkš neh¡F¡fhš ghÂ¥ghš thªÂia c©lh¡»Í« nehæid Ãw¥Ã¡F«.

czÎ:

F‹knuhf g¤Âa«:

 xU tUl« br‹w Áf¥ò mçÁ, f‰f©L, bfhŸS fh£LäUfkhär«, fŸS, gR«ghš, M£L¥ghš, Âuh£ir, vYä¢r¥gH«, f®Éu§fŸ, khJs«gH«, bešèt‰wš, R¡F, bfh‹id¥òë, nkh®, Mkz¡F v©bzŒ, btŸisó©L, ÏsKŸs§», KU§if¡fhŒ, vth¢rhu«, fL¡fhŒ, bgU§fha«, bfhokhJs«gH«, ÂçfLF, nfh_¤Âu«, cZzyF Ôgd«, M»ait g¤Âa§fshF«.

 X¤jlälš, éa®it th§fš, énurd«, t°ÂfUk«, Fjt®¤Â, m¥Ãa§fd«, Ánefghz«, M»ait nehŒ¡nf‰wthW brŒJ bfhŸsyh«.

 vëš brç¡F« czÎfis bfhŸs nt©L«.

 brç¥ig¤ ju¡Toajhd ÏŠÁ, Ãu©il Ïit ngh‹wt‰iw¤ Jitaš, CWfhŒ Kjèat‰iw nr®¤J bfhŸsyh«.

mg¤Âa§fŸ:

 thŒÎr«kªjkhd rfygjh®¤j§fŸ, tU¤jkhd m‹dghdhÂfŸ, cy®ªj khär§fŸ, KŸs§», Û‹fŸ, ¡¥ghd gh©l§fŸ, òÂa jhåa§fŸ, kygªjkhd m‹d§fŸ, äfΫ ry« mUªjš.

 ky«, _¤Âu«, thªÂ Ït‰iw ml¡Fjš, Ïitfns F‹knuh» élnt©oaJ.

3.5 MODERN ASPECT OF THE DISEASE

Peptic ulcer:

Peptic ulcer, also known as PUD or peptic ulcer disease is the most common ulcer of an area of the gastrointestinal tract that is usually acidic and thus extremely painful. A peptic ulcer

is a hole in the lining of the stomach, duodenum, or oesophagus. An ulcer is a sore or erosion that forms when the lining of the digestive tract is corroded by acidic digestive juices.

Ethymologies of peptic ulcer:

The English word peptic comes from the Latin word pepticus which comes from the greek word peptikus meaning“to digest”.

The English word ulcer comes from the Latin word ulcusmeaning “a sore” or “a wound”.

The term peptic ulcer literally means a wound in the digestive system.

Incidence:

It is estimated that between 5%-10% of adults globally are affected by peptic ulcers at least once in their lifetimes. The prevalence of helicobacter pylori roughly matches age. Peptic ulcer disease had a tremendous effect

on morbidity and mortality until the last decades of the 20^thcentury.

India ranks fifth place in the acid peptic disease. About four million people have active peptic ulcers and about 350,000 new cases are diagnosed each year. Four times as duodenal ulcers as gastric ulcers are diagnosed. Approximately 3,000 deaths per year are due to peptic ulcer.

Classification:

 Duodenum(called duodenal ulcer)

 Oesophagus (called oesophageal ulcer)

 Stomach (called gastric ulcer)

 Meckel’s diverticulum(called meckel’s diverticulum ulder, is very tender with palpitation)

Modified Johnson classification of peptic ulcers:

 Type I- ulcer along the body of the stomach, most common along the lesser curve at incisura angularis

 Type II- ulcer in the body in combination with duodenal ulcers. Associated with acid over secretion.

 Type III- in the pyloric channel within 3cm of pylorus. Associated with acid over secretion.

 Type IV- proximal gastrooesophagal ulcer.

 Type V- can occur throughout the stomach. Associated with chronic NSAID use.(such as asprin).

Aetiology:

NSAIDs: These are medications for pains. It is the most common cause of ulcer. Non- steroidal anti inflammatory drugs lower the stomach’s ability to make a protective layer of mucus, making it more susceptible to damage by stomach acids. NSAIDs can also affect the flow of blood to the stomach, undermining the body’s ability to repair cells.

Genetics: A significant number of individuals with peptic ulcers have close relatives with the same problem, suggesting the genetic factors may also be involved.

Smoking: People who regularly smoke tobacco are more likely to defelop peptic ulcers compared to non-smokers.

Alcohol abuse: Regular heavy drinks of alcohol have a higher risl of developing peptic ulcers.

Mental stress: People with mental stress will experience sustained and worse symptoms.

Helicobacter pylori: H.pylori, a type of bacteria is responsible for most ulcers. This organism weakens the protective coating of the stomach and first part of the intestine and allows damaging digestive juices to eat away at the sensitive lining below.

Risk factors:

 Hereditary

 Older age

 Chronic pain causing long term use of asprin or NSAIDs

 Alcohol abuse

 Diabetes may increase your risk of having H.pylori

 Lifestyle factors, including chronic stress, coffee drinking, drinking beverages and smoking, may make you more susceptible to damage of the gastr intestinal tract.

Signs and symptoms:

 Abdominal pain- classically epigastric with severity relating to mealtimes after around three hours of taking a meal.

 Bloating and abdominal fullness.

 Waterbrash (rush of saliva after an episode of regurgitation to dilute the acid in oesophagus.

 Nausea, and copious vomiting

 Indigestion

 Belching

 Pain is often made worse by an empty stomach.

 Loss of appetite and weight loss

 Hematemisis(vomiting of blood) this can occur due to bleeding directly from a gastric ulcer, or from damage to the oesophagus from severe vomiting.

 Melena(tarry foul smelling feaces due to oxidized iron from hemoglobin)

 Rarely, an ulcer can lead to a gastric or duodenal perforation, which leads to acute peritonitis.

 Heart burn and gastroesophageal reflux disease.

Complications:

The risk of complication is much greater if the ulcer is left untreated, or if treatment was not completed.

 Internal bleeding- slow blood loss can lead to anemia, while severe blood loss requires hospitalization and blood transfusions.

 Infection–a peptic ulcer can bore a hole through the wall of the stomach or small intestine, significantly increasing the risk of infection in the abdominal cavity- peritonitis- can be very painful and causes chills and fever, nausea, vomiting and a hard feeling in the abdomen. Individuals with peritonitis should seek medical attention as soon a symptoms are felt.

 Scar tissue- scar tissue caused by peptic ulcers can obstruct the passage of food through the digestive tract, making the patient feel full more easily. Scarring may also cause vomiting and weight loss.

 Pyloric stenosis- chronic inflammation in the lining of the stomach or duodenum caused by a peptic ulcer can result in a narrowing of the pulorus. Pyloric stenosis is the narrowing of the pylorus. Food will not pass through to the intestines, causing vomiting and weight loss.

 Recurrence of peptic ulcers- people with the highest risk of developing peptic ulcers are those who have had the before. Somebody who had a peptic ulcer caused by H.pylori infection runs a 5% risk of having another one during their lifetime. Even after their original ulcer was successfully treated and healed.

Lab investigations:

 Complete blood count

 Upper gastro intestinal X-ray

 Video endoscopy

 Test for H.pylori, which include Urease breath test Stool antigen test Life style modifications:

 Eat a diet rich in fiber, especially from fruits and vegetables. This may reduce your risk of developing an ulcer in the first place and speed your recovery if you have already.

 Foods containing flavonoids, like apples, berries, garlic, and tea may inhibit the growth of H.pylori.

 Eat antioxidants, foods, including fruits and vegetables.

 Eat foods high in vitamin-B and calcium such as almonds, beans, whole grains.

 Avoid refined foods, such as white breads, pastas e.t.c

 Eat fewer red meats and more lean meats, Eat fish or beans for protein.

 Use healthy oils, such as olive oil or vegetable oil.

 Reduce or eliminate trans-fatty acids, found in commercially baked goods.

 Avoid beverages that may irritate the stomach lining or increase acid production including coffee, alcohol, and carbonated drinks.

 Drink 6-8 glasses of filtered water daily

 Avoid taking too spicy foods and fast foods.

 Quit smoking and reduce stress with regular use of relaxation techniques

3.6 LATERAL RESEARCH WORKS.

The following pharmacological studies have been done.

1. Anti inflammatory activity of pongamia pinnata root extracts in albino rats.(singh RK, Nantha et al, Beneras University)

Sequential petroleum ether, benzene, chloroform, acetone and ethanolic extracts of pongamia pinnata root extracts are administered in dose of 50mg/kg intra peritoneal in rats was found to have anti-inflammatory while pentobarbitone induced “sleep time” was reduced by all the extracts except pet ether extract.

Possible mechanism of action could e inhibitor of prostaglandin synthesis and decreased capillary permeability. All extracts of root shows significant anti inflammatory activity in carragenin and PGE1induced oedema models compared to phenylbutazone.

2. Antinociceptive activity of pongamia pinnata root extracts in albino rats( ref:srinivasan et al)

Srinivasan et al, evaluated the analgesic activity of the various root extracts of pongamia pinnata, the petroleum ether extract, n-butanol extract, and ethanolic extract of the roots of pongamia pinnata showed significant analgesic effect in the tail flick test.

3. Influence of circadian variation an lipid per oxidation products and antioxidants.( (ref:

A comprehensive review, SR Arote, P.G yoele)

Effects of pongamia pinnata on lipid per oxidation products and antioxidants in hyperammonemic rats with reference to circadian variations are evaluate by Essa and subramaniam.

They suggested that these alterations could be modulated b pongamia pinnata during hyperammonemic condition which plays a crucial role in disease development. The root extracts showed significant reduction in lipid per oxidation and significant increase in antioxidant levels.

4. Determination of in vitro sunscreen activity of pongamia pinnata root extracts.(ref: drug invention today ISSN 0975-7619)

Various sequential root extracts of pongamia pinnnata root showed significant lowering of melanin formation and exhibited better sunscreening effect against ultraviolet induced radiations.

5. Ethnolic extract of pongamia pinnata root exhibit role in ischaemia reperfusion injury and cerebro vascular insufficiency.(ref: Journal of pharmacology and pharmaceutical sciences) 6. Anti plasmodial activity of pongamia pinnata.(ref: Intenational journal of and pharmaceutical sciences review and research, vol 9, issue 2)

Probably all the parts of pongamia pinnata exhibited anti-plasmodial activity.

The ethanolic extracts of pongamia were examined in vitro for anti-plasmodial property against plasmodium falciparum. The ethanolic extract of pongamia pinnata roots shows significant anti-plasmodial activity.

4. MATERIALS AND METHODS:

4.1 Preperation of pungam ver choornam.

The herbal drug pungam ver choornam is selected from famous classical text Gunapadam porut pambu nool-mooligai vaguppu, by vaidiya rathnam k.s. murugesa mudhaliyar, 4^th edition (2003), page no: 692.

Main ingredient:

Pungam ver-2kg

Collection of the herb:

The plant material used in this study was collected from Theni district, Tamil nadu, India.

Authentication of drug:

The identity and authenticity of the drugs was confirmed by head of the department of post graduate department of Gunapadam and by the botanist.,

purification of the drug:

After the collection of the root, it is rinsed well in the running water, and then the root’s were finely cut into small pieces and then dried in shade.

Preperation of choornam.

The well dried roots of Pongamia pinnata plant were made into fine powder. To make the drug to the finest form the powder is sieved with the help of cotton cloth classically known as vasthirakayam.

Purification of choornam:

The choornam was purified by pittaviyal method using Avi Iyandiram.

Preservation of the drug:

The purified pungam ver choornam was stored in clean glass air tight containers.

Form of medicine : choornam

Dose : 1gm/BD/after food

Vehicle : hot water.

Route of administration :enteral route.

4.2 STANDARDIZATION OF THE DRUG

4.2.1 PHYSIOCHEMICAL ANALYSIS OF PUNGAM VER CHOORNAM

S.No Parameter I II Mean

1. Loss on Drying at 105°C :6.021%, 6.214%, 6.118%

2. Total Ash :4.320%, 4.048%, 4.184%

3. Acid insoluble Ash :0.633%, 0.718%, 0.676%

4. Water Soluble Extractive :13.775%, 13.888%, 14.832%

5. Alcohol Soluble Extractive :10.288%, 10.238%, 10.263%

6. Ph :6.36

7. Thin Layer Chromatography

Thin layer chromatography

TLC of chloroform extract of Pungam ver chooranam

Rf value of Thin layer chromatography:

TABLE 4.1 The Rf values of thin layer chromatography are given below.

Sl.No Rf value Colour of the spot

1 0.10 Purple

2 0.28 Purple

3 0.36 Purple

4 0.48 Pink

5 0.51 Purple

6 0.69 Purple

7 0.73 Yellow

8 0.81 Yellow

9 0.84 Orange

10 0.93 Orange

Solvent system:

Toluene : Ethyl acetate (5:1.5)

TLC plate

:

Aluminium plate precoated with silica gel 60F₂₅₄of 0.2 mm thickness (Merck).

Developing chamber:

Camag’s twin trough chamber.

Visualizing reagent

:

Vanillin-sulphuric acid reagent.

Extract Preparation

:

4 g of the chooranam was soaked overnight in chloroform. Boiled on a water bath for 10 mins, filtered and concentrated to 10 ml.

Procedure:

The extract was applied on the TLC using capillary and developed in the solvent system.

The developed TLC plate was air dried, dipped in vanillin-sulphuric acid reagent and heated in an oven at 105°C until the development of coloured spots.

PHARMACOGNOSTICAL STUDY Pongamia pinnata (L) pierre- Pungu Material and methods

Roots were collected from Theni Dt., Chennai and identified by the Botanist, Siddha Central Research Institute, Chennai–106.

Fresh roots were fixed in FAA solution (70% ethyl alcohol, formalin and acetic acid in the ratio of 90 ml: 5 ml: 5 ml) in the field itself. The materials were left in the fluid for three days, after which they were washed in water and dehydrated with tertiary butyl alcohol. Paraffin wax was infiltered and the specimens were embedded in wax for sectioning. 20 – 30 μm thickness sections were taken and double stained.

Alcoholic safranin (0.5%) counter stained with 0.25 % fast green. All sides, after staining in safranin were dehydrated by employing graded series of ethyl alcohol (30 %, 50%, 70 %, 90

% and absolute alcohol) and stained fast green in clove oil and xylol-alcohol (50-50) and passed through xylol and mounted in DPX mountant (Johansen, 1940). Maceration of the root was prepared by employing Jeffrey’s maceration fluid.

Photomicrographs were taken with the help of Nikon Eclipse E200 Microscope.

Macroscopic

Root externally dull brown in colour, internally pale yellow, hard, fibrous, taste bitter, no characteristic odour.

Microscopic

The structural details of root about 1.1 mm thickness are described here. The cross section of the root is nearly circular in outline (Fig. 2 A). The outermost cork tissue consisting of 7 to 12 or more rows of rectangular, tangentially elongated thin walled cells (Fig. 2 B,C). It is followed by wide secondary cortex, composed of polygonal, tangentially elongated thin walled cells. Most of the cells contain both simple and compound starch grains consisting of 2 or 3 components, rounded to oval in shape, 3 - 11 μm in dia. Some cells contain yellowish brown

contents and prismatic calcium oxalate crystals. Single or groups of stone cells in varying shapes and size are scattered in this region.

The inner bark or bast is composed of tangential groups of bast fibres alternating with the regular phloem elements and the radially extending medullary rays (Fig. 2 B). The medullary rays are mostly straight, 1 or 2 seriate, consisting of radially elongated, thin walled cells towards inner region, tangentially elongated towards the outer region. The phloem parenchyma cells are small, oval or polygonal in shape. Starch grains and crystals similar to those of cortical cells, also present in phloem parenchyma and phloem rays. Secondary xylem consisting of vessels, tracheids, fibres and parenchyma. Vessels found scattered throughout secondary xylem region in singles or in groups of 2 or 3 (Fig. 2 D, E). Fibres thick walled arranged in tangential bands traversed by xylem rays. Xylem parenchyma cells thin walled, rounded to oval in shape. Xylem rays uni to triseriate consisting of radially elongated cells. Starch grains and calcium oxalate crystals are similar to those present in cortical cells and also found scattered in xylem parenchyma and xylem ray cells.

Maceration:

Fibres in single or in group, short with blunt end, xylem vessels with reticulate thickenings, parenchyma, cork cells and stone cells.

4.2.2 BIOCHEMICAL ANALYSIS OF PUNGAM VER CHOORNAM:

Preparation of the extract:

2gm of pungamver choornam was added with 5gm of sodium carbonate and taken in 100ml beaker and 20ml of distilled water was added. The solution was boiled for 10 minutes cooled and then filtered. The extract is called sodium carbonate extract.Then the following tests for the presence of acid radicals and basic radicals were done.

TABLE-4.2.2.1

S.N EXPERIMENT OBSERVATION INFERENCE

1. Test for sulphide.

a) 2ml of the above prepared extract is taken in a test tube.2ml of 4%

ammonium oxalate solution is added.

b) 2ml of sodium carbonate extract is added with 2ml of dilute hydrochloric acid until the effervescence ceases off. 2ml of barium chloride solution is added.

No cloudy appearance/

white precipitate is obtained.

No white precipitate insoluble in concentrated hydrochloric acid is obtained.

Absence of sulphate

Absence of sulphtae

2. Test for chloride.

2ml of sodium carbonate extract is added with dilute nitric acid till the effervescence ceases. Then 2ml of silver nitrate solution is added

Cloudy white precipitate completely soluble in excess of ammonium hydroxide solution is obtained

Presence of chloride.

3. Test for phosphate.

2ml of the extract is treated with 2 ml of ammonium molybdate solution and 2 ml of concentrated nitric acid

Presence of yellow precipitate/cloudy appearance with yellow colour is obtained

Presence of phosphate 4. Test for carbonate.

2 ml of extract is treated with 2 ml of magnesium sulphate solution

cloudy appearance/white precipitate is obtained

Presence of carbonate 5. Test for sulphide.

1 gm of the substance is treated with 2 ml of concentrated hydrochloric acid

Absence of colourless rotten egg smelling gas turning lead acetate paper black on warming.

Absence of sulphide

6. Test of nitrate.

1 gm of substance is heated with copper turning and concentrated sulphuric acid the test tube is viewed vertically down

No copious evolution of reddish brown gas

Absence of nitrate

7. Test for fluoride and oxalate.

a) 2ml of the extract is added with 2ml of dilute acetic acid and 2ml of calcium chloride solution and heated.

b) 5 drops of clear solution is added with 2ml of dilute sulphuric acid and dilute potassium permanganate solution is added

Absence of cloudy white precipitate.

Potassium permanganate solution is not

decolourised.

Absence of fluoride.

Absence of oxalate.

8. Test for nitrate.

3 drops of the extract is placed on a filter paper. On that, 2 drops of acetic acid and drops of benzidine solution

No yellowish red colour is obtained.

Absence of nitrate

9 Test for borate.

2 pinches of the substance is made into paste by using sulphuric acid and alcohol (95%) and introduced into the blue flame.

Absence of green tinged flame.

Absence of borate.

Test for basic radicals

10. Test for lead.

2ml of the extract is added with 2ml of potassium iodide solution

No yellow precipitate is obtained.

Absence of lead.

11. Test for copper.

a) One pinch of substance is made into paste with concentrated hydrochloric acid in a watch glass and introduced into the non-luminous part of the flame.

b) 2ml of the extract is added with excess of ammonia solution.

Absence of bluish green colour flame.

Absence of bluish precipitate.

Absence of copper.

Absence of copper.

12. Test for aluminium.

To the 2ml of the extract sodium hydroxide solution is added in drops to excess.

No white precipitate is obtained.

Absence of aluminium.

13. Test for iron.

a) To the 2ml of extract 2ml of ammonium thiocyanate solution is added.

b) To the 2ml of extract 2ml of ammonium thiocyanate solution and 2ml of concentrated nitric acid is added.

Absence of blood red colour.

Absence of blood red colour.

Absence of ferric iron.

Absence of ferric iron.

14. Test for zinc.

To the 2ml of the extract, sodium hydroxide solution is added in drops to excess.

A white precipitate solution is not obtained.

Presence of zinc

15. Test for calcium

2ml of the extract is added with 2ml of 4% ammonium oxalate solution.

No cloudy/white precipitate is obtained.

Absence of calcium.

16. Test for magnesium.

To the 2ml of the extract sodium hydroxide solution is added in drops to excess.

No white precipitate is obtained.

Absence of magnesium.

17. Test for ammonium.

To 2ml of extract few ml of Nesssler’s regeant and excess of sodium

hydroxide solution are added.

No reddish brown precipitate is obtained.

Absence of ammonium.

18. Test for potassium.

A pinch of substance is treated with 2ml of sodium nitrate solution, treated with 2ml of cobalt nitrite in 30%

glacial acetic acid.

Yellowish precipitate is not obtained.

Absence of potassium.

19. Test for sodium.

2 pinches of the substance is made into a paste by using hydrochloric acid and introduced into the blur flame.

No yellow colour flame is seen.

Absence of sodium.

20. Test for mercury.

2ml of the extract is treated with 2ml

of sodium hydroxide solution. No yellow precipitate is obtained.

Absence of mercury.

21. Test for arsenic.

2ml of the extract is treated with 2ml of silver nitrate solution.

No brownish red precipitate is obtained.

Absence of arsenic.

The results of this table will be illusterated later.

PRELIMINARY PHYTOCHEMICAL ANALYSIS:

Phyto chemical analysis were carried out to find out the presence of secondary metabolites in a plant. Secondary metabolites are substances which are not used by the plants and are considered as a waste products by the plants, these substances are stored in different parts of the plant body. Those substances which are not used by the plants are of great therapeutic value to humans. They possess disease curing principles. So the aim of this phyto chemical study is to evaluate the different kinds of secondary metabolites in roots of pongamia pinnata(linn) in fresh form as well as dry form

Preperation of plant extracts

:

The pungam ver choornam(10gm) was extracted with solvents namely methanol, ethyl acetate and chloroform. The filtrates were tested for the following phytoconstituents. These were the secondary metabolites which are not used by the plants but are more rich in therapeutic value.

TABLE-4.2.1.1

S NO

EXPERIMENT INFERENCE

I. Test for Tannins:

A plant sample dried powder 0.5 gm is boiled in 20 ml of water and filtered.The filterate 2 ml is taken and 3-5 drops of Fecl2 (0.1%) is slowly added to it.

Forms a brownish-green or bluish- black colour.

II. Test for Phlobatannins:

An aqueous 2 ml of plant sample is boiled in a hot water bath with 1 ml of aqueous Hcl

A red precipitate is deposited

III. Test for Saponin:

A powdered 2 gm of plant sample is boiled with 20 ml of distilled water, then filtered, the filterate is added with fresh 5 ml of distilled water and shaken vigorously.

Permenant or persistant froth is not formed.The froth is not turned in to emulsion by adding three drops of olive oil.

IV. Test for Flavonoids:

An aqueous filterate of plant sample is mixed with 5 ml of dilute Ammonium solution and few drops of concentrated H2SO4 is slowly added through the sides of the test tube.

Yellow colour formed and disappears on standing.When 1%

Alluminium solution is added in this mixture re-formation of yellow colour.

V. Test for steroids:

An ethonolic extract of plant sample 2ml is mixed with 2 ml H2SO4and 0.5 gm Acetic anhydride.

The solution turns in to blue to green colour

VI. Test for Cardiac glycosides:

In 5 ml of plant Ethanolic extract, 2 ml of Glacial acetic acid, a drop of Fecl₂and 1 ml of H₂SO₄(slowly on the sides of the test tube) is added.

A brown ring indicates deoxy sugar of cardenolides/violet ring appears below brown ring/ in acetic acid layer a green ring is formed

VII. Test for Terpenoids:

In 5 ml of Ethanolic plant extract, 2 ml of chloroform and 3 ml of concentrated H₂SO₄ (slowly) is added.

A reddish brown interface layer is formed

VIII. Test for Carbohydrates:

An aqueous plant extract is boiled in a water bath with Benedict’s solution.

A green or brick red or red precipitate shows the presence of reducing sugar

IX. Test for Alkaloids:

Alkaloids are identified by precipitate method (a) Mayer’s reagent:

An aqueous plant extract of 2 ml is mixed with 2 ml of mayer’s reagent

(b)Wagner’s reagent:

An aqueous plant extract of 2 ml is mixed with 2 ml of wagner’s reagent

(c) Dragendroff’s reagent:

An aqueous plant extract of 2 ml is mixed with 2 ml of Dragendroff’s reagent.

Forms whitish or yellowish cream colour precipitate

Forms a brown or dark reddish precipitate

Forms reddish brown precipitate

X. Test for Glycosides:

An aqueous plant extract of 2 ml is added with 1 ml of concentrated Hcl and boiled in a water bath for about 2 hours. The filtrate is separated and 3 ml of chloroform is added, then 10% of 1ml Ammonium solution is added.

Forms pink colour

XI. Test for Protein:

An aqueous extract /alcoholic extract of 2 ml is added with few drops of Biuret reagent and kept in hot water bath for 10 minutes.

Formation of light blue or Pale violet colour is absent

XII. Test for Phytosterols:

An ethanolic or a methonolic plant extract 2 ml is mixed with 2 ml of Acetic anhydride stirred well and heated for 2 minutes in hot water bath then allowed to cool.1 or 2 drops of H2SO4is added with the mixture slowly through the sides of the wall .

Forms greenish blue layer on the upper surface

XIII. Test for Phenolic compounds:

About 2 ml of aqueous plant extract is mixed with 2 ml of Fecl₃solution.

Formation of deep bluish green colour is absent

XVI. Test for Volatile oil:

An ethanolic plant extract of 2 ml is mixed with one or two drops of tincture in warm water bath

in a screwed cap test tube.

Red colour is not appeared

XV. Test for Fixed oil:

One ml of ethanolic extract of plant sample is mixed with 1 ml of 1% copper sulphate solution and 5 drops of 10% sodium Hydroxide solution

Formation of a clear blue solution is absent

4.2.3 EVALUATION OF ACUTE TOXICITY AND ANTIULCER PROPERTY OF PUNGAM VER CHOORNAM

INTRODUCTION:

Gastric and duodenal ulcers are illnesses that affect a considerable number of people in the world. Some of the causes of these disorders are: stress, smoking, nutritional deficiencies and ingestion of non-steroidal anti-inflammatory drugs. The pathogenesis of gastro duodenal ulcers is influenced by various aggressive and defensive factors, such as acid-pepsin secretion, mucosal barrier, mucus secretion, blood flow, cellular regeneration and endogenous protective agents (Prostaglandin and epidermal growth factor). In the present study an attempt was made to investigate the anti-ulcerogenic effects of Pungam Ver Choornam in animal models of gastric ulcers induced by acid-alcohol in rats. The acute toxicity of the drug was also investigated in mice.

MATERIALS AND METHODS Stock solution preparation:

The powdered form of Pungam Ver Choornam was filtered through cheesecloth and was mixed uniformly in 2% CMC solution to achieve 100mg/ml as main stock solution and used in this study.

Animals:

Mice of either sex weighing 25-30g and male Wistar rats weighing 150-200g were obtained from the animal house of Vels University. Animals were fed on conventional diets and water ad libitum and they were maintained under standard conditions of humidity, temperature (20-24°C) and light (12-h light: 12-h dark cycle). The rats were randomly assigned to control and different treatment groups, six animals per group. The study was conducted in accordance with CPCSEA (Committee for the Purpose of control and supervision of Experiment on Animals) guidelines and the study was approved by the Institutional Animal Ethical Committee. The animals were acclimatized for one week under laboratory conditions. All animal experiments were carried out in accordance with institutional Ethical Committee acts.

Acute toxicity study:

Acute oral toxicity test for the Pungam Ver Choornam was carried out as per OECD Guidelines 425. Animals are observed individually after dosing at least once during the first 30 minutes, periodically during the first 24 hours, with special attention given during the first 4 hours, and daily thereafter, for a total of 14 days, except where they need to be removed from the study and humanely killed for animal welfare reasons or are found dead.

All observations are systematically recorded and Observations include changes in skin and fur, eyes and mucous membranes, and also respiratory, circulatory, autonomic and central nervous systems, and somatomotor activity and behaviour pattern. Attention was directed to observations of tremors, convulsions, salivation, diarrhoea, lethargy, sleep and coma. The principles and criteria summarised in the Humane Endpoints Guidance Document taken into consideration. Animals found in a moribund condition and animals showing severe pain or enduring signs of severe distress was humanely killed. When animals are killed for humane reasons or found dead, the time of death was recorded.

4.2.4 Pharmacological evaluation of Anti-ulcer activity Hcl-ethanol-induced Gastric ulceration

:

In Hcl-ethanol-induced gastric ulcer protocols, rats were starved of food but not for water 24 hours. Negative control group received saline and test group pretreated with Pungam Ver Choornam at 500mg/kg, p.o for seven days and before receiving Hcl-ethanol and positive control group received ranitidine orally at 60 mg/kg 30 min prior were administered Hcl-ethanol orally to these five groups at 1ml/200g. The volume of the saline, Pungam Ver Choornam in suspension and ranitidine was adjusted to not more than 0.5ml/100g of body weight.

The ulcer index score for intensity of the gastric lesions was measured where score 0 = no ulcer, 1 = superficial mucosal erosion, 2 = deep ulcer or transdermal necrosis, and 3 = perforated or penetrated ulcer. Ulcer index = 1 O/X where,

Total area of stomach mucosa

X=. --- Total ulcerated area

Gastric Secretion

The gastric juice was collected 4hrs after ulcerogen administration and centrifuged for 5 minutes at 2000 rpm and the volume of supernatant was noted. The pH of the gastric juice was recorded by the pH meter. Then the contents were subjected to analysis for free and total acidity.

Free acidity and total acidity were determined using 0.01N NaoH and Topfer’s reagent containing phenolphthalein as indicator.

The acidity level was calculated by using the following formula:

Volume of NaoH X Normality X 100 Acidity = ---

0.1

Statistical analysis:

The statistical analysis was carried out using one-way ANOVA followed by Dunnett's multiple comparison test. All the results obtained in the study were compared with the vehicle control group. P values <0.05 were considered statistically significant.

The results of acute toxicity study and anti ulcer activity and the statistical results will be discussed later.

4.3 CLINICAL ASSESSMENT

Aim:

Peptic ulcer disease is one of the striking problem in the developing countries and also in the developed countries due to lifestyle modifications. Even though lot of medications are available for the treatment of this disease, still there is a need for a newer drug discovery which should have minimal side effects and cost effective. Hence to over come this lacuna I have selected a hebal drug pungam ver choornam for the treatment of peptic ulcer disesease

The aim of this clinical trial is to evaluate the potency of the trial drug clinically.

Objectives:

Primary objective- The primary objective of this study is to evaluate the safety of the drug.

Secondary objective- The secondary objective of this clinical study is to elaborate the efficacy of this trial drug clinically.

Trial method:

Phase II-Open clinical trial method was followed for this clinical study.

Selection of patients:

50 patients were selected for the clinical trial. This selection was based on the inclusion and exclusion criteria. Patients of different age group and different socio economic status were selected, 40 patients were selected out patient treatment and 10 patients were selected for inpatient treatment.

Study centre:

The clinical study was carried out in Gunapadam post graduate out patient and inpatient department, Government Aringar Anna Hospital for Indian Medicine and Homoeopathy, Chennai–106.

Criteria for inclusion

:

 Age group from 20-75

 Tenderness in the epigastric region

 Indigestion

 Belching

 Heart burn

 Nausea/vomiting

 Waterbrash

 Regurgitation

 Loss of appetite

 Constipation

Criteria for exclusion:

 Carcinoma of the stomach

 Pancreatitis

 Hepatic congestion

 Gastroesophagal reflux disease

 Cholecystitis

 Biliary colic

 Inferior myocardial infection

 Referred pain due to pleurisy, pericarditis

Criteria for withdrawal:

 Acute conditions such as hemorrhage, perforations.

 Irregular follow up

 Sudden increase in abdominal pain.

 Alcohol abuse

 Biphasic treatment method

Investigation :

All the 50 patients were investigated symptomatically.

They are also assesses for,

 Blood–TC, DC, ESR, Hb, sugar (random)

 Serum- urea creatinine cholesterol.

 Upper gastro intestinal endoscopy.

Criteria for assessment of response to therapy

:

1) Marked response- 100% complete relief in signs and symptoms, complete normalcy of the pathological conditions.

2) Moderate response-75% relief in the presenting signs and symptoms, marked normalcy in pathological conditions.

3) Mild response- 60-75% relief of signs and symptoms, moderate normalcy of the pathological conditions.

4) Poor responser- 50% relief of signs and symptoms with no marked change in pathological investigation.

Line of treatment:

Medicine : Pungam ver choornam

Route : Enteral

Dose : 1gm/BD/after food

Vehicle : Hot water

Duration of treatment : 7 weeks