Thesis for Master of Philosophy (M.

Phil) 2011

DIURETIC ACTIVITY AND STUDY OF BIOCHEMICAL PARAMETERS IN THE METHANOL EXTRACT OF HIBISCUS ESCULENTUS (Okra) FRESH FRUITS

B DANAMMA

SRI KRISHNA DEVARAYA UNIVERSITY, ANANTAPUR

In partial fulfillment for the award of the degree of

MASTER OF PHILOSOPHY IN Biochemistry
Submitted under the Research Supervision of

Prof. Chitta Suresh Kumar, M.Sc. Ph.D.,

Department of Biochemistry, Sri Krishnadevarya University, Anantapur- 515 003

DIURETIC ACTIVITY AND STUDY OF BIOCHEMICAL PARAMETERS IN THE METHANOL EXTRACT OF HIBISCUS ESCULENTUS (Okra) FRESH FRUITS

B DANAMMA

SRI KRISHNA DEVARAYA UNIVERSITY

INDIA 2011

DECLARATION

I declare that the thesis Diuretic Activity and Study of Biochemical Parameters in the Methanol Extract of Hibiscus Esculentus (Okra) Fresh Fruits has been written by me and that it has not previously formed the basis for the award of any degree or other similar title.

Place: Date:

Anantapur

(B. DANAMMA)

SRI KRISHNADEVARAYA UNIVERSITY ANANTAPUR. 515 003, A.P, INDIA.

Dr. Chitta Suresh Kumar Professor, Department of Biochemistry Phone:08554 . 255056 (off) +919849669899 E-Mail: chitta34c@gmail.com

CERTIFICATE
This is to certify that the thesis entitled Diuretic Activity and study of Biochemical Parameters in the Methanol extract of Hibiscus Esculentus (Okra) Fresh fruits. is the bonafide research work carried out by Smt B Danamma under my supervision and guidance for the degree of Master of Philosophy in Biochemistry to Sri Krishnadevaraya University, Anantapur and it has not been submitted in part or in whole for any degree or diploma of any other University.

Research Director Date: Prof. Chitta Suresh Kumar, PhD., Department of Biochemistry Sri Krishnadevaraya University, Anantapur - 515003. (AP), India.

ACKNOWLEDGEMENT
Any work cannot be the outcome of one persons endeavors. This work owes much to many persons and it gives me pleasure in acknowledging the help of all those who have made it possible. Gratitude is said to be short lived but when it is placed on record, one can hope that it sustains. It is with great pleasure, I record my humble gratitude to our beloved Almighty by his lavish love and blessings, that we are able to complete our study successfully. Hitherto and present his piece of work for which we are eternally indebted. It is my pleasure to express my deep sense of gratitude and indebtedness to my revered guide Dr. CHITTA SURESH KUMAR Professor Department of Biochemistry, Sri Krishna Devaraya University, Anantapur, for his special interest, kindness, co operation and valuable guidance extended to me for the successful completion of this work. The entire valuable advises given by him will remain as permanent treasure. I am sincerely indebted to him. I take this opportunity to express my profound sense of gratitude to Mr. S. Nizamuddin Basha Asst. Professor, Department of Biotechnology, Safa College of Pharmacy, Kurnool, for his valuable suggestions and moral support throughout the work. I owe my gratitude to Dr. S. Ashok Reddy, M. Pharm Ph. D, Principal, Safa college of Pharmacy, Kurnool for providing all the facilities during my work and for his encouragement and support. And I am thankful for the permission to use the lab for the research work. I express my thanks to Asst. Prof. Mr. Murali and Mohd Dada. Department of Pharmacology, Safa college of Pharmacy, for extending the work and look after my experimental rats during the work It gives me immense pleasure to thank Mrs. K. Aruna Kumari, Lecturer in Biochemistry, SJDC, Kurnool, for her timely help. I extend my thanks to Dr. Meera bai, Dept of Botany, Rayalaseema University, Kurnool, for helping in the procurement and maintenance of the plant extract. I express my sincere thanks to Babajan, Rajasekar, Chaitanya, Madhusudan for their support and encouragement. I acknowledge my parents, husband and brother in - law for all the encouragement, love and support Finally I express my sincere appreciation and gratitude to my parents and well wishers, who have encouraged me throughout my career

AUTHENTICATION OF PLANT

This is to certified that the plant name Hibiscus Esculentus (Okra) has been used by B. Danamma for the M. Phil work under the guidance of Prof. Chitta Suresh Kumar, Department of Biochemistry, S, K. University, Anantapur

Place: Date:

Anantapur

Signature

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ABSTRACT
Hibiscus esculentus (Family: Malvaceae) is a most common weed in India. The whole plant is used as medicine. According to Ayurveda, Hibiscus esculentus is used as in the treatment of constipation, inflammation, anemia, abdominal disorders, urinary tract diseases, general debility. The present study was undertaken to investigate diuretic effect of methanol extract of the Hibiscus esculentus (MEHE) in albino rats. Acute oral toxicity study was performed as per OECD guidelines. In acute oral toxicity study, mortality was not observed up to 2000 mg/kg bodyweight. MEHE were administered at the doses of 250 and 500 mg/kg, p.o. Furosemide 20mg per kg body weight was used as positive control in study. The diuretic effect of the extract was evaluated by measuring urine volume, sodium and potassium content. Urine volume is significantly increased at two doses of MEHE 250 & 500 mg/kg body wt in treated rats. The excretion of sodium, Potassium levels was also increased by the MEHE. The diuretic effect of the extract was similar to furosemide. The MEHE had the additional advantage of chloride conserving effect. This study concludes that MEHE produced notable diuretic effect which appeared to be comparable to that produced by the standard diuretic furosemide. The present study provides a quantitative basis for explaining the folkloric use of Hibiscus esculentus as a diuretic agent. In addition to this different Biochemical parameters were also evaluated.

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LIST OF PUBLICATIONS
1. DIURETIC ACTIVITY AND STUDY OF BIOCHEMICAL PARAMETERS IN THE METHANOL EXTRACT OF HIBISCUS ESCULENTUS (OKRA) FRESH FRUITS B Danamma 1*, K Aruna kumari 2 , B Jayasimha Goud 3, S Nizamuddin Basha 1 International Journal of Pharmacy and Biological Sciences (eISSN: 2230-7605)
IJPBS |Volume 1| Issue 3 |JULY-SEPT |2011| Pg No: 160-169

2. EVALUATION OF ANTIULCER ACTIVITY IN THE METHANOL EXTRACT OF ACANTHUS ILICIFOLIUS LEAVES IN EXPERIMENTAL RATS *Nizamuddin Basha S1, Chitta Suresh1, Danamma B1, Mohd Dada2, Abdul Maajid3 Int. J. Pharm & Ind. Res Vol 01 Issue 01 Jan - Mar 2011Pg No: 57 - 62

3. HYPO GLYCEMIC ACTIVITY OF A METHANOL EXTRACT OF ARTEMISIA ABSINTHIUM LEAVES IN EXPERIMENTAL RATS *1B Jayasimha Goud, B Danamma2, , K.S.Dayananda3, B.K.Chikka swamy1 IJAPR / July 2011/ Vol. 2 / Issue. 7 / Pg No: 307 - 312 ISSN: 2230 7583

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TABLE OF CONTENTS

ABSTRACT LIST OF PUBLICATIONS TABLE OF CONTENTS LIST OF TABLES LIST OF FIGURES LIST OF ABBREVIATIONS

TITLE OF CHAPTER 1. INTRODUCTION 2. METHODOLOGY 3. RESULTS & DISCUSSION 4. SUMMARY AND CONCLUSION 5. REFERENCES

Page No 1 19 20 3 38 - 55 56 - 57 58 - 63

LIST OF TABLES
1. Table A: Classification of common diuretics and their mechanisms of action

2. Table1: Effect of oral administration of methanol extracts of H. Esculentus and Furosemide on urine volume, diuretic index and pH. 3. Table 2: Effect Of MEHE On Sodium Ion Concentration In Rats Urine

4. Table 3: Effect Of MEHE On Sodium Ion Concentration In Rats Serum 5. Table 4: Effect of MEHE on Potassium Ion Concentration in Rats Urine

6. Table 5: Effect of MEHE on Potassium Ion Concentration in Rats Serum

7. Table 6: Effect of MEHE on Na+ / K+ ratio in Rats Urine 8. Table 7: Effect of MEHE on Chloride Ion Concentration in Rats Urine

9. Table 8: Effect of MEHE on Chloride Ion Concentration in Rats Serum

10. Table 9: Effect Of MEHE On Urea Concentration In Rats Urine

11. Table 10: Effect Of MEHE On Urea Concentration In Rats Serum

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LIST OF FIGURES

Fig: A Fig B Fig C Fig D Fig E Fig F Fig G Fig 1

Hibiscus Esculentus Hibiscus esculentus fruit Sodium reabsorption in early PCT Sodium reabsorption in the thick ascending limb of loop of henle Sodium reabsorption in the DCT Sodium reabsorption in the collecting duct Sites of action of diuretics Effect Of Oral Administration Of MEHE And Furosemide On Urine Volume, Diuretic Index and PH

Fig 2

Effect Of Oral Administration Of Methanol Extracts Of H. Esculentus And Furosemide On Urine Volume, Diuretic Index & PH

Fig 3 Fig 4 Fig 5 Fig 6 Fig 7 Fig 8 Fig 9

Effect Of MEHE On Sodium Ion Concentration In Rats Urine Effect Of MEHE On Sodium Ion Concentration In Rats Serum Effect of MEHE On Potassium Ion Concentration In Rats Urine Effect of MEHE On Potassium Ion Concentration In Rats Urine Effect Of MEHE On Potassium Ion Concentration In Rats Serum Effect Of MEHE On Na+ / K+ Ratio In Rats Urine Effect Of MEHE On Na+ / K+ Ratio In Rats Urine

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Fig 10 Fig 11 Fig 12 Fig 13 Fig 14 Fig 15

Effect Of MEHE On Chloride Ion Concentration In Rats Urine Effect Of MEHE On Chloride Ion Concentration In Rats Serum Effect Of MEHE On Urea Concentration In Rats Urine Effect Of MEHE On Urea Concentration In Rats Serum Effect Of MEHE On Urea Concentration In Rats Urine Effect Of MEHE On Urea Concentration In Rats Serum

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LIST OF ABBREVIATIONS

MEHE CHF UTI OECD DI LV mg ml dl g SEM ANOVA GFR CC CPCSEA TLC AgNO3 HCO3 Na+ K+ ClEDTA nm kg m eq

Methanol Extract of Hibiscus Esculentus Congestive Heart Failure Urinary Tract Infections Organization of Economic co operation and development Diuretic Index Lipschitz value milligram milli litre Deci litre Micro grams Standard Error Mean Analysis Of Variance Glomerular Filtration Rate Creatinine Clearance Control and Supervision of Experiments on Animals Thin Layer Chromatography Silver Nitrate Bi Carbonate Sodium Potassium Chloride Ethylene Diamine Tetra Acetate nano meters kilo gram milli equivalents