hepatoprotective activity against ethanol induced oxidative stress and in-vitro antioxidant activities

91 Pages·2016·2.13 MB·English

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HEPATOPROTECTIVE ACTIVITY AGAINST
ETHANOL INDUCED OXIDATIVE STRESS AND
IN-VITRO ANTIOXIDANT ACTIVITIES OF
FUNCTIONAL GRAPE WINES

KAVYA M. V.

DEPARTMENT OF POSTHARVEST TECHNOLOGY
COLLEGE OF HORTICULTURE, BAGALKOT
UNIVERSITY OF HORTICULTURAL SCIENCES,
BAGALKOT- 587 104

JUNE, 2016
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HEPATOPROTECTIVE ACTIVITY AGAINST
ETHANOL INDUCED OXIDATIVE STRESS AND
IN-VITRO ANTIOXIDANT ACTIVITIES OF
FUNCTIONAL GRAPE WINES

Thesis submitted to the
University of Horticultural Sciences, Bagalkot
in partial fulfilment of the requirements for the award of the
Degree of

Master of Science (Horticulture)
in
Post Harvest Technology

By
KAVYA M. V.
UHS14PGM484

DEPARTMENT OF POSTHARVEST TECHNOLOGY
COLLEGE OF HORTICULTURE, BAGALKOT
UNIVERSITY OF HORTICULTURAL SCIENCES,
BAGALKOT- 587 104

JUNE, 2016
iii
DEPARTMENT OF POSTHARVEST TECHNOLOGY
COLLEGE OF HORTICULTURE, BAGALKOT
UNIVERSITY OF HORTICULTURAL SCIENCES,
BAGALKOT - 587104

CERTIFICATE
This is to certify that, the thesis entitled “Hepatoprotective activity against
ethanol induced oxidative stress and in-vitro antioxidant activities of functional grape
wine” submitted by Miss. KAVYA, M.V., I. D. NO. UHS14PGM484 for the award
of the degree of MASTER OF SCIENCE (HORTICULTURE) in POST
HARVEST TECHNOLOGY of the University of Horticultural Sciences, Bagalkot,
is a record of research work carried out by her during the period of her study in this
university under my guidance and supervision. The data of this thesis has not
previously formed the basis for the award of any degree, diploma, associateship,
fellowship or other similar titles.

Place: BAGALKOT (E. RAJASHEKHARA)
Date: JUNE, 2016 MAJOR ADVISOR
Associate Professor
Department of Agriculture Microbiology,
College of Horticulture,
Bagalkot – 587 104
Approved by:
Chairman:
___________________________
(E. RAJASHEKHARA)

Members: 1. ________________________
(A. B. PATIL)

2. __________________________
(S. L. JAGADEESH)

3. ___________________________
(S. N. PATIL)

4. __________________________
(MALLIKARJUNA AWATI)

5. __________________________
(D. L. RUDRESH)

6. ________________________
(CHANDRASHEKARA V. M.)
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ACKNOWLEDGMENT
With heartfelt memories…
The beatitude, bliss and euphoria that accompany successful completion of
any task would be incomplete without the expression of appreciation of simple
certitude to the people who made it possible to achieve the goal by their encouraging
guidance and proper steering. It is still great at this juncture to recall all the faces and
spirit in the form of parents, teachers, friends, near and dear ones.
Words are not enough to express my deep sense of gratitude, pride and
indebtedness to Dr. E. Rajashekhara, Associate professor and head, Department of
Agriculture Microbiology, College of Horticulture, Bagalkot, Chairman of my
advisory committee. I consider myself most fortunate to have worked under his
guidance and also for his expert, generous, valuable suggestion, constructive
criticism, critical analysis, meticulous correction of the manuscript and affectionate
dealings throughout the course of investigation. These kinds of consideration and
thoughtfulness have meant lot to me and all made it possible successful completion of
my thesis.
Iam greatly indebted to Dr. Chandrashekara, V.M., Professor and head,
Department of Pharmacology and Toxicology, Hangal Shri Kumareshwar College of
Pharmacy, Bagalkot, for his guidance, intellectual suggestions and for providing a
great opportunity to conduct in-vivo studies and for bountiful co-operation throughout
my investigation.
It also gives me an immense pleasure to express my deep sense of heartfelt
gratitude and sincere thanks to Dr. S. L. Jagadeesh, Professor and Head, Department
of Post Harvest Technology, College of Horticulture, Bagalkot, for his constant
support, encouragement and for providing opportunity to study under guidance of
Rajashekhar sir. I also extend my regards to Dr. D. L. Rudresh, Dr. A. B. Patil,
Dr. S. N. Patil, Dr. Mallikarjuna Awati members of my advisory committee for
their valuable advice and outstanding support during period of investigation.
With immense pleasure I would like to express my deepest gratitude to
Dr. Chidanand P. Mansur, Mr. Kirankumar Gorabal, Dr. Prabhakar E., Dr. Sathish
Patepur, Dr. Shankar Meti, Dr. Nagaraj M. S., Dr. Prasanna, Dr. Bhuvaneshwari G.,
Dr. Jameel Jhalegar M., Dr. Ragavendra S. and Miss. Netravati for their timely
support, valuable suggesstions and encouragement to reach this valuable goal and
completion of my PG programme.
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One uses the choicest words to acknowledge the invaluable help rendered by
someone very close to heart. But I find such a measure inadequate to quantify the
sacrifices, love, affection, support and concern shown to me by my father
Sri. Vishwanath, mother Smt. Pramila and my whole family members. Their
unselfish struggle towards my wellbeing and attainment of perfection in career goals
has been boundless and my gratitude is beyond words.
I solemonly dedicate heartfelt regards and gratitude to Dr. Shivakumar
Halgatti and Mr. Vivekananda kamble, without their moral, cheerful support and
help during in-vivo studies, its highly impossible for me to complete my investigation
successfully.
I fail in words to express my heartfelt gratitude towards my friends Ranjitha ,
Jhanavi, Deepa konti, Divya, Madukara, Nithinkumar, Ashwini, Shubhangini,
Chandana, Rigzen Tsewang, Charan, Gajanan, Govind and my junior friends who
stood beside me for my support all the time, Chiranjeevi, Siddharth, Preethi, Asha and
Deepa hugar for their outstanding support and relentless encouragement throughout
my investigation successfully.
Iam also very thankfull to lab assistants Hiremat, Jakir, Deepak, Akkamma,
Chaitali, Adivappa, Mallappa, Shakuntala and all labour friends who helped me a lot
during my research work.
I thankfully acknowledge the help rendered by Elite Vintage Vinery, India
Pvt, Ltd. and Mr. Anand Salimat and also all my senior and junior friends of Hangal
Shri Kumareshwar College of Pharmacy, Bagalkot for their valuable support.
Last but not least I would like to thank my welwishers Smt. Radhika,
Sangamitra, Chandrashekar, Lalithya, Rashmi Hegde, Reethu, Sathya, Spoorthi,
Gururaj, Kavyashree, Manjula, Pooja, Asha, Sakkubai and also all those who helped
me directly or indirectly during period of my stay in this campus.
As I sacrificed many rats during my in-vivo studies for the sake of completion
of my investigation, God please forgive me.
In case of any omission or deletion doesnot mean lack of gratitude.

JUNE, 2016
BAGALKOT (KAVYA M. V.)
vi

Affectionately dedicated
to
my family and my guide
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CONTENTS

Chapter Page
Chapter particulars
No. No.
CERTIFICATE iii
ACKNOWLEDGMENT iv
LIST OF ABBREVIATIONS ix
LIST OF TABLES xi
LIST OF FIGURES xii
LIST OF PLATES xiii
LIST OF APPENDIX xiv
1. INTRODUCTION 1-3
2. REVIEW OF LITERATURE 4-13
2.1 Studies on physico-chemical, phytochemicals and in- 4
vitro antioxidant activities of ginger and pepper extracts
of different media
2.2 Studies on in-vitro antioxidant activities of grape wines 6
fortified with ginger and pepper extracts
2.3 Evaluation of hepato-protective activity of selected 8
fortified wines against ethanol induced oxidative stress
3. MATERIAL AND METHODS 14-29
3.1 Materials 14
3.2 Studies on physico-chemicals, phytochemicals and in- 18
vitro antioxidant properties of ginger and pepper extracts
of different media
3.3 Studies on in-vitro antioxidant activities of grape wines 20

fortified with ginger and pepper extracts
3.4 Evaluation of hepato-protective activity of ginger or 23
pepper extracts fortified wines against ethanol induced
oxidative stress
4. EXPERIMENTAL RESULTS 30-48
4.1 Studies on physico-chemicals, phytochemicals and in- 30
vitro antioxidant properties of ginger and pepper extracts
of different media
4.2 Studies on in-vitro antioxidant activities of grape wines 35
fortified with ginger and pepper extracts
4.3 Evaluation of hepato-protective activity of ginger or 39
pepper extracts fortified wines against ethanol induced
oxidative stress

Contd.....
viii
Chapter Page
Chapter particulars
No. No.
5. DISCUSSION 49-59
5.1 Studies on physico-chemicals, phytochemicals and in – 49
vitro antioxidant properties of ginger and pepper extracts
of different media
5.2 Studies on in-vitro antioxidant activities of grape wines 52
fortified with ginger and pepper extracts
5.3 Evaluation of hepato-protective activity of ginger or 58
pepper extracts fortified wines against ethanol induced
oxidative stress
6. SUMMARY AND CONCLUSIONS 60-62
REFERENCES 63-74
APPENDIX 75
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LIST OF ABBREVIATIONS

ABBREVIATIONS
AAE Ascorbic Acid Equivalent
ABTS 2,2’-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)
AEAC Acid Equivalent Antioxidant Capacity
ALP Alkaline Phosphatase
ALT Alanine Transaminase
AST Aspartate Transaminase
CBS Columbia Broadcasting System
CCl Carbon Tetrachloride
4
CE Catechin Equivalent
CUPRAC Cupric Ion Reducing Antioxidant Capacity
DNPH 2,4-Dinitrophenylhydrazine
DPPH 2,2- diphenyl-1-picrylhydrazyl
FRAP Ferric Reducing Ability of Plasma
GAE Gallic Acid Equivalent
GSH Glutathione
LDH Lactate Dehydrogenase
LPO Lipid Peroxidation
MDA Malondialdehyde
ORAC Oxygen Radical Absorbance Capacity
SD Sprague dawley
SGOT Serum Glutamic Oxaloacetic Transaminase
SGPT Serum Glutamic Pyruvic Transaminase
TG Triglyceride
TBARS Thiobarbituric Acid Reactive Substance
x
LIST OF TABLES

Table Page
Title
No. No.
1. Total sugar content (g/L) of aqueous, ethanolic and wine extracts 31
of ginger and pepper.
2. Total acidity (%) of aqueous, ethanolic and wine extracts of 31
ginger and pepper.
3. Total phenol content (mg/100ml) of aqueous, ethanolic and wine 33
extracts of ginger and pepper.
4. Tannin content (mg/100ml) of aqueous, ethanolic and wine 33
extracts of ginger and pepper.
5. Total flavonoid content (mg/100ml) of aqueous, ethanolic and 34
wine extracts of ginger and pepper.
6. Total antioxidant content (mg/100ml) of aqueous, ethanolic and 34
wine extracts of ginger and pepper.
7. DPPH radical scavenging activity (% scavenging activity) of 36
aqueous, ethanolic and wine extracts of ginger and pepper.
8A. Total sugars (g/L) content of wines fortified with ginger and 37
pepper extracts.
8B. Total acidity (%) of wines fortified with ginger and pepper 37
extracts.
8C. Total alcohol (%) content of wines fortified with ginger and 37
pepper extracts.
9A. Total phenols (mg GAE /100 ml) content of wines fortified with 38
ginger and pepper extracts.
9B. Tannin contents (mg/100ml) of wines fortified with ginger and 38
pepper extracts.
9C. Total flavonoids (mg/ml) content of wines fortified with ginger 38
and pepper extracts.
10A. Total flavonoids (mg/ml) content of wines fortified with ginger 40
and pepper extracts.
10B. DPPH radical scavenging activity (%) of wines fortified with 40
ginger and pepper extracts.
Contd.....