The aim of present study was to investigate the Hepatoprotective activity of aqueous extract of Hibiscus sabdariffa (Malvaceace) leaves in albino rats on alcohol induced hepatotoxic activity. .

1. HEPATOPROTECTIVE ACTIVITY OF AQUEOUS
EXTRACT OF HIBISCUS SABDARIFFA ON ALCOHOL
INDUCED HEPATOTOXITY IN RATS
Submitted By
B.Krupa jeeva, Bhavana.G*, G.Ram Jagan Mohan
M.sandhya and R.Rajani,
UNDER THE SUPERVISION OF
Dr.D.Jeevan Mani Babu Mpharm,Ph.D
DEPARTMENT OF PHARMACOLOGY

2. AIM AND OBJECTIVE
AIMOF STUDY
The aim of present study was to investigate the Hepatoprotective activity of
aqueous extract of Hibiscus sabdariffa (Malvaceace) leaves in albino rats on
alcohol induced hepatotoxic activity .
OBJECTIVES OF STUDY
The objective of the study is
 Induction of Hepatotoxicity in animals and its evaluation by-
a) Ethyl Alcohol,
b) Silymarin with diet and
c) Ethyl Alcohol induced hepatotoxicity models.

3. INTRODUCTION TO LIVER
The liver is the largest organ, present inside the body just below the
right lung and the diaphragm. The liver lies close to the body colon,
the intestines and the right kidney. It roughly weighs about 1.5 kg and
is deep red in colour due to a rich blood supply.
Important functions:
1. Metabolic process.
2. stores various nutrients such as fat, carbohydrates, proteins.
3. It also plays an important role in the detoxification process and
clears the body of harmful substances.
TYPES OF LIVER DISEASES
 HEPATITIS: Inflammation of liver.
 ALCOHOLIC LIVER DISEASE: Hepatic manifestation of alcohol
consumption. Includes Fatty liver, Alcoholic Hepatitis.
 FATTY LIVER DISEASE (hepatic statuses) : is a reversible condition
where large vacuoles of triglyceride fat accumulate in liver
cells,etc.

4. USEOF HERBALMEDICINESFOR
LIVERDISEASE
Herbal medicines are being used by about 80% of the world population
primarily in the developing countries for primary health care.
They have stood the test of time for their safety, efficacy, cultural
acceptability and lesser side effects. The chemical constituents present in them
are a part of the physiological functions of living flora and hence they are
believed to have better compatibility with the human body.
Herbal medicines have been used in the treatment of liver disease for a Slong
time.

5. Botanical Classification
1) Kingdom : Plantae (Plants)
2) Subkingdom : Tracheobionta
3) Super division: Spermatophyte
4) Division : Magnoliophyta
5) Class :Magnoliopsida (diCotyledons)
6) Subclass :Dilleniidae
7) Order :Malvales
8) Family :Malvaceae (Mallow family)
9) Genus : Hibiscus L. (Rose mallow)
10) Species : Hibiscus sabdariffa

6. PHYTOCHEMISTRY
 The leaf is reported to contain protein, fat, carbohydrate, fibre, ash,
calcium, phosphorus, iron, thiamine, β –carotene, riboflavin, niacin and
ascorbic acid.
 The flower yields a yellow dye; the major pigment identified is
daphniphylline. The plant contains flavonoids such as hibiscitrin and
hibiscetin and dried calyces contain the flavonoids gossypetine,
hibiscetine and Sabdaretine. It also contains alkaloids, β – sitosterol.
 The calyces are rich in acid and pectin. Analysis of calyces has shown the
presence of crude protein and minerals such as iron, phosphorus, calcium,
manganese, aluminium, magnesium, sodium and potassium.

7. PHARMACOLOGY
The drug has various actions like-
1. Antihypertensive
2. Hepato protective
3. Anti-lipidemic
4. Anti-oxidant
5. Anti-cancer.
MATERIALS AND METHODS
Preparation of extract
The leaves were collected and washed and shade dried for 3 weeks.
They were made into powder and the weight was taken as 330.5gm.The
coarse form was then macerated with water and methanol is added as
preservative and was placed in a conical flask. The extract was left to stand
for specific time and the extract was filtered out with the help of a clean filter
cloth and funnel. The resulting aqueous extract was then air dried until
complete evaporation.

8. EXPERIMENTAL INDUCTION OF
HEPATOTOTOXICITY
Ethanol (3g/kg body weight) was dissolved in water and injected to
rats through oral feeding needles, for specific period.
EXPERIMENTAL DESIGN
 In the experiment, total of 36 rats were used in the study. The rats were
divided into 6 groups of 6 rats in each group.
 The animals were allowed to acclimatise for 2 weeks prior to distribution
into different groups of 6 rats each and treated for 6 week as follows:
 GROUP I: Received water (5ml/kg p o) for 21 days once daily, and
serves as a control.
 GROUP II: Received water (5ml/kg p. o) for 21 days once daily and in
addition to normal diet received 40% of alcohol solution (v/v, 2ml/100g)
for 21 days.
 GROUP III: Received standard drug silymarin (100mg/kg p o) for 21
days once daily and addition to normal diet received 40% of alcohol
solution (v/v, 2ml/100g) for 21 days.
 GROUP IV& V: Received AELHS (200mg/kg, 400mg/kg) 21 days once
daily and addition to normal diet received 40% of alcohol solution (v/v,
2ml/100g) for 21 days.

9. RESULTS
PRELIMINARY PHYTOCHEMICAL SCREENING
The revealed results of the preliminary phytochemical screening of the
alcoholic extract of H.sabdariffa eaves were shown below:
S.NO Phytochemical Tests Results
1 Test for Carbohydrates +
2 Test for Glycosides +
3 Test for Tannins +
4 Test for Alkaloids +
5 Test for Flavonoids +
6 Test for Phenols +

10. ACUTE TOXICITY AND GROSS BEHAVIOUR CHANGES
In acute toxicity studies there were no any behavioural changes,
signs of toxicity as well as any mortality were observed with limit test
dose of 2000 mg/kg body according to the OECD guidelines.
Effect of pre-treatment with H.Sabdariffa aqueous leaf extract on
serum level changes in chronic alcohol fed rats (g/week )
Serum
levels
Normal Toxicant Standard 200mg/kg+
alc
400mgS/kg+alc
AST
83.27±0.25 337.53±0.4 87.73±0.52 249.20±0.6
3
92.91±0.72***
ALT 37.22±0.30 257.43±0.5 47.27±0.56 186.54±0.7
7
52.23±4.26***
ALP 0.12.20±35 391.42±0.3 132.82±0.6
2
253.24±0.6
6
139.65±0.77***
Bilurubin 0.58±0.2 3.48±1.20 0.98±1 2±1.5 0.97±1.2***

12. EVALUATION OF EXPERIMENTAL DATA ANALYSIS
OF HEPATOPROTCTIVE ACTIVITY
The results of serum parameters revealed that ALT ALP, AST are
significantly increased several times in case of disease control group
compared to normal control group. The plant extract at a dose of 400 mg/kg
showed significant hepato protective activity compared to the disease control
group. This is manifested by decreased levels of AST, ALP and ALT etc. In
serum of experimental animals and restores towards its normal value.
Standard drug silymarin treated group animals exhibited its usual significant
hepato protective activity compared to disease control group along with serum
parameters.

13. DECLARATION
Therefore ethanol treatment to albino rats for 8 weeks induced a
fair degree of de-arrangement of liver structure and function. There
was a significant elevation in the levels of serum marker enzymes like
SGPT, SGOT, etc. content of ethanol in toxicated animals. In contrast,
pre-treatment with AELHS (200,400mg/kg) and silymarin49
(100mg/kg p.o) exhibited an ability to counter act the hepatotoxicity
by decreased serum marker enzymes.
In the ethanol treated groups, there was a significant increase in
total bilirubin content. Whereas, pre -treatment with AELHS caused
significant reduction in total bilirubin.

14. END OF PRESENTATION