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World Journal of Science and Technology | wwwworldjournalofsciencecom | 2011 | 1(10) 01-05

World Journal of Science and Technology 2011 1(10) 01-05ISSN 2231 ndash 2587

wwwworldjournalofsciencecom

EVALUATION OF ANTIMICROBIAL AND ANTI-INFLAMMATORY

ACTIVITIES OF BARK OF JATROPHA GOSSYPIFOLIA

Purohit MC and Purohit Reena

Department of Chemistry HNB Garhwal University Campus Pauri Uttarakhand

India

Corresponding author e-mail mcpurohit123gmailcom

Abstract

Antimicrobial and Anti-inflammatory potential of extracts of Jatropha gossypifolia were investigated The anti-inflammatory activities of methanol and petroleum ether extracts of dried bark extracts of Jatropha gossypifolia

were determined by oral administration of extracts at dose of 200 mgkg of body weight to healthy animals The

extracts were studied for their anti-inflammatory activity in carrageenan-induced hind paw oedema in rats and the

paw volume was measured plethysmometrically at 0 and 4 h after injection The methanol and petroleum ether

extracts of Jatropha gossypifolia significantly (plt005) reduced 75 and 56 carrageenan-induced paw oedema

in rats The methanol extract showed a potent anti-inflammatory activity in comparison to the standard drug

Diclofenac sodium (65 reduction in paw oedema) The antimicrobial activity was determined by well diffusion

method against the pathogenic microbial strains The methanol extracts of bark of the plant showed prominent

antimicrobial activity in comparison to petroleum ether extracts at specific dose 200 microg100 microl The petroleum ether

extracts showed no antimicrobial activity These results indicated that the methanol extract of Jatropha gossypifolia

exhibited more significant activity in the treatment of pain inflammation and microbial infections

Keywords J gossypifolia Antimicrobial activity Anti-inflammatory activity Methanol extractsPetroleum ether extracts

Introduction

There is a worldwide agreement over thepresent need to develop novel agents to treatbacterial and fungal infections that have becomeincreasingly unresponsive to standard antimicrobialtherapy The plant Jatropha gossypifolia (FamilyEuphorbiaceae) is a bushy gregarious shrub growswildly almost throughout India It possesses

significant anticancer and pesticidal activity (Hartwell1969 Chatterjee et al 1980 Panda et al 2009) Theleaf decoction of Jatropha gossypifolia is used forbathing wounds The stem sap stops bleeding anditching of cuts and scratches (Morton 1968 a Morton1980 b) A decoction of the bark is used as anemmenagogue and leaves for stomachache venerealdisease and as blood purifier (Kirtikar and Basu1996 Banerjee and Das 1993) The bark contains

the alkaloid jatrophine and a lignan jatrodien is foundin its stems (Matsuse et al 1999 Omoregbe et al 1996) The latex of Jatropha gossypifolia yielded twocyclic octapeptides ie cyclogossineA and B Theaerial parts contain a new lignan gossypiline Theantimicrobial and anti-inflammatory activities of someplants were reported (Mathur et al 2010 a 2011 b)The present study was undertaken to find out thepossible actions of bark extract of Jatrophagossypifolia for its medicinal property in favour ofoedema and against some pathogenic microbes

Materials and Methods

Chemicals and ReagentsAll materials and reagents used were of

Analytical grade and were obtained from Ranchemand CDH India The media used for the growth of

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bacterial and fungal cultures were procured from Hi-Media Pvt Ltd Bombay IndiaMicrobial Cultures

The pure bacterial and fungal cultures wereobtained from NCL Pune India and were revived forfurther use

Animal Model for determination of Anti-inflammatory activity

Carrageenan-induced Swiss albino rats wereused as animal models in Pharmacology Lab SBS(PG) Institute of Biomedical Sciences BalawalaDehradun Uttarakhand India

Collection of Plant MaterialBark of Jatropha gossypifolia Linn were

collected in the month of October fromSahashradhara and Raipur region of Dehradun

Uttarakhand India and were dried under shade for 15days The plant was identified with the help ofavailable literature and authenticated by BSI

Preparation of extractsDried barks powdered material was soaked

separately methanol and petroleum ether for 72 hThe mixture was stirred after every 24 h and wasfiltered after 72 h The filtrates obtained wereconcentrated in vacuum using rotary evaporator at30oC (Alade and Irobi 1993)

Determination of Antimicrobial activityCulture Media used for growth of bacterial

and fungal cultures were soybean casein digestagarbroth and Sabouraudrsquos dextrose agarbrothrespectively The bacteria were inoculated intosoybean casein digest agarbroth and incubated at370C for 4 h Similar procedure was done for fungalstrains by inoculating in Sabouraudrsquos dextrose brothfor 6 h The test organisms used were Escherichiacoli K-1 Staphylococcus aureus and S pyogenes

Aniger Calbicans P notatum and Scerevisae Thesolvent extracts were introduced separately in each of

the bacterial and fungal cultures By well diffusionmethod (Perez et al 1990) the wells were puncturedand the extracts were introduced in order todetermine the antimicrobial activity by the diameter ofzone of inhibition observed The MIC and MLC valuesof the potent extracts were determined according to

the method adopted by Vollekova et al 2001 andUsman et al 2007 with some modificationsDetermination of Anti-inflammatory activityAnimals

Male albino rats (180ndash200 g) were usedtaking into account international principles and local

regulations concerning the care and use of laboratoryanimals (Olfert et al 1993) The animals had freeaccess to a standard commercial diet and water adlibitum and were kept in rooms maintained at 22 plusmn1ordmC with a 12 h lightdark cycle The institutionalanimal ethical committee has approved the protocolof the study

Carrageenan-induced oedema in ratsFor screening in vivo anti-inflammatory

activity for each of the extracts 4 groups of fiveanimals each were used Paw swelling was induced

by sub-plantar injection of 01 ml 1 sterilecarrageenan in N-saline into the right hind paw Thesolvent extracts of plant at dose of 200 mgkg wereadministered orally 60 minutes before carrageenaninjection Diclofenac sodium (10 mgkg) was used asreference drug Control group received the vehicleonly (10 mlkg) The inflammation was quantified bymeasuring the volume displaced by the paw using aplethysmometer at time 0 1 2 and 4 h aftercarrageenan injection The difference between the leftand the right paw volumes (indicating the degree ofinflammation) was determined and the percent

inhibition of oedema was calculated in comparison tothe control animals

Results

Antimicrobial activity The methanol and petroleum ether extracts

of plant Jatropha gossypifolia were tested forantimicrobial activity using agar well diffusion methodat sample concentration 200 microg100 983221l The methanolextracts of plants showed strong and broad spectrumantibacterial activity against Staphylococcus aureus

S pyogenes and Escherichia coli and antifungalactivity against A niger C albicans P notatum Scerevisae at 200 983221g100 983221l while petroleum etherextracts showed no potency against any of themicrobial culture The results are shown in Table 1and Figure 1

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Table 1Antimicrobial activity of extracts of bark of Jatropha gossypifolia

Diameter of zone of inhibition (mm)

Test

organism

Chloramphenicol

mgml

PC (bacteria)

Fucanazole

mgml

PC (fungus)

Methanol

NC

Jatropha

gossifolia

(methanolextract)

MIC

(mgml)

E coli 26 -- NA 35 0039

S aureus 33 -- NA 31 0039

S pyogenes 28 -- NA 26 0625

A niger -- 26 NA 26 0625

C albicans -- 28 NA 25 0625

P notatum -- 27 NA 32 0039

S cerevisae -- 30 NA 25 0625PC Positive control NC Negative control NA No activity

Fig 1 Antimicrobial activity of methanol extracts of bark of Jatropha gossypifolia

Anti-inflammatory activity

The anti-inflammatory activities of thesolvent extracts of the bark of the plant Jatrophagossypifolia were determined by carrageenaninduced albino rats model The anti-inflammatoryactivities of both the extracts were found to haveeffect in dose-dependent manner Administration ofCarrageenan (01 ml of 1 N-saline) in the subplantar region of the left hind paw of each rat caused

a significant increase in the paw volume of controlgroup animals after 2 hours of administrationMethanol extract and petroleum ether extract at doselevel of 200 mgKg showed 75 and 56 reductionin paw oedema respectively which was significant(plt005) as compared to control Standard drugDiclofenac sodium (50 mgKg) showed 65 reduction in paw oedema The values of reduction inpaw volume at 4 h after carrageenan administration

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viz 010 plusmn 0002 and 016 plusmn 0002 were foundsignificantly of methanol extract and petroleum etherextract respectively of bark of Jatropha gossypifolia

Diclofenac sodium (50 mgkg) showed 014 plusmn 0002reduction in paw volume at 4 h after carrageenanadministration The results are shown in Table 2

Table 2 Anti-inflammatory activities of different extracts of bark of Jatropha gossypifolia

Paw volume (ml)

plusmn SD Standard Deviation plt005 level of significance

Discussion

In the present study methanol and barkextracts of the plant Jatropha gossypifolia wereevaluated for antimicrobial and anti-inflammatorypotential The results suggest that methanol extractsof bark of the plant are potent antimicrobial (Dhaleand Birari 2010 Seth and Sarin 2010) and anti-inflammatory agent (Misra and Misra 2010 Khyadeand Vaikos 2011) Although petroleum ether extractsshowed anti-inflammatory activity while noantimicrobial activity was investigated The resultsvalidate the use of bark extracts as potent

antimicrobial and anti-in flammatory agent

Acknowledgement

The author is thankful to the Head of DepartmentChemistry Pauri campus for supporting me timely andsincere thanks to all other staff members

Sincere thanks for the cooperation to DrVarsha Parcha Professor Dept of Pharmacology

SBS (PG) College Dehradun (UK) India for

providing us the animal house approved by ethicalcommittee for carrying out the research workMy heartily thanks to the Department of

Biochemistry SIPAS Dehradun for allowing me toperforming the anti-microbial activities

References

1 Alade PI and Irobi ON (1993) Antimicrobialactivities of crude leaf extracts of Acalyphawilkensiana Journal of Ethnopharmacology 39171-174

2 Banerjee J and Das B (1993) MAPA Deptof Chemistry University College of ScienceCalcutta India 15 1002-1017

3 Chatterjee A Das BN Chaudhary A andDabkirtaniya S (1980) Note on the insecticidalproperties of the seeds of Jatropha gossypifoliaLinn Indian J Agri Sci 50 637-638

Experiment Control Diclofenac

Sodium

(50mgkg

orally)

Methanol

extract

(200 mgkg)

Petroleum

ether extract

(200mgkg)

1h after

treatment

025plusmn003 021plusmn

003

023plusmn 003 028plusmn003

2h after

treatment


003


4h aftertreatment

025plusmn003 014plusmn002

65

reduction inpaw

oedema

010plusmn 002

75

reduction inpaw oedema

016plusmn002

56


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4 Dhale DA and Birari AR (2010) Preliminary screening of antimicrobial andphytochemical studies of Jatropha gossypifoliaLinn Recent Research in Science andTechnology 2(7) 24-28

5 Hartwell JL (1969) Plants used againstcancer A survey Lloydia 32 153-205

6 Khyade MS and Vaikos NP (2011) Pharmacognostical and PhytochemicalEvaluation of leaf of Jatropha gossypifolia LInternational Journal of Research in Ayurvedaand Pharmacy 2(11) 177-180

7 Kirtikar KR and Basu BD (1996) IndianMedicinal Plants III International BookDistributors Allahabad pp 22-47

8 Mathur A Prasad GBKS Dua VK (2010) Screening of some Indian plants for theirantibacterial and antifungal properties Flora andFauna 16(2) 166-170

9 Mathur A Purohit R Mathur D PrasadGBKS Dua VK (2011) Pharmacologicalinvestigation of methanol extract of Syzigumcuminii seeds and Crateva nurvula bark on thebasis of antimicrobial antioxidant and anti-inflammatory properties Der Chemica Sinica2(1) 174-181

10 Misra M and Misra AN (2010) JatrophaThe Biodiesel Plant Biology Tissue Culture andGenetic Transformation A Review Int J PureAppliSci Technol 1(1) 11-24

11 Morton JF (1968 a) A survey of medicinalplants Curacao Economic Botany 22 87-102

12 Morton JF (1980 b) Caribbean and LatinAmerican folk medicine and its influence in theUnited States Quarterly Journal of Crude Drug

Research 18 57-75

13 Matsuse IT Lim YA Hattori M CorreaM and Gupta MP (1999)Search of anti-viralproperties in Panamanian medicinal plants theeffects on HIV and its essential enzymes Journalof Ethnopharmacology 64 15-22

14 Olfert ED Cross BM and McWilliamsAA (1993) Canadian Council of Animal Careguide to the care and use of experimentalanimals 2nd edition Vol1

15 Omoregbe RE Ikuebe OM and IhimireIG (1996) Antimicrobial activity of somemedicinal plants extracts on Escherichia coliSalmonella paratyphi and Shigella dysenteriaeAfr J Med Med Sci 25 373-375

16 Panda BB Gaur K Nema RK Sharma

CS Jain AK and Jain CP (2009) Hepatoprotective activity of Jatropha gossypifolia1against carbon tetrachloride- induced hepaticinjury in rats Asian Journal of Pharmaceuticaland Clinical Research 2(1) 50-54

17 PerezC and Anesini C (1993) In vitro antimicrobial activity of Argentine folk medicinalplants against Salmonella typhii Journal ofEthnopharmacology 44 41-46

18 Seth R and Sarin R (2010) Analysis of thephytochemical content and antimicrobial activity

of Jatropha gossypifolia L 2(5) 285-291

19 UsmanH AbdulrahmanFI and Ladan AH(2007) Phytochemical and antimicrobialevaluation of Tribulus terrestris L(Zygophylaceae) Growing in Nigeria Res JBio Sci Medwell Journals 2(3) 244-247

20 Vollekova AD Kostalova Sochorova R(2001)Alkaloids from Mahonia aquifolium stembark is active against Malassezia sp FoliaMicrobiol 46 107-111

8132019 9791-27264-1-PB



bacterial and fungal cultures were procured from Hi-Media Pvt Ltd Bombay IndiaMicrobial Cultures

The pure bacterial and fungal cultures wereobtained from NCL Pune India and were revived forfurther use

Animal Model for determination of Anti-inflammatory activity

Carrageenan-induced Swiss albino rats wereused as animal models in Pharmacology Lab SBS(PG) Institute of Biomedical Sciences BalawalaDehradun Uttarakhand India

Collection of Plant MaterialBark of Jatropha gossypifolia Linn were

collected in the month of October fromSahashradhara and Raipur region of Dehradun

Uttarakhand India and were dried under shade for 15days The plant was identified with the help ofavailable literature and authenticated by BSI

Preparation of extractsDried barks powdered material was soaked

separately methanol and petroleum ether for 72 hThe mixture was stirred after every 24 h and wasfiltered after 72 h The filtrates obtained wereconcentrated in vacuum using rotary evaporator at30oC (Alade and Irobi 1993)

Determination of Antimicrobial activityCulture Media used for growth of bacterial

and fungal cultures were soybean casein digestagarbroth and Sabouraudrsquos dextrose agarbrothrespectively The bacteria were inoculated intosoybean casein digest agarbroth and incubated at370C for 4 h Similar procedure was done for fungalstrains by inoculating in Sabouraudrsquos dextrose brothfor 6 h The test organisms used were Escherichiacoli K-1 Staphylococcus aureus and S pyogenes

Aniger Calbicans P notatum and Scerevisae Thesolvent extracts were introduced separately in each of

the bacterial and fungal cultures By well diffusionmethod (Perez et al 1990) the wells were puncturedand the extracts were introduced in order todetermine the antimicrobial activity by the diameter ofzone of inhibition observed The MIC and MLC valuesof the potent extracts were determined according to

the method adopted by Vollekova et al 2001 andUsman et al 2007 with some modificationsDetermination of Anti-inflammatory activityAnimals

Male albino rats (180ndash200 g) were usedtaking into account international principles and local

regulations concerning the care and use of laboratoryanimals (Olfert et al 1993) The animals had freeaccess to a standard commercial diet and water adlibitum and were kept in rooms maintained at 22 plusmn1ordmC with a 12 h lightdark cycle The institutionalanimal ethical committee has approved the protocolof the study

Carrageenan-induced oedema in ratsFor screening in vivo anti-inflammatory

activity for each of the extracts 4 groups of fiveanimals each were used Paw swelling was induced

by sub-plantar injection of 01 ml 1 sterilecarrageenan in N-saline into the right hind paw Thesolvent extracts of plant at dose of 200 mgkg wereadministered orally 60 minutes before carrageenaninjection Diclofenac sodium (10 mgkg) was used asreference drug Control group received the vehicleonly (10 mlkg) The inflammation was quantified bymeasuring the volume displaced by the paw using aplethysmometer at time 0 1 2 and 4 h aftercarrageenan injection The difference between the leftand the right paw volumes (indicating the degree ofinflammation) was determined and the percent

inhibition of oedema was calculated in comparison tothe control animals

Results

Antimicrobial activity The methanol and petroleum ether extracts

of plant Jatropha gossypifolia were tested forantimicrobial activity using agar well diffusion methodat sample concentration 200 microg100 983221l The methanolextracts of plants showed strong and broad spectrumantibacterial activity against Staphylococcus aureus

S pyogenes and Escherichia coli and antifungalactivity against A niger C albicans P notatum Scerevisae at 200 983221g100 983221l while petroleum etherextracts showed no potency against any of themicrobial culture The results are shown in Table 1and Figure 1

8132019 9791-27264-1-PB





Test

organism

Chloramphenicol

mgml

PC (bacteria)

Fucanazole

mgml

PC (fungus)

Methanol

NC

Jatropha

gossifolia

(methanolextract)

MIC

(mgml)

E coli 26 -- NA 35 0039

S aureus 33 -- NA 31 0039


A niger -- 26 NA 26 0625


P notatum -- 27 NA 32 0039






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Paw volume (ml)


Discussion



Acknowledgement






References





Sodium

(50mgkg

orally)

Methanol

extract

(200 mgkg)

Petroleum

ether extract

(200mgkg)

1h after

treatment


003


2h after

treatment


003


4h aftertreatment


65

reduction inpaw

oedema

010plusmn 002

75


016plusmn002

56


8132019 9791-27264-1-PB












Research 18 57-75











8132019 9791-27264-1-PB





Test

organism

Chloramphenicol

mgml

PC (bacteria)

Fucanazole

mgml

PC (fungus)

Methanol

NC

Jatropha

gossifolia

(methanolextract)

MIC

(mgml)

E coli 26 -- NA 35 0039

S aureus 33 -- NA 31 0039


A niger -- 26 NA 26 0625


P notatum -- 27 NA 32 0039






8132019 9791-27264-1-PB






Paw volume (ml)


Discussion



Acknowledgement






References





Sodium

(50mgkg

orally)

Methanol

extract

(200 mgkg)

Petroleum

ether extract

(200mgkg)

1h after

treatment


003


2h after

treatment


003


4h aftertreatment


65

reduction inpaw

oedema

010plusmn 002

75


016plusmn002

56


8132019 9791-27264-1-PB












Research 18 57-75











8132019 9791-27264-1-PB






Paw volume (ml)


Discussion



Acknowledgement






References





Sodium

(50mgkg

orally)

Methanol

extract

(200 mgkg)

Petroleum

ether extract

(200mgkg)

1h after

treatment


003


2h after

treatment


003


4h aftertreatment


65

reduction inpaw

oedema

010plusmn 002

75


016plusmn002

56


8132019 9791-27264-1-PB












Research 18 57-75











8132019 9791-27264-1-PB












Research 18 57-75











9791-27264-1-PB

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