Phytochemical Investigations of Zingiber officinale (Ginger) in relation

to its Anti Arthritic Activity

SYNOPSIS FOR

DOCTOR OF PHILOSOPHY

IN

BOTANY (LIFE SCIENCES)

SUBMITTED TO

By

ARIF BASHIR

(GOVT COLEEGE OF SCIENCE AND COMMERCE BENAZEER

BHOPAL)

Under The Guidance Of

DR JAGRATI TRIPATHI

Asstt. Professor Deptt Of Biotechnology

Unique College Bhopal

Under The Co Guidance Of

DR DEWEEDI

Asstt Professor Deptt Of Chemistry

Govt College Of Science And Commerce Benazer Bhopal

SYNOPSIS

1.TITLE:

Phytochemical Investigations Of Zingiber officinale (Ginger ) In Relation

To Its Anti Arthritic Activity.

2.INTRODUCTION:

The plant kingdom is abundant in species that act as anti arthritic and anti

inflammatory to animal tissue. Most of the biological needs of humanity

and other animals are met by our environment. Inflammation , a process

unpleasantly familiar to everyone occurs in response to a range of

traumas from sunburn and wounds and to infection and auto immune

conditions. The most important inflammatory conditions to affect

humanity are the varieties of arthritis and rheumatism. The herbal

medicine is used in its treatment throughout the world. Arthritis is a

general term used for approximately 100 named diseases, that produce

either inflammation of connective tissue , particularly in joints or non

inflammatory degeneration of these tissues. Rheumatoid arthritis (RA) is

a chronic disability and auto immune disorder that affects about 1% of

population. About one out of seven Americans exhibit some form of

arthritis . immune system is vital to survive , because a hyper active

immune system may cause fatal disease due to over whelming allergic

reaction leading to series of derangements , loss of normal capacity to

distinguish self from nonself resulting in immune reactions against ones

own tissues and cells called auto immune diseases. A common auto

immune form of arthritis is Rheumatoid Arthritis (RA). Blood tests often

reveal the presence of rheumatoid factors, protein produced by the

immune system in response to the rheumatic process.

Medicinal plants are a part of social life. It is said that medicinal plants

cure almost all diseases. These herbal remedies obtained from various

medicinal plants are less expensive. The medicinal plants are the only

easy accessible health care alternative for most of our population.

Herblists around the world know of many plants with inflammation

inhibiting and anti arthritic activities. In a well known study, 163 species

of plants and fungi are tested to determine antiinflamatory activity and

among 500 sps only 50 sps. Are tested to determine antiarthritic activity.

Unfortunately chemical identification of these activities is given much

more attention then therapeutic use. This is because plants are viewed as

sources of potentially valuable drugs rather than living inherent value in

themselves. The attention of scientists all over the world is focused

towards the alternative system of medicine in the recent past. There is a

growing importance in traditional health systems in providing health care

for a wider population across the globe, especially in developing

countries. WHO currently encourages , recommends and promotes

traditional as well as natural remedies in national health care programmes

as they are easily available at low cost, comparatively safe and are

culturally acceptable.

The drugs commonly used for the treatment of inflammation and

rheumatoid arthritis include glucocortocids e,g; cortisone and

prednisone , Ibuprofen and Naproxen etc. disease modifying anti

inflammatory and anti rheumatic drugs e,g; Methotrexate (MTX) and

Leflunomide etc. biological response modifiers e,g; Tumor Necrosis

factors , alpha blocking agents. Besides their high cost, severe adverse

reactions and toxicity, including some risks of infection in sub sets of

patients being treated with biological response modifiers . as aresult

alternative treatments based on natural plant products and herbal mixtures

are used. These herbal formulations are becoming increasingly popular in

india , US and other countries. The herbs which have been used to treat

joint pains or arthritis include Medicago sativa (Alfa Alfa), Angelica

archangelica (Angelica), Larrea tridentate ( Chaparral), Zingiber

officinale ( Ginger), Brassica alba ( Mustard Plaster) etc.

3. REVIEW OF LITERATURE:

Mohammad Rahmatullah et al reported that arthritis is a chronic,

systemic inflammatory disorder that principally affects synovial joints .

Modern allopathic medicine has no complete cure for the disease ;most

medicines deal with treatment of accompanying symptoms like pain and

inflammation . It was the objective of the present study to conduct a

survey among Kavirajaes of six districts of Bangladesh. A total of 32

medicinal plants distributed into 23 families . The Araceae and

Solanaceae family contributed 03 plants each. The leaves constituted the

plant part most frequently used (28.8%) followed by fruits (16.9%). The

medicinal plants obtained in present survey offer considerable potential

for further scientific studies to be carried out.

S Tripathy et al studied the effect of alcoholic and aqueous extracts of

Hybanthus enneaspermus (Violaceae) on Freund’s adjuvant arthritis. The

percentage of yield was found to be 12.8 and 10.6% for alcoholic and

aqueous extracts respectively. Both the extracts significantly (p<0.001)

decreased the paw thickness at the end of 30 days treatment. At the end of

the studies the alcoholic extract shows more pronounce effect (59.4%) as

compared to aqueous extract (57.4%). The phytochemical analysis

reveals the presence of alkaloids, flavonoids, glycosides, phenols,

carbohydrates and tannins in the extracts. This result support the folkore

use of this plant against the inflammatory conditions like arthritis.

S.Jubae et al studied the anti arthritic activity of stem barks of Allangium

salvifolium belonging to family Alangiaceae was studied in rats. The

barks of Allangium salvifolium wang were collected and dried in shade

and subject for successive extraction with petroleum ether ,ethyl acetate ,

choloroform, methanol using Soxhlet apparatus and distilled water by

maceration. Each extracts were then subjected for preliminary

phytochemical studies and pharmacological investigation. The acute

toxicity studies were carried out according to the up and down method of

CPCSEA guidelines no 425 and anti arthritic activity by Freund’s

adjuvant arthritis model . All the extracts have exhibited significant anti

arthritic activity.

M. Kamalutheen et al studied that the various extracts of Merremia

tridentata were investigated for anti inflammatory and anti arthritic

activity in male albino rats. The anti arthritic activity was carried out

using complete Freund’s adjuvant induced arthritis model. Indomethacin

(10mg/kg bw) was used as standard drug . The ethanol extract of

Merremia tridentata exhibited significant dose dependent activity in

acute inflammation and the doses of 100mg/kg bw and 200mg/kg bw

produced 38.3% and 42.8% inhibition respectively after 3h as compared

with that of standard drug which showed 48.5% inhibition. In arthritis

model the dose of 100mg/kg bw and 200mg/kg bw of the ethanol extract

produced 49.0% and 51.7% inhibition respectively after 19 days when

compared to that of standard drug(55.5%) . The doses of ethanol extract

of M.tridentata exhibit significant anti arthritic activities.

A.K.Chakarborty and H.K.Roy showed that various extracts of Cleome

rutidosperma were investigated for its anti arthritic activity in male albino

rats . The evaluation of anti arthritic activity was carried out using cotton

pellet granuloma method and Freund’s adjuvant induced arthritis model.

Prednisolone (5mg/kg bw) was used as a standard drug. The ethanolic

extract of C.rutidosperma exhibited significant anti arthritic activity as

compared to other extracts. The doses of 200mg/kg bw of ethanolic

extract of C.rutidosperma , in chronic model of granuloma pouch in rats

produced 48.0% and in arthritis model produced 44.0% inhibition

respectively with that of the standard drug Prednisolone(5mg/kg) which

produced 58.5% and 59% inhibition.

Rekha Rajendran and Ekambaram Krishnakumar studied that

adjuvant induced arthritis is a chronic crippling , skeleton muscular

disorder having nearest approximation to human rheumatoid arthritis. The

wood of Premna serratifolia is reported to be useful in the treatment of

arthritis. The anti arthritic activity of ethanol extract of Premna

serratifolia Linn., wood is done by Freund’s adjuvant induced arthritis

model. The ethanol extract at the dose of 300mg/kg body weight

inhibited the raw paw edema by 68.32% which is comparable with

standard drug indomethacin 74.87% inhibition of rat paw edema after 21

days. The results of the current investigation concluded that ethanol

extract of P.serratifolia Linn.,wood possess significant anti arthritic

activity due to the presence of phytoconstituents such as irridoid

glycosides, alkaloids, phenolic compounds and flavonoids.

Otari,K.V, et al studied the anti arthritic and anti inflammatory activity

of ethanolic extract of seeds of Vernonia anthelmintica . The seeds were

extracted by Soxhlet method using ethanol (99.9%) and subjected to

preliminary phytochemical and acute toxicity studies. The effects of EVA

was evaluated for acute inflammation in carrageenan induced rat paw

edema and for chronic inflammationin complete Freund’s adjuvant

(CFA) induced arthritis in rats . Further the biochemical,

histopathological and radiographic evaluation was performed. The

phytochemical evaluation revealed the presence of alkaloids, flavonoids,

steroids, triterpenes and polyphenols in EVA. The results of present study

were revealed the effectiveness of EVA in acute as well as a chronic

inflammatory conditions without ulcerogenic potential.

Uma Chandur et al studied the extracts of Zingiberaceae family

A.galanga was evaluated for anti arthritic activity. The petroleum ether,

chloroform, alcoholic extracts of the A.galanga rhizomes in the presence

of chemically active compounds by standard methods. The results

showed the presence of flavonoids, glycosides, phenolic compounds,

saponins and tannins. The application of all the three extracts exhibited

statistically significant edema inhibition. The results obtained suggest that

A.galanga rhizome extracts is endowed with effective anti arthritic

activity.

Mangesh S Bansod et al aimed to assess the anti arthritic nature of poly

herbal formulation containing Cissampelos Pereira Linn, Pongamia

pinnata (Linn) and Pierre and Vitex negundo Linn. Against Freund’s

adjuvant induced arthritis in rats. Polyherbal formulation (200mg/kg,

400mg/kg and 600 mg/kg) reduces hind paw swelling and body weight

alongwith significant improvement in haematological study ,while

histopathology reveals the significant reduction in mono nuclear in

filtration, pannus formation and bone erosion. The radiological pictures

of the joints particularly metatarsal, phalanges and ankle joint space of

polyherbal formulation treated group animals showed protective effect

against adjuvant induced arthritis.

M.Sangeetha et al studied that Clerodendron inerme belongs to family

Verbenaceae is used for the treatment of veneral diseases, rheumatism,

elephantiasis and intermitlant fever. The qualitative phytochemical

screening showed the presence of alkaloid, steroids, phenols, flavonoids,

tannins, carbohydrates, fixed oils and volatile oils. The petroleum ether,

choloroform, ethyl acetate, ethanol and water fractions of the leaves of

Clerodendron inerme were subjected to in vitro anti inflammatory

activity of HRBC membrane stabilization method in various

concentrations. The petroleum ether, choloroform, ethyl acetate, ethanol

and water fractions of the leaves of Clerodendron inerme were subjected

to in vitro anti arthritic activity by protein denaturation method. All the

extracts showed positive response. The effect was represented as follows

Ethyl acetate > Chloroform > Ethanol > Water > Petroleum ether.

4. NOTEWORTHY CONTRIBUTIONS:

Jitender K Malik et al described that the leaf extracts of Gymnema

sylvestre have an anti arthritic activity using albino rats. He studied the

anti arthritic action of leaves of G.sylvestre using Freund’s adjuvant

arthritis in rats. The standard drug used in his study was diclofenac

sodium. He revealed that the petroleum ether (40-60) extract and aqueous

extract of G.sylvestre possessed significant anti arthritic activityin alla the

parameters of his study. He also showed that the more potent anti arthritic

activity of leaves of G.sylvestre is due to nature of steroids, triterpenoids

and saponin glycosides.

Havagiray R Chitme and Nitin P Patel described that Aristolachia

bracteata is well known for its anti arthritis activity in Indian system of

medicine. They analysed its anti arthritic activity using Freund’s

complete adjuvant in rats. The course of treatment was followed for over

and 4 weeks post inoculation period using health parameters , clinical and

behavioural methods of study. The results indicate that regular treatment

of adjuvant induced arthritic rats with A.bracteata extracts improves

ESR,Hb value and restores body weight. Sugnificant (p<0.01) inhibitory

effect was observed with A.bracteata extract on Freund’s complete

adjuvant induced paw edema throughout the study (p<0.001). They also

analysed that treatment of FCA induced rats with A.bracteata extracts

shown (p<0.05) increase in pain threshold, weight bearing ability and

also decline in scratching, defecation and urination. They obtained results

which showed promising effect on FCA modulated health status, clinical

observations and behavioural changes.

R.Lavanya et al described that the phytochemical analysis of methanolic

extracts of A.malabarica and indicated the presence of steroid. Flavonoid

and terpenoid type of compounds in it. They showed the anti arthritic

activity of A.malabarica by inhibition of protein denaturation method.

The methanolic extracts of the plant exhibited remarkable anti arthritic

activity. The maximum membrane stabilization of A.malabarica was

found to be 98.34% at a dose of 1000mcg/0.5ml and that of protein

denaturation was found to be 97.47% at a dose of 250mcg/ml. They also

showed that the isolation and use of active constituents from A.

malabarica treating rheumatism and inflammation.

5. MATERIALS AND METHODS:

The present study has been undertaken to study the anti arthritic activity

of bioactive compounds isolated from Zingiber officinale (Ginger) .

1. Drugs and Chemicals.

2. Identification and collection of plant material.

3. Drying and grinding process of plant parts using sophisticated

instruments.

4. Preparation of extracts:

(a) Maceration process for aqueous extract preparation.

(b) Extraction with various solvents using Soxhlet apparatus.

5. Phytochemical screening:

(a) Preliminary phytochemical screening.

(b) Purification procedure using chromatographic purification.

(TLC,HPLC)

(c) Spectrum analysis (IR,NMR,Mass Spectroscopy)

6. Pharmacological Screening:

(a) Experimental animals.

(b) Acute toxicity studies.

(c) Anti arthritic activity: In this the Freund’s adjuvant arthritis

model is followed.

(d) Statistical analysis.

7. Compilation of data.

6. EXPECTED OUTCOME:

Arthritis is one of the most common auto immune inflammatory

conditions of unknown etiology. The presently available pharmacological

treatments in the market are not only causing economical exploitation but

also associated with severe adverse effects. It has been predicted that near

about 500 plants have been indicated in treatment of arthritis ,however,

only few plants have been scientifically evaluated (<50).So the present

study has been undertaken to study the anti arthritic activity of Zingiber

officinale and to develop a proper medication which will be eco friendly

and has very less side effects that can be used for prophylactic and

therapeutic purpose to control this dangerous disease. About 60-90% of

dissatisfied arthritis patients are likely to seek CAM therapy i.e

physiotherapy, yoga and extract of medicinal plants.

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