8
Research Article Abrus precatorius Leaves: Antioxidant Activity in Food and Biological Systems, pH, and Temperature Stability Vanitha Reddy Palvai, Sowmya Mahalingu, and Asna Urooj Department of Studies in Food Science and Nutrition, University of Mysore, Manasagangotri, Mysore 570006, India Correspondence should be addressed to Asna Urooj; [email protected] Received 31 August 2013; Revised 19 November 2013; Accepted 3 December 2013; Published 30 January 2014 Academic Editor: Maria Cristina Breschi Copyright © 2014 Vanitha Reddy Palvai et al. is is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Natural antioxidants present in foods and other biological materials have attracted considerable interest because of their presumed safety and potential nutritional and therapeutic effects. Antioxidant constituents of plant materials act as radical scavengers and convert the radicals to less reactive species. Abrus precatorius (AP) was analyzed for its proximate and phytochemical composition. e leaves were extracted with methanol (ME) and analyzed for antioxidant activity by radical scavenging method, reducing power, ferric reducing capacity, and in vitro inhibition of Fenton’s reagent-induced oxidation in oil emulsion and microsomes. In addition, the effect of temperature (100 C, 15, and 30min) and pH (4.5, 7, and 9) C on the antioxidant activity of ME was investigated. e leaves were rich in total polyphenols, flavonoids, -carotene, glutathione, -tocopherol, and ascorbic acid. e ME exhibited varying degree of antioxidant activity in a dose-dependent manner. e AP exhibited more inhibition of oxidation in microsomes (73%) than compared to oil emulsion (21%). Heat treatment resulted in an increase of radical scavenging activity of extract (28% to 43%). At pH 4.5 the extract exhibited more antioxidant activity and stability compared to pH 7 and 9. Data indicates that potential exists for the utilization of Abrus precatorius as a natural antioxidant. 1. Introduction Natural antioxidants present in food and other biological materials have attracted considerable interest because of their presumed safety and potential nutritional and therapeutic effects. Because extensive and expensive testing of food additives is required to meet safety standards, synthetic antioxidants have generally been eliminated from many food applications. e increasing interest in the search for natural replacements for synthetic antioxidants has led to the antioxidant evaluation of a number of plant sources [1] especially spices and herbs [2]. A large number of plants have been screened as viable sources of natural antioxidants including tocopherol, vitamin C, carotenoids, and phenolic compounds which are responsible for maintenance of health and protection from coronary heart diseases and cancer [3, 4]. In present time, medicinal plants as rich source of natural bioactive components are given priority to study their antioxidant activity and explore their utilization in treatment of diabetes mellitus, dyslipidemia, and cardio- vascular diseases. Our team had explored some medicinal plants, namely, Aegle marmelos [5], Morus indica [6], Moringa oleifera [7], Raphanus sativus [8], and Psidium guajava [9] for their antioxidant activity and stability. Before exploring a medicinal plant, there is a need to analyze the plant for its phytochemical composition, antioxidant activity, and its stability. In the present experiment medicinal plant, namely, Abrus precatorius (common name: Rosary pea), was selected to study the proximate, phytochemical composition, antioxi- dant potency, and stability in its methanol extract. 2. Materials and Methods 2.1. Plant Material. e selected plant material Abrus pre- catorius (AP) leaves were collected from Western Ghats of Karnataka. e samples were identified by Dr. Janardhan, Department of Studies in Botany, University of Mysore, Hindawi Publishing Corporation International Journal of Medicinal Chemistry Volume 2014, Article ID 748549, 7 pages http://dx.doi.org/10.1155/2014/748549

Research Article Abrus precatorius Leaves: Antioxidant ...downloads.hindawi.com/journals/ijmc/2014/748549.pdf · Research Article Abrus precatorius Leaves: Antioxidant Activity in

  • Upload
    vannhi

  • View
    220

  • Download
    3

Embed Size (px)

Citation preview

Page 1: Research Article Abrus precatorius Leaves: Antioxidant ...downloads.hindawi.com/journals/ijmc/2014/748549.pdf · Research Article Abrus precatorius Leaves: Antioxidant Activity in

Research ArticleAbrus precatorius Leaves Antioxidant Activity in Food andBiological Systems pH and Temperature Stability

Vanitha Reddy Palvai Sowmya Mahalingu and Asna Urooj

Department of Studies in Food Science and Nutrition University of Mysore Manasagangotri Mysore 570006 India

Correspondence should be addressed to Asna Urooj asnauroojfoodsciuni-mysoreacin

Received 31 August 2013 Revised 19 November 2013 Accepted 3 December 2013 Published 30 January 2014

Academic Editor Maria Cristina Breschi

Copyright copy 2014 Vanitha Reddy Palvai et al This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Natural antioxidants present in foods and other biological materials have attracted considerable interest because of their presumedsafety and potential nutritional and therapeutic effects Antioxidant constituents of plant materials act as radical scavengers andconvert the radicals to less reactive species Abrus precatorius (AP) was analyzed for its proximate and phytochemical compositionThe leaves were extracted withmethanol (ME) and analyzed for antioxidant activity by radical scavengingmethod reducing powerferric reducing capacity and in vitro inhibition of Fentonrsquos reagent-induced oxidation in oil emulsion andmicrosomes In additionthe effect of temperature (100∘C 15 and 30min) and pH (45 7 and 9) C on the antioxidant activity of ME was investigatedThe leaves were rich in total polyphenols flavonoids 120573-carotene glutathione 120572-tocopherol and ascorbic acid The ME exhibitedvarying degree of antioxidant activity in a dose-dependent manner The AP exhibited more inhibition of oxidation in microsomes(73) than compared to oil emulsion (21) Heat treatment resulted in an increase of radical scavenging activity of extract (28 to43) At pH 45 the extract exhibited more antioxidant activity and stability compared to pH 7 and 9 Data indicates that potentialexists for the utilization of Abrus precatorius as a natural antioxidant

1 Introduction

Natural antioxidants present in food and other biologicalmaterials have attracted considerable interest because of theirpresumed safety and potential nutritional and therapeuticeffects Because extensive and expensive testing of foodadditives is required to meet safety standards syntheticantioxidants have generally been eliminated from manyfood applications The increasing interest in the search fornatural replacements for synthetic antioxidants has led tothe antioxidant evaluation of a number of plant sources [1]especially spices and herbs [2] A large number of plantshave been screened as viable sources of natural antioxidantsincluding tocopherol vitamin C carotenoids and phenoliccompounds which are responsible for maintenance of healthandprotection fromcoronary heart diseases and cancer [3 4]

In present time medicinal plants as rich source ofnatural bioactive components are given priority to studytheir antioxidant activity and explore their utilization in

treatment of diabetes mellitus dyslipidemia and cardio-vascular diseases Our team had explored some medicinalplants namelyAeglemarmelos [5]Morus indica [6]Moringaoleifera [7] Raphanus sativus [8] and Psidium guajava [9]for their antioxidant activity and stability Before exploringa medicinal plant there is a need to analyze the plant forits phytochemical composition antioxidant activity and itsstability

In the present experiment medicinal plant namelyAbrusprecatorius (common name Rosary pea) was selected tostudy the proximate phytochemical composition antioxi-dant potency and stability in its methanol extract

2 Materials and Methods

21 Plant Material The selected plant material Abrus pre-catorius (AP) leaves were collected from Western Ghats ofKarnataka The samples were identified by Dr JanardhanDepartment of Studies in Botany University of Mysore

Hindawi Publishing CorporationInternational Journal of Medicinal ChemistryVolume 2014 Article ID 748549 7 pageshttpdxdoiorg1011552014748549

2 International Journal of Medicinal Chemistry

and voucher specimen was retained in the laboratory forfuture reference The leaves were washed dried in the ovenovernight at 50∘C powdered passed through 60mesh andstored at 4∘C till further use

22 Proximate Composition In the dry powder moisturecontent was determined by using moisture analyser (MetlerToledoMJ33 Lab systems Bangalore India) Fat protein ashtotal fiber (soluble and insoluble fiber) iron calcium andphosphorus were estimated as per the AOAC [10]

23 Determination of Phytochemical Components In the drysample different antioxidant components were estimatedusing standard methods Ascorbic acid was determinedaccording to the titrimetric method using 26-dichloro-phenol-indophenol dye [11] 120572-Tocopherol was extracted bydirect saponification of dried sample and estimated basedon formation of a red complex from reaction of 1205721205721015840-bipyridyl with ferrous ion due to reduction of ferric ion bytocopherol [12] 120573-Carotene was quantified by open columnchromatography followed by measuring the absorbance ofelute at 450 nm against standard 120573-carotene [11] Reducedglutathione was determined based on the development of ayellow compound due to reaction of DTNB (551015840minusdithiobis-2-nitrobenzoic acid) with compounds containing sulfhydrylgroups [13] Total phenolswere extracted fromaweighed por-tion (50ndash500mg) of dried sample with 5mL of 12M HCl in50 aqueous methanol for 2 h and with 70 acetone shakenfor 2 h and analyzed by Folin-Ciocalteau micromethod [14]Results were expressed as mg or g of gallic acid equivalentThe flavonoid content was determined by pharmacopoeiamethod using rutin as a reference compound The amountof flavonoids in plant extracts in rutin equivalents (RE) wascalculated by the following formula

119883 =119860 sdot 119898119900sdot 10

(119860119900sdot 119898119900) (1)

where119883-flavonoid content mgg119860mdashthe absorption of plantextract 119860

119900mdashabsorption of standard Rutin solution 119898mdash

weight of plant extract (g) 119898119900mdashweight of the Rutin in the

solution (g) [15]

24 Extraction of Antioxidants A 15 g sample was extractedwith 50mL methanol for 6 h in a mechanical shaker Theextracts were filtered and filtrates were evaporated at 40∘Cunder reduced pressure to dryness in a rotary evaporator(Superfit India) The residue of each extract was stored inairtight container at 4∘C until used

25 Radical Scavenging Activity (RSA) Theability of extractsto scavenge 22-diphenyl-1-picrylhydrazyl (DPPH) radicalswas determined according to the method of Blois 1958 [16]

26 Reducing Power Assay (RPA) The ability of extracts toreduce iron (III) to iron (II) was determined as per Yildirimet al 2003 [17]

27 Ferric Reducing Antioxidant Power (FRAP) Measure-ment of ferric reducing antioxidant power of the herbalextract was carried out based onBenzie and Strain procedure1999 [18]

28 Inhibition of Lipid Oxidation The antioxidant activity ofthe above extracts (300ndash500 120583g) was determined in an edibleoil emulsion and liver microsomes by modified method ofthiobarbituric acid-reactive-substances TBARSThis methoddetermines the resistance of lipid or lipid emulsions to oxida-tion in the presence of the antioxidantThemalondialdehyde(MDA) or TBARS assay has been used extensively since the1950s to estimate the peroxidation of lipids in membrane andbiological systems [19]

281 In Oil Emulsion Five grams of oil (sunflower) wasweighed and an emulsion was prepared in phosphate bufferof 001M 7pH and Tween 20 To different concentrations(300ndash500 120583g) of extracts 300 120583L of emulsion and 450 120583L ofFentonrsquos reagent were added and volume was made up to2mL and incubated at 50∘C for 6 hrs A control was runwithout extract After incubation 1mL of thiobarbituric acid(TBA) was added and heated in boiling water bath for 30minand cooled immediately The inhibition of lipid peroxidationin sun flower oil was determined by TBA in which thesecondary oxidation products (TBARS expressed as MDAequivalents) formed by oxidation of oil were determined bymeasuring the absorbance at 532 nm [20]

282 In Microsomes A healthy male adult rat was obtainedfrom the Central Animal House Department of ZoologyUniversity of Mysore after availing clearance from the Uni-versity Animal Ethics Committee of University of Mysore(no MGZ26202011-12 dated 31012012) The rat was sac-rificed and liver was immediately removed from the rat andplaced in cold triethanolamine HCl buffer (0ndash4∘C) at pH74 The liver was thoroughly chilled and homogenized Thehomogenate was centrifuged in ultra-centrifuge at 60000 gfor 60min The 60000 g microsomal pellet was then rinsedwith buffer and frozen in a freezer (minus20∘C) The suspendedmicrosomes to be used for the assay were diluted with bufferto give a protein concentration of 5ndash10mgmL [21]

To an aliquot of liver microsomes (1mg protein concen-tration) Fentonrsquos reagent and methanol extract (at varyingconcentration 300ndash500120583g)were added and incubated at 50∘Cfor 2 hrs After incubation 1mL of TCA (10) and 1mL ofTBA were added and heated in boiling water bath (15min)and cooled in ice bath immediately After cooling 2mL ofbutanol was added and the pink color developed was read at535 nm A control was run without plant extract [22]

Antioxidant activity

=Absorbance of control minus Absorbance of sample

Absorbance of controltimes 100

(2)

29 Heat and pH Stability The extracts were heated ina boiling water bath for 15 to 30min and the residual

International Journal of Medicinal Chemistry 3

Table 1 Proximate and phytochemical composition of the Abrus precatorius

Parameter g100 g dry basis Phytochemicals g100 g dry basisMoisture 10485 plusmn 0049 Polyphenols 106 plusmn 006

Protein 5875 plusmn 008 Flavonoidslowast 1325 plusmn 0085

Fat 440 plusmn 014 Glutathionelowastlowast 375 plusmn 17677

Soluble fiber 6 plusmn 009 120572-Tocopherollowast 366 plusmn 305

Insoluble fiber 2 plusmn 015 Tanninslowast 1160 plusmn 0076

Ash 8968 plusmn 097 120573-Carotenelowastlowastlowast 12600 plusmn 005

Calciumlowast 10395 plusmn 5 Alkaloidslowast 11000 plusmn 003

Phosphorouslowast 18320 plusmn 1031 Total saponins 0244 plusmn 001

Ironlowast 5458 plusmn 058 Steroidal saponins 0186 plusmn 0028

Vitamin C (fresh basis) 0443 plusmn 0011

Vitamin C (dry basis) 0643 plusmn 004

lowastmg100 g lowastlowastmmol100 g and lowastlowastlowast120583g100 g dry basisValues are mean of triplicates (119899 = 3)

antioxidant activity was determined by RSA using DPPHas described previously For pH stability the extracts wereincubated for 72 h at pH 45 70 and 90 and the residualantioxidant activity was determined during the incubationperiod at different time intervals (0min 24 48 and 72 h)using RSA method [23]

3 Statistical Analysis

Mean values of triplicates (119899 = 3) were subjected to one-wayANOVA and Tukeyrsquos multiple comparison tests using SPSSsoftware (version 11) (119875 lt 005)

4 Results and Discussion

41 Proximate and Phytochemical Composition Abrus preca-torius (AP) was found to be good source of nutrients and phy-tochemicals such as 120573-carotene glutathione 120572-tocopherolascorbic acid and phytochemicals namely total polyphenolsand flavonoids (Table 1)

42 Radical Scavenging Activity The radical scavengingactivity of AP was measured at 100ndash500 120583g (Figure 1) Fromthe figure it is evident that the scavenging activity of sampleswas dose dependent that is 100ndash500120583g

43 Reducing Power Assay The reducing power of a com-pound is related to its electron transfer ability andmay there-fore serve as a significant indicator of its potential antioxidantactivityThe electron donating capacity wasmeasured at 100ndash500120583g (Figure 2) The reducing power of the samples wasdose dependent which had ranged between In the presentstudy the ability of extract to reduce iron (III) to iron (II)was compared with that of a standard (ascorbic acid) and thereducing capacity was inferior to that of ascorbic acid

44 Ferric Reducing Antioxidant Power (FRAP) The changein absorbance at 593 nm owing to the formation of a bluecolored Fe (II)-tripyridyltriazine compound from colorless

aa

a

b

b

0

5

10

15

20

25

30

35

RSA

()

100 200 300 400 500

(120583g)

Figure 1 Radical scavenging activity of Abrus precatoriusmethanolextract Values are expressed as mean of triplicates (119899 = 3) (119875 le005)

oxidized Fe (III) is formed by the action of electron donatingantioxidants This represents an electron exchange reactionSample showed high reducing power at lower concentration(100 120583g) and increased with the concentration (Figure 3)

45 Inhibition of Lipid Peroxidation Lipid oxidation is a pro-cess in which PUFA undergoes oxidative damage resultingin the formation of lipid-derived radicals such as alkoxyand peroxyl radicals further causing membrane damage andcellular injury In biological systems antioxidants are capableof stabilizing or deactivating free radicals before they attackcells [24] The sample exhibited high activity in case ofmicrosomes than the oil emulsion (Figures 4 and 5) Therate of inhibition of the oxidation by AP in microsomes was300 120583g 4343 plusmn 9 400 120583g 6058 plusmn 479 500120583g 7329 plusmn 684

Plant-derived antioxidants have been shown to func-tion as singlet and triplet oxygen quenchers free radicalscavengers peroxide decomposers enzyme inhibitors andsynergists The most current research on antioxidant actionfocuses on phenolic compounds such as flavonoids Fruitsand vegetables contain different antioxidant compounds

4 International Journal of Medicinal Chemistry

a a

a

b

b

0

005

01

015

02

025

03

100 200 300 400 500

RPA-

OD700

nm

(120583g)

Figure 2 Reducing power assay of Abrus precatorius methanolextract Values are expressed as mean of triplicates (119899 = 3) (119875 le005)

a

b

ccd

d

0

1

2

3

4

5

6

7

FRA

P

100 200 300 400 500

(120583g)

Figure 3 Ferric reducing antioxidant power (FRAP) of Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119899 = 3) (119875 le 005)

such as vitamin C vitamin E and carotenoids whose activi-ties have been well established [25]

Glutathione tocopherol 120573 carotenoids flavonoids andso forth come under second line defense antioxidants 120573-carotene is an excellent scavenger of singlet oxygen VitaminC directly interacts with radicals like O

2

minus and OH∙ Glu-tathione is a good scavenger of many free radicals like O

2

minus

and OH∙ and various lipid hydroperoxides and may help todetoxify many inhaled oxidizing pollutants like ozone NO

2

and cigarette smoke in respiratory tract Vitamin E scavengesperoxyl radicals intermediates in lipid peroxidation and isresponsible for protecting PUFA present in cell membraneand LDL against lipid peroxidation [26] On the whole thesephytochemicals and antioxidants bestow high medicinalactivities of AP

Results presented here clearly indicate that the methanolextract of AP with some percentage of polyphenols exhibitedantioxidant activity in different in vitromodels In the processof oxidation free radicals are considered to play a cardinalrole in numerous chronic pathologies The RSA of AP iscomparatively less than the antioxidant activity of few othermedicinal plants such asAegle marmelos [7]Moringa oleifera

0102030405060708090

Inhi

bitio

n of

oxi

datio

n (

)

300 400 500

a

b

c

(120583g)

Figure 4 Inhibition of oxidation in edible oil emulsion by Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119875 le 005)

Inhi

bitio

n of

oxi

datio

n (

)

0102030405060708090

300 400 500

a

b

c

(120583g)

Figure 5 Inhibition of oxidation in microsomes by Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119875 le 005)

[5]Morus indica [27] and in case of Moringa oleifera corre-lation between the polyphenol content and RSAwas reported[5] However the polyphenol content of APmethanol extractis less than the polyphenol content of above medicinal plants

Binding of metal ions such as iron in vivo is anantioxidant action of itself preventing metal ion catalyzedgeneration of reactive species [24] In reducing assay theyellow color of the test solution changes to various shadesof green and blue depending on the reducing power ofeach compoundThe presence of reducers (ie antioxidants)causes the reduction of the Fe3+ferricyanide complex to theferrous form Therefore measuring the formation of PerlrsquosPrussian blue at 700 nm can monitor the Fe2+ concentrationThe ability of APmethanol extract to reduce iron (III) to iron(II) is inferior than the standard (ascorbic acidmdash0973)Aeglemarmelos [7] andMorus indica [6] and superior to Raphanussativus [8] The reducing power of a sample is due to theirhydrogen-donating ability Accordingly AP might containhigher amounts of reductone which could react with freeradicals to stabilize and block radical chain reactions

Processed foods containing fats and oils oxidize slowlyduring storage various oxidation products cause rancidity

International Journal of Medicinal Chemistry 5

and deterioration of the sensory and nutritional property ofthe food products Auto-oxidation of fats and oil in processedfoods may be prevented by use of oxidation inhibitors orantioxidants To protect the cells and organ systems of thebody against reactive oxygen species humans have evolveda highly sophisticated and complex antioxidant protectionsystem that functions interactively and synergistically toneutralize free radicals Thus antioxidants are capable ofstabilizing or deactivating free radicals before they attack cell[28] Vitamin E scavenges peroxyl radicals intermediates inlipid peroxidation and is responsible for protecting PUFApresent in cell membrane and LDL against lipid peroxidationThe polarity of phytochemicals plays a key role in exhibitingantioxidant role at lipid phase especially at unsaturation sitewhich influences the chain breaking reaction [24] Presentlyin methanol extract of AP antioxidants with high partitioncoefficientmay be distributed hydrophobic compartments forthe protection of lipids AP was more effective in inhibitingthe oxidation in microsomes compared to oil emulsion Sim-ilarly results were found in case ofMorus indica [6] Here theuse of two lipid systems was helpful in studying the inhibitionpotency of different extracts in food and biological level

46 Stability of AP Extracts to High Temperature (100∘C 15and 30min) It is well known that many factors such asantioxidant concentration temperature and pH of themediaprocessing treatments influence the antioxidant activity Rad-ical scavenging activity of the heat treated sample extract isshown in Figure 6 After 15min of heat treatment a slightdecline in the activity was observed at all the concentrationsexcept at 300 120583g which showed an increase in the activityA significant increase was seen at the end of 30min atconcentrations greater than 100 120583g This may be due to theactivation of antioxidant components after 30min of heattreatment In another study extracts of drumstick leavesmint leaves and carrot were subjected to heat treatment at100∘C (15min) which resulted in a significant decrease inantioxidant activity (AOA) in drumstick leaves (DL) extractwhile no difference was observed in AOA of carrot extractThe significant increase in AOA due to thermal processingoccurred in mint leaves extract Thermal processing caninduce the formation of novel compounds with antioxidantproperties or improve theAOAof naturally occurring antiox-idants [29] Another study reports that WE (water) ME(methanol) and EE (ethanol) extracts of Aegle marmelosL treated at 100∘C for 15min showed varied stability WEshowed maximum stability as measured by RSA comparedto ME and EE after heat treatment [7] Similar results werefound in case of different extracts of Raphanus sativus [8] andMoringa oleifera [5]

47 Stability of AP Extract to Different pH The antioxidantstability of sample extract to different pH (45 70 and 90)was analysed by radical scavenging activity (Figure 7(a)mdashpH45 and Figure 7(b)mdashpH 70) At pH 45 and 70 the extractat different concentrations and time intervals has shownvarying antioxidant activity The influence of pH 45 on thestability of AP methanol extract is shown in Figure 7(a) and

05

101520253035404550

0 100 200 300 400 500 600

RSA

()

01530

Figure 6 Stability of Abrus precatorius extract to high temperature(100∘C 15 and 30min) Values are expressed as mean of triplicates

at the end of 72 h a decrease in radical scavenging activitywas observed However the decline in the activity was notlinear The maximum antioxidant activity observed was 50at 500120583g

At pH 70 after 72 h a significant increase in RSA wasobserved up to 300 120583g and subsequently the RSA decreased athigher concentrations (400 and 500120583g)However the declinein RSA was not significant compared to the zero min At pH9 the extract did not exhibit RSA and the absorbance wasmore than that of control this may be due to the structuraldenaturation of natural antioxidant components and similarresults were found in fenugreek seeds and ginger rhizomeextracts [30] In Aegle marmelos [7] and Rapahnus sativus[8] the antioxidant activity decreased by increasing the pHof media The antioxidant activity of different extracts fromcocoa byproducts was found to be higher at alkaline pH [31]In an earlier study it was observed that extracts from mintand carrot leaves showed higher antioxidant activity at pH 9than pH 4 [32] These differences might be due to differentsamples used various compounds being extracted in eachcase and the method used to evaluate antioxidant potency

5 Conclusion

Results of the present study reveal that the Abrus precatoriusis a potential source of antioxidants which are responsiblefor the antioxidant activity The stability to heat and pHof the different extracts of AP leaves extracts with strongantioxidant activity indicates its scope for utilization in foodand biological systems In addition to being consumed ashealthy antioxidants the compounds present in AP that areresponsible for antioxidant activity could be isolated and thenused as food additives to delay the oxidative deteriorationof foods and as nutraceutical in food formulations againstdegenerative diseases

Conflict of Interests

The authors declare that there is no conflict of interests

6 International Journal of Medicinal Chemistry

0

10

20

30

40

50

60

0 100 200 300 400 500 600

RSA

()

024

4872

(a)

05

1015202530354045

0 100 200 300 400 500 600

RSA

()

4872

024

(b)

Figure 7 (a) Stability of Abrus precatorius extract to pH 45 values are expressed as mean of triplicates (b) Stability of Abrus precatoriusextract at pH 7 values are expressed as mean of triplicates

Acknowledgment

The authors acknowledge UGC-IOE (Institute of Excellence)for providing financial assistance for this project

References

[1] A Lugasi J Hovari K V Sagi and L Bıro ldquoThe role ofantioxidant phytonutrients in the prevention of diseasesrdquo ActaBiologica Szegediensis vol 47 no 1ndash4 pp 119ndash125 2003

[2] N Nakatani ldquoAntioxidants from spices and herbsrdquo in NaturalAntioxidants ChemistryHealth Effects andApplications pp 64ndash73 AOCS Press Champaign Ill USA 1997

[3] J J M Castenmiller J P H Linssen I M Heinonen AL Hopia K Schwarz and P C H Hollmann ldquoAntioxidantproperties of differently processed spinach productsrdquo NahrungFood Journal vol 46 pp 290ndash293 2002

[4] C Kaur and H C Kapoor ldquoAntioxidants in fruits andvegetablesmdashthe millenniumrsquos healthrdquo International Journal ofFood Science and Technology vol 36 no 7 pp 703ndash725 2001

[5] V P Reddy and A Urooj ldquoAntioxidant properties and stabilityof aegle marmelos leaves extractsrdquo Journal of Food Science andTechnology vol 50 no 1 pp 135ndash140 2013

[6] P V Reddy and A Urooj ldquoProximate phytochemical profileand antioxidant activity (in vitro and ex vivo) of Morus indicavarietiesrdquo International Journal of Pharmaceutical Sciences andResearch vol 4 no 4 pp 1626ndash1634 2013

[7] A Urooj and P V Reddy ldquoMoringa oleifera antioxidant prop-erties and stability of various solvent extractsrdquo InternationalJournal of Pharmaceutical Sciences and Biotechnology vol 1 no1 pp 1ndash6 2010

[8] P V Reddy S Desai and F A A Urooj ldquoAntioxidant propertiesand stability of Raphanus sativus extractsrdquo Journal of PharmacyResearch vol 3 no 3 pp 658ndash661 2011

[9] P V Reddy N Sahana and A Urooj ldquoAntioxidant activityof Aegle marmelos and Psidium guajava leavesrdquo InternationalJournal of Medicinal and Aromatic Plants vol 2 no 1 pp 155ndash160 2012

[10] AOAC Official Methods of Analysis AOAC InternationalWashington DC USA 11th edition 1970

[11] S Ranganna Handbook of Analysis and Quality Control forFruits and Vegetables Products McGrow-Hill NewDelhi India2nd edition 1999

[12] M Freed Methods of Vitamins Assay Association of VitaminChemists Interscience New York NY USA 3rd edition 1966

[13] E Beutler and B M Kelly ldquoThe effect of sodium nitrite on redcell GSHrdquo Experientia vol 19 no 2 pp 96ndash97 1963

[14] K Slinkard and V L Singleton ldquoTotal phenol analysis automa-tion and comparison with manual methodsrdquo American Journalof Enology and Viticare vol 28 pp 49ndash55 1967

[15] GMiliauskas P R Venskutonis andT A VanBeek ldquoScreeningof radical scavenging activity of some medicinal and aromaticplant extractsrdquo Food Chemistry vol 85 no 2 pp 231ndash237 2004

[16] M S Blois ldquoAntioxidant determinations by the use of a stablefree radicalrdquo Nature vol 181 no 4617 pp 1199ndash1200 1958

[17] A Yildirim A Mavi and A A Kara ldquoAntioxidant and antimi-crobial activities of Polygonum cognatum Meissn extractsrdquoJournal of the Science of Food and Agriculture vol 83 no 1 pp64ndash69 2003

[18] I F F Benzie and J J Strain ldquoFerric reducingantioxidant powerassay direct measure of total antioxidant activity of biologicalfluids and modified version for simultaneous measurementof total antioxidant power and ascorbic acid concentrationrdquoMethods in Enzymology vol 299 pp 15ndash27 1998

[19] H Tamura and A Yamagami ldquoAntioxidative activity of monoa-cylated anthocyanins isolated from Muscat Bailey A graperdquoJournal of Agricultural and Food Chemistry vol 42 no 8 pp1612ndash1615 1994

[20] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[21] D J Shapiro and V W Rodwell ldquoRegulation of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase and choles-terol synthesisrdquo Journal of Biological Chemistry vol 246 no 10pp 3210ndash3216 1971

[22] J A Buege and SD Aust ldquo[30]Microsomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978

[23] S Arabshahi-D D Vishalakshi Devi and A Urooj ldquoEvaluationof antioxidant activity of some plant extracts and their heat pHand storage stabilityrdquo Food Chemistry vol 100 no 3 pp 1100ndash1105 2007

International Journal of Medicinal Chemistry 7

[24] L J Lizcano M Viloria-Bernal F Vicente et al ldquoLipid oxi-dation inhibitory effects and phenolic composition of aqueousextracts from medicinal plants of colombian amazoniardquo Inter-national Journal of Molecular Sciences vol 13 no 5 pp 5454ndash5467 2012

[25] S Mandal S Yadav S Yadav and R K Nema ldquoAntioxidants areviewrdquo Journal of Chemical and Pharmaceutical Research vol1 no 1 pp 102ndash104 2009

[26] V K Gupta and S K Gupta ldquoPlants as natural antioxidantsrdquoNatural Product Radiance vol 5 no 6 pp 326ndash334 2006

[27] S Arabshahi-Delouee and A Urooj ldquoAntioxidant properties ofvarious solvent extracts of mulberry (Morus indica L) leavesrdquoFood Chemistry vol 102 no 4 pp 1233ndash1240 2007

[28] S M Zachariah N A Aleykutty V Viswanad S Jacob andV Prabhakar ldquoIn-vitro antioxidant potential of methanolicextracts of Mirabilis jalapa Linnrdquo Free Radicals and Antioxi-dants vol 1 no 4 pp 82ndash86 2011

[29] V Reddy A Urooj and A Kumar ldquoEvaluation of antioxidantactivity of some plant extracts and their application in biscuitsrdquoFood Chemistry vol 90 no 1-2 pp 317ndash321 2005

[30] E H Mansour and A H Khalil ldquoEvaluation of antioxidantactivity of some plant extracts and their application to groundbeef pattiesrdquo Food Chemistry vol 69 no 2 pp 135ndash141 2000

[31] A H Azizah NM Nik Ruslawati and T Swee Tee ldquoExtractionand characterization of antioxidant from cocoa by-productsrdquoFood Chemistry vol 64 no 2 pp 199ndash202 1999

[32] M G Miguel ldquoAntioxidant activity of medicinal and aromaticplants A reviewrdquo Flavour and Fragrance Journal vol 25 no 5pp 291ndash312 2010

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

Page 2: Research Article Abrus precatorius Leaves: Antioxidant ...downloads.hindawi.com/journals/ijmc/2014/748549.pdf · Research Article Abrus precatorius Leaves: Antioxidant Activity in

2 International Journal of Medicinal Chemistry

and voucher specimen was retained in the laboratory forfuture reference The leaves were washed dried in the ovenovernight at 50∘C powdered passed through 60mesh andstored at 4∘C till further use

22 Proximate Composition In the dry powder moisturecontent was determined by using moisture analyser (MetlerToledoMJ33 Lab systems Bangalore India) Fat protein ashtotal fiber (soluble and insoluble fiber) iron calcium andphosphorus were estimated as per the AOAC [10]

23 Determination of Phytochemical Components In the drysample different antioxidant components were estimatedusing standard methods Ascorbic acid was determinedaccording to the titrimetric method using 26-dichloro-phenol-indophenol dye [11] 120572-Tocopherol was extracted bydirect saponification of dried sample and estimated basedon formation of a red complex from reaction of 1205721205721015840-bipyridyl with ferrous ion due to reduction of ferric ion bytocopherol [12] 120573-Carotene was quantified by open columnchromatography followed by measuring the absorbance ofelute at 450 nm against standard 120573-carotene [11] Reducedglutathione was determined based on the development of ayellow compound due to reaction of DTNB (551015840minusdithiobis-2-nitrobenzoic acid) with compounds containing sulfhydrylgroups [13] Total phenolswere extracted fromaweighed por-tion (50ndash500mg) of dried sample with 5mL of 12M HCl in50 aqueous methanol for 2 h and with 70 acetone shakenfor 2 h and analyzed by Folin-Ciocalteau micromethod [14]Results were expressed as mg or g of gallic acid equivalentThe flavonoid content was determined by pharmacopoeiamethod using rutin as a reference compound The amountof flavonoids in plant extracts in rutin equivalents (RE) wascalculated by the following formula

119883 =119860 sdot 119898119900sdot 10

(119860119900sdot 119898119900) (1)

where119883-flavonoid content mgg119860mdashthe absorption of plantextract 119860

119900mdashabsorption of standard Rutin solution 119898mdash

weight of plant extract (g) 119898119900mdashweight of the Rutin in the

solution (g) [15]

24 Extraction of Antioxidants A 15 g sample was extractedwith 50mL methanol for 6 h in a mechanical shaker Theextracts were filtered and filtrates were evaporated at 40∘Cunder reduced pressure to dryness in a rotary evaporator(Superfit India) The residue of each extract was stored inairtight container at 4∘C until used

25 Radical Scavenging Activity (RSA) Theability of extractsto scavenge 22-diphenyl-1-picrylhydrazyl (DPPH) radicalswas determined according to the method of Blois 1958 [16]

26 Reducing Power Assay (RPA) The ability of extracts toreduce iron (III) to iron (II) was determined as per Yildirimet al 2003 [17]

27 Ferric Reducing Antioxidant Power (FRAP) Measure-ment of ferric reducing antioxidant power of the herbalextract was carried out based onBenzie and Strain procedure1999 [18]

28 Inhibition of Lipid Oxidation The antioxidant activity ofthe above extracts (300ndash500 120583g) was determined in an edibleoil emulsion and liver microsomes by modified method ofthiobarbituric acid-reactive-substances TBARSThis methoddetermines the resistance of lipid or lipid emulsions to oxida-tion in the presence of the antioxidantThemalondialdehyde(MDA) or TBARS assay has been used extensively since the1950s to estimate the peroxidation of lipids in membrane andbiological systems [19]

281 In Oil Emulsion Five grams of oil (sunflower) wasweighed and an emulsion was prepared in phosphate bufferof 001M 7pH and Tween 20 To different concentrations(300ndash500 120583g) of extracts 300 120583L of emulsion and 450 120583L ofFentonrsquos reagent were added and volume was made up to2mL and incubated at 50∘C for 6 hrs A control was runwithout extract After incubation 1mL of thiobarbituric acid(TBA) was added and heated in boiling water bath for 30minand cooled immediately The inhibition of lipid peroxidationin sun flower oil was determined by TBA in which thesecondary oxidation products (TBARS expressed as MDAequivalents) formed by oxidation of oil were determined bymeasuring the absorbance at 532 nm [20]

282 In Microsomes A healthy male adult rat was obtainedfrom the Central Animal House Department of ZoologyUniversity of Mysore after availing clearance from the Uni-versity Animal Ethics Committee of University of Mysore(no MGZ26202011-12 dated 31012012) The rat was sac-rificed and liver was immediately removed from the rat andplaced in cold triethanolamine HCl buffer (0ndash4∘C) at pH74 The liver was thoroughly chilled and homogenized Thehomogenate was centrifuged in ultra-centrifuge at 60000 gfor 60min The 60000 g microsomal pellet was then rinsedwith buffer and frozen in a freezer (minus20∘C) The suspendedmicrosomes to be used for the assay were diluted with bufferto give a protein concentration of 5ndash10mgmL [21]

To an aliquot of liver microsomes (1mg protein concen-tration) Fentonrsquos reagent and methanol extract (at varyingconcentration 300ndash500120583g)were added and incubated at 50∘Cfor 2 hrs After incubation 1mL of TCA (10) and 1mL ofTBA were added and heated in boiling water bath (15min)and cooled in ice bath immediately After cooling 2mL ofbutanol was added and the pink color developed was read at535 nm A control was run without plant extract [22]

Antioxidant activity

=Absorbance of control minus Absorbance of sample

Absorbance of controltimes 100

(2)

29 Heat and pH Stability The extracts were heated ina boiling water bath for 15 to 30min and the residual

International Journal of Medicinal Chemistry 3

Table 1 Proximate and phytochemical composition of the Abrus precatorius

Parameter g100 g dry basis Phytochemicals g100 g dry basisMoisture 10485 plusmn 0049 Polyphenols 106 plusmn 006

Protein 5875 plusmn 008 Flavonoidslowast 1325 plusmn 0085

Fat 440 plusmn 014 Glutathionelowastlowast 375 plusmn 17677

Soluble fiber 6 plusmn 009 120572-Tocopherollowast 366 plusmn 305

Insoluble fiber 2 plusmn 015 Tanninslowast 1160 plusmn 0076

Ash 8968 plusmn 097 120573-Carotenelowastlowastlowast 12600 plusmn 005

Calciumlowast 10395 plusmn 5 Alkaloidslowast 11000 plusmn 003

Phosphorouslowast 18320 plusmn 1031 Total saponins 0244 plusmn 001

Ironlowast 5458 plusmn 058 Steroidal saponins 0186 plusmn 0028

Vitamin C (fresh basis) 0443 plusmn 0011

Vitamin C (dry basis) 0643 plusmn 004

lowastmg100 g lowastlowastmmol100 g and lowastlowastlowast120583g100 g dry basisValues are mean of triplicates (119899 = 3)

antioxidant activity was determined by RSA using DPPHas described previously For pH stability the extracts wereincubated for 72 h at pH 45 70 and 90 and the residualantioxidant activity was determined during the incubationperiod at different time intervals (0min 24 48 and 72 h)using RSA method [23]

3 Statistical Analysis

Mean values of triplicates (119899 = 3) were subjected to one-wayANOVA and Tukeyrsquos multiple comparison tests using SPSSsoftware (version 11) (119875 lt 005)

4 Results and Discussion

41 Proximate and Phytochemical Composition Abrus preca-torius (AP) was found to be good source of nutrients and phy-tochemicals such as 120573-carotene glutathione 120572-tocopherolascorbic acid and phytochemicals namely total polyphenolsand flavonoids (Table 1)

42 Radical Scavenging Activity The radical scavengingactivity of AP was measured at 100ndash500 120583g (Figure 1) Fromthe figure it is evident that the scavenging activity of sampleswas dose dependent that is 100ndash500120583g

43 Reducing Power Assay The reducing power of a com-pound is related to its electron transfer ability andmay there-fore serve as a significant indicator of its potential antioxidantactivityThe electron donating capacity wasmeasured at 100ndash500120583g (Figure 2) The reducing power of the samples wasdose dependent which had ranged between In the presentstudy the ability of extract to reduce iron (III) to iron (II)was compared with that of a standard (ascorbic acid) and thereducing capacity was inferior to that of ascorbic acid

44 Ferric Reducing Antioxidant Power (FRAP) The changein absorbance at 593 nm owing to the formation of a bluecolored Fe (II)-tripyridyltriazine compound from colorless

aa

a

b

b

0

5

10

15

20

25

30

35

RSA

()

100 200 300 400 500

(120583g)

Figure 1 Radical scavenging activity of Abrus precatoriusmethanolextract Values are expressed as mean of triplicates (119899 = 3) (119875 le005)

oxidized Fe (III) is formed by the action of electron donatingantioxidants This represents an electron exchange reactionSample showed high reducing power at lower concentration(100 120583g) and increased with the concentration (Figure 3)

45 Inhibition of Lipid Peroxidation Lipid oxidation is a pro-cess in which PUFA undergoes oxidative damage resultingin the formation of lipid-derived radicals such as alkoxyand peroxyl radicals further causing membrane damage andcellular injury In biological systems antioxidants are capableof stabilizing or deactivating free radicals before they attackcells [24] The sample exhibited high activity in case ofmicrosomes than the oil emulsion (Figures 4 and 5) Therate of inhibition of the oxidation by AP in microsomes was300 120583g 4343 plusmn 9 400 120583g 6058 plusmn 479 500120583g 7329 plusmn 684

Plant-derived antioxidants have been shown to func-tion as singlet and triplet oxygen quenchers free radicalscavengers peroxide decomposers enzyme inhibitors andsynergists The most current research on antioxidant actionfocuses on phenolic compounds such as flavonoids Fruitsand vegetables contain different antioxidant compounds

4 International Journal of Medicinal Chemistry

a a

a

b

b

0

005

01

015

02

025

03

100 200 300 400 500

RPA-

OD700

nm

(120583g)

Figure 2 Reducing power assay of Abrus precatorius methanolextract Values are expressed as mean of triplicates (119899 = 3) (119875 le005)

a

b

ccd

d

0

1

2

3

4

5

6

7

FRA

P

100 200 300 400 500

(120583g)

Figure 3 Ferric reducing antioxidant power (FRAP) of Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119899 = 3) (119875 le 005)

such as vitamin C vitamin E and carotenoids whose activi-ties have been well established [25]

Glutathione tocopherol 120573 carotenoids flavonoids andso forth come under second line defense antioxidants 120573-carotene is an excellent scavenger of singlet oxygen VitaminC directly interacts with radicals like O

2

minus and OH∙ Glu-tathione is a good scavenger of many free radicals like O

2

minus

and OH∙ and various lipid hydroperoxides and may help todetoxify many inhaled oxidizing pollutants like ozone NO

2

and cigarette smoke in respiratory tract Vitamin E scavengesperoxyl radicals intermediates in lipid peroxidation and isresponsible for protecting PUFA present in cell membraneand LDL against lipid peroxidation [26] On the whole thesephytochemicals and antioxidants bestow high medicinalactivities of AP

Results presented here clearly indicate that the methanolextract of AP with some percentage of polyphenols exhibitedantioxidant activity in different in vitromodels In the processof oxidation free radicals are considered to play a cardinalrole in numerous chronic pathologies The RSA of AP iscomparatively less than the antioxidant activity of few othermedicinal plants such asAegle marmelos [7]Moringa oleifera

0102030405060708090

Inhi

bitio

n of

oxi

datio

n (

)

300 400 500

a

b

c

(120583g)

Figure 4 Inhibition of oxidation in edible oil emulsion by Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119875 le 005)

Inhi

bitio

n of

oxi

datio

n (

)

0102030405060708090

300 400 500

a

b

c

(120583g)

Figure 5 Inhibition of oxidation in microsomes by Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119875 le 005)

[5]Morus indica [27] and in case of Moringa oleifera corre-lation between the polyphenol content and RSAwas reported[5] However the polyphenol content of APmethanol extractis less than the polyphenol content of above medicinal plants

Binding of metal ions such as iron in vivo is anantioxidant action of itself preventing metal ion catalyzedgeneration of reactive species [24] In reducing assay theyellow color of the test solution changes to various shadesof green and blue depending on the reducing power ofeach compoundThe presence of reducers (ie antioxidants)causes the reduction of the Fe3+ferricyanide complex to theferrous form Therefore measuring the formation of PerlrsquosPrussian blue at 700 nm can monitor the Fe2+ concentrationThe ability of APmethanol extract to reduce iron (III) to iron(II) is inferior than the standard (ascorbic acidmdash0973)Aeglemarmelos [7] andMorus indica [6] and superior to Raphanussativus [8] The reducing power of a sample is due to theirhydrogen-donating ability Accordingly AP might containhigher amounts of reductone which could react with freeradicals to stabilize and block radical chain reactions

Processed foods containing fats and oils oxidize slowlyduring storage various oxidation products cause rancidity

International Journal of Medicinal Chemistry 5

and deterioration of the sensory and nutritional property ofthe food products Auto-oxidation of fats and oil in processedfoods may be prevented by use of oxidation inhibitors orantioxidants To protect the cells and organ systems of thebody against reactive oxygen species humans have evolveda highly sophisticated and complex antioxidant protectionsystem that functions interactively and synergistically toneutralize free radicals Thus antioxidants are capable ofstabilizing or deactivating free radicals before they attack cell[28] Vitamin E scavenges peroxyl radicals intermediates inlipid peroxidation and is responsible for protecting PUFApresent in cell membrane and LDL against lipid peroxidationThe polarity of phytochemicals plays a key role in exhibitingantioxidant role at lipid phase especially at unsaturation sitewhich influences the chain breaking reaction [24] Presentlyin methanol extract of AP antioxidants with high partitioncoefficientmay be distributed hydrophobic compartments forthe protection of lipids AP was more effective in inhibitingthe oxidation in microsomes compared to oil emulsion Sim-ilarly results were found in case ofMorus indica [6] Here theuse of two lipid systems was helpful in studying the inhibitionpotency of different extracts in food and biological level

46 Stability of AP Extracts to High Temperature (100∘C 15and 30min) It is well known that many factors such asantioxidant concentration temperature and pH of themediaprocessing treatments influence the antioxidant activity Rad-ical scavenging activity of the heat treated sample extract isshown in Figure 6 After 15min of heat treatment a slightdecline in the activity was observed at all the concentrationsexcept at 300 120583g which showed an increase in the activityA significant increase was seen at the end of 30min atconcentrations greater than 100 120583g This may be due to theactivation of antioxidant components after 30min of heattreatment In another study extracts of drumstick leavesmint leaves and carrot were subjected to heat treatment at100∘C (15min) which resulted in a significant decrease inantioxidant activity (AOA) in drumstick leaves (DL) extractwhile no difference was observed in AOA of carrot extractThe significant increase in AOA due to thermal processingoccurred in mint leaves extract Thermal processing caninduce the formation of novel compounds with antioxidantproperties or improve theAOAof naturally occurring antiox-idants [29] Another study reports that WE (water) ME(methanol) and EE (ethanol) extracts of Aegle marmelosL treated at 100∘C for 15min showed varied stability WEshowed maximum stability as measured by RSA comparedto ME and EE after heat treatment [7] Similar results werefound in case of different extracts of Raphanus sativus [8] andMoringa oleifera [5]

47 Stability of AP Extract to Different pH The antioxidantstability of sample extract to different pH (45 70 and 90)was analysed by radical scavenging activity (Figure 7(a)mdashpH45 and Figure 7(b)mdashpH 70) At pH 45 and 70 the extractat different concentrations and time intervals has shownvarying antioxidant activity The influence of pH 45 on thestability of AP methanol extract is shown in Figure 7(a) and

05

101520253035404550

0 100 200 300 400 500 600

RSA

()

01530

Figure 6 Stability of Abrus precatorius extract to high temperature(100∘C 15 and 30min) Values are expressed as mean of triplicates

at the end of 72 h a decrease in radical scavenging activitywas observed However the decline in the activity was notlinear The maximum antioxidant activity observed was 50at 500120583g

At pH 70 after 72 h a significant increase in RSA wasobserved up to 300 120583g and subsequently the RSA decreased athigher concentrations (400 and 500120583g)However the declinein RSA was not significant compared to the zero min At pH9 the extract did not exhibit RSA and the absorbance wasmore than that of control this may be due to the structuraldenaturation of natural antioxidant components and similarresults were found in fenugreek seeds and ginger rhizomeextracts [30] In Aegle marmelos [7] and Rapahnus sativus[8] the antioxidant activity decreased by increasing the pHof media The antioxidant activity of different extracts fromcocoa byproducts was found to be higher at alkaline pH [31]In an earlier study it was observed that extracts from mintand carrot leaves showed higher antioxidant activity at pH 9than pH 4 [32] These differences might be due to differentsamples used various compounds being extracted in eachcase and the method used to evaluate antioxidant potency

5 Conclusion

Results of the present study reveal that the Abrus precatoriusis a potential source of antioxidants which are responsiblefor the antioxidant activity The stability to heat and pHof the different extracts of AP leaves extracts with strongantioxidant activity indicates its scope for utilization in foodand biological systems In addition to being consumed ashealthy antioxidants the compounds present in AP that areresponsible for antioxidant activity could be isolated and thenused as food additives to delay the oxidative deteriorationof foods and as nutraceutical in food formulations againstdegenerative diseases

Conflict of Interests

The authors declare that there is no conflict of interests

6 International Journal of Medicinal Chemistry

0

10

20

30

40

50

60

0 100 200 300 400 500 600

RSA

()

024

4872

(a)

05

1015202530354045

0 100 200 300 400 500 600

RSA

()

4872

024

(b)

Figure 7 (a) Stability of Abrus precatorius extract to pH 45 values are expressed as mean of triplicates (b) Stability of Abrus precatoriusextract at pH 7 values are expressed as mean of triplicates

Acknowledgment

The authors acknowledge UGC-IOE (Institute of Excellence)for providing financial assistance for this project

References

[1] A Lugasi J Hovari K V Sagi and L Bıro ldquoThe role ofantioxidant phytonutrients in the prevention of diseasesrdquo ActaBiologica Szegediensis vol 47 no 1ndash4 pp 119ndash125 2003

[2] N Nakatani ldquoAntioxidants from spices and herbsrdquo in NaturalAntioxidants ChemistryHealth Effects andApplications pp 64ndash73 AOCS Press Champaign Ill USA 1997

[3] J J M Castenmiller J P H Linssen I M Heinonen AL Hopia K Schwarz and P C H Hollmann ldquoAntioxidantproperties of differently processed spinach productsrdquo NahrungFood Journal vol 46 pp 290ndash293 2002

[4] C Kaur and H C Kapoor ldquoAntioxidants in fruits andvegetablesmdashthe millenniumrsquos healthrdquo International Journal ofFood Science and Technology vol 36 no 7 pp 703ndash725 2001

[5] V P Reddy and A Urooj ldquoAntioxidant properties and stabilityof aegle marmelos leaves extractsrdquo Journal of Food Science andTechnology vol 50 no 1 pp 135ndash140 2013

[6] P V Reddy and A Urooj ldquoProximate phytochemical profileand antioxidant activity (in vitro and ex vivo) of Morus indicavarietiesrdquo International Journal of Pharmaceutical Sciences andResearch vol 4 no 4 pp 1626ndash1634 2013

[7] A Urooj and P V Reddy ldquoMoringa oleifera antioxidant prop-erties and stability of various solvent extractsrdquo InternationalJournal of Pharmaceutical Sciences and Biotechnology vol 1 no1 pp 1ndash6 2010

[8] P V Reddy S Desai and F A A Urooj ldquoAntioxidant propertiesand stability of Raphanus sativus extractsrdquo Journal of PharmacyResearch vol 3 no 3 pp 658ndash661 2011

[9] P V Reddy N Sahana and A Urooj ldquoAntioxidant activityof Aegle marmelos and Psidium guajava leavesrdquo InternationalJournal of Medicinal and Aromatic Plants vol 2 no 1 pp 155ndash160 2012

[10] AOAC Official Methods of Analysis AOAC InternationalWashington DC USA 11th edition 1970

[11] S Ranganna Handbook of Analysis and Quality Control forFruits and Vegetables Products McGrow-Hill NewDelhi India2nd edition 1999

[12] M Freed Methods of Vitamins Assay Association of VitaminChemists Interscience New York NY USA 3rd edition 1966

[13] E Beutler and B M Kelly ldquoThe effect of sodium nitrite on redcell GSHrdquo Experientia vol 19 no 2 pp 96ndash97 1963

[14] K Slinkard and V L Singleton ldquoTotal phenol analysis automa-tion and comparison with manual methodsrdquo American Journalof Enology and Viticare vol 28 pp 49ndash55 1967

[15] GMiliauskas P R Venskutonis andT A VanBeek ldquoScreeningof radical scavenging activity of some medicinal and aromaticplant extractsrdquo Food Chemistry vol 85 no 2 pp 231ndash237 2004

[16] M S Blois ldquoAntioxidant determinations by the use of a stablefree radicalrdquo Nature vol 181 no 4617 pp 1199ndash1200 1958

[17] A Yildirim A Mavi and A A Kara ldquoAntioxidant and antimi-crobial activities of Polygonum cognatum Meissn extractsrdquoJournal of the Science of Food and Agriculture vol 83 no 1 pp64ndash69 2003

[18] I F F Benzie and J J Strain ldquoFerric reducingantioxidant powerassay direct measure of total antioxidant activity of biologicalfluids and modified version for simultaneous measurementof total antioxidant power and ascorbic acid concentrationrdquoMethods in Enzymology vol 299 pp 15ndash27 1998

[19] H Tamura and A Yamagami ldquoAntioxidative activity of monoa-cylated anthocyanins isolated from Muscat Bailey A graperdquoJournal of Agricultural and Food Chemistry vol 42 no 8 pp1612ndash1615 1994

[20] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[21] D J Shapiro and V W Rodwell ldquoRegulation of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase and choles-terol synthesisrdquo Journal of Biological Chemistry vol 246 no 10pp 3210ndash3216 1971

[22] J A Buege and SD Aust ldquo[30]Microsomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978

[23] S Arabshahi-D D Vishalakshi Devi and A Urooj ldquoEvaluationof antioxidant activity of some plant extracts and their heat pHand storage stabilityrdquo Food Chemistry vol 100 no 3 pp 1100ndash1105 2007

International Journal of Medicinal Chemistry 7

[24] L J Lizcano M Viloria-Bernal F Vicente et al ldquoLipid oxi-dation inhibitory effects and phenolic composition of aqueousextracts from medicinal plants of colombian amazoniardquo Inter-national Journal of Molecular Sciences vol 13 no 5 pp 5454ndash5467 2012

[25] S Mandal S Yadav S Yadav and R K Nema ldquoAntioxidants areviewrdquo Journal of Chemical and Pharmaceutical Research vol1 no 1 pp 102ndash104 2009

[26] V K Gupta and S K Gupta ldquoPlants as natural antioxidantsrdquoNatural Product Radiance vol 5 no 6 pp 326ndash334 2006

[27] S Arabshahi-Delouee and A Urooj ldquoAntioxidant properties ofvarious solvent extracts of mulberry (Morus indica L) leavesrdquoFood Chemistry vol 102 no 4 pp 1233ndash1240 2007

[28] S M Zachariah N A Aleykutty V Viswanad S Jacob andV Prabhakar ldquoIn-vitro antioxidant potential of methanolicextracts of Mirabilis jalapa Linnrdquo Free Radicals and Antioxi-dants vol 1 no 4 pp 82ndash86 2011

[29] V Reddy A Urooj and A Kumar ldquoEvaluation of antioxidantactivity of some plant extracts and their application in biscuitsrdquoFood Chemistry vol 90 no 1-2 pp 317ndash321 2005

[30] E H Mansour and A H Khalil ldquoEvaluation of antioxidantactivity of some plant extracts and their application to groundbeef pattiesrdquo Food Chemistry vol 69 no 2 pp 135ndash141 2000

[31] A H Azizah NM Nik Ruslawati and T Swee Tee ldquoExtractionand characterization of antioxidant from cocoa by-productsrdquoFood Chemistry vol 64 no 2 pp 199ndash202 1999

[32] M G Miguel ldquoAntioxidant activity of medicinal and aromaticplants A reviewrdquo Flavour and Fragrance Journal vol 25 no 5pp 291ndash312 2010

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

Page 3: Research Article Abrus precatorius Leaves: Antioxidant ...downloads.hindawi.com/journals/ijmc/2014/748549.pdf · Research Article Abrus precatorius Leaves: Antioxidant Activity in

International Journal of Medicinal Chemistry 3

Table 1 Proximate and phytochemical composition of the Abrus precatorius

Parameter g100 g dry basis Phytochemicals g100 g dry basisMoisture 10485 plusmn 0049 Polyphenols 106 plusmn 006

Protein 5875 plusmn 008 Flavonoidslowast 1325 plusmn 0085

Fat 440 plusmn 014 Glutathionelowastlowast 375 plusmn 17677

Soluble fiber 6 plusmn 009 120572-Tocopherollowast 366 plusmn 305

Insoluble fiber 2 plusmn 015 Tanninslowast 1160 plusmn 0076

Ash 8968 plusmn 097 120573-Carotenelowastlowastlowast 12600 plusmn 005

Calciumlowast 10395 plusmn 5 Alkaloidslowast 11000 plusmn 003

Phosphorouslowast 18320 plusmn 1031 Total saponins 0244 plusmn 001

Ironlowast 5458 plusmn 058 Steroidal saponins 0186 plusmn 0028

Vitamin C (fresh basis) 0443 plusmn 0011

Vitamin C (dry basis) 0643 plusmn 004

lowastmg100 g lowastlowastmmol100 g and lowastlowastlowast120583g100 g dry basisValues are mean of triplicates (119899 = 3)

antioxidant activity was determined by RSA using DPPHas described previously For pH stability the extracts wereincubated for 72 h at pH 45 70 and 90 and the residualantioxidant activity was determined during the incubationperiod at different time intervals (0min 24 48 and 72 h)using RSA method [23]

3 Statistical Analysis

Mean values of triplicates (119899 = 3) were subjected to one-wayANOVA and Tukeyrsquos multiple comparison tests using SPSSsoftware (version 11) (119875 lt 005)

4 Results and Discussion

41 Proximate and Phytochemical Composition Abrus preca-torius (AP) was found to be good source of nutrients and phy-tochemicals such as 120573-carotene glutathione 120572-tocopherolascorbic acid and phytochemicals namely total polyphenolsand flavonoids (Table 1)

42 Radical Scavenging Activity The radical scavengingactivity of AP was measured at 100ndash500 120583g (Figure 1) Fromthe figure it is evident that the scavenging activity of sampleswas dose dependent that is 100ndash500120583g

43 Reducing Power Assay The reducing power of a com-pound is related to its electron transfer ability andmay there-fore serve as a significant indicator of its potential antioxidantactivityThe electron donating capacity wasmeasured at 100ndash500120583g (Figure 2) The reducing power of the samples wasdose dependent which had ranged between In the presentstudy the ability of extract to reduce iron (III) to iron (II)was compared with that of a standard (ascorbic acid) and thereducing capacity was inferior to that of ascorbic acid

44 Ferric Reducing Antioxidant Power (FRAP) The changein absorbance at 593 nm owing to the formation of a bluecolored Fe (II)-tripyridyltriazine compound from colorless

aa

a

b

b

0

5

10

15

20

25

30

35

RSA

()

100 200 300 400 500

(120583g)

Figure 1 Radical scavenging activity of Abrus precatoriusmethanolextract Values are expressed as mean of triplicates (119899 = 3) (119875 le005)

oxidized Fe (III) is formed by the action of electron donatingantioxidants This represents an electron exchange reactionSample showed high reducing power at lower concentration(100 120583g) and increased with the concentration (Figure 3)

45 Inhibition of Lipid Peroxidation Lipid oxidation is a pro-cess in which PUFA undergoes oxidative damage resultingin the formation of lipid-derived radicals such as alkoxyand peroxyl radicals further causing membrane damage andcellular injury In biological systems antioxidants are capableof stabilizing or deactivating free radicals before they attackcells [24] The sample exhibited high activity in case ofmicrosomes than the oil emulsion (Figures 4 and 5) Therate of inhibition of the oxidation by AP in microsomes was300 120583g 4343 plusmn 9 400 120583g 6058 plusmn 479 500120583g 7329 plusmn 684

Plant-derived antioxidants have been shown to func-tion as singlet and triplet oxygen quenchers free radicalscavengers peroxide decomposers enzyme inhibitors andsynergists The most current research on antioxidant actionfocuses on phenolic compounds such as flavonoids Fruitsand vegetables contain different antioxidant compounds

4 International Journal of Medicinal Chemistry

a a

a

b

b

0

005

01

015

02

025

03

100 200 300 400 500

RPA-

OD700

nm

(120583g)

Figure 2 Reducing power assay of Abrus precatorius methanolextract Values are expressed as mean of triplicates (119899 = 3) (119875 le005)

a

b

ccd

d

0

1

2

3

4

5

6

7

FRA

P

100 200 300 400 500

(120583g)

Figure 3 Ferric reducing antioxidant power (FRAP) of Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119899 = 3) (119875 le 005)

such as vitamin C vitamin E and carotenoids whose activi-ties have been well established [25]

Glutathione tocopherol 120573 carotenoids flavonoids andso forth come under second line defense antioxidants 120573-carotene is an excellent scavenger of singlet oxygen VitaminC directly interacts with radicals like O

2

minus and OH∙ Glu-tathione is a good scavenger of many free radicals like O

2

minus

and OH∙ and various lipid hydroperoxides and may help todetoxify many inhaled oxidizing pollutants like ozone NO

2

and cigarette smoke in respiratory tract Vitamin E scavengesperoxyl radicals intermediates in lipid peroxidation and isresponsible for protecting PUFA present in cell membraneand LDL against lipid peroxidation [26] On the whole thesephytochemicals and antioxidants bestow high medicinalactivities of AP

Results presented here clearly indicate that the methanolextract of AP with some percentage of polyphenols exhibitedantioxidant activity in different in vitromodels In the processof oxidation free radicals are considered to play a cardinalrole in numerous chronic pathologies The RSA of AP iscomparatively less than the antioxidant activity of few othermedicinal plants such asAegle marmelos [7]Moringa oleifera

0102030405060708090

Inhi

bitio

n of

oxi

datio

n (

)

300 400 500

a

b

c

(120583g)

Figure 4 Inhibition of oxidation in edible oil emulsion by Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119875 le 005)

Inhi

bitio

n of

oxi

datio

n (

)

0102030405060708090

300 400 500

a

b

c

(120583g)

Figure 5 Inhibition of oxidation in microsomes by Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119875 le 005)

[5]Morus indica [27] and in case of Moringa oleifera corre-lation between the polyphenol content and RSAwas reported[5] However the polyphenol content of APmethanol extractis less than the polyphenol content of above medicinal plants

Binding of metal ions such as iron in vivo is anantioxidant action of itself preventing metal ion catalyzedgeneration of reactive species [24] In reducing assay theyellow color of the test solution changes to various shadesof green and blue depending on the reducing power ofeach compoundThe presence of reducers (ie antioxidants)causes the reduction of the Fe3+ferricyanide complex to theferrous form Therefore measuring the formation of PerlrsquosPrussian blue at 700 nm can monitor the Fe2+ concentrationThe ability of APmethanol extract to reduce iron (III) to iron(II) is inferior than the standard (ascorbic acidmdash0973)Aeglemarmelos [7] andMorus indica [6] and superior to Raphanussativus [8] The reducing power of a sample is due to theirhydrogen-donating ability Accordingly AP might containhigher amounts of reductone which could react with freeradicals to stabilize and block radical chain reactions

Processed foods containing fats and oils oxidize slowlyduring storage various oxidation products cause rancidity

International Journal of Medicinal Chemistry 5

and deterioration of the sensory and nutritional property ofthe food products Auto-oxidation of fats and oil in processedfoods may be prevented by use of oxidation inhibitors orantioxidants To protect the cells and organ systems of thebody against reactive oxygen species humans have evolveda highly sophisticated and complex antioxidant protectionsystem that functions interactively and synergistically toneutralize free radicals Thus antioxidants are capable ofstabilizing or deactivating free radicals before they attack cell[28] Vitamin E scavenges peroxyl radicals intermediates inlipid peroxidation and is responsible for protecting PUFApresent in cell membrane and LDL against lipid peroxidationThe polarity of phytochemicals plays a key role in exhibitingantioxidant role at lipid phase especially at unsaturation sitewhich influences the chain breaking reaction [24] Presentlyin methanol extract of AP antioxidants with high partitioncoefficientmay be distributed hydrophobic compartments forthe protection of lipids AP was more effective in inhibitingthe oxidation in microsomes compared to oil emulsion Sim-ilarly results were found in case ofMorus indica [6] Here theuse of two lipid systems was helpful in studying the inhibitionpotency of different extracts in food and biological level

46 Stability of AP Extracts to High Temperature (100∘C 15and 30min) It is well known that many factors such asantioxidant concentration temperature and pH of themediaprocessing treatments influence the antioxidant activity Rad-ical scavenging activity of the heat treated sample extract isshown in Figure 6 After 15min of heat treatment a slightdecline in the activity was observed at all the concentrationsexcept at 300 120583g which showed an increase in the activityA significant increase was seen at the end of 30min atconcentrations greater than 100 120583g This may be due to theactivation of antioxidant components after 30min of heattreatment In another study extracts of drumstick leavesmint leaves and carrot were subjected to heat treatment at100∘C (15min) which resulted in a significant decrease inantioxidant activity (AOA) in drumstick leaves (DL) extractwhile no difference was observed in AOA of carrot extractThe significant increase in AOA due to thermal processingoccurred in mint leaves extract Thermal processing caninduce the formation of novel compounds with antioxidantproperties or improve theAOAof naturally occurring antiox-idants [29] Another study reports that WE (water) ME(methanol) and EE (ethanol) extracts of Aegle marmelosL treated at 100∘C for 15min showed varied stability WEshowed maximum stability as measured by RSA comparedto ME and EE after heat treatment [7] Similar results werefound in case of different extracts of Raphanus sativus [8] andMoringa oleifera [5]

47 Stability of AP Extract to Different pH The antioxidantstability of sample extract to different pH (45 70 and 90)was analysed by radical scavenging activity (Figure 7(a)mdashpH45 and Figure 7(b)mdashpH 70) At pH 45 and 70 the extractat different concentrations and time intervals has shownvarying antioxidant activity The influence of pH 45 on thestability of AP methanol extract is shown in Figure 7(a) and

05

101520253035404550

0 100 200 300 400 500 600

RSA

()

01530

Figure 6 Stability of Abrus precatorius extract to high temperature(100∘C 15 and 30min) Values are expressed as mean of triplicates

at the end of 72 h a decrease in radical scavenging activitywas observed However the decline in the activity was notlinear The maximum antioxidant activity observed was 50at 500120583g

At pH 70 after 72 h a significant increase in RSA wasobserved up to 300 120583g and subsequently the RSA decreased athigher concentrations (400 and 500120583g)However the declinein RSA was not significant compared to the zero min At pH9 the extract did not exhibit RSA and the absorbance wasmore than that of control this may be due to the structuraldenaturation of natural antioxidant components and similarresults were found in fenugreek seeds and ginger rhizomeextracts [30] In Aegle marmelos [7] and Rapahnus sativus[8] the antioxidant activity decreased by increasing the pHof media The antioxidant activity of different extracts fromcocoa byproducts was found to be higher at alkaline pH [31]In an earlier study it was observed that extracts from mintand carrot leaves showed higher antioxidant activity at pH 9than pH 4 [32] These differences might be due to differentsamples used various compounds being extracted in eachcase and the method used to evaluate antioxidant potency

5 Conclusion

Results of the present study reveal that the Abrus precatoriusis a potential source of antioxidants which are responsiblefor the antioxidant activity The stability to heat and pHof the different extracts of AP leaves extracts with strongantioxidant activity indicates its scope for utilization in foodand biological systems In addition to being consumed ashealthy antioxidants the compounds present in AP that areresponsible for antioxidant activity could be isolated and thenused as food additives to delay the oxidative deteriorationof foods and as nutraceutical in food formulations againstdegenerative diseases

Conflict of Interests

The authors declare that there is no conflict of interests

6 International Journal of Medicinal Chemistry

0

10

20

30

40

50

60

0 100 200 300 400 500 600

RSA

()

024

4872

(a)

05

1015202530354045

0 100 200 300 400 500 600

RSA

()

4872

024

(b)

Figure 7 (a) Stability of Abrus precatorius extract to pH 45 values are expressed as mean of triplicates (b) Stability of Abrus precatoriusextract at pH 7 values are expressed as mean of triplicates

Acknowledgment

The authors acknowledge UGC-IOE (Institute of Excellence)for providing financial assistance for this project

References

[1] A Lugasi J Hovari K V Sagi and L Bıro ldquoThe role ofantioxidant phytonutrients in the prevention of diseasesrdquo ActaBiologica Szegediensis vol 47 no 1ndash4 pp 119ndash125 2003

[2] N Nakatani ldquoAntioxidants from spices and herbsrdquo in NaturalAntioxidants ChemistryHealth Effects andApplications pp 64ndash73 AOCS Press Champaign Ill USA 1997

[3] J J M Castenmiller J P H Linssen I M Heinonen AL Hopia K Schwarz and P C H Hollmann ldquoAntioxidantproperties of differently processed spinach productsrdquo NahrungFood Journal vol 46 pp 290ndash293 2002

[4] C Kaur and H C Kapoor ldquoAntioxidants in fruits andvegetablesmdashthe millenniumrsquos healthrdquo International Journal ofFood Science and Technology vol 36 no 7 pp 703ndash725 2001

[5] V P Reddy and A Urooj ldquoAntioxidant properties and stabilityof aegle marmelos leaves extractsrdquo Journal of Food Science andTechnology vol 50 no 1 pp 135ndash140 2013

[6] P V Reddy and A Urooj ldquoProximate phytochemical profileand antioxidant activity (in vitro and ex vivo) of Morus indicavarietiesrdquo International Journal of Pharmaceutical Sciences andResearch vol 4 no 4 pp 1626ndash1634 2013

[7] A Urooj and P V Reddy ldquoMoringa oleifera antioxidant prop-erties and stability of various solvent extractsrdquo InternationalJournal of Pharmaceutical Sciences and Biotechnology vol 1 no1 pp 1ndash6 2010

[8] P V Reddy S Desai and F A A Urooj ldquoAntioxidant propertiesand stability of Raphanus sativus extractsrdquo Journal of PharmacyResearch vol 3 no 3 pp 658ndash661 2011

[9] P V Reddy N Sahana and A Urooj ldquoAntioxidant activityof Aegle marmelos and Psidium guajava leavesrdquo InternationalJournal of Medicinal and Aromatic Plants vol 2 no 1 pp 155ndash160 2012

[10] AOAC Official Methods of Analysis AOAC InternationalWashington DC USA 11th edition 1970

[11] S Ranganna Handbook of Analysis and Quality Control forFruits and Vegetables Products McGrow-Hill NewDelhi India2nd edition 1999

[12] M Freed Methods of Vitamins Assay Association of VitaminChemists Interscience New York NY USA 3rd edition 1966

[13] E Beutler and B M Kelly ldquoThe effect of sodium nitrite on redcell GSHrdquo Experientia vol 19 no 2 pp 96ndash97 1963

[14] K Slinkard and V L Singleton ldquoTotal phenol analysis automa-tion and comparison with manual methodsrdquo American Journalof Enology and Viticare vol 28 pp 49ndash55 1967

[15] GMiliauskas P R Venskutonis andT A VanBeek ldquoScreeningof radical scavenging activity of some medicinal and aromaticplant extractsrdquo Food Chemistry vol 85 no 2 pp 231ndash237 2004

[16] M S Blois ldquoAntioxidant determinations by the use of a stablefree radicalrdquo Nature vol 181 no 4617 pp 1199ndash1200 1958

[17] A Yildirim A Mavi and A A Kara ldquoAntioxidant and antimi-crobial activities of Polygonum cognatum Meissn extractsrdquoJournal of the Science of Food and Agriculture vol 83 no 1 pp64ndash69 2003

[18] I F F Benzie and J J Strain ldquoFerric reducingantioxidant powerassay direct measure of total antioxidant activity of biologicalfluids and modified version for simultaneous measurementof total antioxidant power and ascorbic acid concentrationrdquoMethods in Enzymology vol 299 pp 15ndash27 1998

[19] H Tamura and A Yamagami ldquoAntioxidative activity of monoa-cylated anthocyanins isolated from Muscat Bailey A graperdquoJournal of Agricultural and Food Chemistry vol 42 no 8 pp1612ndash1615 1994

[20] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[21] D J Shapiro and V W Rodwell ldquoRegulation of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase and choles-terol synthesisrdquo Journal of Biological Chemistry vol 246 no 10pp 3210ndash3216 1971

[22] J A Buege and SD Aust ldquo[30]Microsomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978

[23] S Arabshahi-D D Vishalakshi Devi and A Urooj ldquoEvaluationof antioxidant activity of some plant extracts and their heat pHand storage stabilityrdquo Food Chemistry vol 100 no 3 pp 1100ndash1105 2007

International Journal of Medicinal Chemistry 7

[24] L J Lizcano M Viloria-Bernal F Vicente et al ldquoLipid oxi-dation inhibitory effects and phenolic composition of aqueousextracts from medicinal plants of colombian amazoniardquo Inter-national Journal of Molecular Sciences vol 13 no 5 pp 5454ndash5467 2012

[25] S Mandal S Yadav S Yadav and R K Nema ldquoAntioxidants areviewrdquo Journal of Chemical and Pharmaceutical Research vol1 no 1 pp 102ndash104 2009

[26] V K Gupta and S K Gupta ldquoPlants as natural antioxidantsrdquoNatural Product Radiance vol 5 no 6 pp 326ndash334 2006

[27] S Arabshahi-Delouee and A Urooj ldquoAntioxidant properties ofvarious solvent extracts of mulberry (Morus indica L) leavesrdquoFood Chemistry vol 102 no 4 pp 1233ndash1240 2007

[28] S M Zachariah N A Aleykutty V Viswanad S Jacob andV Prabhakar ldquoIn-vitro antioxidant potential of methanolicextracts of Mirabilis jalapa Linnrdquo Free Radicals and Antioxi-dants vol 1 no 4 pp 82ndash86 2011

[29] V Reddy A Urooj and A Kumar ldquoEvaluation of antioxidantactivity of some plant extracts and their application in biscuitsrdquoFood Chemistry vol 90 no 1-2 pp 317ndash321 2005

[30] E H Mansour and A H Khalil ldquoEvaluation of antioxidantactivity of some plant extracts and their application to groundbeef pattiesrdquo Food Chemistry vol 69 no 2 pp 135ndash141 2000

[31] A H Azizah NM Nik Ruslawati and T Swee Tee ldquoExtractionand characterization of antioxidant from cocoa by-productsrdquoFood Chemistry vol 64 no 2 pp 199ndash202 1999

[32] M G Miguel ldquoAntioxidant activity of medicinal and aromaticplants A reviewrdquo Flavour and Fragrance Journal vol 25 no 5pp 291ndash312 2010

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

Page 4: Research Article Abrus precatorius Leaves: Antioxidant ...downloads.hindawi.com/journals/ijmc/2014/748549.pdf · Research Article Abrus precatorius Leaves: Antioxidant Activity in

4 International Journal of Medicinal Chemistry

a a

a

b

b

0

005

01

015

02

025

03

100 200 300 400 500

RPA-

OD700

nm

(120583g)

Figure 2 Reducing power assay of Abrus precatorius methanolextract Values are expressed as mean of triplicates (119899 = 3) (119875 le005)

a

b

ccd

d

0

1

2

3

4

5

6

7

FRA

P

100 200 300 400 500

(120583g)

Figure 3 Ferric reducing antioxidant power (FRAP) of Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119899 = 3) (119875 le 005)

such as vitamin C vitamin E and carotenoids whose activi-ties have been well established [25]

Glutathione tocopherol 120573 carotenoids flavonoids andso forth come under second line defense antioxidants 120573-carotene is an excellent scavenger of singlet oxygen VitaminC directly interacts with radicals like O

2

minus and OH∙ Glu-tathione is a good scavenger of many free radicals like O

2

minus

and OH∙ and various lipid hydroperoxides and may help todetoxify many inhaled oxidizing pollutants like ozone NO

2

and cigarette smoke in respiratory tract Vitamin E scavengesperoxyl radicals intermediates in lipid peroxidation and isresponsible for protecting PUFA present in cell membraneand LDL against lipid peroxidation [26] On the whole thesephytochemicals and antioxidants bestow high medicinalactivities of AP

Results presented here clearly indicate that the methanolextract of AP with some percentage of polyphenols exhibitedantioxidant activity in different in vitromodels In the processof oxidation free radicals are considered to play a cardinalrole in numerous chronic pathologies The RSA of AP iscomparatively less than the antioxidant activity of few othermedicinal plants such asAegle marmelos [7]Moringa oleifera

0102030405060708090

Inhi

bitio

n of

oxi

datio

n (

)

300 400 500

a

b

c

(120583g)

Figure 4 Inhibition of oxidation in edible oil emulsion by Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119875 le 005)

Inhi

bitio

n of

oxi

datio

n (

)

0102030405060708090

300 400 500

a

b

c

(120583g)

Figure 5 Inhibition of oxidation in microsomes by Abrusprecatorius methanol extract Values are expressed as mean oftriplicates (119875 le 005)

[5]Morus indica [27] and in case of Moringa oleifera corre-lation between the polyphenol content and RSAwas reported[5] However the polyphenol content of APmethanol extractis less than the polyphenol content of above medicinal plants

Binding of metal ions such as iron in vivo is anantioxidant action of itself preventing metal ion catalyzedgeneration of reactive species [24] In reducing assay theyellow color of the test solution changes to various shadesof green and blue depending on the reducing power ofeach compoundThe presence of reducers (ie antioxidants)causes the reduction of the Fe3+ferricyanide complex to theferrous form Therefore measuring the formation of PerlrsquosPrussian blue at 700 nm can monitor the Fe2+ concentrationThe ability of APmethanol extract to reduce iron (III) to iron(II) is inferior than the standard (ascorbic acidmdash0973)Aeglemarmelos [7] andMorus indica [6] and superior to Raphanussativus [8] The reducing power of a sample is due to theirhydrogen-donating ability Accordingly AP might containhigher amounts of reductone which could react with freeradicals to stabilize and block radical chain reactions

Processed foods containing fats and oils oxidize slowlyduring storage various oxidation products cause rancidity

International Journal of Medicinal Chemistry 5

and deterioration of the sensory and nutritional property ofthe food products Auto-oxidation of fats and oil in processedfoods may be prevented by use of oxidation inhibitors orantioxidants To protect the cells and organ systems of thebody against reactive oxygen species humans have evolveda highly sophisticated and complex antioxidant protectionsystem that functions interactively and synergistically toneutralize free radicals Thus antioxidants are capable ofstabilizing or deactivating free radicals before they attack cell[28] Vitamin E scavenges peroxyl radicals intermediates inlipid peroxidation and is responsible for protecting PUFApresent in cell membrane and LDL against lipid peroxidationThe polarity of phytochemicals plays a key role in exhibitingantioxidant role at lipid phase especially at unsaturation sitewhich influences the chain breaking reaction [24] Presentlyin methanol extract of AP antioxidants with high partitioncoefficientmay be distributed hydrophobic compartments forthe protection of lipids AP was more effective in inhibitingthe oxidation in microsomes compared to oil emulsion Sim-ilarly results were found in case ofMorus indica [6] Here theuse of two lipid systems was helpful in studying the inhibitionpotency of different extracts in food and biological level

46 Stability of AP Extracts to High Temperature (100∘C 15and 30min) It is well known that many factors such asantioxidant concentration temperature and pH of themediaprocessing treatments influence the antioxidant activity Rad-ical scavenging activity of the heat treated sample extract isshown in Figure 6 After 15min of heat treatment a slightdecline in the activity was observed at all the concentrationsexcept at 300 120583g which showed an increase in the activityA significant increase was seen at the end of 30min atconcentrations greater than 100 120583g This may be due to theactivation of antioxidant components after 30min of heattreatment In another study extracts of drumstick leavesmint leaves and carrot were subjected to heat treatment at100∘C (15min) which resulted in a significant decrease inantioxidant activity (AOA) in drumstick leaves (DL) extractwhile no difference was observed in AOA of carrot extractThe significant increase in AOA due to thermal processingoccurred in mint leaves extract Thermal processing caninduce the formation of novel compounds with antioxidantproperties or improve theAOAof naturally occurring antiox-idants [29] Another study reports that WE (water) ME(methanol) and EE (ethanol) extracts of Aegle marmelosL treated at 100∘C for 15min showed varied stability WEshowed maximum stability as measured by RSA comparedto ME and EE after heat treatment [7] Similar results werefound in case of different extracts of Raphanus sativus [8] andMoringa oleifera [5]

47 Stability of AP Extract to Different pH The antioxidantstability of sample extract to different pH (45 70 and 90)was analysed by radical scavenging activity (Figure 7(a)mdashpH45 and Figure 7(b)mdashpH 70) At pH 45 and 70 the extractat different concentrations and time intervals has shownvarying antioxidant activity The influence of pH 45 on thestability of AP methanol extract is shown in Figure 7(a) and

05

101520253035404550

0 100 200 300 400 500 600

RSA

()

01530

Figure 6 Stability of Abrus precatorius extract to high temperature(100∘C 15 and 30min) Values are expressed as mean of triplicates

at the end of 72 h a decrease in radical scavenging activitywas observed However the decline in the activity was notlinear The maximum antioxidant activity observed was 50at 500120583g

At pH 70 after 72 h a significant increase in RSA wasobserved up to 300 120583g and subsequently the RSA decreased athigher concentrations (400 and 500120583g)However the declinein RSA was not significant compared to the zero min At pH9 the extract did not exhibit RSA and the absorbance wasmore than that of control this may be due to the structuraldenaturation of natural antioxidant components and similarresults were found in fenugreek seeds and ginger rhizomeextracts [30] In Aegle marmelos [7] and Rapahnus sativus[8] the antioxidant activity decreased by increasing the pHof media The antioxidant activity of different extracts fromcocoa byproducts was found to be higher at alkaline pH [31]In an earlier study it was observed that extracts from mintand carrot leaves showed higher antioxidant activity at pH 9than pH 4 [32] These differences might be due to differentsamples used various compounds being extracted in eachcase and the method used to evaluate antioxidant potency

5 Conclusion

Results of the present study reveal that the Abrus precatoriusis a potential source of antioxidants which are responsiblefor the antioxidant activity The stability to heat and pHof the different extracts of AP leaves extracts with strongantioxidant activity indicates its scope for utilization in foodand biological systems In addition to being consumed ashealthy antioxidants the compounds present in AP that areresponsible for antioxidant activity could be isolated and thenused as food additives to delay the oxidative deteriorationof foods and as nutraceutical in food formulations againstdegenerative diseases

Conflict of Interests

The authors declare that there is no conflict of interests

6 International Journal of Medicinal Chemistry

0

10

20

30

40

50

60

0 100 200 300 400 500 600

RSA

()

024

4872

(a)

05

1015202530354045

0 100 200 300 400 500 600

RSA

()

4872

024

(b)

Figure 7 (a) Stability of Abrus precatorius extract to pH 45 values are expressed as mean of triplicates (b) Stability of Abrus precatoriusextract at pH 7 values are expressed as mean of triplicates

Acknowledgment

The authors acknowledge UGC-IOE (Institute of Excellence)for providing financial assistance for this project

References

[1] A Lugasi J Hovari K V Sagi and L Bıro ldquoThe role ofantioxidant phytonutrients in the prevention of diseasesrdquo ActaBiologica Szegediensis vol 47 no 1ndash4 pp 119ndash125 2003

[2] N Nakatani ldquoAntioxidants from spices and herbsrdquo in NaturalAntioxidants ChemistryHealth Effects andApplications pp 64ndash73 AOCS Press Champaign Ill USA 1997

[3] J J M Castenmiller J P H Linssen I M Heinonen AL Hopia K Schwarz and P C H Hollmann ldquoAntioxidantproperties of differently processed spinach productsrdquo NahrungFood Journal vol 46 pp 290ndash293 2002

[4] C Kaur and H C Kapoor ldquoAntioxidants in fruits andvegetablesmdashthe millenniumrsquos healthrdquo International Journal ofFood Science and Technology vol 36 no 7 pp 703ndash725 2001

[5] V P Reddy and A Urooj ldquoAntioxidant properties and stabilityof aegle marmelos leaves extractsrdquo Journal of Food Science andTechnology vol 50 no 1 pp 135ndash140 2013

[6] P V Reddy and A Urooj ldquoProximate phytochemical profileand antioxidant activity (in vitro and ex vivo) of Morus indicavarietiesrdquo International Journal of Pharmaceutical Sciences andResearch vol 4 no 4 pp 1626ndash1634 2013

[7] A Urooj and P V Reddy ldquoMoringa oleifera antioxidant prop-erties and stability of various solvent extractsrdquo InternationalJournal of Pharmaceutical Sciences and Biotechnology vol 1 no1 pp 1ndash6 2010

[8] P V Reddy S Desai and F A A Urooj ldquoAntioxidant propertiesand stability of Raphanus sativus extractsrdquo Journal of PharmacyResearch vol 3 no 3 pp 658ndash661 2011

[9] P V Reddy N Sahana and A Urooj ldquoAntioxidant activityof Aegle marmelos and Psidium guajava leavesrdquo InternationalJournal of Medicinal and Aromatic Plants vol 2 no 1 pp 155ndash160 2012

[10] AOAC Official Methods of Analysis AOAC InternationalWashington DC USA 11th edition 1970

[11] S Ranganna Handbook of Analysis and Quality Control forFruits and Vegetables Products McGrow-Hill NewDelhi India2nd edition 1999

[12] M Freed Methods of Vitamins Assay Association of VitaminChemists Interscience New York NY USA 3rd edition 1966

[13] E Beutler and B M Kelly ldquoThe effect of sodium nitrite on redcell GSHrdquo Experientia vol 19 no 2 pp 96ndash97 1963

[14] K Slinkard and V L Singleton ldquoTotal phenol analysis automa-tion and comparison with manual methodsrdquo American Journalof Enology and Viticare vol 28 pp 49ndash55 1967

[15] GMiliauskas P R Venskutonis andT A VanBeek ldquoScreeningof radical scavenging activity of some medicinal and aromaticplant extractsrdquo Food Chemistry vol 85 no 2 pp 231ndash237 2004

[16] M S Blois ldquoAntioxidant determinations by the use of a stablefree radicalrdquo Nature vol 181 no 4617 pp 1199ndash1200 1958

[17] A Yildirim A Mavi and A A Kara ldquoAntioxidant and antimi-crobial activities of Polygonum cognatum Meissn extractsrdquoJournal of the Science of Food and Agriculture vol 83 no 1 pp64ndash69 2003

[18] I F F Benzie and J J Strain ldquoFerric reducingantioxidant powerassay direct measure of total antioxidant activity of biologicalfluids and modified version for simultaneous measurementof total antioxidant power and ascorbic acid concentrationrdquoMethods in Enzymology vol 299 pp 15ndash27 1998

[19] H Tamura and A Yamagami ldquoAntioxidative activity of monoa-cylated anthocyanins isolated from Muscat Bailey A graperdquoJournal of Agricultural and Food Chemistry vol 42 no 8 pp1612ndash1615 1994

[20] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[21] D J Shapiro and V W Rodwell ldquoRegulation of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase and choles-terol synthesisrdquo Journal of Biological Chemistry vol 246 no 10pp 3210ndash3216 1971

[22] J A Buege and SD Aust ldquo[30]Microsomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978

[23] S Arabshahi-D D Vishalakshi Devi and A Urooj ldquoEvaluationof antioxidant activity of some plant extracts and their heat pHand storage stabilityrdquo Food Chemistry vol 100 no 3 pp 1100ndash1105 2007

International Journal of Medicinal Chemistry 7

[24] L J Lizcano M Viloria-Bernal F Vicente et al ldquoLipid oxi-dation inhibitory effects and phenolic composition of aqueousextracts from medicinal plants of colombian amazoniardquo Inter-national Journal of Molecular Sciences vol 13 no 5 pp 5454ndash5467 2012

[25] S Mandal S Yadav S Yadav and R K Nema ldquoAntioxidants areviewrdquo Journal of Chemical and Pharmaceutical Research vol1 no 1 pp 102ndash104 2009

[26] V K Gupta and S K Gupta ldquoPlants as natural antioxidantsrdquoNatural Product Radiance vol 5 no 6 pp 326ndash334 2006

[27] S Arabshahi-Delouee and A Urooj ldquoAntioxidant properties ofvarious solvent extracts of mulberry (Morus indica L) leavesrdquoFood Chemistry vol 102 no 4 pp 1233ndash1240 2007

[28] S M Zachariah N A Aleykutty V Viswanad S Jacob andV Prabhakar ldquoIn-vitro antioxidant potential of methanolicextracts of Mirabilis jalapa Linnrdquo Free Radicals and Antioxi-dants vol 1 no 4 pp 82ndash86 2011

[29] V Reddy A Urooj and A Kumar ldquoEvaluation of antioxidantactivity of some plant extracts and their application in biscuitsrdquoFood Chemistry vol 90 no 1-2 pp 317ndash321 2005

[30] E H Mansour and A H Khalil ldquoEvaluation of antioxidantactivity of some plant extracts and their application to groundbeef pattiesrdquo Food Chemistry vol 69 no 2 pp 135ndash141 2000

[31] A H Azizah NM Nik Ruslawati and T Swee Tee ldquoExtractionand characterization of antioxidant from cocoa by-productsrdquoFood Chemistry vol 64 no 2 pp 199ndash202 1999

[32] M G Miguel ldquoAntioxidant activity of medicinal and aromaticplants A reviewrdquo Flavour and Fragrance Journal vol 25 no 5pp 291ndash312 2010

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

Page 5: Research Article Abrus precatorius Leaves: Antioxidant ...downloads.hindawi.com/journals/ijmc/2014/748549.pdf · Research Article Abrus precatorius Leaves: Antioxidant Activity in

International Journal of Medicinal Chemistry 5

and deterioration of the sensory and nutritional property ofthe food products Auto-oxidation of fats and oil in processedfoods may be prevented by use of oxidation inhibitors orantioxidants To protect the cells and organ systems of thebody against reactive oxygen species humans have evolveda highly sophisticated and complex antioxidant protectionsystem that functions interactively and synergistically toneutralize free radicals Thus antioxidants are capable ofstabilizing or deactivating free radicals before they attack cell[28] Vitamin E scavenges peroxyl radicals intermediates inlipid peroxidation and is responsible for protecting PUFApresent in cell membrane and LDL against lipid peroxidationThe polarity of phytochemicals plays a key role in exhibitingantioxidant role at lipid phase especially at unsaturation sitewhich influences the chain breaking reaction [24] Presentlyin methanol extract of AP antioxidants with high partitioncoefficientmay be distributed hydrophobic compartments forthe protection of lipids AP was more effective in inhibitingthe oxidation in microsomes compared to oil emulsion Sim-ilarly results were found in case ofMorus indica [6] Here theuse of two lipid systems was helpful in studying the inhibitionpotency of different extracts in food and biological level

46 Stability of AP Extracts to High Temperature (100∘C 15and 30min) It is well known that many factors such asantioxidant concentration temperature and pH of themediaprocessing treatments influence the antioxidant activity Rad-ical scavenging activity of the heat treated sample extract isshown in Figure 6 After 15min of heat treatment a slightdecline in the activity was observed at all the concentrationsexcept at 300 120583g which showed an increase in the activityA significant increase was seen at the end of 30min atconcentrations greater than 100 120583g This may be due to theactivation of antioxidant components after 30min of heattreatment In another study extracts of drumstick leavesmint leaves and carrot were subjected to heat treatment at100∘C (15min) which resulted in a significant decrease inantioxidant activity (AOA) in drumstick leaves (DL) extractwhile no difference was observed in AOA of carrot extractThe significant increase in AOA due to thermal processingoccurred in mint leaves extract Thermal processing caninduce the formation of novel compounds with antioxidantproperties or improve theAOAof naturally occurring antiox-idants [29] Another study reports that WE (water) ME(methanol) and EE (ethanol) extracts of Aegle marmelosL treated at 100∘C for 15min showed varied stability WEshowed maximum stability as measured by RSA comparedto ME and EE after heat treatment [7] Similar results werefound in case of different extracts of Raphanus sativus [8] andMoringa oleifera [5]

47 Stability of AP Extract to Different pH The antioxidantstability of sample extract to different pH (45 70 and 90)was analysed by radical scavenging activity (Figure 7(a)mdashpH45 and Figure 7(b)mdashpH 70) At pH 45 and 70 the extractat different concentrations and time intervals has shownvarying antioxidant activity The influence of pH 45 on thestability of AP methanol extract is shown in Figure 7(a) and

05

101520253035404550

0 100 200 300 400 500 600

RSA

()

01530

Figure 6 Stability of Abrus precatorius extract to high temperature(100∘C 15 and 30min) Values are expressed as mean of triplicates

at the end of 72 h a decrease in radical scavenging activitywas observed However the decline in the activity was notlinear The maximum antioxidant activity observed was 50at 500120583g

At pH 70 after 72 h a significant increase in RSA wasobserved up to 300 120583g and subsequently the RSA decreased athigher concentrations (400 and 500120583g)However the declinein RSA was not significant compared to the zero min At pH9 the extract did not exhibit RSA and the absorbance wasmore than that of control this may be due to the structuraldenaturation of natural antioxidant components and similarresults were found in fenugreek seeds and ginger rhizomeextracts [30] In Aegle marmelos [7] and Rapahnus sativus[8] the antioxidant activity decreased by increasing the pHof media The antioxidant activity of different extracts fromcocoa byproducts was found to be higher at alkaline pH [31]In an earlier study it was observed that extracts from mintand carrot leaves showed higher antioxidant activity at pH 9than pH 4 [32] These differences might be due to differentsamples used various compounds being extracted in eachcase and the method used to evaluate antioxidant potency

5 Conclusion

Results of the present study reveal that the Abrus precatoriusis a potential source of antioxidants which are responsiblefor the antioxidant activity The stability to heat and pHof the different extracts of AP leaves extracts with strongantioxidant activity indicates its scope for utilization in foodand biological systems In addition to being consumed ashealthy antioxidants the compounds present in AP that areresponsible for antioxidant activity could be isolated and thenused as food additives to delay the oxidative deteriorationof foods and as nutraceutical in food formulations againstdegenerative diseases

Conflict of Interests

The authors declare that there is no conflict of interests

6 International Journal of Medicinal Chemistry

0

10

20

30

40

50

60

0 100 200 300 400 500 600

RSA

()

024

4872

(a)

05

1015202530354045

0 100 200 300 400 500 600

RSA

()

4872

024

(b)

Figure 7 (a) Stability of Abrus precatorius extract to pH 45 values are expressed as mean of triplicates (b) Stability of Abrus precatoriusextract at pH 7 values are expressed as mean of triplicates

Acknowledgment

The authors acknowledge UGC-IOE (Institute of Excellence)for providing financial assistance for this project

References

[1] A Lugasi J Hovari K V Sagi and L Bıro ldquoThe role ofantioxidant phytonutrients in the prevention of diseasesrdquo ActaBiologica Szegediensis vol 47 no 1ndash4 pp 119ndash125 2003

[2] N Nakatani ldquoAntioxidants from spices and herbsrdquo in NaturalAntioxidants ChemistryHealth Effects andApplications pp 64ndash73 AOCS Press Champaign Ill USA 1997

[3] J J M Castenmiller J P H Linssen I M Heinonen AL Hopia K Schwarz and P C H Hollmann ldquoAntioxidantproperties of differently processed spinach productsrdquo NahrungFood Journal vol 46 pp 290ndash293 2002

[4] C Kaur and H C Kapoor ldquoAntioxidants in fruits andvegetablesmdashthe millenniumrsquos healthrdquo International Journal ofFood Science and Technology vol 36 no 7 pp 703ndash725 2001

[5] V P Reddy and A Urooj ldquoAntioxidant properties and stabilityof aegle marmelos leaves extractsrdquo Journal of Food Science andTechnology vol 50 no 1 pp 135ndash140 2013

[6] P V Reddy and A Urooj ldquoProximate phytochemical profileand antioxidant activity (in vitro and ex vivo) of Morus indicavarietiesrdquo International Journal of Pharmaceutical Sciences andResearch vol 4 no 4 pp 1626ndash1634 2013

[7] A Urooj and P V Reddy ldquoMoringa oleifera antioxidant prop-erties and stability of various solvent extractsrdquo InternationalJournal of Pharmaceutical Sciences and Biotechnology vol 1 no1 pp 1ndash6 2010

[8] P V Reddy S Desai and F A A Urooj ldquoAntioxidant propertiesand stability of Raphanus sativus extractsrdquo Journal of PharmacyResearch vol 3 no 3 pp 658ndash661 2011

[9] P V Reddy N Sahana and A Urooj ldquoAntioxidant activityof Aegle marmelos and Psidium guajava leavesrdquo InternationalJournal of Medicinal and Aromatic Plants vol 2 no 1 pp 155ndash160 2012

[10] AOAC Official Methods of Analysis AOAC InternationalWashington DC USA 11th edition 1970

[11] S Ranganna Handbook of Analysis and Quality Control forFruits and Vegetables Products McGrow-Hill NewDelhi India2nd edition 1999

[12] M Freed Methods of Vitamins Assay Association of VitaminChemists Interscience New York NY USA 3rd edition 1966

[13] E Beutler and B M Kelly ldquoThe effect of sodium nitrite on redcell GSHrdquo Experientia vol 19 no 2 pp 96ndash97 1963

[14] K Slinkard and V L Singleton ldquoTotal phenol analysis automa-tion and comparison with manual methodsrdquo American Journalof Enology and Viticare vol 28 pp 49ndash55 1967

[15] GMiliauskas P R Venskutonis andT A VanBeek ldquoScreeningof radical scavenging activity of some medicinal and aromaticplant extractsrdquo Food Chemistry vol 85 no 2 pp 231ndash237 2004

[16] M S Blois ldquoAntioxidant determinations by the use of a stablefree radicalrdquo Nature vol 181 no 4617 pp 1199ndash1200 1958

[17] A Yildirim A Mavi and A A Kara ldquoAntioxidant and antimi-crobial activities of Polygonum cognatum Meissn extractsrdquoJournal of the Science of Food and Agriculture vol 83 no 1 pp64ndash69 2003

[18] I F F Benzie and J J Strain ldquoFerric reducingantioxidant powerassay direct measure of total antioxidant activity of biologicalfluids and modified version for simultaneous measurementof total antioxidant power and ascorbic acid concentrationrdquoMethods in Enzymology vol 299 pp 15ndash27 1998

[19] H Tamura and A Yamagami ldquoAntioxidative activity of monoa-cylated anthocyanins isolated from Muscat Bailey A graperdquoJournal of Agricultural and Food Chemistry vol 42 no 8 pp1612ndash1615 1994

[20] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[21] D J Shapiro and V W Rodwell ldquoRegulation of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase and choles-terol synthesisrdquo Journal of Biological Chemistry vol 246 no 10pp 3210ndash3216 1971

[22] J A Buege and SD Aust ldquo[30]Microsomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978

[23] S Arabshahi-D D Vishalakshi Devi and A Urooj ldquoEvaluationof antioxidant activity of some plant extracts and their heat pHand storage stabilityrdquo Food Chemistry vol 100 no 3 pp 1100ndash1105 2007

International Journal of Medicinal Chemistry 7

[24] L J Lizcano M Viloria-Bernal F Vicente et al ldquoLipid oxi-dation inhibitory effects and phenolic composition of aqueousextracts from medicinal plants of colombian amazoniardquo Inter-national Journal of Molecular Sciences vol 13 no 5 pp 5454ndash5467 2012

[25] S Mandal S Yadav S Yadav and R K Nema ldquoAntioxidants areviewrdquo Journal of Chemical and Pharmaceutical Research vol1 no 1 pp 102ndash104 2009

[26] V K Gupta and S K Gupta ldquoPlants as natural antioxidantsrdquoNatural Product Radiance vol 5 no 6 pp 326ndash334 2006

[27] S Arabshahi-Delouee and A Urooj ldquoAntioxidant properties ofvarious solvent extracts of mulberry (Morus indica L) leavesrdquoFood Chemistry vol 102 no 4 pp 1233ndash1240 2007

[28] S M Zachariah N A Aleykutty V Viswanad S Jacob andV Prabhakar ldquoIn-vitro antioxidant potential of methanolicextracts of Mirabilis jalapa Linnrdquo Free Radicals and Antioxi-dants vol 1 no 4 pp 82ndash86 2011

[29] V Reddy A Urooj and A Kumar ldquoEvaluation of antioxidantactivity of some plant extracts and their application in biscuitsrdquoFood Chemistry vol 90 no 1-2 pp 317ndash321 2005

[30] E H Mansour and A H Khalil ldquoEvaluation of antioxidantactivity of some plant extracts and their application to groundbeef pattiesrdquo Food Chemistry vol 69 no 2 pp 135ndash141 2000

[31] A H Azizah NM Nik Ruslawati and T Swee Tee ldquoExtractionand characterization of antioxidant from cocoa by-productsrdquoFood Chemistry vol 64 no 2 pp 199ndash202 1999

[32] M G Miguel ldquoAntioxidant activity of medicinal and aromaticplants A reviewrdquo Flavour and Fragrance Journal vol 25 no 5pp 291ndash312 2010

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

Page 6: Research Article Abrus precatorius Leaves: Antioxidant ...downloads.hindawi.com/journals/ijmc/2014/748549.pdf · Research Article Abrus precatorius Leaves: Antioxidant Activity in

6 International Journal of Medicinal Chemistry

0

10

20

30

40

50

60

0 100 200 300 400 500 600

RSA

()

024

4872

(a)

05

1015202530354045

0 100 200 300 400 500 600

RSA

()

4872

024

(b)

Figure 7 (a) Stability of Abrus precatorius extract to pH 45 values are expressed as mean of triplicates (b) Stability of Abrus precatoriusextract at pH 7 values are expressed as mean of triplicates

Acknowledgment

The authors acknowledge UGC-IOE (Institute of Excellence)for providing financial assistance for this project

References

[1] A Lugasi J Hovari K V Sagi and L Bıro ldquoThe role ofantioxidant phytonutrients in the prevention of diseasesrdquo ActaBiologica Szegediensis vol 47 no 1ndash4 pp 119ndash125 2003

[2] N Nakatani ldquoAntioxidants from spices and herbsrdquo in NaturalAntioxidants ChemistryHealth Effects andApplications pp 64ndash73 AOCS Press Champaign Ill USA 1997

[3] J J M Castenmiller J P H Linssen I M Heinonen AL Hopia K Schwarz and P C H Hollmann ldquoAntioxidantproperties of differently processed spinach productsrdquo NahrungFood Journal vol 46 pp 290ndash293 2002

[4] C Kaur and H C Kapoor ldquoAntioxidants in fruits andvegetablesmdashthe millenniumrsquos healthrdquo International Journal ofFood Science and Technology vol 36 no 7 pp 703ndash725 2001

[5] V P Reddy and A Urooj ldquoAntioxidant properties and stabilityof aegle marmelos leaves extractsrdquo Journal of Food Science andTechnology vol 50 no 1 pp 135ndash140 2013

[6] P V Reddy and A Urooj ldquoProximate phytochemical profileand antioxidant activity (in vitro and ex vivo) of Morus indicavarietiesrdquo International Journal of Pharmaceutical Sciences andResearch vol 4 no 4 pp 1626ndash1634 2013

[7] A Urooj and P V Reddy ldquoMoringa oleifera antioxidant prop-erties and stability of various solvent extractsrdquo InternationalJournal of Pharmaceutical Sciences and Biotechnology vol 1 no1 pp 1ndash6 2010

[8] P V Reddy S Desai and F A A Urooj ldquoAntioxidant propertiesand stability of Raphanus sativus extractsrdquo Journal of PharmacyResearch vol 3 no 3 pp 658ndash661 2011

[9] P V Reddy N Sahana and A Urooj ldquoAntioxidant activityof Aegle marmelos and Psidium guajava leavesrdquo InternationalJournal of Medicinal and Aromatic Plants vol 2 no 1 pp 155ndash160 2012

[10] AOAC Official Methods of Analysis AOAC InternationalWashington DC USA 11th edition 1970

[11] S Ranganna Handbook of Analysis and Quality Control forFruits and Vegetables Products McGrow-Hill NewDelhi India2nd edition 1999

[12] M Freed Methods of Vitamins Assay Association of VitaminChemists Interscience New York NY USA 3rd edition 1966

[13] E Beutler and B M Kelly ldquoThe effect of sodium nitrite on redcell GSHrdquo Experientia vol 19 no 2 pp 96ndash97 1963

[14] K Slinkard and V L Singleton ldquoTotal phenol analysis automa-tion and comparison with manual methodsrdquo American Journalof Enology and Viticare vol 28 pp 49ndash55 1967

[15] GMiliauskas P R Venskutonis andT A VanBeek ldquoScreeningof radical scavenging activity of some medicinal and aromaticplant extractsrdquo Food Chemistry vol 85 no 2 pp 231ndash237 2004

[16] M S Blois ldquoAntioxidant determinations by the use of a stablefree radicalrdquo Nature vol 181 no 4617 pp 1199ndash1200 1958

[17] A Yildirim A Mavi and A A Kara ldquoAntioxidant and antimi-crobial activities of Polygonum cognatum Meissn extractsrdquoJournal of the Science of Food and Agriculture vol 83 no 1 pp64ndash69 2003

[18] I F F Benzie and J J Strain ldquoFerric reducingantioxidant powerassay direct measure of total antioxidant activity of biologicalfluids and modified version for simultaneous measurementof total antioxidant power and ascorbic acid concentrationrdquoMethods in Enzymology vol 299 pp 15ndash27 1998

[19] H Tamura and A Yamagami ldquoAntioxidative activity of monoa-cylated anthocyanins isolated from Muscat Bailey A graperdquoJournal of Agricultural and Food Chemistry vol 42 no 8 pp1612ndash1615 1994

[20] H Ohkawa N Ohishi and K Yagi ldquoAssay for lipid peroxidesin animal tissues by thiobarbituric acid reactionrdquo AnalyticalBiochemistry vol 95 no 2 pp 351ndash358 1979

[21] D J Shapiro and V W Rodwell ldquoRegulation of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase and choles-terol synthesisrdquo Journal of Biological Chemistry vol 246 no 10pp 3210ndash3216 1971

[22] J A Buege and SD Aust ldquo[30]Microsomal lipid peroxidationrdquoMethods in Enzymology vol 52 pp 302ndash310 1978

[23] S Arabshahi-D D Vishalakshi Devi and A Urooj ldquoEvaluationof antioxidant activity of some plant extracts and their heat pHand storage stabilityrdquo Food Chemistry vol 100 no 3 pp 1100ndash1105 2007

International Journal of Medicinal Chemistry 7

[24] L J Lizcano M Viloria-Bernal F Vicente et al ldquoLipid oxi-dation inhibitory effects and phenolic composition of aqueousextracts from medicinal plants of colombian amazoniardquo Inter-national Journal of Molecular Sciences vol 13 no 5 pp 5454ndash5467 2012

[25] S Mandal S Yadav S Yadav and R K Nema ldquoAntioxidants areviewrdquo Journal of Chemical and Pharmaceutical Research vol1 no 1 pp 102ndash104 2009

[26] V K Gupta and S K Gupta ldquoPlants as natural antioxidantsrdquoNatural Product Radiance vol 5 no 6 pp 326ndash334 2006

[27] S Arabshahi-Delouee and A Urooj ldquoAntioxidant properties ofvarious solvent extracts of mulberry (Morus indica L) leavesrdquoFood Chemistry vol 102 no 4 pp 1233ndash1240 2007

[28] S M Zachariah N A Aleykutty V Viswanad S Jacob andV Prabhakar ldquoIn-vitro antioxidant potential of methanolicextracts of Mirabilis jalapa Linnrdquo Free Radicals and Antioxi-dants vol 1 no 4 pp 82ndash86 2011

[29] V Reddy A Urooj and A Kumar ldquoEvaluation of antioxidantactivity of some plant extracts and their application in biscuitsrdquoFood Chemistry vol 90 no 1-2 pp 317ndash321 2005

[30] E H Mansour and A H Khalil ldquoEvaluation of antioxidantactivity of some plant extracts and their application to groundbeef pattiesrdquo Food Chemistry vol 69 no 2 pp 135ndash141 2000

[31] A H Azizah NM Nik Ruslawati and T Swee Tee ldquoExtractionand characterization of antioxidant from cocoa by-productsrdquoFood Chemistry vol 64 no 2 pp 199ndash202 1999

[32] M G Miguel ldquoAntioxidant activity of medicinal and aromaticplants A reviewrdquo Flavour and Fragrance Journal vol 25 no 5pp 291ndash312 2010

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

Page 7: Research Article Abrus precatorius Leaves: Antioxidant ...downloads.hindawi.com/journals/ijmc/2014/748549.pdf · Research Article Abrus precatorius Leaves: Antioxidant Activity in

International Journal of Medicinal Chemistry 7

[24] L J Lizcano M Viloria-Bernal F Vicente et al ldquoLipid oxi-dation inhibitory effects and phenolic composition of aqueousextracts from medicinal plants of colombian amazoniardquo Inter-national Journal of Molecular Sciences vol 13 no 5 pp 5454ndash5467 2012

[25] S Mandal S Yadav S Yadav and R K Nema ldquoAntioxidants areviewrdquo Journal of Chemical and Pharmaceutical Research vol1 no 1 pp 102ndash104 2009

[26] V K Gupta and S K Gupta ldquoPlants as natural antioxidantsrdquoNatural Product Radiance vol 5 no 6 pp 326ndash334 2006

[27] S Arabshahi-Delouee and A Urooj ldquoAntioxidant properties ofvarious solvent extracts of mulberry (Morus indica L) leavesrdquoFood Chemistry vol 102 no 4 pp 1233ndash1240 2007

[28] S M Zachariah N A Aleykutty V Viswanad S Jacob andV Prabhakar ldquoIn-vitro antioxidant potential of methanolicextracts of Mirabilis jalapa Linnrdquo Free Radicals and Antioxi-dants vol 1 no 4 pp 82ndash86 2011

[29] V Reddy A Urooj and A Kumar ldquoEvaluation of antioxidantactivity of some plant extracts and their application in biscuitsrdquoFood Chemistry vol 90 no 1-2 pp 317ndash321 2005

[30] E H Mansour and A H Khalil ldquoEvaluation of antioxidantactivity of some plant extracts and their application to groundbeef pattiesrdquo Food Chemistry vol 69 no 2 pp 135ndash141 2000

[31] A H Azizah NM Nik Ruslawati and T Swee Tee ldquoExtractionand characterization of antioxidant from cocoa by-productsrdquoFood Chemistry vol 64 no 2 pp 199ndash202 1999

[32] M G Miguel ldquoAntioxidant activity of medicinal and aromaticplants A reviewrdquo Flavour and Fragrance Journal vol 25 no 5pp 291ndash312 2010

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of

Page 8: Research Article Abrus precatorius Leaves: Antioxidant ...downloads.hindawi.com/journals/ijmc/2014/748549.pdf · Research Article Abrus precatorius Leaves: Antioxidant Activity in

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Inorganic ChemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

International Journal ofPhotoenergy

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Carbohydrate Chemistry

International Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Advances in

Physical Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom

Analytical Methods in Chemistry

Journal of

Volume 2014

Bioinorganic Chemistry and ApplicationsHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

SpectroscopyInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Medicinal ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Chromatography Research International

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Applied ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Theoretical ChemistryJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Spectroscopy

Analytical ChemistryInternational Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Quantum Chemistry

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Organic Chemistry International

ElectrochemistryInternational Journal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

CatalystsJournal of