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.
8. REFERENCES:
1. Aggarwal R B ,Rangari V D, Indian J.Exp Bio ,Vol. 41,2003 Aug,
890-894.
2. Arya Vaidya Shala ,Indian Medicinal Plants,A Compendium of
500 Species,Vol.3,Orient Longmann Ltd.,Madras1997,107-109.
3. Bansod S Mangesh et al., International Journal of Pharmacy and
Pharmaceutical Sciences, Vol.3,Suppl 2,2011, 186-192.
4. Chandur Uma et al.,International Journal of Pharmaceutical
Sciences., May Aug 2010;2(2);593-597.
5. Chitme R Havagiray and Patel P Nitin .,The Open Natural
Products Journal,2009.2.6-15.
6. Chakarborty A.K,Roy H K; Journal of Pharmaceutical Science
and Technology;Vol.2(10),2010,330-332.
7. Islam,N and M.Kahn,1981.,Rheumatic diseases other than rheu
matic fever.Bangladesh Medical Research Council Bulletin,7:18-
21.
8. Jubie S et al ;Anti arthritic activity of Bark extracts of Alangium
salvifolium wang,Vol. 1 ,No 3,433-436.
9. Kamalutheen.M et al ;E Journal of Chemistry 6(4)-943-948.
10. Kristina J S,Robert W,Ankie B,Petra M,Britta T R,Sonja C O,Azar
G,Phytochemistry,2005,66,1448-1464.
11. Kirtikar K R and Basu B D;Indian Medicinal Plants,Oriental
Enterprises,Second Edition,2003.
12. Lavanya R et al,Research Journal of Pharmaceutical,Biological
and Chemical Sciences,Oct-Dec 2010,1(4),P.no745-752.
13. Malik K Jitender et al .,Der Pharmacia Lettre,2010:2(1) ,336-341.
14. Otari,K.V.,et al.,Journal of Cell and Tissue
Research,Vol.10(2),2269-2280.
15. Rahmatullah M et al;Advances In Natural and Applied
Sciences ,2010,4(2);124-127.
16. Rahmatullah M et al;An Ethonobotanical Survey of Dhamrai sub
Distt. In Dhaka Bangladesh,American Eurasian Journal of
Sustainable Agriculture,3(4);881-888.
17. Rajendran Rekha and Krishna Kumar Ekambaram;Avicenna
Journal of Medical Biotechnology,Vol.2,No 2,April June 2010.
18. Singh S S,Pandey S C,Srivastava S,Gupta V S,Patro B,Ghosh A C;
Chemistry and Medicinal Properties of Tinospora cordifolia
(Guduchi).Indian J Pharma.,35:83-91.
19. Sangeetha M et al.,Research Journal of Pharmaceutical,Biological
and Chemical Sciences,Jan Mar 2011,Vol 2 P.no822-827.
20. Tripathy S et al,2009,African Journal of Pharmacy and
Pharmacology Vol.3(12).pp611-614.
21. William J Arthritis and allied condition;A Textbook of
Rheumatology(Vol.1),3rd ed,Baltimore Tokyo;A waverly
Company:1996,207-226.