PHARMACOLOGICAL EVALUATION OF WHOLE PLANT EXTRACT FROM CROTON BONPLANDIANUM BAILL

PHARMACOLOGICAL EVALUATION OF WHOLE PLANT EXTRACT FROM CROTON BONPLANDIANUM BAILL

A Dissertation submitted to

THE TAMILNADU DR. M.G.R. MEDICAL UNIVERSITY CHENNAI- 600 032

In partial fulfillment of the requirement for the award of the Degree of MASTER OF PHARMACY

IN

BRANCH IX - PHARMACOLOGY

Submitted By G. RAMACHANDRAN

Reg. No: 261725151

Under the guidance of

Mr. K.A.S MOHAMMED SHAFEEQ, M. Pharm., Associate Professor, Department of Pharmacology

PERIYAR COLLEGE OF PHARMACEUTICAL SCIENCES TIRUCHIRAPALLI-620 021

(An ISO 9001: 2015 Certified Institution) MAY 2019

Mr. K.A.S. Mohammed Shafeeq, M. Pharm., Associate Professor, Department of Pharmacology Periyar College of Pharmaceutical Sciences Trichirappalli - 620 021.

CERTIFICATE

This is to certify that the dissertation entitled

“PHARMACOLOGICAL EVALUATION OF WHOLE PLANT EXTRACT FROM CROTON

BONPLANDIANUM BAILL.”

Submitted by

Mr. G.RAMACHANDRAN [Reg. No: 261725151]

for the award of the degree of

“MASTER OF

PHARMACY”

is a bonafide research work done by him in the Department of Pharmacology, Periyar College of Pharmaceutical Sciences, Trichirappalli during the academic year 2017-2019 under my direct guidance and supervision.

Place: Trichirappalli

Date:

(K. A. S. Mohammed Shafeeq)

Forwarded

Prof. Dr. R. Senthamarai, M. Pharm., Ph.D., Principal

Periyar College of Pharmaceutical Sciences Trichirappalli - 620 021.

CERTIFICATE

This is to certify that the dissertation entitled

“PHARMACOLOGICAL EVALUATION OF WHOLE PLANT EXTRACT FROM CROTON

BONPLANDIANUM BAILL.”

done by

Mr. G.RAMACHANDRAN [Reg. No: 261725151]

for the award of the degree of

“MASTER OF

PHARMACY”

under The Tamilnadu Dr. M.G.R. Medical University, Chennai is a bonafide research work performed by him in the Department of Pharmacology, Periyar College of Pharmaceutical Sciences, Trichirappalli. The work was performed under the guidance and supervision of Mr. K.A.S. Mohammed Shafeeq, M. Pharm., Associate Professor, Department of Pharmacology, Periyar College of Pharmaceutical Sciences, Trichirappalli.

This dissertation is submitted for acceptance as project for partial fulfillment of the

degree of

“MASTER OF PHARMACY”

in Pharmacology, of The Tamilnadu Dr. M.G.R. Medical University, during May 2019.

Place: Trichirappalli

Date: (Dr. R. Senthamarai)

ACKNOWELEDGEMENT

Though words are seldom sufficient to express gratitude and feelings, it somehow give us an opportunity to thank those who helped us during the tenure of our study.

I consider it as my grater privilege to express my ardent thanks and ineffable sense of gratitude to my guide Mr. K. A. S. Mohammed Shafeeq, M. Pharm., Associate Professor, Department of Pharmacology, Periyar College of Pharmaceutical Sciences, Trichirappalli. It is my foremost duty to express my sincere independence to his constant help, affection and valuable guidance during the course of present investigation.

I feel to honor to owe my profund sense of gratitude and heartfelt thanks to Dr. R. Senthamarai, M. Pharm., Ph.D., Principal, Periyar College of Pharmaceutical

Sciences, Trichirappalli for her whole hearted co-operation in rendering facilities to proceed with this study.

I express my profund thanks to Dr. A.M. Ismail, M. Pharm., Ph.D., Professor Emeritus and Prof. Dr. G. Krishnamoorthy, B.Sc., M. Pharm., Ph.D., Vice Principal and Head, Department of Pharmaceutical Chemistry. Periyar College of Pharmaceutical Sciences, Trichirappalli for their moral support to complete my project work and have always propelled me to perform better.

My heartful and deep sense of gratitude to Dr. K. Veeramani, M.A., B.L., honurable Chairperson, Periyar College of Pharmaceutical Sciences, Trichirappalli for providing all infrastructural facilities to carryout this work during my studies.

I submit my sincere thanks and respectful regards to Thiru. Gnana Sebastian, Correspondent, Periyar College of Pharmaceutical Sciences, Trichirappalli for his constant support and encouragement to carry out work and my studies.

I offer my warmest acknowledgement to Dr. S. Karpagam Kumara Sundari, M. Pharm., Ph.D., Head, Department of Pharmacology and Dean (UG) Periyar College of

Pharmaceutical Sciences, Trichirappalli for her continuous guidance and moral support from date of admission to complete my project work and have always propelled me to perform better.

I gratefully to acknowledge Dr. T. Shri Vijaya Kirubha, M. Pharm., Ph.D., Head, Department of Pharmacognosy for her constant support during extraction and Phytochemical screening of the Selected plant.

I express my earnest thanks to Dr. V. Nandagobalan, M.Sc., M.Phil., Ph.D., SLST., Dean of Sciences, Associate Professor, National College, Trichirappalli for his valuable help in authentication of plant.

I extend my heartfelt thanks to all the Staff Members of Periyar College of Pharmaceutical Sciences, Trichirappalli for their valuable support.

I thank sincerely the Librarian and Assistant Librarian for the reference to the resource of knowledge and wisdom.

Not as words but from the depth I thank my parents for giving me unconditional support and motivation to pursue my interest even it went beyond the boundaries.

Finally I convey my thanks to everyone for their help in the completion of this research work successfully.

G.RAMACHANDRAN

PERIYAR COLLEGE OF PHARMACEUTICAL SCIENCES DEPARTMENT OF PHARMACOLOGY

INSTITUTIONAL ANIMAL ETHICAL COMMITTEE (IAEC) CENTRAL ANIMAL HOUSE REGISTRATION NUMBER: 265/2000/CPCSEA

Title of the project : Pharmacological Evaluation of Whole Plant Extract From Croton Bonplandianum Baill

Authors : G. Ramachandran & Mr. K.A.S. Mohammed Shafeeq Proposal number : PCP/IAEC/001/2019

Date first received : 06.11.2018 Date received after

modification (if any) : 12.12.2018 Date received after second

modification (if any) : 02.01.2019

Approval date : 03.01.2019

Expiry date : 03.01.2020

Name of IAEC/CPCSEA

Chairperson : The HoD

Department of Pharmacology

Periyar College of Pharmaceutical Sciences Trichy – 21

Date : 03.01.2019 CHAIRMAN

INSTITUTIONAL ANIMAL ETHICS COMMITTEE PERIYAR COLLEGE OF PHARMACEUTICAL

SCIENCES

CONTENTS

S.NO CONTENTS Pg. No.

1 INTRODUCTION 1

2 LITERATURE REVIEW 25

3 OBJECTIVE 44

4 PLAN OF THE WORK 45

5 PLANT PROFILE 46

6 METHODOLOGY 52

7 RESULTS AND DISCUSSION 60

8 CONCLUSION 79

9 BIBLIOGRAPHY 81

LIST OF TABLES

Table

No. Particulars Page

No.

1. Herbs used in ulcer and anthelmintic 24

2. Preliminary Phytochemical Studies 60

3. Data Showing the HPTLC of Croton bonplandianium Rf Value at 254 nm 61 4. Data Showing the HPTLC of Croton bonplandianium Rf Value at 366 nm 61 5. Behavioral changes in Acute Oral Toxicity in Albino rats 66 6. Effect of Test compound on Body Weight in Acute oral toxicity in Albino rats 68 7. Effect of Test compound on Biochemical parameters in Acute oral toxicity in

Albino rats 68

8. Effect of Ethanolic extract of Croton bonplandianum on aspirin - induced

ulcer in rats 70

9. Anthelmintic activity of Ethanolic extract of Croton bonplandianum against

Pheretima posthuma 73

LIST OF FIGURES

Figure

No. Particulars

1. Healthy and Ulcerated Stomach

2. Pathophysiology of ulcer

3. Mechanism Action of Histamine Antagonist

4. Mechanism Action of Proton Pumping Inhibitors

5. Mechanism Action of Anti-H. Pylori Drugs

6. Mechanism Action of Anti-Ulcer Drugs

7. Stastices of Ulcer

8. Plant of Croton bonplandianum baill

9. HPTLC Finger Printing Chromatogram (5µl & 10 µl)

10. HPTLC Finger Printing Chromatogram (15µl)

11. HPTLC under UV Light

12. HPTLC under visible light & 3D Display Chromatogram

13. Effect of Test compound on Body Weight in Acute oral toxicity in Albino rats 14. Effect of Test compound on Biochemical parameters in Acute oral toxicity in

Albino rats

15. Effect of Aspirin induced Gastri Ulcer.

16. Effect of Drugs on Ranitidine

17. Effect of test drug on Croton bonplandianum

18. Antiulcer activity of Ethanolic Extract of Croton bonplandianum against Aspirin induced ulcer in Albino rats

19. Control - 1% of CMC in 10ml of Normal saline

20. Standard – Albendazole

21. Test drug – 100mg/ml of Ethanolic extract of Croton bonplandianum 22. Anthelmintic activity of Ethanolic extract of Croton bonplandianum against

Phertima Posthuma

ABBREVIATIONS

WHO World Health Organization

CSIR Council for Scientific and Industrial Research SOP Standard Operating Procedure

GAP Good Agricultural Practice GLP Good Laboratory Practice

GSP Good Supply Practice

GMP Good Manufacturing Practice

BMI Body Mass Index

CT Computed Tomography Scanning

MRI Magnetic Resonance Index

ADR Adverse Drug Reaction

OD Once a Day

SSRI Selective Serotonin Reuptake Inhibitor MAOI Mono Amino Oxidase Inhibitor

CNS Central Nervous System

NSAIDs Non - Steroidal Anti-Inflammtory Drugs IC Inhibitory Concentration

LD Lethal Dose

HPTLC High Performance Thin Layer Chromatography

TLC Thin Layer Chromatography

MIC Minimum Inhibitory Concentration

OECD Organisation for Economic Cooperation and Development

IAEC Institutional Animal Ethical Committee ANOVA Analysis of Variance

CPCSEA Committee for the Purpose of Control and Supervision of Experimental animals

kg Kilogram

mg Milligram

ml Milli Litres

Mmol Milli Mole

pH Negative Logarithm of Hydrogen Ion Concentration

p.o. Per Oral

S.E.M. Standared Error Mean ZES Zollinger Ellison Syndrome

IM Inner Membrane

OM Outer Membrane

PG Prostaglandin

PPIs Proton Pump Inhibitors GIT Gastro Intestinal Tract LC50 Lethal Concentration CRS Cold Resistant Stress

PL Pylorus Ligation

LPO Lipid Peroxidation

AgNPs Silver Nanoparticles

SDS Sodium Dodecyl Sulphate

MDA Melon Dialdehyde

PUD Peptic Ulcer Disease

UI Ulcer Index

NS Normal Saline

OTC Over the Counter Drug

FDA Food and Drug Administration

LD50 Lethal Dose

V/V Volume /Volume

W/V Weight/Volume

BW Body Weight

1. INTRODUCTION

1.1 HERBAL MEDICINES

Indian traditional systems of medicines always played important role in meeting the global health care needs. They are continuing to do so at present and shall play major role in future also. The system of medicines which are considered to be Indian in origin or the systems of medicine, which have come to India from outside and got assimilated in to Indian culture are known as Indian Systems of Medicine. India has the unique distinction of having six recognized systems of medicine in this category. They are Ayurveda, Siddha, Unani and Yoga, Naturopathy and Homoeopathy. Though homoeopathy came to India in 18^th Century it completely assimilated in to the Indian culture and got enriched like any other traditional system hence it is considered as part of Indian Systems of Medicine. Apart from these systems there are large number of healers in the folklore stream who have not been organized under any category. In the present review, attempt would be made to provide brief profile of three systems to familiarize the readers about them so as to facilitate acquisition of further information.

Most of the traditional systems of India including ayurveda have their roots in folk medicine. However what distinguishes ayurveda from other systems is that it has a defined conceptual framework that is consistent throughout the ages. In conceptual base, it was perhaps highly evolved and far ahead of its time. It was among the first medical systems to advocate an integrated approach towards matters of health and disease. Another important distinguishing feature of ayurveda is that unlike other medical systems, which developed their conceptual framework based on the results obtained with the use of drugs and therapy, it first provided philosophical framework that determined the therapeutic practice with good effects. It is philosophical base is partly derived from ‘Samkhya’ and ‘Nyaya vaisheshika’ streams of Indian philosophy. This enabled it to evolve into rational system of medicine quite early in its evolution and to get detached from religious influence. It laid great emphasis on the value of evidence of senses and human reasoning.

Herbal medicine is an interdisciplinary branch between herbal medicine and ayurveda and it covers all the fields of herbal medicine related to botany, medicinal plant research Pharmacognosy, Phytochemistry, Phytotherapy, botanical medicines, ayurveda and

natural chemistry, agriculture science, unani medicine, biotechnology and biochemistry.

Herbal medicine is an open access journal and the main aim of this journal to provide a platform for scientists, researchers in herbal medicine and Ayurveda fields all over the world to present their new ideas discuss new strategies and promote developments in all areas of herbal medicine¹.

1.2. MODERN HERBAL MEDICINE²

The World Health Organization (WHO) estimates that 80 percent of the population of some Asian and African countries presently use herbal medicine for some aspect of primary health care. Pharmaceuticals are prohibitively expensive for most of the world's population, half of whom lived on less than $2 U.S. per day in 2002. In comparison, herbal medicines can be grown from seed or gathered from nature for little or no cost.

Many of the pharmaceuticals currently available to physicians have a long history of use as herbal remedies including opium, aspirin, digitalisand quinine. According to the World Health Organization approximately 25% of modern drugs used in the United States have been derived from plants. At least 7,000 medical compounds in the modern pharmacopoeia are derived from plants. Among the 120 active compounds currently isolated from the higher plants and widely used in modern medicine today, 80% show a positive correlation between their modern therapeutic use and the traditional use of the plants from which they are derived.

1.3. METHODS OF HERBAL SYSTEM

 Siddha

 Ayurveda

 Unani

 Homeopathy.

Siddha Medicine

Siddha medicine is a system of traditional medicine originating in ancient tamilnadu in south India and sri lanka. Traditionally it is taught that the siddhars laid the foundation for this system of medication. Siddhars were spiritual adepts who possessed the ashta siddhisor the eight supernatural powers. Agastyar is considered the first siddha and the guru of all siddhars the siddha system is believed to have been handed over to him

by muruganson of shiva and parvati. Siddha is focused on "Ashtamaha siddhi" the eight super natural power. Those who attained or achieved these powers are known as siddhars.

There were 18 important siddhars in olden days and they developed this system of medicine. Hence it is called siddha medicine.

Concept of Disease and Cause³

It is assumed that when the normal equilibrium of the three humors vaadham, pitham and kapamis disturbed disease is caused. The factors assumed to affect this equilibrium are environment, climatic conditions, diet, physical activities and stress. Under normal conditions the ratio between vaadham, pitham and kapam are 4:2:1 respectively.

According to the Siddha medicine system diet and lifestyle play a major role in health and in curing diseases. This concept of the Siddha medicine is termed as pathiyam and apathiyam which is essentially a list of "do's and don'ts".

Diagnosis

 Varnam (colour)

 Kural(voice)

 Kan (eyes)

 Thodal(touch)

 Malam (stool)

 Neer (urine)

 Naadi (pulse) Ayurveda Medicine⁴

The main classical ayurveda texts begin with accounts of the transmission of medical knowledge from the Gods to sages and then to human physicians. In Sushruta Samhita, sushrutawrote thatdhanvantarihindu god of ayurveda in carnated himself as a king of varanasi and taught medicine to a group of physicians including sushruta. Ayurveda therapies have varied and evolved over more than two millennia.Therapies are typically based on complex herbal compounds minerals and metal substances. Ancient ayurveda texts also taught surgical techniques including kidney stone extractions sutures and the extraction of foreign objects.

Eight components in Ayurveda

 Kayacikitsa

 Kaumara

 Salyatantra

 Salakyatantra

 Bhutavidya

 Agadatantra

 Rasayanatantra

 Vajikaranatantra Diagnosis

Ayurveda has eight ways to diagnose illness called Nadi (pulse), Mootra (urine), Mala (stool), Jihva (tongue), Shabda (speech), Sparsha (touch), Drunk (vision) and Aakruti (appearance). Ayurvedic practitioners approach diagnosis by using the five senses.

For example hearing is used to observe the condition of breathing and speech.

Treatment and prevention

Two of the eight branches of classical ayurveda deal with surgery but temporary ayurveda tendsto stress attaining vitality by building a healthy metabolic system and maintaining good digestion and excretion. Ayurveda also focuses on exercise, yoga and meditation. One type of prescription is a sattvic diet. Ayurveda follows the concept says thatnatural cycles. Are important for health hygiene, including regular bathing cleaning of teeth, skin care and eye washing is also a central practice.

Unani Medicine

Unanimedicineis the term for Arabic traditional medicine as practiced in mughal India and in Muslim culture in south Asia and modern day central Asia. The Unani medicine is considered to be a product of pseudoscience by several skeptics. The term means "Greek"as the Perso-arabic system of medicine was based on the teachings of the Greek physicians Hippocrates and Galen. The hellenistic origin of unani medicine is still visible in its being based on the classical Phlegm blood, (Dam), Yellow bile and Black bile but it has also been influenced by indian and chinesetraditional systems.

Diagnosis and Treatment

Unani classical literature consists of thousands of books. According to unani medicine management of any disease depends upon the diagnosis of disease. In the diagnosis clinical features such as signs, symptoms, laboratory feature are important.

According to unanipractitioners any cause or factor is countered by the failing of which may lead to quantitatively or qualitatively derangement of the normal equilibrium of body which constitute the tissues and organs. This abnormal humor is believed to lead to pathological changes in the tissues anatomically and physiologically at the affected site and exhibits the clinical manifestations.

After diagnosing the disease is determined on the basis of etiology in the following pattern:

 Elimination of cause.

 Normalization of humors.

 Normalization of tissues/organs.

 Regimental therapy - The disease may be treated by the modification of six essential pre-requisites of health. May be modified by the use of one or more regimens and alteration of food.

 Pharmacotherapy - For this purpose prescription is formulated which contain the single or compound unani drugs having desired actions as per requirements.

As an alternative form of medicine unani has found favor in India where popular products like Roghan Baiza Murghare commonly used for hair care. Unani practitioners can practice as qualified doctors in India as the government approves their practice. Unani medicine has similarities to ayurveda. Both are based on theory of the presence of the elements in the human body. According to followers of unani medicinethese elements are present in fluids and their balance leads to health and their imbalance leads to illness.

The theory postulates the presence of blood, phlegm, yellow bile and black bile in the human body. Each person's unique mixture of these substancesare determine. Predominance of blood gives a sanguine temperament of phlegm makes one phlegmatic yellow bile, bilious and black bile, melancholic.

Homeopathy⁵

Homeopathy is a system of alternative medicine created in 1796 by Samuel Hahnemannbased on his doctrine oflike cures like, a claim that a substance that causes the symptoms of a disease in healthy people would cure similar symptoms in sick people. Homeopathy is a pseudoscience a belief that is incorrectly presented as scientific.

Homeopathic preparations are not effective for treating any condition. Large scale studies have found homeopathy to be no more effective than a placebo, indicating that any positive effects that follow treatment are due to factors such as normal recovery from illness or regression toward the mean.

Hahnemann believed the underlying causes of disease were phenomena that he termed that homeopathic preparations addressed the preparations are manufactured using a process of homeopathic dilutionin which a chosen substance is repeatedly diluted in alcohol or distilled water each time with the containing vessels being struck against an elastic material commonly a leather bound book. Dilution typically continues well past the point where no molecules of the original substance remain homeopathic by consulting reference books known as repertories and by considering the totality of the patient's symptoms, personal traits, physical and psychological state and life history.

Homeopathy is not a plausible system of treatment as its dogmas about how drugsillness the human body, liquids and solutions operate are contradicted by a wide range of discoveries across biology, psychology, physics and chemistry made in the two centuries since its invention. Although some clinical trials produce positive results, multiple systematic reviews have indicated that this is because of chance, flawed research methods and reporting bias. Homeopathic practice has been criticized as unethical because it discourages the use of effective treatments with the World Health Organization warning against using homeopathy to try to treat severe diseases such as HIV and malaria. The continued practice of homeopathy despite a lack of evidence of efficacy has led to it being characterized within the scientific and medical communities as nonsense quackery and a sham.

History

The concept of cures like may have been suggested by Hippocrates around 400 BC when he prescribed a small dose of mandrake root to treat mania, knowing it produces mania in much larger doses. Similarly in the 16th century often translated as "what makes a

man also cures him". In the late 18th and 19^thcenturies mainstream medicine used methods like bloodletting and purging and administered complex mixtures such as which was made from 64 substances including these treatments often worsened symptoms and sometimes proved fatal. Hahnemann rejected these practices which had been extolled for centuriesas irrational and inadvisable instead he advocated the use of single drugs at lower doses and promoted an immaterial, view of how living organisms function, believing that diseases have spiritual, as well as physical causes.

2. PEPTIC ULCER

An ulcer is a sore, which means it is an open, painful wound. Peptic ulcers are ulcers that form in the stomach or the upper part of the small intestine, called the duodenum.

Peptic ulcers are actually very common. Peptic ulcers are open sores that develop on the inside lining of stomach and the upper portion of small intestine. The most common symptom of a peptic ulcer is stomach pain⁶.

Fig. No.1:Healthy and Ulcerated Stomach

FACTORS INVOLVED IN ULCER⁷ Prostaglandin Role in Ulcer

Prostaglandins are important in the pathophysiology of peptic ulcer disease and possibly in its prevention and treatment as well. Prostaglandins have been shown to inhibit gastric secretion, stimulate bicarbonate secretion and increase gastric blood volume.

Pretreatment with prostaglandins has recently been found to promote rapid restitution of the superficial epithelium of the gastric mucosa and preservation of the deeper layers.

Histamine Role in Ulcer⁸

Ulcer have an increased vagal drive leading to augmented histamine release and a decreased mucosal histamine vagotomy abolished the vagal drive,

decreased histamine release and increased histamine content which led to a reduction in stimulated gastric acid secretion.

Gastrin Role in Ulcer

Duodenal ulcer patients are characterized by an antrum-predominant, body-sparing, non-atrophic Helicobacter pylori (H. pylori) gastritis, which results in increased gastrin release and increased acid secretion. The increased gastrin release is caused by the infection impairing the acid-mediated inhibitory control of gastrin release. The elevated levels of the gastrin stimulate the healthy uninflamed, non-atrophic acid-secreting region of the stomach to secrete excess amounts of acid. The increased gastrin also exerts trophic effects on the oxyntic mucosa, causing hyperplasia of both the Enterochromaffin-like cells and the parietal cells. These trophic changes in the mucosa further enhance its ability to secrete acid. The increased acid secretion results in an increased duodenal acid load, causing gastric metaplasia of the duodenal bulb and eventually the development of ulceration.

Acetylcholine Role in Ulcer

The role of gastrin-releasing peptide (GRP) and acetylcholine in the secretion of gastrin which plays a major role in gastric acid secretion and the relationship between gastrin secretion and stomach pH. Bombesin, which has a structure analogous to that of GRP, was used in the experiment. We also investigated whether acetylcholine has muscarine-like or nicotine-like action. Our findings pointed to the presence of an alternative, GRP-mediated, route for stimulating gastric secretion from G cells other than the acetylcholine-mediated route. This secretion was not inhibited by atropine. The results suggested that there are two routes for inducing gastrin secretion from G cells an acetylcholine-mediated route and a GRP-mediated route intramural peptide neurons. As GRP induced gastrin secretion, regardless of stomach pH, GRP was considered to be more closely related to gastrin secretion. The results also suggested that a muscarine-like action, particularly in the M3 receptor-mediated route, plays a significant role in acetylcholine- mediated gastrin secretion and that nicotine-like action is not involved in gastrin secretion.

Adenylcyclase Role in Ulcer⁹

Adenylcyclase also commonly known as adenylcyclase is an enzyme with key regulatory roles in essentially all cells. Human gastric mucosal adenylatecyclase activation by histamine H2-receptor stimulation and inhibition by cimetidine in vitro and in peptic ulcer patients. The G- protein associates with adenylyl cyclase, which converts ATP to cAMP, spreading the signal.

Types of peptic ulcer

 Gastric ulcers that occur on the inside of the stomach.

 Duodenal ulcers that occur on the inside of the upper portion of the small intestine.

 The most common causes of peptic ulcers are infection with the bacterium of H.

pylori and long-term use of aspirin and non-steroidal anti-inflammatory drugs.

 Stress and spicy foods do not cause peptic ulcers. However, they can make symptoms worse.

2.1 CAUSES OF ULCER

Peptic ulcers occur when acid in the digestive tract eats away at the inner surface of the stomach or small intestine. The acid can create a painful open sore that may bleeding.

Digestive tract is coated with a mucous layer that normally protects against acid. But if the amount of acid is increased or the amount of mucus is decreased that could develop an ulcer. In 1982, two doctors Barry Marshall and Robin Warren discovered a certain kind of bacteria that can live and grow in the stomach. Both doctors went on to win the Nobel Prize for their discovery. The medical name for these bacteria is Helicobacter pylori. Today doctors know that most peptic ulcers are caused by an infection from H. pylori. Experts believe that 90% of all people with ulcers are infected with H. pylori.

 H. pylori bacteria commonly live in the mucous layer that covers and protects tissues that line the stomach and small intestine. Often, the H. pylori bacterium causes no problems but it can cause inflammation of the stomach's inner layer, producing an ulcer. H. pylori infection spreads it may be transmitted from person to person by close contact, such as kissing. People may also contract H. pylori through food and water.

 Regular use of certain pain relievers taking aspirin, as well as certain over-the- counter and prescription pain medications called non-steroidal anti-inflammatory drugs (NSAIDs) can irritate or inflame the lining of your stomach and small intestine. These medications include ibuprofen, naproxen sodium etc.

 Other medication staking certain other medications along with NSAIDs, such as steroids, anticoagulants, low-dose aspirin, selective serotonin reuptake inhibitors (SSRIs) Alendronate and Risedronate can greatly increase the chance of developing ulcers.

 Smoking may increase the risk of peptic ulcers in people who are infected with H.

pylori.

 Alcohol can irritate the mucous lining of stomach and it increases the amount of stomach acid that produced ulcer.

OTHER WAY OF ULCER FORMATION

Stress: Numerous studies have presented conflicting conclusions about the role of Psychological stress in peptic ulcer disease. Stress may worsen ulcers not cause them.

However in patients with duodenal ulcers, stress has been shown to increase acid secretion.

Physical stress meanwhile may increase the risk of stomach ulcers. People with severe burns or those undergoing major surgery often require treatment to prevent ulcers.

Zollinger Ellison Syndrome (ZES): This rare disorder causes tumors in the pancreas and duodenum and ulcers in the stomach and duodenum. The tumors secrete a hormone called gastrin that causes the stomach to produce too much acid.

COMPLICATIONS¹¹

 Internal bleeding can occur as slow blood loss that leads to anemia or as severe blood loss that may require hospitalization or a blood transfusion. Severe blood loss may cause black or bloody vomit or black or bloody stools.

 Infectionpeptic ulcers can eat a hole through the wall of your stomach or small intestine, putting you at risk of serious infection of abdominal cavity.

 Obstructionpeptic ulcers can block passage of food through the digestive tract, causing you to become full easily, to vomit and to lose weight through either swelling from inflammation or scarring.

2.2 SIGNS AND SYMPTOMS

Stomach pain is the most common symptom of an ulcer. It usually feels like sharp aches between the breast bone and the belly button. This pain often comes a few hours after eating. It can also happen during the night or early in the morning, when the stomach is empty. Eating something or taking an antacid medication sometimes makes the pain go away for a while. Anyone who thinks he or she may have an ulcer needs to see a doctor.

Over time, untreated ulcers grow larger and deeper and can lead to other problems, such as bleeding in the digestive system or a hole in the wall of the stomach or duodenum, which can make someone very sick.

 Loss of appetite.

 Sudden, sharp stomach pains.

 Nausea.

 Frequent burning.

 Weight loss.

 Vomiting.

 Blackish bowel movements.

2.3 PATHOPHYSIOLOGY OF ULCER¹²

Heterogeneity is the most important consideration in the pathophysiology of peptic ulcer disease. Acute ulcers and erosions present clinically with gastrointestinal bleeding or perforation. If they heal there is no predictable recurrence. Factors concerned with mucosal defense are relatively more important than aggressive factors such as acid and pepsin. Local ischemia is the earliest recognizable gross lesion. The gastric mucosa is at least as vulnerable as the duodenal mucosa and probably more so. Most drug-induced ulcers occur in the stomach. Chronic or recurrent true peptic ulcers usually present with abdominal pain.

Many duodenal ulcer patients report that the pain occurs when the stomach is empty or is relieved by food and follows a pattern of relatively long periods of freedom from symptoms

between recurrences. Approximately 50% of patients experience a recurrence within a year if anti-ulcer medication is stopped. In most western countries recurrent duodenal ulcer is more common than gastric ulcer. Peptic ulcer disease is also more common in men. Recent evidence indicates genetic and familial factors in duodenal ulcer and increased acid-pepsin secretion in response to a variety of stimuli. However, it is also becoming clear that of all the abnormal functions noted, few are present in all subjects and many are clustered in subgroups. In chronic gastric ulcer of the corpus, defective defense mechanisms, such as duodenogastric reflux and atrophic gastritis, seem to be more important than aggressive factors. Nevertheless, anti-secretory medications accelerate the healing of such ulcers. It remains to be seen whether prostaglandins, mucus secretion or gastric mucosal blood flow are impaired in chronic ulcer disease.

Fig. No. 2: Pathophysiology of ulcer

DIAGNOSIS¹³

In addition to doing a physical examination, the doctor will take a medical history by asking about any concerns and symptoms you have your past health, health any medications you are taking any allergies you may have and other issues. If you have stomach pain or other symptoms of an ulcer the doctor will perform some tests to help make the diagnosis. One test is called an upper gastrointestinal (GI) series this is a type of X-ray of the stomach, duodenum and esophagus the muscular tube that links the mouth to the stomach. Person drinks a whitish liquid called barium while getting an X-ray and if he or she has an ulcer it should be outlined on the X-ray.

Another common procedure to look for an ulcer is called an endoscopy. During this test the doctor uses an endoscope, a skinny, lighted tube with a special camera on the end. A person getting an endoscopy is given anesthesia and will have no memory of the procedure.

For an endoscopy the doctor gently guides the endoscope into the throat and down into the esophagus and finally into the stomach and upper intestines. The doctor is able to look at the inner lining of these organs from the camera on a television screen and can even take pictures. Tissue can be removed during an endoscopy and then tested for H. pylori bacteria.

Laboratory tests for H. Pylori your doctor may recommend tests to determine whether the bacterium H. pylori is present in your body. He or she may look for H. pylori using a blood, stool or breath test. The breath test is the most accurate. Blood tests are generally inaccurate and should not be routinely used.

Endoscopy

Doctor may use a scope to examine your upper digestive system. During endoscopy your doctor passes a hollow tube Equipped with a lens down throat and into your esophagus, stomach and small intestine. Using the endoscope, doctor looks for ulcers. If your doctor detects an ulcer, small tissue samples may be removed for examination in a lab.

A biopsy can also identify whether H. pylori is in your stomach lining. Doctor is more likely to recommend endoscopy if you are older, have signs of bleeding or have experienced recent weight loss or difficulty eating and swallowing. If the endoscopy shows an ulcer in your stomach, a follow-up Endoscopy should be performed after treatment to show that it has healed even if your symptoms improve.

 Upper gastrointestinal series sometimes called a barium swallow, this series of X-rays of your upper digestive system creates images of your esophagus, stomach and small intestine. During the X-ray, you swallow white liquid containing barium that coats digestive tract and makes an ulcer more visible. Usually two antibiotics to kill the H.

pylori bacteria are taken every day for about 2 weeks.

 Antacid acid blockers or proton pump inhibitors are given for 2 months or longer to lessen the amount of acid in the stomach and help protect the lining of the stomach so the ulcer can heal.

2.4 TREATMENT OF ULCER¹⁴

Treatment for peptic ulcers depends on the cause usually treatment will involve killing the H. pylori bacterium if present, eliminating or reducing use of NSAIDs if possible and helping your ulcer to heal with medication

Medications can include

 Antibiotic medications to kill H. pylori is found in digestive tract, doctor may recommend a combination of antibiotics to kill the bacterium. These may include amoxicillin, clarithromycin, metronidazole, tinidazole, tetracycline and levofloxacin.

The antibiotics used will be determined by where you live and current antibiotic resistance rates. You'll likely need to take antibiotics for two weeks, as well as additional medications to reduce stomach acid, including a proton pump inhibitor and possibly bismuth subsalicylate.

 Medications that block acid production and promote healing proton pump inhibitors also called PPIs reduce stomach acid by blocking the action of the parts of cells that produce acid. These drugs include the prescription and over-the-counter medications Omeprazole, Lansoprazole, Rabeprazole, Esomeprazole and Pantoprazole.

Long-term use of proton pump inhibitors, particularly at high doses, may increase your risk of hip, wrist and spine fracture. Ask your doctor whether a calcium supplement may reduce this risk.

 Medications to reduce acid production acid blockers also called histamine H2

blockers reduce the amount of stomach acid released into your digestive tract, which relieves ulcer pain and encourages healing. Available by prescription or over-the- counter, acid blockers include the medications Ranitidine, famotidine, cimetidine and Nizatidine.

 Antacids that neutralize stomach acidyour doctor may include an antacid in your drug regimen. Antacids neutralize existing stomach acid and can provide rapid pain relief. Side effects can include constipation or diarrhea, depending on the main ingredients. Antacids can provide symptom relief, but generally aren't used to heal your ulcer.

 Medications that protect the lining of your stomach and small intestinein some cases your doctor may prescribe medications called Cytoprotective agents that help protect the tissues that line your stomach and small intestine. Options include the prescription medications sucralfate and misoprostol.

2.5 CLASSIFICATION OF DRUGS

Selective Histamine Type 2 Receptor Antagonists

 Cimetidine

 Famotidine

 Nizatidine

Fig.No.3: Mechanism Action of Histamine Antagonist

Proton Pumping Inhibitor

 Dexlansoprazole

 Esomeprazole

 Lansoprazole

 Omeprazole

 Pantoprazole

 Rabeprazole

Fig. No. 4: Mechanism Action of Proton Pumping Inhibitors

Anti-H.Pylori Drugs¹⁵

 Tetracycline

 Clarithromycin

 Metronidazole

 Tinidazole

 Amoxicillin

First and second line broad spectrum antibiotics used in H. pylori treatment target DNA synthesis and replication, transcription and translation of new proteins and membrane and cell wall synthesis. Novel broad-spectrum targets under investigation for H. pylori treatment include bacterial metabolic or synthetic pathways, electron transport chains and carriers. In addition, a number of pathways are targeted that are expected to result in inhibitors with narrow-spectrum activity aimed at H. pylori specifically. These include pH control pathways, H. pylori adherence and virulence factor secretion and activity.

Fig. No. 5: Mechanism Action of Anti-H. Pylori Drugs

IM: Inner membrane; OM: Outer membrane; PG: Peptidoglycan.

Fig. No. 6 : Mechanism Action of Anti-Ulcer Drugs

PREVENTION OF ULCER

Doctors are not totally certain how H. pylori bacteria are transmitted from person to person. The bacteria have been found in saliva, so kissing may be one way. They also may be spread through food, water or contact with vomit that has been infected with the bacteria.

The best advice in ulcer prevention is to always wash your hands after you use the bathroom and before you eat and to take good care of your body by exercising regularly and not smoking or drinking. Peptic ulcer is a disorder of the upper gastrointestinal tract that results when gastric acid and bacteria, drugs or other things cause breaks in the mucosa the moist tissue that lines the stomach, duodenum and other digestive organs. Ulcers can develop in the esophagus, stomach, duodenum or other areas of the GI tract.

2.6 STASTICES OF PEPTIC ULCER DISEASE¹⁶

Approximately 500,000 new cases of peptic ulcer are reported each year. At any given time, as many as 5 million people are affected in the United States alone. Patients with peptic ulcer disease may experience a range of symptoms from mild abdominal pain and burning to bleeding, vomiting catastrophic perforation of the organ lining a life threatening condition requiring emergency surgery. Thirty-five percent of patients diagnosed with gastric ulcers will suffer serious complications such as bleeding and perforation. Although mortality rates from peptic ulcer disease are low the high prevalence of ulcers and the resulting pain, suffering and expense are very costly. Ulcer treatment has improved significantly in the past two decades thanks to the discovery that a chief cause of ulcers is the bacterium helicobacter pylori one possible reason ulcers seem to run in families. In patients with ulcers due to H. pylori eradication of the bacterium has been shown to heal ulcers and prevent their reoccurrence. In addition, new acid reducing drugs are now available and have made a dramatic difference for patients.

Fig. No. 7: Stastices of Ulcer

The annual incidence per 1000 population varies from about 1 in Japan to 1.5 in Norway, 1.8 in USA and 2.7 in Scotland and the frequency also varies within many individual countries, such as Australia, China and India and among races such as a higher prevalence among whites than blacks in USA and among Chinese than Javanese in Indonesia. Most places report a rise of ulcer rates among the elderly in recent decades. The male to female ratio also varies geographically, for example from 1:1 in USA to 18:1 in India, and with time such as moving from 2:1 to 1:1 in the last two decades in USA, and the duodenal ulcer to gastric ulcer ratio varies widely from place to place, for example from 0.8 in Japan to 19:1 in Africa and 32:1 in India. Placebo healing rates also differ geographically, ranging from 5% in Philippines to 78% in Mexico. These epidemiological data can only be explained by the presence of multiple aetiological factors, including analgesics, society stress, cigarette smoking, Helicobacter pylori, dietary factors and genetic factors¹⁷.

3. ANTHELMINTIC DRUGS¹⁸

Worm infestation is one of the major global public health problems, more so in tropical countries. Besides the environmental conditions peculiar to tropics, poverty, illiteracy, lack of adequate sanitary facilities and of pure water supply make total eradication of this problem very difficult. The commonest parasites observed are round worms, hookworms, thread worms, tape worms, filarial worms.

Worms can cause various GI and general symptoms. In addition, some of them cause blood loss, nutritional deficiencies, urticarial and other allergic manifestations, intestinal obstruction and hepatosplnomegaly. Round worms have been implicated in the pathogenesis of bronchial spasm in endemic areas. Peripheral blood eosinophilia occurs in all nematode infestations. Careful examinations of stool may often spare the unnecessary removal of teeth and tonsils, usually blamed for spectic foci, in case with resistant urticarial.

An anthelmintic drug which kills the worm is called vermicidal, while that which affects the worm in such a way that it is easily expelled is known as vermifuge.

Drug Therapy of Roundworms

Mebendazole - This broad spectrum anthelmintic is benzimidazole derivative.

Given orally it is poorly absorbed. It inhibits microtubule polymerization by binding to beta tubulin.

It is highly effective in ascariasis, enterobiasis, trichuriasis and in hook worm infestation It is the drug of choice in enterobiasis and in trichuriasis. It also has some action against S.stercoralis. The drug is slow acting and it may take 2-3 days for parasitic clearance from the gut. It also adversely affects the ova of the trichuris and the hook worms.

It is effective invivo against the larve of Trichinella spiralis and exerts a lethal effect on the germinal membrane of the larvae of Echinococcus granulosus.

Adverse drug Reactions

These are usually mild and consist of abdominal pain, nausea and diarrhoea. Large oral dose may cause veritigo, dizziness, headache and arthralgia. Benzimidazole are embryo toxic and terartogenic in animals and should be avoided in pregnancy.

Preparation and Dosage

It is available as 100mg tablets and liquid suspension.

Therapeutic uses

Enterobiasis: A single dose of 100mg repeated after one week.

Hookworm and round worm infestation: 100mg bid for 3 days.

Taenia infsestation: 300mg tid for 3 days.

Albendazole

This broad spectrum benzimidazole has actions similar to those of mebendazole. It also has larvicidal actions in hydatid disease, ascariasis and ankylostomiasis and ovicidal properties in ascariasis, ankylostomiasis and trichuriasis.

Given orally, it is rapidly absorbed and has better bioavailability than mebendazole.

After absorption, it undergoes first pass metabolism in the liver to it is active metabolite,

albendazole sulfoxide. It is well distributed in tissues, bile CSF and hydatid cyst. It has plasma t1/2 8-12 hours. It I major advantage is that its effective against many common intestinal worms in a single dose and is cost effective.

The drug is well tolerated and adverse reactions are mild, mainly GI disturbances.

When used in hydatid diseases for long term therapy it may cause alopecia, liver damage and bone marrow depression.

Therapeutic Uses

 Ascariasis ankylostomiasis and trichuriasis - Usually 400mg single dose hookworm and trichuriasis may need a repeat dose.

 Enterobiasis - 400mg single dose, repeated after 4 weeks.

 Hydatid disease - 400mg bid for one months, repeated if necessary.

 Trichinella sprialis - 400mg bid for 8-14 days.

Piperazine

This drug was extensively used in the therapy of ascariasis and enterobiasis.

Mechanism of action

It act as a GABA agonist and cause hyperpolarization of ascariasis muscle resulting in paralysis of the worms which are then easily expelled by peristaltic movements. This eliminate the danger of worm migration.

Absorption, fate and excretion

Piperazine is absorbed from the gut, to the extent of about 30% the drug is partly metabolized in the body and partly excreted unchanged in the urine.

Adverse reactions

Piperazine has a wide margin of safety. Adverse effects are uncommon. They include nausea, vomiting, diarrhoea and urticaria. Neurotoxic effects, observed rarely, include veritigo, muscular incoordination, hypotonia and ataxia.

4. HERBS USED IN ULCER AND ANTHELMINTIC

Table No. 1: Herbs Used in Ulcer and Anthelmintic¹⁹

Plant Plant

Acacia Arabica Linn Cassia kleinii Arn Adansonia digitata Linn Kountze Casearia esculenta Roxb Aegle marmelos Linn Catharanthus roseus Linn

Allium sativum Linn Careya arborea Roxb

Aloe vera Linn Burm Carica papaya Linn

Annona squamosal Linn Cocculus hirsutus Linn Anacardium occidentale Linn Dioscorea dumetorum Pax Annona squamosa Linn Euphorbia neriifolia Linn Annona muricata Linn Ficus hispida Linn Balsamodendron mukul Arn Galega purpurea Linn Bauhinia variegate Linn Hydrocotyle asiatica Linn Berberis aristata Baill HibiscusrosasinensisLinn Betavulgaris Subsp.Martima Indigofera tinctoria Linn Boerhaavia diffusa Linn Mangifera indica Linn Bougainvillea spectabilis Linn Mimosa pudica Linn Bridelia ndellensis Beille Moringa oleifera Linn

Myrtus communis Linn Ocimum sanctum Linn

Murraya koenigii Linn Panax ginseng Linn

Peucedanum grande Linn Phyllanthus niruri Linn Pinus longifolia Linn Plantago ispagula Linn

Psidium guyava Linn Rhus coriaria Linn

Sesbania grandiflora Linn Shorea robusta Roth

Solanum nigrum Linn Tamarindus indica Linn

Terminalia chebula Retz Terminalia catapa Linn

2. LITERATURE REVIEW

1. K. Abedulla Khan et al., (2018)²⁰ reported real for the discovery of new compounds are antibacterial activities, possibly acting through mechanism of actions, which are distinct from those of wellknown classes of antimicrobial agents to which many clinically relevant pathogens are now resistant. Croton bonplandianum is a species in the genus Croton.

2. M.K. Biswas et al., (2017)²¹ reported antimicrobial activity of ethyl acetate, benzene, hexane, chloroform, methanol extracts of leaves of Croton bonplandianum against bacteria such as Staphylococcus haemolyticus MTCC and Salmonella enterica. The process was carried out by agar well diffusion method. The extracts were poured into the wells at different concentrations like 25mg/ml, 50mg/ml, 150mg/ml and 300mg/ml. After incubation zones of inhibition were observed. As the concentrations of extracts increased the activity also increased and thus the zone of inhibition too increased. Among five extracts zone of inhibition was best in ethyl acetate extract. . In case of Salmonella enterica ser. typhi (MTCC 8767) the ethyl acetate extract (300 mg/ml) showed maximum zone of inhibition 20.6±1mm, while chloroform extract (25mg/ml) showed minimum zone of inhibition 11.0±00. In case of Staphylococcus haemolyticus (MTCC 3383) the ethyl acetate extract (300 mg/ml) showed maximum zone of inhibition 18.0±1mm, while benzene extract (25mg/ml) showed minimum zone of inhibition 11.0±2.0.Hence Croton bonplandianum can be used in developing drugs and medicines against various activities of bacteria. Study has also been shown the presence of various phytochemical constituents such as alkaloid, tannin, saponin thiamine, ascorbic acid, phenolic content in the leaf of Croton bonplandianum.

3. K. Abedulla Khan et al., (2017)²² reported anti-diabetic activity of Croton bonplandianum baill in Streptozotocin induced diabetes on wistar rats. The method of anti-diabetic effect of ethanol leaf extract of Croton bonplandianum Baill (50, 100, 200mg/kg body weight) were administered orally in diabetic rats. After the oral administration of such ethanol leaf extract the blood glucose levels were monitored at specific intervals. In consequence it was observe that extract was significantly reduced blood glucose level. Simultaneously the action of the extract on diabetes induced hyperlipidemia was examined where it notably decreased the elevated total

cholesterol, triglyceride, low density lipoprotein (LDL) and very low density lipoprotein (VLDL) level while increased the high density lipoprotein (HDL).

Glibenclamide was used as a standard drug at a dose of 4 mg/kg.

4. Rajeswari Rajaram et al., (2017)²³ reported the biogenesis of zinc oxide (ZnO) nanoparticles using aqueous extracts of the leaves, stem, root, flowers and fruits of Croton bonplandianum baill at room temperature. Croton bonplandianum is an important medicinal plant used to cure many pathological conditions in the traditional systems of Indian medicines due to the presence of important and specific bioactive compounds in the plant parts of this plant. Aqueous solution of Zinc Nitrate hexahydrate [Zn(NO3)2•6H2O] was used as a precursor and the various plant extracts played as reducing agents. The formation of ZnO nanoparticles was monitored by UV-Visible spectrophotometric analysis. The leaf extract showed strong absorbance peak at 302 nm, stem and fruit at 293 nm, root at 290 nm and flowers at 305 nm.

5. Tanmay Ghosh et al., (2017)²⁴ reported antimicrobial activity of ethyl acetate, benzene, hexane, chloroform, methanol extracts of leaves of Croton bonplandianum against bacteria such as Staphylococcus haemolyticus. MTCC 3383 and Salmonella enterica ser. typhi MTCC 8767. The process was carried out by agar well diffusion method. The extracts were poured into the wells at different concentrations like 25mg/ml, 50mg/ml, 150mg/ml and 300mg/ml. After incubation zones of inhibition were observed. As the concentrations of extracts increased the activity also increased and thus the zone of inhibition to increased. Among five extracts zone of inhibition was best in ethyl acetate extract. In case of Salmonella enterica ser. typhi (MTCC 8767) the ethyl acetate extract (300mg/ml) showed maximum zone of inhibition 20.6±1mm while chloroform extract (25mg/ml) showed minimum zone of inhibition 11.0±00. In case of Staphylococcus haemolyticus (MTCC 3383) the ethyl acetate extract (300 mg/ml) showed maximum zone of inhibition 18.0±1mm, while benzene extract (25mg/ml) showed minimum zone of inhibition 11.0±2.0.Hence Croton bonplandianum can be used in developing drugs and medicines against various activities of bacteria. Study has also been shown the presence of various

phytochemical constituents such as alkaloid, tannin, saponin thiamine, ascorbic acid and phenolic content in the leaf of Croton bonplandianum.

6. K. Khanra et al., (2016)²⁵ reported silver nanoparticles and to evaluate the bactericidal and cytotoxicity properties of silver nanoparticles synthesized from root extract of Croton bonplandianum baill.

7. J. Bhavana et al., (2016)²⁶ reported acetone extract of the Croton bonplandianus baill an exotic weed of the euphorbiaceae family, was studied for cytotoxicity, apoptosis, cell cycle arrest in A549 cell line and antioxidant capacities using MTT assay, acridine orange ethidium bromide (AO/EB staining), cell cycle analysis and DPPH radical scavenging assay, respectively. Based on the cytotoxic activity, the extract was tested for the apoptotic effect using AO/EB and Hoechst 33258 staining. The apoptosis was characterized by chromatin condensation and DNA fragmentation. Further to determine the stage of cell death, cell cycle analysis was performed by flow cytometry and acetone extract was found to arrest G2/M phase in a dose dependent manner. The number of cells in G2/M phase increases with concurrent accumulation of cells in sub G0/G1 phase indicates the induction of apoptosis at G2M phase. The free radical scavenging activity of the acetone extract against DPPH was considerably significant. The cytotoxic, apoptotic and antioxidant effect of the acetone extract could be well correlated with the presence of potent free radical scavenging secondary metabolites such as phenols (43±0.05 μg/mL), flavonoids (3.5±0.07 μg/mL) and tannin (0.36±0.1 μg/mL). Our study has shown that A549 cells were more sensitive to acetone extract with an IC50 of 15.68±0.006 µg/mL compared to the standard drug 2.20±0.008 µg/mL. The results suggest that Croton bonplandianum could serve as a potential source of alternative therapeutic agent for treating cancer. Further research is required to isolate the active principle compound and determination of its anticancer property.

8. Nagaraju K. et al., (2015)²⁷ reported whole plant extract of acalyphaindica and croton bonplandianum obtained cold extraction of mixture of ethanol and water was chosen for pharmacological screening. The Swiss albino mice are subjected to extract at 125 mg/kg to check the CNS depressant activity by actophotometer test. The test and standard were given orally. After 60 min. the animal are placed in to the

actophotometer and observation were recorded at the interval of 90, 120 and 180 min.

acalypha indica showed CNS depressant effect but croton bonplandianum has no depressant activity.

9. J. Gokulakrishnan et al., (2015)²⁸ reported croton bonplandianus, Ficus religiosa and Coix lacryma was tested against Aedes aegypti mosquito. The present research showed that the highest LC50 values of methanol extracts of F. microcarpa against aegypti larvae were 91.63 ppm followed by, LC50 values of C. lacryma, P.

graveolens, C. berryi, and M. dubia extracts against. aegypti larvae were 92.77, 95.65, 96.52 and 100.12 ppm, respectively.

10. Narendra kumar singh et al., (2014)²⁹ reported pharmacognostical standardization parameters for C.bonplandianum methods Physicochemical and pharmacognostical standardization parameters for C.bonplandianum was developed as per the methods provided in World Health Organization (WHO) guidelines. Quantitative estimation of flavonoid total phenolics, tannins and total saponines contents were estimated by UV- spectrophotometry. Total alkaloids content was estimated by gravimetric analysis.

Results Morphological observations revealed that leaves of C. bonplandianum are simple, petiolate, alternate, 3–5 cm long, oval to lanceolate in shape, serrated margin with acute apex. The stem is cylindrical, branched, woody, light brown in color, odourless and bitter in taste. Roots are small dark brown to black in color, cylindrical, odourless and bitter in taste. Histological observations revealed the presence of discontinuous layer of lignified sclerenchymatous cells (stone cells), paracytic stomata, stellate trichomes, xylem with scalariform thickenings and libriform fibres.

Qualitative and quantitative study of phytoconstituents revealed the presence of flavonoids (0.57% w/w, equivalent to rutin), phenolics (4.25% w/w, equivalent to tannic acid) tannins (2.15% w/w equivalent tannic acid) and saponins (7.4 % w/w, equivalent to diosgenin) and gravimetric estimation of alkaloids content was found to be 2.5 % w/w.

11. S. Satyanarayana et al., (2014)³⁰ reported croton bonplandianum is one such plant which is famed for its therapeutic efficiency in different diseases globally in the complementary and alternative medicinal systems. Therefore, this review

comprehensively covers the medicinal and pharmacological activities of different parts of the plant C. bonplandianum.

12. Tapas Kumar Chaudhuri et al., (2014)³¹ reported croton bonplandianum standard biochemical and spectrophotometric methods were employed. All the analysis were performed in multiple sets. The total alkaloid, flavonoid and phenolic content were found to be 59.44±0.28 g/100 g, 3.86±0.12 mg/g and 67.37±0.46 mg/g respectively besides the presence of a certain amount of tannin (51.94±0.38 mg/100 g) and saponin (16.10±0.05 g/100 g). Among the three vitamins, total riboflavin (0.35±0.02 mg/100 g) and ascorbic acid (0.91±0.02 mg/100 g) content were found to be higher, followed by thiamine content of 0.31±0.02 mg/100 g. From the present study, it may be concluded that the stem of C. bonplandianus contains very high amount of various important phytochemicals. These phytochemicals are chiefly responsible for various medicinal properties of a plant. Therefore, the presence of the high quantity of phytocompounds may lead to the potent medicinal capacity of C. bonplandianus stem.

13. R. Govindarajan et al., (2014)³² reported the bark of Anogeissus latifolia wall. ex Guill. & Perr.(Combretaceae) has been to be used in the treatment of various disorders including stomach and skin diseases. We studied the antiulcer potential and antimicrobial activity of the 50% aqueous alcoholic extract in order to validate ethnobotanical claims regarding the plant use in the above-mentioned disorders.

Gastro protective potential of the Anogeissus latifolia extract (ALE) (100 and 200mg/kg/body weight) was studied on aspirin, cold-resistant stress (CRS), pylorus ligated (PL) and ethanol-induced ulcers. Status of the antioxidant enzymes superoxide dismutase (SOD) and catalase along with lipid peroxidation (LPO) was also studied in CRS-induced ulcers. The results of the present study showed for the first time that the ALE possessed gastroprotective activity as evidenced by its significant inhibition in the formation of ulcers induced by physical and chemical agents with a maximum of 84.16% curation (200 mg/kg body weight) in CRS- induced ulcers. ALE decreased LPO and SOD with concomitant increase in catalase activity in CRS-induced ulcers. Moderate antibacterial activity and antifungal activity was also observed. High performance thin layer chromatography (HPTLC) showed the presence of gallic acid and ellagic acid (0.95%, w/w, 0.25%, w/w, respectively) in

the plant. These findings could justify, at least partially, the inclusion of this plant in the management of gastric disorders in traditional medicine.

14. Priyankardey et al., (2013)³³ reported methanolic extract of Croton bonplandianum root was studied to preliminary screen and quantitate the presence of phytochemicals like tannin phlobatannin, cholesterol, glycoside, terpinoids, phenolics, flavonoid, steroid, anthraquinone, saponin, carbohydrate, alkaloid and protein. Root extract of C. bonplandianum may provide the protection of various diseases and may also develop the resistance against some diseases because of the presence of different phytochemicals. Method spectrophotometric and other standard biochemical methods provided a rapid and easy approach for the detection and quantification of the phytochemicals in roots of the plant. Principal Component Analysis (PCA) based on the correlation matrix was performed to correlate among these phytochemicals available in the root. Result High percentages of alkaloid (55.71 ± 0.11g/100g), saponin (14.25 ± 0.11 g/100g), phenolic content (64.36 ± 6.82 mg/g), protein (45.71

± 6.82 mg/g), lipid (17.05 ± 1.43 mg/g), tannin (26.68 ± .94 mg/100g), thiamine (0.37 ± 0.01 mg/100g) and also certain quantities of riboflavin or vitamin B2 (0.25 ± 0.03 mg/100g), ascorbic acid (0.35 ± 0.03 mg/100g) have been detected in the root of this plant conclusion.

15. Bhaumik Dave et al., (2013)³⁴ reported croton bonplandianum sedges and Balanites aegyptiaca amongst the all exhibit superior potential of heavy metal accumulation.

This is the first report to the heavy metal accumulation property of three different plant species which can be exploited for the bioremediation of heavy metals.

16. Arnab Kumar et al., (2013)³⁵ reported diabetes mellitus a carbohydrate metabolic disorder occurs due to absolute or relative deficiency of insulin. Current treatment strategies involve either preventing or delaying the intestinal absorption of glucose to lower the levels of postprandial hyperglycemia (PPHG). Herbal remedies have been since ancient times for treating diabetes mellitus. Therefore identifying novel phytocompounds with α-amylase and α-glucosidase inhibitory activity that would reduce the glucose absorption as well as the rise in postprandial blood glucose level is vital. Consequently, the present study was aimed to investigate the anti- hyperglycemic activity of Croton bonplandianum against these pancreatic enzymes.

17. Deepika Patel et al., (2013)³⁶ reported croton bonplandianum biologically active plant extracts have been well documented for evolving an ecologically sound and environmentally acceptable mosquito control programmes.

18. G.Arumugam et al., (2013)³⁷ reported various medicinal plants used for antidiabetic activity. Diabetes mellitus is one of the most common non- communicable diseases globally. It is the fourth leading causes of death in the most developed countries and there in substantial evendiced that it in epidemic in many developing and newly industrialized nations. This posing a serious threat to be met within 21st century. Since ancient time plants have been exemplary source of medicine. Ayurveda and other Indian literature mentioned the used of plants in treatment of various ailments. Out of an estimated 2,50,000 higher plants, less than 1% have been screened pharmacologically and very few in regard to diabetes mellitus. Systematic studies on the folklore medicinal plants that combat diabetes mellitus are scanty.

19. Mannan Hajimahmoodi et al., (2013)³⁸ reported Punica granatum L study has been conducted to evaluate this therapeutic property study the antiulcerogenic effects of pomegranate peel methanol extract, was tested on male Wistar albino rats. Oral pretreatment with peel extracts (25, 50 and 100 mg/kg) for 15 days protected the gastric mucosa against the damage induced by indomethacin (50 mg/kg). The incidence of ulceration in the control group was 100%. The best results were found in a dosage of 50 mg/kg in sour summer cultivar which inhibited the peptic ulcerin comparison with indomethacin induced gastric ulcer group. Lowest ulcer index (5.4 ± 0.55), an apparent decrease in the infiltration of polymorphs nuclear leukocytes and hemorrhage were observed after administration of sour summer extracts (50 mg/kg).

In conclusion present study showed that pomegranate peel extract, especially sour summer, has curative potential as an antiulcer, possibly via its high antioxidant activity. These results from pomegranate peel extract can provide an extra income and may contribute about good nutritional values of this product.