free radicals scavenging and inhibition of lipid peroxidation by salvadora persica

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Research Article Pharmacology

International Journal of Pharma and Bio SciencesISSN

0975-6299

FREE RADICALS SCAVENGING AND INHIBITION OF LIPID PEROXIDATION BY

SALVADORA PERSICA LINN

MANGAL SAIN HOODA1

AND JANARDHAN SINGH2

1Scholar of Jodhpur National University Jodhpur Rajasthan India

2Pt BD Sharma PGIMS Rohtak124001 Haryana India

ABSTRACT

Salvadora persica ( Salvadoraceae) is widely used in Unani system of medicine Its rootand stem were extracted using solvents of different polarities and explored for in-vitro free radical scavenging activity and lipid per-oxidation assay in rat brain homogenatePreliminary assays of six different extracts of Salvadora persica scavenges 11-diphenyl-2-picrylhydrazyl stable free radicals in concentration dependent manner Allstudied extracts possess electron donating ability and reduce ferric ion to ferrous in acell free system at pH-74 Also 250 and 500microgml alcoholic and hydro alcoholicextracts reduced the activated lipid per-oxidation in rat brain homogenate Further bothextracts reduced the lipid per-oxidation to the extended time period (1 to 12 hour afterinitiation of lipid per-oxidation) Root extracts were found more effective when comparedto stem in scavenging 11-diphenyl-2-picrylhydrazyl reducing ferric ion and inhibitingthe lipid per-oxidation Hydro-alcoholic extracts were found more effective in scavengingthe 11-diphenyl-2-picrylhydrazyl and reducing the ferric ions The significantcorrelations exist between extract concentrations and percentage scavenging activity ofradicals in all models These results clearly indicate that Salvadora persica is effectivefree radical scavenger and chain breaking antioxidant

KEY WORDS Antioxidant DPPH Free Radical Scavenging Miswak Salvadora persica LToothbrush tree

Corresponding author

MANGAL SAIN HOODA

Scholar of Jodhpur National University Jodhpur Rajasthan India



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INTRODUCTION

Free radicals may be designated as molecularsharks that damage molecules in cellmembranes mitochondria DNA and are veryunstable tends to rob electrons from the

molecules in the immediate surroundings inorder to replace their own losses

1 Reactive

oxygen species (ROS) is a collective termwhich includes not only the oxygen radicalsbut also some non-radical derivatives ofoxygen these include hydrogen peroxidehypochlorous acid and ozone2 Numerousdisorders like rheumatoid arthritiscardiovascular disorders cystic fibrosismetabolic disorders neurodegenerativediseases and gastrointestinal ulcerogenesis

are reported as ROS mediated34 The roleplayed by ROS in stress induced gastric ulcerand inflammatory bowel diseases have beenwell established as well as their involvementin the process of ageing56 The oxygenintermediates differ significantly in theirinteractions and can cause extensive cellulardamage such as nucleic acid strand scissionmodification of polypeptides and lipid per-oxidation etc

7 Screening of compounds which

scavenge the free radicals could lead to

promising compounds Most of theantioxidants used in therapy are derived fromnatural sources About 28 drugs approvedby the FDA between 1981 and 2002 are eithernatural products or chemicals derived fromthem

89 Many synthetic antioxidant

components have shown toxic andormutagenic effects Hence the attention hasbeen given to naturally occurring antioxidantsTherefore identification of effectiveantioxidants and free radical scavengers from

plants origin is an ideal strategy for new drugdevelopment Hence present study wasdesign to explore the antioxidant and freeradical scavenging activity of Salvadora persica in a prospective waySalvadora persica ( SP) also known asmiswak toothbrush tree and mustard treedistributed mainly in tropical and sub tropicalAsia The plant is a large evergreen profusely

branched shrub or a small tree up to 4-6 mtall SP has been used commonly astoothbrush to strengthen the gums10 Thefresh root barks and leaves have been used in

folk medicine for the treatment of a wide rangeof medical problems such as cough asthmascurvy piles rheumatism leprosygonorrohoea headache and hepaticdisorders

11 Various phytochemical studies on

Salvadora persica reported the presence ofalkaloids salvadorine trimethylaine andsalvadoricine

12 Essential oil from the roots of

SP contain benzyl isothiocyanate (70) withother components such as α-pinenecamphene benzaldehyde β-pinene myrcene

δ-3-carene limonene terpinolene benzylnitrile umbellulone β-elemene γ-muurolenemyristicin β-caryophyllene and longifolene13Proteinstannins steroids flavonoids andterpenoids in Tagetes erecta flower extractswhich therefore encourages antioxidantstudies

14 Various pharmacological activities

on SP including in-vivo antimicrobial activityespecially on lactobacilli and streptococcusmutans

15with moderate secretory activity

significantly high acetyl cholinesterase

inhibiting ability antifertility activity in malerats16 The aqueous extract of SP leavespossesses analgesic activity and reducescarrageenan induced inflammation in ratpaw17 Aqueous and alcoholic extract fromleaves of SP reduce elevated urinary oxalatelevels and deposition of stone-formingconstituents in the kidneys of calculogenicrats

18 In our previous study we showed root

hydroalcoholic extract of SP reducescarrageenan induced inflammation in rats and

scavenges ABTS [22-azino bis (3-ethylbenzo-thiazoline-6-sulphonic acid)] andsuperoxide radicals in a cell free system19 Inpresent study we aimed to explore theantioxidant potential of different extracts (rootand stem) of SP using solvent of differentpolarity (alcoholic hydro alcoholic aqueous)



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MATERIALS AND METHODS

Chemicals Details1 1-diphenyl-2-picryl hydrazyl (DPPH) o-phenanthroline ferric chloride ascorbic acidand EDTA were procured from HaryanaScientific amp Engg Corp Rohtak Root andstem of Salvadora persica were collected inMarch- April month from Kharainti MehamTeh Rohtak District Haryana (India) theplant material was authenticated by Dr AshokKumar Sharma MD (Dravyaguna Vigyan)Prof amp Head of Department Shri BabaMastnath Ayurvedic Degree College AsthalBohar Rohtak

Extraction processThe shed dried plant material (either root orstem) were grinded and powdered material(100 g) was used for extraction The sixdifferent extracts three from each root andstem were prepared by hot continuouspercolation method in a Soxhlet apparatusFollowing six extractions were collectedseparately and dried in vacuo [1] Rootextracted with 99 ethanol (root-AA yield0915) [2] Root extracted with 70 ethanol

(root-HA yield 1165) [3] Root extractedwater (root-W yield 1225) [4] Stemextracted with 99 ethanol (stem-AA yield1005) [5] Stem extracted with 70 ethanol(stem-HA yield 1145) and [6] Stemextracted with water (stem-W yield 1195)

These extracts were condensed by re-distillation and dried in vacuum desiccators toobtain a final extract residue

DPPH radical scavenging activity 20 The free radical scavenging activity wasmeasured in terms of hydrogen donating orradical scavenging ability by using stable freeradical DPPH (100 microM) in ethanol The equalvolumes of different extracts were incubatedwith DPPH in graded concentrations of (10 to

500 microgml) for 20 min at room temperatureand by using a digital UVVISspectrophotometer (model 371E) absorbancewas recorded at 517 nm The experiment wasperformed in triplicate Scavenging ability wasmeasured by decreased DPPH absorbance intest sample when compared to standardDPPH solution Results were expressed aspercentage inhibition of DPPH by comparingwith blank

Absorbance of control sample- Absorbance of test sampleage Scavenging = ---------------------------------------------------------------------------- X 100

Absorbance of control sample

Reduction of ferric-ions21 The electron donating capability (reducingability) was studied by ferric chloride reductionin cell free system The reaction mixturecontained 10 ml of phosphate buffer (pH 74)100 microM ferric chloride and 05 ml of test

compounds of different concentrations (10-500 microgml) After 3 min of incubation EDTA(100 microM) and ortho-phenanthroline (300 microM)were added reaction was allowed for 10 min

at room temperature and by using a digitalUVVIS spectrophotometer (model 371E)absorbance was recorded at 510 nm Ascorbicacid was used as standard as equivalent to100 reduction of ferric ions comparativereduction of Fe3+ by Salvadora persica extract

was calculated Percentage reduction of Ferric ion by testcompound

Absorbance of test sampleage Reduction = --------------------------------------- X 100

Absorbance of ascorbic acid



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Lipid per-oxidation22 The brain isolated from healthy albino rat(180-200 gm) was used as lipid source Brainhomogenate (10 wv) was prepared in 150mM KCl and centrifuged at 800 g for 10

minutes The supernatant was collected andused immediately to study in-vitro lipid per-oxidation Briefly the reaction mixturecontained 03 ml of brain homogenate KCl(100 microM) ascorbic acid (100 microM) ferric-chloride (100 microM) 05 ml of gradedconcentrations of extracts and final volumewas made with buffer After incubating at roomtemperature for 20 minutes 10 ml ofthiobarbituric acid-trichloroacetic acid (TBA-

TCA) reagent was added The resultedmixtures were heated at 800C for 20 minutescool and centrifuged for 10 minutes at 1000rpm and by using a digital UVVISspectrophotometer (model 371E) recorded the

absorbance at 532 nm Control and standard(curcumin 10 microM) were carried out at similarmanner Percentage inhibition of thiobarbituricacid reactive substance (TBARS) formation byextractstandard drug (curcumin) wascalculated by comparing with control Allexperiments were carried out in triplicate andresults are the means of one such individualexperiment Percentage inhibition of lipid per-oxidation by test compound

Absorbance of control sample-Absorbance of test sample age inhibition = ----------------------------------------------------------------------- X 100


RESULTS

DPPH radical scavenging activityRoot and stem extracts of SP scavenged theDPPH stable free radicals in a concentrationdependent manner (10-500 microgml) All theextracts showed maximum scavenging activity

at 400microgml beyond this biphasic effect wasobserved with reduced scavenging activityThe root-HA showed maximum activity(8815) root-AA (730) while root-W

showed a maximum activity (7185) atconcentration 400 microgml of root extracts andstem extracts also be in same manner Thestem-HA showed maximum activity (8772)stem-AA (640) while stem-W showed amaximum activity (530) at concentration 400microgml In all the cases biphasic effect was

observed above concentration 400 microgml Theinteraction of Salvadora persica root and stemextracts with DPPH radicals presented in Taband Fig1a

Table 1aInteraction of Salvadora persicaa root extracts with DPPH Radical

DrugConcugml

Root- HA Root-AA Root-W Stem-HA Stem-AA Stem-W

Ab Sca Ab Sca Ab Sca Ab Sca Ab Sca Ab Sca

500 107plusmn003 8414 204plusmn005 6991 210plusmn003 6900 148plusmn002 7810 277plusmn003 5900 352plusmn003 4800

400 080plusmn005 8815 183plusmn004 7300 190plusmn001 7185 083plusmn001 8772 246plusmn003 6400 322plusmn003 5300200 237plusmn007 6490 263plusmn003 6121 275plusmn003 5925 275plusmn004 5931 406plusmn001 4011 475plusmn003 2994





Control 675plusmn004 0000 678plusmn002 0000 675plusmn006 0000 676plusmn004 0000 678plusmn002 0000 678plusmn002 0000a All values are mean (n = 3) Data are expressed as Mean plusmn SD



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Figure1aInteraction of Salvadora persica root extracts with DPPH Radical

Reduction of ferric ionReduction of ferric ion ie Fe2+- Fe3+ couple isknown to be involved in various free radicalreactions All of these extracts of root-HAroot-AA and stem-HA reduce Fe3+ in to Fe2+atpH 74 In this study reduction of Fe3+ intoFe

2+by ascorbate was taken as 100 The

root-HA and stem-HA (500 microgml) caused a

significant reduction of Fe3+

to (7683) and(7108) respectively while the root-AA (500microgml) produced (6675) The stem-AA androot-W (500 microgml) caused a significantreduction of (60) and (5136) respectivelywhile stem-W (500 microgml) produced areduction of 4129 as shown in Tab and Fig1b

Table 1b

Reduction of ferric ions by Salvadora persicaa root extracts

Drugconcugml


Ab Red Ab Red Ab Red Ab Red Ab Red Ab Red

AscAStd300 microM

410plusmn002 100 406plusmn003 100 405plusmn003 100 408plusmn001 100 410plusmn003 100 402plusmn003 100







10 057plusmn001 1390 018plusmn001 443 006plusmn001 148 045plusmn002 1103 009plusmn002 220 003plusmn001 75a All values are mean (n = 3) Data are expressed as Mean plusmn SD



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Figure1bReduction of ferric ions by Salvadora persica root extracts

Lipid peroxidation in rat brain homogenate The amount of thiobarbituric acid reactivesubstance (TBARS) was calculated andpercentage inhibition of TBARS formed wascompared with control and standard drug(curcumin) The root-AA and root-HA(500microgml) inhibited 6435 and 4507formation of TBARS when compared tocontrol respectively The stem-AA and stem-HA (500microgml) inhibited 6404 and 2788

formation of TBARS respectively Waterextracts of root or stem doesnrsquot inhibit the

TBARS formation Alcoholic and hydro-alcoholic extracts (root-AA root-HA stem-AAand stem-HA) significantly inhibited the lipidper-oxidation at concentrations 250 and 500microgml Further alcoholic and hydro-alcoholicextracts (root-AA root-HA stem-AA and stem-HA) inhibited the formation of lipid per-oxidation for a prolong time after initiation oflipid per-oxidation Root-AA reduced theformation of TBARS from 6425 to 5979 at

1hour and 12 hours post-initiation to lipid per-oxidation as shown in Tab and Fig 1c amp 1d

Table 1cLipid peroxidation inhibition by Salvadora persicaa root extracts

Drugconcugml

Root- AA Root-HA Root-W Stem-AA Stem-HA Stem-W

Ab Inh Ab Inh Ab Inh Ab Inh Ab Inh Ab Inh


Control 965plusmn006 0000 965plusmn006 0000 965plusmn006 0000 965plusmn006 0000 965plusmn006 0000 965plusmn006 0000







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DISCUSSION

Free radicals have been a subject ofsignificant interest among scientists in the pastdecade and their possible role in humandiseases has gained importance now in

days2324 Antioxidants neutralize toxin andvolatile free radicals that are defined as atomsor groups of atoms having an unpairedelectron6 These also include related reactiveoxygen species (ROS) that leads to freeradical generation causes the cascadingchain reaction in biological system In anormal healthy organism or human body thegeneration of pro-oxidants in the form of ROSis effectively kept in check by various levels ofantioxidant defense Antioxidants present in

various dietary supplements offered theirbeneficial effects by neutralizing these ROSduring various disease conditions Lipidsproteins and DNA are all susceptible to attackby free radicals and cellular damage inducedby oxidative stress has been implicated in theetiology of numerous diseases

DPPH radical is widely used as amodel to investigate the scavenging potentialof several natural compounds such asphenolic and anthocyanins or crude extract of

plants25 The ability of stem and root extractsof SP to reduce DPPH radicals supports itsfree radical scavenging activity Our studyindicates the proton donating property may beresponsible for free radical scavenging activityof SP Antioxidant compounds for examplesesamol gallic acid poly-phenols reduce theFe3+ to Fe2+ and are considered as chainbreaking antioxidant for their proton donatingactivity

26 SP reduces ferric ion at pH 74

which indicates its proton donating ability and

supports its free radical scavenging activityThe reducing capacity of a compound mayserve as a significant indicator of its potentialantioxidant activity It was therefore to beexpected that SP has potent antioxidant andradical scavenging ability This activity isbelieved to be mainly due to their redoxproperties which play an important role inadsorbing and neutralizing free radicals

quenching singlet and triplet oxygen ordecomposing peroxides Hence present studyrevealed antioxidant property of SP root andstem extracts

Lipid per-oxidation is an oxidativealteration of polyunsaturated fatty acids in the

cell membranes that generates a number ofdegradation products

27 TBARS one of the

products of lipid per-oxidation has beenstudied widely as an index of lipid per-oxidation and as a marker of oxidativestress28 We observed incubation of SPextracts with brain homogenate reduced thelipid per-oxidation at great extent whichindicates the protecting effect of SP againstlipid per-oxidation and TBARS formation Ithas been proved that free radicals initiated

lipid per oxidation play an important role in thepathogenesis of certain diseases and ageing5Reducing the formation of TBARS improvesmany pathological conditions Hence SP couldbe a promising plant to neutralizing freeradicals and reducing the lipid per-oxidation inmany pathological conditions There are manyreports that support the use of antioxidantsupplementation in reducing the level ofoxidative stress and in slowing or preventingthe development of complications associated

with diseases Numerous plant constituentshave shown free radical scavenging orantioxidant activity

CONCLUSIONS

Our study showed that hydro alcoholic rootextract of Salvadora persica is most effectiveantioxidant when compared to other extractsin various in vitro assay systems SP has beenreported to contain indole alkaloids

flavonoids the sulphur-containing compoundstropaedoin triterpenes and phytosterols

2930

of these compounds the alkaloids andflavonoids are probably responsible for its freeradical and reducing property observed in thisstudy As compared to previous reports ourstudy showed that Salvadora persica exerteda potent effect Further studies are required tobetter understand these compounds and their



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effects on cellular function Antioxidantproperties of Salvadora persica could be

beneficial in pathological condition involvingoxidative stress

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[1] Halliwell B The wanderings of a freeradical Free Radic Biol Med 46 531-542 (2009)

[2] Toyokuni S Reactive oxygen species-induced molecular damage and itsapplication in pathology Pathol Int 4991-102 (1999)

[3] Muller FL Lustgarten MS Jang YRichardson A Van Remmen H Trendsin oxidative ageing theories Free RadicBiol Med 43 477-503 (2007)

[4] Ischiropoulos H al-Mehdi AB Fisher ABReactive species in ischemic rat lunginjury contribution of peroxynitrite AmJ Physiol 269 L158-L164 (1995)

[5] McCord J M Oxygen-derived freeradicals in post ischemic tissue injury NEngl J Med 312 159-163 (1985)

[6] Finkel T Holbrook NJ Oxidantsoxidative stress and the biology ofageing Natur e 408 239-247 (2000)

[7] Pryor WA Porter NA Suggestedmechanisms for the production of 4-

hydroxy-2-nonenal from the autoxidationof polyunsaturated fatty acids FreeRadic Biol Med 8 541-543 (1990)

[8] Clardy J Walsh C Lessons from naturalmolecules Nature 432 829-837 (2004)

[9] Schmidt BM Ribnicky DM Lipsky PERaskin I Revisiting the ancient conceptof botanical therapeutics Nat Chem

Biol 3 360-366 (2007)[10] Sofrata AH Claesson RL Lingstrom PK

Gustafsson AK Strong antibacterial

effect of miswak against oralmicroorganisms associated withperiodontitis and caries J Periodontol79 1474 -1479 (2008)

[11] Kirtikar KR Basu BD Blatter E IndianMedicinal Plants Periodical ExpertsVol-2 Delhi 1537-1539 (1975)

[12] Malik S Ahmad SS Haider SI MuzaffarA Salvadoricine A new indole alkaloid

from the leaves of Salvadora PersicaTetrahedron Letters 28 163-164 (1987)

[13] Bader A Flamini G The composition ofthe root oil of Salvadora persica Linn JEssent Oil Res 14 128-129 (2002)

[14] Basavaraj V Chivde Karnakumar VBiradar Rajabhau ShiramaneKamshetty V Manoj In-vitro antioxidantactivity studies on the flower of TagetesErectal L Int J Pharma and Bio Sci2(3) 223-229 (2011)

[15] Almas K Al-Zeid Z The immediateantimicrobial effect of a toothbrush andmiswak on cariogenic bacteria A clinicalstudy J Contemp Dent Pract 5 105-114 (2002)

[16] Darmani H AI-Hiyasat AS Elbetieha AMAlkofahi A The effect of an extract ofSalvadora persica (miswak chewingstick) on fertility of male and female micePhytomedicine 10 63-65 (2003)

[17] Parihar VK Ganesh S Kumar SMehrotra A Reddy M Sreedhara Pai

KR Unnikrishnan MK Rao MCAnalgesic anti-inflammatory and in-vitro antioxidant activities of Salvadora persicaLinn XXXIX Annual Conference of Indianpharmacological Society held at NIPERMohali Chandigarh Nov 1-3 2007 PP-14

[18] Geetha K Manavalan R VenkappayyaD Control of urinary risk factors of stoneformation by Salvadora persica inexperimental hyperoxaluria Methods

Find Exp Clin Pharmacol 32 623-629(2010)

[19] Hooda MS Parihar VK Mor VAntioxidant and anti-inflammatory activityof crude root extract of SalvadoraPersica Linn Indian J Pharm 38 AA-6S18-19 (2006)

[20] Narla RS Rao MNA Scavenging of freeradicals and inhibition of lipid



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peroxidation by 3-phenyl syndone JPharm Pharmacol 47 623-625 (1995)

[21] Parihar VK Dhawan J Kumar SManjula SN Subramanian GUnnikrishnan MK Rao MC Free radicalscavenging and radioprotective activity of

dehydrozingerone against whole bodygamma radiation in Swiss albino miceChemico biological Interactions 170 49-58 (2007)

[22] Rajakumar DV Rao MNA Antioxidantproperties of dehydrozingerone andcurcumin in rat brain homogenates Molecular and cellular Biochemistry 14073-79 (1994)

[23] Maxwell SR Prospects for the use ofantioxidant therapies Drugs 49 345-361

(1995)[24] Jovanovic SV Simic MG Antioxidants in

nutrition Ann N Y Acad Sci 899 326-634 (2000)

[25] Veerapur VP Prabhakar KR Parihar VKKandadi MR Ramakrishana S Mishra BSatish Rao SB Srinivasan KKPriyadarsini KI Unnikrishnan MK Ficusracemosa stem bark extract a potentantioxidant and a probable natural

radioprotector Evid Based ComplementAlternat Med 6 317-324 (2009)

[26] Parihar VK Prabhakar KR Veerapur VPSudheer KM Rosi Reddy Y Joshi RUnnikrishnan MK Rao MC Effect ofsesamol on radiation induced cytotoxicity

in Swiss albino mice Mutation Research611 1-9 (2006)

[27] Pratt DA Tallman KA Porter NA Freeradical oxidation of polyunsaturatedlipids New mechanistic insights and thedevelopment of peroxyl radical clocksAcc Chem Res 44 458-467(2011)

[28] Prabhakar KR Veerapur VP Parihar VKPriyadarsini KI Rao BSS UnnikrishnanMK Evaluation and optimization ofradioprotective activity of Coronopus

didymus Linn in gamma-irradiated miceInt J Radiat Biol 82 525-536 (2006)

[29] Kamel MS Ohtani K Assaf MH Kasai REl-Shanawany MA Yamasaki K Lignanglycosides from stems of Salvadora persica Phytochemistry 31 2469-2471(1992)

[30] Akhtar MS Ajmal M Significance ofchewing-sticks (miswak) in oral hygienefrom a pharmacological view- point JPak Med Assoc 31 89-95 (1981)



C o p y r i g h t o f I n t e r n a t i o n a l J o u r n a l o f P h a r m a amp B i o S c i e n c e s i s t h e p r o p e r t y o f I n t e r n a t i o n a l

J o u r n a l o f P h a r m a amp B i o S c i e n c e s a n d i t s c o n t e n t m a y n o t b e c o p i e d o r e m a i l e d t o m u l t i p l e

s i t e s o r p o s t e d t o a l i s t s e r v w i t h o u t t h e c o p y r i g h t h o l d e r s e x p r e s s w r i t t e n p e r m i s s i o n

H o w e v e r u s e r s m a y p r i n t d o w n l o a d o r e m a i l a r t i c l e s f o r i n d i v i d u a l u s e



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983120 983085 983093983090983090

INTRODUCTION

Free radicals may be designated as molecularsharks that damage molecules in cellmembranes mitochondria DNA and are veryunstable tends to rob electrons from the

molecules in the immediate surroundings inorder to replace their own losses

1 Reactive

oxygen species (ROS) is a collective termwhich includes not only the oxygen radicalsbut also some non-radical derivatives ofoxygen these include hydrogen peroxidehypochlorous acid and ozone2 Numerousdisorders like rheumatoid arthritiscardiovascular disorders cystic fibrosismetabolic disorders neurodegenerativediseases and gastrointestinal ulcerogenesis

are reported as ROS mediated34 The roleplayed by ROS in stress induced gastric ulcerand inflammatory bowel diseases have beenwell established as well as their involvementin the process of ageing56 The oxygenintermediates differ significantly in theirinteractions and can cause extensive cellulardamage such as nucleic acid strand scissionmodification of polypeptides and lipid per-oxidation etc

7 Screening of compounds which

scavenge the free radicals could lead to

promising compounds Most of theantioxidants used in therapy are derived fromnatural sources About 28 drugs approvedby the FDA between 1981 and 2002 are eithernatural products or chemicals derived fromthem

89 Many synthetic antioxidant

components have shown toxic andormutagenic effects Hence the attention hasbeen given to naturally occurring antioxidantsTherefore identification of effectiveantioxidants and free radical scavengers from

plants origin is an ideal strategy for new drugdevelopment Hence present study wasdesign to explore the antioxidant and freeradical scavenging activity of Salvadora persica in a prospective waySalvadora persica ( SP) also known asmiswak toothbrush tree and mustard treedistributed mainly in tropical and sub tropicalAsia The plant is a large evergreen profusely

branched shrub or a small tree up to 4-6 mtall SP has been used commonly astoothbrush to strengthen the gums10 Thefresh root barks and leaves have been used in

folk medicine for the treatment of a wide rangeof medical problems such as cough asthmascurvy piles rheumatism leprosygonorrohoea headache and hepaticdisorders

11 Various phytochemical studies on

Salvadora persica reported the presence ofalkaloids salvadorine trimethylaine andsalvadoricine

12 Essential oil from the roots of

SP contain benzyl isothiocyanate (70) withother components such as α-pinenecamphene benzaldehyde β-pinene myrcene

δ-3-carene limonene terpinolene benzylnitrile umbellulone β-elemene γ-muurolenemyristicin β-caryophyllene and longifolene13Proteinstannins steroids flavonoids andterpenoids in Tagetes erecta flower extractswhich therefore encourages antioxidantstudies

14 Various pharmacological activities

on SP including in-vivo antimicrobial activityespecially on lactobacilli and streptococcusmutans

15with moderate secretory activity

significantly high acetyl cholinesterase

inhibiting ability antifertility activity in malerats16 The aqueous extract of SP leavespossesses analgesic activity and reducescarrageenan induced inflammation in ratpaw17 Aqueous and alcoholic extract fromleaves of SP reduce elevated urinary oxalatelevels and deposition of stone-formingconstituents in the kidneys of calculogenicrats

18 In our previous study we showed root

hydroalcoholic extract of SP reducescarrageenan induced inflammation in rats and

scavenges ABTS [22-azino bis (3-ethylbenzo-thiazoline-6-sulphonic acid)] andsuperoxide radicals in a cell free system19 Inpresent study we aimed to explore theantioxidant potential of different extracts (rootand stem) of SP using solvent of differentpolarity (alcoholic hydro alcoholic aqueous)



983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983091


















983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983092







RESULTS






DrugConcugml












983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983093







Table 1b


Drugconcugml



AscAStd300 microM











983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983094







Drugconcugml











983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983096

DISCUSSION












27 TBARS one of the




CONCLUSIONS



2930




983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983097



REFERENCES





























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983091983088

























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983091


















983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983092







RESULTS






DrugConcugml












983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983093







Table 1b


Drugconcugml



AscAStd300 microM











983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983094







Drugconcugml











983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983096

DISCUSSION












27 TBARS one of the




CONCLUSIONS



2930




983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983097



REFERENCES





























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983091983088

























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983092







RESULTS






DrugConcugml












983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983093







Table 1b


Drugconcugml



AscAStd300 microM











983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983094







Drugconcugml











983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983096

DISCUSSION












27 TBARS one of the




CONCLUSIONS



2930




983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983097



REFERENCES





























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983091983088

























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983093







Table 1b


Drugconcugml



AscAStd300 microM











983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983094







Drugconcugml











983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983096

DISCUSSION












27 TBARS one of the




CONCLUSIONS



2930




983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983097



REFERENCES





























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983091983088

























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983094







Drugconcugml











983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983096

DISCUSSION












27 TBARS one of the




CONCLUSIONS



2930




983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983097



REFERENCES





























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983091983088



























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983096

DISCUSSION












27 TBARS one of the




CONCLUSIONS



2930




983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983097



REFERENCES





























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983091983088

























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983090983097



REFERENCES





























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983091983088

























983113983150983156 983114 983120983144983137983154983149 983106983145983151 983123983139983145 983090983088983089983090 983119983139983156983099 983091983080983092983081983098 983080983120983081 983093983090983089 983085 983093983091983088


983120 983085 983093983091983088























free radicals scavenging and inhibition of lipid peroxidation by salvadora persica

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