nii korley kortei (Antioxidants)

Research ArticleEvaluating the Effect of Gamma Radiation on the Total PhenolicContent Flavonoids and Antioxidant Activity ofDried Pleurotus ostreatus ((Jacq ex Fr) Kummer) Stored inPackaging Materials

Nii Korley Kortei1 George Tawia Odamtten12 Mary Obodai3

Victoria Appiah1 Felicia Akuamoa4 Afua Kobi Adu-Bobi4 Sylvester Nana Yao Annan5

Jonathan Nii Okai Armah5 and Stanley Akwesi Acquah5

1 Graduate School of Nuclear and Allied Sciences Department of Nuclear Agriculture and Radiation ProcessingPO Box LG 80 Legon Ghana

2Department of Botany University of Ghana PO Box LG 55 Legon Ghana3 CSIR-Food Research Institute PO Box M20 Accra Ghana4Applied Radiation Biology Centre Ghana Atomic Energy Commission PO Box AE 1 Atomic Accra Ghana5 Radiation Technology Centre Ghana Atomic Energy Commission PO Box AE 1 Atomic Accra Ghana

Correspondence should be addressed to Nii Korley Kortei nii korley 1yahoocom

Received 30 April 2014 Revised 22 July 2014 Accepted 31 July 2014 Published 24 August 2014

Academic Editor Maria Cristina Bonferoni

Copyright copy 2014 Nii Korley Kortei 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

DriedPleurotus ostreatusmushrooms stored in polythene and polypropylene packswere exposed to gamma radiation froma cobalt-60 source at doses of 0 05 1 15 and 2 kGy at a dose rate of 17 kGyhr and stored for a period of 1 month Total phenolic contentsflavonoids and free radical scavenging activity DPPH (221015840-diphenyl-1-picrylhydrazyl) were determined using aqueous ethanolandmethanol extracts by Folin-Ciocaultaeumethod as a source of potential natural antioxidants Total phenol content ranged 056plusmn001ndash1096plusmn 17mgGAE flavonoids ranged 164plusmn 005ndash892plusmn 06mgQEDPPHradical scavenging activity also ranged 702plusmn 010ndash1303 plusmn 004 and IC

50values also ranged 008ndash016mgmL Statistical differences (P lt 005) were recorded for the extracts and the

treatment doses ofmushrooms stored in polythene andpolypropylene packsA significant linear correlationwas confirmedbetweenvalues for the total phenolic content and antioxidant activity of mushroom extracts The high contents of phenolic compoundsindicated that these compounds contribute to high antioxidant activityPleurotus ostreatus can be regarded as a promising candidatefor natural mushroom sources of antioxidants with high value The use of low dose gamma radiation by the local food industrycould improve the hygienic quality extend shelf-life and preserve nutrients and antinutrients

1 Introduction

Phenolic compounds are secondary metabolites that arederivatives of the pentose phosphate shikimate and phenyl-propanoid pathways in living organisms [1ndash3] They actin defense against pathogens animal mycophage or fun-givore aggression and as response to various abiotic stress

conditions such as rainfall and ultraviolet radiation [4]Polyphenols have protective activity which has been previ-ously attributed to free radical scavenging metal chelatingproperties capability of inhibiting or reducing differentenzymes such as telomerase [5] cyclooxygenase [6 7] orlipoxygenase [8 9] and thenmost importantly as antioxidantcompounds with the ability to trap free radicals and thus

Hindawi Publishing CorporationAdvances in PharmaceuticsVolume 2014 Article ID 262807 8 pageshttpdxdoiorg1011552014262807

2 Advances in Pharmaceutics

inhibit the oxidative mechanisms However in more recentyears they play roles such as interacting with signal trans-duction pathways and cell receptors [10ndash12]

Naturally occurring antioxidants can be found in wholegrains fruits vegetables teas spices and herbs Mushroomshave also been reported as living organisms with antioxidantactivity which is correlated with their phenolic and polysac-charide compounds [13ndash15] Their global economic value isnow incredible and the reason for the rise in consumption is acombination of their value as food [16 17] and theirmedicinalor nutraceutical properties [18ndash21]

Harmful effects of disturbed antioxidant-prooxidant bal-ance can be largely prevented by intake of antioxidantsubstances [22 23] Antioxidants have already been found inmushroom materials and supplements [24 25] Due to theirnatural origin the antioxidants obtained from living organ-isms are of greater benefit in comparison to synthetic ones[26 27] The use of natural antioxidants from mushroomsdoes not induce side effects while synthetic antioxidantswerefound to have genotoxic effect [28 29]

Free radicals reactive nitrogen species (RNS) and otherreactive oxygen species (ROS) such as superoxide anionshydroxyl radicals and hydrogen peroxide are an entireclass of highly reactive species derived from the normalmetabolism of oxygen or from exogenous factors and agentssuch as pollutants and ultraviolet (UV) radiation Accordingto [30 31] oxidative damage to crucial cellular moleculesinduced by ROS has been implicated as a possible factorin the etiology of several human diseases such as cancercardiovascular diseases atherosclerosis Alzheimer diseaseand ageing [32] reported that irradiated flavonoid solutionsproduce chemical reactions between the solute and reactivespecies which are the result of the radiolyzed solvent Thesereactions according to [32] take place in radiolyzed alcoholsolutions a medium that is very rich in many differenthighly reactive species and that hosts specific reactionsThe reactivity of the solute (flavonoid) with the alkoxyradicals CH

3Olowast and CH

3CH2Olowast formed in methanol and

ethanol respectively and their carbon-centered isomersthe 1-hydroxy-methyl lowastCH

2OH and the 1-hydroxy-ethyl

CH3

lowastCHOH radicals operate to quickly repair the chemicalmodifications to biomolecules

Published studies by several researchers [33 34] showhow polyphenol compounds in our diets help to improveendothelial function which is a critical factor in preventingatherosclerosis They have also been shown to inhibit theabnormal blood platelet aggregation that cause most suddenheart attacks and strokes while fighting inflammation andsupporting healthy blood lipids They have the potential toreduce allergic conditions [35] by blocking the release ofhistamine (an irritating substance causing inflammation anditching) from the mast cells that mediate allergic reactionsLetenneur [36] reported that polyphenols from differentnatural sources may work synergistically when consumedtogether with benefits from the combination resulting inmore than the sum of the parts

Antimicrobial activity of polyphenols occurring in veg-etable foods medicinal plants and mushrooms has been

extensively investigated against a wide range of microor-ganisms Among polyphenols flavan-3-ols flavonols andtannins received most attention due to their wide spectrumand higher antimicrobial activity in comparison with otherpolyphenols and to the fact that most of them are able tosuppress a number of microbial virulence factors (such asinhibition of biofilm formation reduction of host ligandsadhesion and neutralization of bacterial toxins) and showsynergism with antibiotics [4] The antimicrobial propertiesof certain classes of polyphenols have been proposed eitherto develop new food preservatives [37] due to the increasingconsumer pressure on the food industry to avoid syntheticpreservatives or to develop innovative therapies for the treat-ment of various microbial infections [38 39] consideringthe increase in microbial resistance against conventionalantibiotic therapy

This study is to investigate the effect of gamma radiationon total phenolic content flavonoids and antioxidant activityin various extracts of the species Pleurotus ostreatus in differ-ent packaging materials using spectrophotometric methods

2 Materials and Methods

21 Chemicals Analytical ethanol methanol and sodiumhydrogen carbonate NAHCO

3were purchased from Sigma-

Aldrich USA Standards of phenolic acids (gallic acid[345-trihydroxybenzoic acid]) and of flavonoids potas-sium acetate quercetin [3310158404101584057-pentahydroxyflavone]and 22-dyphenyl-1-picrylhydrazyl (DPPH) were obtainedfrom Sigma Chemicals Co St Louis MO USA The Folin-Ciocalteursquos phenol reagent and Aluminium chloride (AlCl

3)

were from Fluka Chemie AG Buchs Switzerland

22 Mushroom Material Pleurotus ostreatus originally fromMauritius was cultivated on Triplochiton scleroxylon sawdustcomposted for 28 days supplemented with 1 CaCO

3and

10 wheat bran [40] at the Mushroom Unit of CSIR-FoodResearch Institute Accra Ghana Cultivation and harvestingwere from the period between September and December2012 Mushroom fruit bodies were solar-dried at temperaturerange of 20ndash40∘C to a moisture content of about 12 Driedmushroom parts were cut up to average size of 2 cm times 3 cmand stored in tight-sealed polythene (P) and polypropylene(Bx) containers at room temperature until needed

23 Irradiation of Mushroom Materials Forty (40) gramsof dried mushrooms were packed into various containersand irradiated at doses of 0 05 1 15 and 2 kGy at adose rate of 17 kGy per hour from a cobalt60 source (SLL515 Hungary) in air Absorbed doses were confirmed usingthe ethanol-chlorobenzene (ECB) dosimetry system at theRadiation Technology Centre of the Ghana Atomic EnergyCommission Accra Ghana

24 Preparation of Mushroom Extracts Mushroom extractswere prepared according to a standard protocol [41] Preparedmushroom material (10 g) was transferred to dark-coloredflasks and mixed with 200mL of solvents with different

Advances in Pharmaceutics 3

polarities (deionized water methanol ethanol) respectivelyand stored at room temperature After 24 h infusions werefiltered through Whatman number 1 filter paper and residuewas reextracted with equal volume of solvents After 48 hthe process was repeated Combined supernatants wereevaporated to dryness under vacuum at 30∘C using a rotaryevaporator The obtained extracts were kept in sterile sampletubes and stored in a refrigerator at 4∘C

25 Determination of Total Phenolic Contents in the Mush-room Extracts The concentration of phenolics in mushroomextracts was determined using spectrophotometric method[42] Methanolic solution of the extract in the concentrationof 1mgmLwas used in the analysisThe reactionmixturewasprepared by mixing 50mL of methanolic solution of extracts25mL of 10 Folin-Ciocalteursquos reagent (vv) dissolved inwater and 25mL 75 NaHCO

3 Blank was concomitantly

prepared containing 50mL methanol 25mL 10 Folin-Ciocalteursquos reagent dissolved in water and 25mL of 75 ofNaHCO

3The sampleswere thereafter incubated in a thermo-

stat at 45∘C for 45minThe absorbance was determined usingspectrophotometer at 120582max = 760 nm The samples wereprepared in triplicate for each analysis and the mean value ofabsorbance was obtained The same procedure was repeatedfor the standard solution of gallic acid and the calibrationline was constructed Based on the measured absorbancethe concentration of phenolics was read (mgmLminus1) from thecalibration line then the content of phenolics in extracts wasexpressed in terms of gallic acid equivalent (mg of GAg ofextract)

26 Determination of Flavonoid Concentrations in the Mush-room Extracts The content of flavonoids in the examinedmushroom extracts was determined using a modified spec-trophotometric method [43] The sample contained 500mLof ethanol solution of the extract in the concentrationof 1mgmL and 100mL of 10 AlCl

3solution dissolved

in 1500mL ethanol The samples were incubated for anhour at room temperature The absorbance was determinedusing spectrophotometer at 120582max = 415 nm The sampleswere prepared in triplicate for each analysis and the meanvalue of absorbance was obtained The same procedure wasrepeated for the standard solution of 500mL of quercetinand 100mL of potassium acetate (10) and the calibrationline was constructed Based on the measured absorbancethe concentration of flavonoids was read (mgmL) on thecalibration line then the content of flavonoids in extractswas expressed in terms of quercetin equivalent (mg of QEgof extract)

27 Evaluation of Antioxidant Activity and IC50 The ability

of the mushroom extract to scavenge DPPH free radicalswas assessed by the standard method [44] adopted withsuitable modifications [45] The stock solutions of extractswere prepared in methanol to achieve the concentration of1mgmLminus1 Dilutions were made to obtain concentrations of001 005 01 015 and 02mgmLminus1 Diluted solutions of

sample (200mL each) weremixed with 3800mL ofmethano-lic solution of DPPH After 30min incubation in darknessat room temperature (23∘C) the absorbance was recordedat 517 nm Control sample contained all the reagents exceptthe extract Percentage inhibition was calculated using (1)the data were presented as mean values plusmn standard deviation(119899 = 2) Consider

inhibition = 119860 of control minus 119860 of sample119860 of control

times 100 (1)

IC50

value is the concentration of sample required toscavenge 50 of DPPH free radical and was calculated byplotting a graph of concentration and inhibition A linearregression was estimated to plot 119909-119910 and fit data with astraight line From linear equation 119910 = 119898119909 + 119888 where 119910 = 50and 119909 is calculated

28 Statistical Analysis All experimental measurements we-re carried out in duplicate and are expressed as average oftwo analyses plusmn standard deviation The magnitude of corre-lation between variables was done using a SPSS (ChicagoIL) statistical software package (SPSS for Windows versionXIV 2004) Significances of differences were conducted witha Duncanrsquos Multiple Range Test (DMRT) Linear regressionto correlate between total phenolics and total flavonoid withantioxidant activity was carried out using Excel 2003

3 Results and Discussions

31 The Total Phenolic Content The major antioxidants fo-und in mushrooms are phenolic compounds while otherpotential antioxidants such as ascorbic acid 120573-carotenelycopene and 120574-tocopherol have only been found in verysmall amounts [46ndash48] Phenolics are important constituentswith scavenging ability due to their hydroxyl groups andhence may contribute directly to the antioxidative action[49] Irradiation dose of 05 kGy apparently influenced theproduction of phenolics and caused a significantly (119875 lt005) higher contents with aqueous and ethanol extracts Thetotal phenolic content was expressed in terms of gallic acidequivalent in milligram (mg) per gram of dry material Thecalibration curve (the standard curve119910 = 1225119909minus0003 1199032 =0988) was used to determine total phenolic content as shownin Figure 1 The total phenolic content of the various extractswas compiled in Table 1 Generally high values of antioxidantactivity were recorded for aqueous 196 plusmn 005ndash1096 plusmn17mgGAEg Methanol and ethanol extracts ranged from056 plusmn 001ndash481 plusmn 04mgGAEg and 132 plusmn 002ndash236 plusmn02mgGAEg respectively Irradiation dose of 05 kGy pro-duced greatest amounts of phenols for all the extracts while2 kGy recorded least for the extracts In terms of packagingmaterial suitability there was no significant (119875 gt 005)difference between polythene and polypropylene The valueswere within the range of total phenolic content of antioxidantactivity of 1538 plusmn 056ndash1680 plusmn 015 gGE100 g dry weightof gamma irradiated pomegranate (Punica granatum) peelsrecorded by [50] The phenol contents of P ostreatus are alsowithin the same range as that of some commonly consumedgreen leafy vegetables in W Africa [51 52]


Table 1 Total phenolic content (mgGAEg) of dried and irradiatedmushrooms of polythene (P) and polypropylene (Bx) packs from 3extracts

TreatmentExtract

Aqueous(mgGAEg)

Ethanol(mgGAEg)

Methanol(mgGAEg)

P 0 384 plusmn 04a 20 plusmn 01b 24 plusmn 01b

P 05 50 plusmn 03ab 236 plusmn 02bc 236 plusmn 02b

P 1 463 plusmn 03a 154 plusmn 0 18b 481 plusmn 04c

P 15 278 plusmn 01a 132 plusmn 002a 283 plusmn 01b


Bx 0 254 plusmn 01a 132 plusmn 002a 193 plusmn 005b

Bx 05 446 plusmn 03a 196 plusmn 005bc 231 plusmn 02b

Bx 1 1096 plusmn 17b 153 plusmn 004b 135 plusmn 003a

Bx 15 446 plusmn 08a 189 plusmn 007bc 056 plusmn 001a

Bx 2 25 plusmn 02a 176 plusmn 005b 14 plusmn 003a

Meanswith same letters in a columnare not significantly different (119875 gt 005)

0

005

01

015

02

025

03

0 005 01 015 02 025

Abso

rban

ce (120582

nm)

Gallic acid concentration (mg mLminus1)

y = 12253x minus 00034

R2= 09888

Figure 1 Calibration curve for the determination of total phenolics

32 Flavonoid Content Flavonoids are a class of secondaryplant metabolites with significant antioxidant and chelatingproperties Antioxidant activity of flavonoids depends on thestructure and substitution pattern of hydroxyl groups [53]The concentration of flavonoids in plant extracts dependson the polarity of solvents used in the extract preparation[54] The flavonoid content was expressed in terms ofquercetin equivalent in milligrams per gram of dry materialThe calibration curve (the standard curve equation 119910 =0005119909 + 000 119903

2= 0986) was used to determine flavonoid

content as shown in Figure 2 The total flavonoid contentof the various extracts was compiled in Table 2 Resultsof flavonoid content are presented in Table 2 There weresignificant differences (119875 le 005) between flavonoid contentsof doses and solvent extracts The extraction concentra-tions followed a descending order of aqueousgt methanolgtethanol Extraction with aqueous ranged 309 plusmn 02ndash842 plusmn06 mgQEg methanol ranged 20 plusmn 01ndash490 plusmn 04mgQEg

Table 2 Flavonoid content (mgQEg) of dried and irradiatedmushrooms of polythene (P) and polypropylene (Bx) packs from 3extracts

TreatmentsExtracts

Aqueous(mgQEg) Ethanol (mgQEg) Methanol

(mgQEg)P 0 767 plusmn 08a 425 plusmn 02ab 393 plusmn 02a

P 05 892 plusmn 06a 727 plusmn 04b 381 plusmn 02a


P 15 37 plusmn 02ab 164 plusmn 005a 351 plusmn 02a

P 2 673 plusmn 05a 788 plusmn 04c 288 plusmn 01a

Bx 0 309 plusmn 02ab 473 plusmn 02ab 418 plusmn 02a


Bx 1 792 plusmn 06a 442 plusmn 02ab 250 plusmn 02a


Bx 2 346 plusmn 03ab 588 plusmn 03b 49 plusmn 04a


and ethanol 164 plusmn 005ndash788 plusmn 04mgQEg There weresignificant differences (119875 lt 005) recorded for aqueous andethanol extracts However no significant differences (119875 gt005) were recorded for methanol extracts Irradiation doseof 05 kGy recorded greatest amounts of flavonoids in generalfor all the extracts while 0 05 and 2 kGy recorded theleast flavonoids In terms of packaging material suitabilitythere was no significant (119875 gt 005) difference betweenpolythene and polypropylene The flavonoid concentrationsof P ostreatus were found to be lower than works ofsome authors [55ndash57] Nonetheless the values recorded werehigher than and were contrary to works of [57 58] whofound no flavonoids in P ostreatus as they investigated thecomparative phytochemical antimicrobial and antioxidantproperties and also contents of vitamins mineral elementsand some phenolic compounds in cultivated mushroomsrespectively

33 DPPH Free Radical Scavenging Activity Free radicalscavenging is one of the mechanisms in inhibiting lipidoxidation commonly used to estimate antioxidant activity[59 60] The extraction pattern was consistent with totalphenolic and flavonoid The DPPH free radical scavengingactivity was expressed in terms of gallic acid equivalent inmilligrams(mg) per gram(g) of dry material The calibrationcurve (the standard curve 119910 = minus3309119909 + 07374 1199032 =08507) as shown in Figure 3 Extractionwith aqueous ranged962 plusmn 010ndash125 plusmn 002mgGAEg methanol ranged 673 plusmn004ndash942 plusmn 013mgGAEg and ethanol 189 plusmn 005ndash365 plusmn022mgGAEg (Table 3) There were significant differences(119875 lt 005) recorded between extracts of aqueous and ethanolMethanol extracts however recorded no significant differ-ences Packaging materials were not significantly different

34 Percentage () Inhibition of DPPH and IC50 The half

maximal inhibitory concentration (IC50) is a measure of the


0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30

y = 00058x + 00005

R2= 09863

Abso

rban

ce (120582

nm)

Quercetin concentration (mg mLminus1)

Figure 2 Calibration curve for the determination of flavonoids

Table 3 DPPH free radical scavenging activity of dried andirradiated mushrooms of polythene (P) and polypropylene (Bx)packs from 3 extracts

TreatmentsExtracts

Aqueous(mgGAEg)

Ethanol(mgGAEg)

Methanol(mgGAEg)

P 0 007 plusmn 001ab 010 plusmn 003bc 011 plusmn 003a

P 05 010 plusmn 003b 011 plusmn 003c 013 plusmn 004a

P 1 012 plusmn 005b 009 plusmn 002bc 013 plusmn 004a


P 2 009 plusmn 002b 008 plusmn 002b 011 plusmn 003a

Bx 0 010 plusmn 003b 009 plusmn 002bc 010 plusmn 003a

Bx 05 008 plusmn 002b 011 plusmn 003c 011 plusmn 003a

Bx 1 011 plusmn 003b 007 plusmn 001a 009 plusmn 002a




effectiveness of a compound in inhibiting a biological orbiochemical function The antioxidant activity with IC

50lt

10 micro gramsmL value corresponds to good antioxidantactivity and is included in the category of powerful antioxi-dants class [61ndash63] The process of free radicals reduction isachieved through themechanism of hydrogen donation fromantioxidants [64] Free radicals used are synthetic DPPH thatreacts with an antioxidant compound through the donationof electrons from an antioxidant compound to get a pairof electrons DPPH radical compound deep purple wouldfade to yellow if it is reduced by antioxidants into nonradicalDPPH [62] when the free electron of DPPH radical has beenpaired with electrons from traps compounds (antioxidants)that would reduce DPPH radical (DPPH-H) and form stablecompounds that are DPP Hydrazine [63]

Aqueous extraction ranged 824 plusmn 003ndash116 plusmn005mgGAEg methanol ranged 879 plusmn 006ndash1303 plusmn004mgGAEg and ethanol ranged 831 plusmn 004ndash1081 plusmn

0

01

02

03

04

05

06

07

08

09

0 005 01 015 02 025

y = minus3309x + 07374

R2= 08507

Abso

rban

ce (120582

nm)


Figure 3 Calibration curve for the determination of DPPH freeradical scavenging activity

025mgGAEg (Table 4) In terms of packaging materialsuitability there was no significant (119875 gt 005) differencebetween polythene and polypropylene Figure 2 shows theamount of each extract needed for 50 inhibition (IC

50)

IC50mgmLminus1 of the extracts was in the order aqueous lt

methanol lt ethanol and corresponded to 0064 103 and103 respectively IC

50of the standard compound quercetin

was 001mgmLminus1 The highest radical scavenging activitywas shown by aqueous extract with IC

50= 0064mgmLminus1

which is lower than that of quercetin (119875 lt 005) All of theextracts showed significant difference (119875 le 005) from thestandard compound (Figure 4)Thehigh contents of phenoliccompounds and significant linear correlation between thevalues of the concentration of phenolic compounds andantioxidant activity indicated that these compounds inmushrooms contribute to the strong antioxidant activity

4 Conclusions

Although the interaction of gamma radiation and polytheneandor polypropylene packaging materials caused some sig-nificant differences (119875 le 005) in total phenolic contentflavonoids and antioxidant activity which eventually causeoxidative damage and impair flavor in food there wasno apparent difference between the packaging materials(Tables 1 2 and 3) However the action of effective andradiostable natural antioxidants may prevent chemical oxi-dation of biomolecules in irradiated food Gamma radiationhad no adverse effect on the dried mushrooms It enhancedshelf-life as it was able to prevent insect and pest infestation


Table 4 Percentage inhibition () of DPPH in dried and irradiatedmushrooms in storage packages

TreatmentsExtracts

Aqueous ()(mgGAEg)

Ethanol ()(mgGAEg)

Methanol ()(mgGAEg)

P 0 702 plusmn 010a 1006 plusmn 013bc 109 plusmn 026b

P 05 965 plusmn 013bc 1047 plusmn 024c 1303 plusmn 004b

P 1 116 plusmn 005d 936 plusmn 012bc 1282 plusmn 003b

P 15 946 plusmn 013bc 936 plusmn 012bc 1030 plusmn 027b

P 2 926 plusmn 016bc 831 plusmn 004a 1060 plusmn 03b

Bx 0 965 plusmn 013bc 895 plusmn 005bc 1040 plusmn 03b

Bx 05 824 plusmn 003a 1081 plusmn 025d 1121 plusmn 004b

Bx 1 1112 plusmn 005c 744 plusmn 016a 909 plusmn 012a

Bx 15 1127 plusmn 002c 884 plusmn 005bc 879 plusmn 006a



minus04 minus02 0 02 04 06 08 1 12 14

Methanol

Ethanol

Quercetin

Aqueous

Extr

acts

IC50 (mg mLminus1)

Figure 4 IC50

(mgmLminus1) values of mushroom extracts for freeradical scavenging activity by DPPH radical (1) Methanol (2)ethanol (3) quercetin (standard) and (4) aqueous

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

Theauthors are grateful to the staff and laboratory techniciansof Applied Radiation Biology Centre (ARBC) of the GhanaAtomic Energy Commission Accra

References

[1] D Tura and K Robards ldquoSample handling strategies for thedetermination of biophenols in food and plantsrdquo Journal ofChromatography A vol 975 no 1 pp 71ndash93 2002

[2] H L Madsen and G Bertelsen ldquoSpices as antioxidantsrdquo Trendsin Food Science and Technology vol 6 no 8 pp 271ndash277 1995

[3] J B Harborne Phytochemical Methods Springer London UK1998

[4] MDaglia ldquoPolyphenols as antimicrobial agentsrdquoCurrent Opin-ion in Biotechnology vol 23 no 2 pp 174ndash181 2012

[5] I Naasani F Oh-hashi T Oh-hara et al ldquoBlocking telomeraseby dietary polyphenols is a major mechanism for limiting thegrowth of human cancer cells in vitro and in vivordquo CancerResearch vol 63 no 4 pp 824ndash830 2003

[6] T Hussain S Gupta V M Adhami and H Mukhtar ldquoGreentea constituent epigallocatechin-3-gallate selectively inhibitsCOX-2 without affecting COX-1 expression in human prostatecarcinoma cellsrdquo International Journal of Cancer vol 113 no 4pp 660ndash669 2005

[7] K A OrsquoLeary S de Pascual-Tereasa P W Needs Y Bao NM OrsquoBrien and G Williamson ldquoEffect of flavonoids and Vita-min E on cyclooxygenase-2 (COX-2) transcriptionrdquo MutationResearch vol 551 no 1-2 pp 245ndash254 2004

[8] C D Sadik H Sies and T Schewe ldquoInhibition of 15-lipoxygenases by flavonoids structure-activity relations andmode of actionrdquo Biochemical Pharmacology vol 65 no 5 pp773ndash781 2003

[9] T Schewe C Sadik L Klotz T Yoshimoto H Kuhn andH Sies ldquoPolyphenols of cocoa inhibition of mammalian 15-lipoxygenaserdquoThe Journal of Biological Chemistry vol 382 no12 pp 1687ndash1696 2001

[10] A T Kong R Yu C Chen S Mandlekar and T PrimianoldquoSignal transduction events elicited by natural products roleof MAPK and caspase pathways in homeostatic response andinduction of apoptosisrdquoArchives of Pharmacal Research vol 23no 1 pp 1ndash16 2000

[11] J P Spencer C Rice-Evans and R J Williams ldquoModulationof pro-survival Aktprotein kinase B and ERK12 signalingcascades by quercetin and its in vivo metabolites underlietheir action on neuronal viabilityrdquo The Journal of BiologicalChemistry vol 278 no 37 pp 34783ndash34793 2003

[12] S Wiseman T Mulder and A Rietveld ldquoTea flavonoidsbioavailability in vivo and effects on cell signaling pathways invitrordquo Antioxidants and Redox Signaling vol 3 no 6 pp 1009ndash1021 2001

[13] S Wei and L J L D van Griensven ldquoPro- and antioxidativeproperties ofmedicinalmushroom extractsrdquo International Jour-nal of Medicinal Mushrooms vol 10 no 4 pp 315ndash324 2008

[14] N J Dubost B Ou and R B Beelman ldquoQuantification ofpolyphenols and ergothioneine in cultivated mushrooms andcorrelation to total antioxidant capacityrdquo Food Chemistry vol105 no 2 pp 727ndash735 2007

[15] J Mau S Tsai Y Tseng and S Huang ldquoAntioxidant propertiesof hot water extracts from Ganoderma tsugae Murrillrdquo LWT -Food Science and Technology vol 38 no 6 pp 589ndash597 2005

[16] P Kalac ldquoChemical composition and nutritional value ofEuropean species of wild growing mushrooms a reviewrdquo FoodChemistry vol 113 no 1 pp 9ndash16 2009

[17] J N K Kortei Determination of optimal growth and yieldparameters of Pleurotusostreatus grown on composted cassavapeel based formulations [MS thesis] Department of Bio-chemistry and Biotechnology Kwame Nkrumah University ofScience and Technology Kumasi Ghana 2008

[18] J N K KorteiGrowing Oyster Mushrooms (Pleurotus ostreatus)onCompostedAgrowastes An EfficientWay ofUtilizing Lignocel-lulosic Materials Lambert Academic Publishing SaarbruckenGermany 2011


[19] I C F R Ferreira J A Vaz M H Vasconcelos and AMartinsldquoCompounds from wild mushrooms with antitumor potentialrdquoAnti-Cancer Agents in Medicinal Chemistry vol 10 no 5 pp424ndash436 2010

[20] I C F R Ferreira L Barros and R M V Abreu ldquoAntioxidantsin wild mushroomsrdquo Current Medicinal Chemistry vol 16 no12 pp 1543ndash1560 2009

[21] V Shlyakhovenko V Kosak and S Olishevsky ldquoApplication ofDNA from mushroom Pleurotus ostreatus for cancer biother-apy a pilot studyrdquo Experimental Oncology vol 28 no 2 pp132ndash135 2006

[22] T Ghosh T K Maity P Sengupta D K Dash and A BoseldquoAntidiabetic and in vivo antioxidant activity of ethanolicextract ofBacopamonnieri Linn Aerial parts a possiblemecha-nism of actionrdquo Iranian Journal of Pharmaceutical Research vol7 no 1 pp 61ndash68 2008

[23] B I Ognjanovic S D Markovic S Z Pavlovic R V Zikic AS Stajn and Z S Saicic ldquoEffect of chronic cadmium exposureon antioxidant defense system in some tissues of rats protectiveeffect of seleniumrdquoPhysiological Research vol 57 no 3 pp 403ndash411 2008

[24] L Barros M Duenas I C F R Ferreira P Baptista andC Santos-Buelga ldquoPhenolic acids determination by HPLC-DAD-ESIMS in sixteen different Portuguese wild mushroomsspeciesrdquo Food and Chemical Toxicology vol 47 no 6 pp 1076ndash1079 2009

[25] B Queiros J C M Barreira A C Sarmento and I C F RFerreira ldquoIn search of synergistic effects in antioxidant capacityof combined edible mushroomsrdquo International Journal of FoodSciences and Nutrition vol 60 no 6 pp 160ndash172 2009

[26] A Rohman S Riyanto N Yuniarti W R Saputra R UtamiandW Mulatsih ldquoAntioxidant activity total phenolic and totalflavaonoid of extracts and fractions of red fruit (PandanusconoideusLam)rdquo International FoodResearch Journal vol 17 no1 pp 97ndash106 2010

[27] W Zheng and S Y Wang ldquoAntioxidant activity and phenoliccompounds in selected herbsrdquo Journal of Agricultural and FoodChemistry vol 49 no 11 pp 5165ndash5170 2001

[28] A Błaszczyk and J Skolimowski ldquoPreparation of ethoxyquinsalts and their genotoxic and antioxidant effects on humanlymphocytesrdquo Arkivoc vol 2007 no 6 pp 217ndash229 2007

[29] F Pourmorad S J Hosseinimehr andN Shahabimajd ldquoAntiox-idant activity phenol and flavonoid contents of some selectedIranian medicinal plantsrdquo African Journal of Biotechnology vol5 no 11 pp 1142ndash1145 2006

[30] I I Koleva TA vanBeek J PH LinssenA deGroot andLNEvstatieva ldquoScreening of plant extracts for antioxidant activitya comparative study on three testing methodsrdquo PhytochemicalAnalysis vol 13 no 1 pp 8ndash17 2002

[31] M Atanassova S Georgieva and K Ivancheva ldquoTotal phenolicand total flavonoid contents antioxidant capacity and biologicalcontaminants in medicinal herbsrdquo Journal of the University ofChemical Technology and Metallurgy vol 46 no 1 pp 81ndash882011

[32] D Kozlowski P Marsal M Steel et al ldquoTheoretical investi-gation of the formation of a new series of antioxidant dep-sides from the radiolysis of flavonoid compoundsrdquo RadiationResearch vol 168 no 2 pp 243ndash252 2007

[33] T Tokura N Nakano T Ito et al ldquoInhibitory effect ofpolyphenol-enriched apple extracts on mast cell degranulation

in vitro targeting the binding between IgE and Fc120576RIrdquo Bio-science Biotechnology and Biochemistry vol 69 no 10 pp 1974ndash1977 2005

[34] T Enomoto Y Nagasako-Akazome T Kanda M Ikeda andY Dake ldquoClinical effects of apple polyphenols on persistentallergic rhinitis a randomized double-blind placebo-controlledparallel arm studyrdquo Journal of Investigational Allergology andClinical Immunology vol 16 no 5 pp 283ndash289 2006

[35] H Akiyama J Sakushima S Taniuchi et al ldquoAntiallergic effectof apple polyphenols on the allergic model mouserdquo Biologicaland Pharmaceutical Bulletin vol 23 no 11 pp 1370ndash1373 2000

[36] L Letenneur C Proust-Lima A Le Gouge J F Dartigues andP Barberger-Gateau ldquoFlavonoid intake and cognitive declineover a 10-year periodrdquo The American Journal of Epidemiologyvol 165 no 12 pp 1364ndash1371 2007

[37] M J Rodrıguez Vaquero P A Aredes Fernandez M C Mancade Nadra and A M Strasser de Saad ldquoPhenolic compoundcombinations on Escherichia coli viability in a meat systemrdquoJournal of Agricultural and Food Chemistry vol 58 no 10 pp6048ndash6052 2010

[38] P Jayaraman M K Sakharkar C S Lim T H Tang andK R Sakharkar ldquoActivity and interactions of antibiotic andphytochemical combinations against pseudomonas aeruginosain vitrordquo International Journal of Biological Sciences vol 6 no6 pp 556ndash568 2010

[39] M J Saavedra A Borges C Dias et al ldquoAntimicrobial activityof phenolics and glucosinolate hydrolysis products and theirsynergy with streptomycin against pathogenic bacteriardquoMedic-inal Chemistry vol 6 no 3 pp 174ndash183 2010

[40] M Obodai J Cleland-Okine and K A Vowotor ldquoComparativestudy on the growth and yield of Pleurotus ostreatusmushroomon different lignocellulosic by-productsrdquo Journal of IndustrialMicrobiology and Biotechnology vol 30 no 3 pp 146ndash149 2003

[41] M S Stankovic ldquoTotal phenolic content flavonoid concen-tration and antioxidant activity of Marrubium peregrinum Lextractsrdquo Kragujevac Journal of Science vol 33 pp 63ndash72 2011

[42] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of folin-ciocalteu reagentrdquo Methods inEnzymology vol 299 pp 152ndash178 1999

[43] C Quettier-Deleu B Gressier J Vasseur et al ldquoPhenolic com-pounds and antioxidant activities of buckwheat (Fagopyrumesculentum Moench) hulls and flourrdquo Journal of Ethnopharma-cology vol 72 no 1-2 pp 35ndash42 2000

[44] T Takao F Kitatani N Watanabe A Yagi and K SakataldquoA simple screening method for antioxidants and isolation ofseveral antioxidants produced by marine bacteria from fish andshellfishrdquo Bioscience Biotechnology and Biochemistry vol 58no 10 pp 1780ndash1783 1994

[45] Y Kumarasamy M Byres P J Cox M Jaspars L Nahar andS D Sarker ldquoScreening seeds of some Scottish plants for freeradical scavenging activityrdquo Phytotherapy Research vol 21 no7 pp 615ndash621 2007

[46] L Barros S Falcao P Baptista C Freire M Vilas-Boasand I C F R Ferreira ldquoAntioxidant activity of Agaricussp mushrooms by chemical biochemical and electrochemicalassaysrdquo Food Chemistry vol 111 no 1 pp 61ndash66 2008

[47] L Barros M Ferreira B Queiros I C F R Ferreira and PBaptista ldquoTotal phenols ascorbic acid120573-carotene and lycopenein Portuguese wild edible mushrooms and their antioxidantactivitiesrdquo Food Chemistry vol 103 no 2 pp 413ndash419 2007


[48] J Yang H Lin and J Mau ldquoAntioxidant properties of severalcommercial mushroomsrdquo Food Chemistry vol 77 no 2 pp229ndash235 2002

[49] C W Choi S C Kim S S Hwang et al ldquoAntioxidantactivity and free radical scavenging capacity between Koreanmedicinal plants and flavonoids by assay-guided comparisonrdquoPlant Science vol 163 no 6 pp 1161ndash1168 2002

[50] S S Lele A B Mali and K Khedkar ldquoEffect of gammairradiation on total phenolic content and in vitro antioxidantactivity of pomegranate (Punica Granatum L) peelsrdquo Food andNutrition Sciences vol 2 pp 428ndash433 2011

[51] F L Ola and G Oboh ldquoFood value of two commonly foundNigerian edible Mushrooms (Termitomycetes striatus and Ter-mitomycetes robustus)rdquo Journal of Technoscience vol 4 pp 1ndash32000

[52] F L Ola and G Oboh ldquoNutrient distribution and zincbioavailability estimation in some tropical edible mushroomsrdquoFoodNahrung vol 45 no 1 pp 67ndash68 2001

[53] F Sharififar G Dehghn-Nudeh and M Mirtajaldini ldquoMajorflavonoids with antioxidant activity from Teucrium polium LrdquoFood Chemistry vol 112 no 4 pp 885ndash888 2009

[54] G Min and L Chun-Zhao ldquoComparison of techniques for theextraction of flavonoids fromcultured cells of SaussureamedusaMaximrdquo World Journal of Microbiology and Biotechnology vol21 pp 1461ndash1463 2005

[55] A M Fernandes de Oliveira L S Pinheiro C K S Pereiraet al ldquoTotal phenolic content and antioxidant activity of someMalvacaea family speciesrdquo Antioxidants vol 1 pp 33ndash43 2012

[56] A Eghdami and F Sadeghi ldquoDetermination of total phenolicand flavonoid contents in methanolic and aqueous extract ofAchillea millefolliumrdquo Organic Chemistry Journal vol 2 pp 81ndash84 2010

[57] B A Iwalokun U A Usen A A Otunba and D K OlukoyaldquoComparative phytochemical evaluation antimicrobial andantioxidant properties of Pleurotus ostreatusrdquoAfrican Journal ofBiotechnology vol 6 no 15 pp 1732ndash1739 2007

[58] P Mattila K Konko M Eurola et al ldquoContents of vitaminsmineral elements and some phenolic compounds in cultivatedmushroomsrdquo Journal of Agricultural and Food Chemistry vol49 no 5 pp 2343ndash2348 2001

[59] K O Interaprichet and P Chirinang ldquoAmino acids antioxidantproperties of the oyster mushrooms Pleurotus ostreatus andPleurotus sajor-cajurdquo ScienceAsia vol 35 pp 326ndash331 2009

[60] J Y Wong and F Y Chye ldquoAntioxidant properties of selectedtropical wild edible mushroomsrdquo Journal of Food Compositionand Analysis vol 22 no 4 pp 269ndash277 2009

[61] S Phongpaicht S Preedana N Rungiondamai et al ldquoBiologyactivities of extracts from endophytic fungi isolated fromGarcinia plantsrdquo Immunology andMedical Microbiology vol 51pp 517ndash525 2007

[62] L Yu Wheat Antioxidants vol 45 John Wiley amp Sons NewYork NY USA 2008

[63] P Molyneux ldquoUse of DPPH to estimate antioxidant activityrdquoJournal of Science and Technology vol 26 p 2 2004

[64] A R Ahmad A M Munrsquoim and B Elya ldquoStudy of antioxidantactivity with reduction of DPPH radical and xanthine oxidaseinhibitor of the extract of Ruella tuberosa L leafrdquo InternationalResearch Journal of Pharmacy vol 3 no 11 pp 66ndash70 2012

Submit your manuscripts athttpwwwhindawicom

PainResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom

Volume 2014

ToxinsJournal of

VaccinesJournal of

Hindawi Publishing Corporation httpwwwhindawicom Volume 2014

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

AntibioticsInternational Journal of

ToxicologyJournal of


StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Drug DeliveryJournal of



Advances in Pharmacological Sciences

Tropical MedicineJournal of


Medicinal ChemistryInternational Journal of


AddictionJournal of



BioMed Research International

Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Autoimmune Diseases


Anesthesiology Research and Practice

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of


Pharmaceutics


MEDIATORSINFLAMMATION

of


inhibit the oxidative mechanisms However in more recentyears they play roles such as interacting with signal trans-duction pathways and cell receptors [10ndash12]

Naturally occurring antioxidants can be found in wholegrains fruits vegetables teas spices and herbs Mushroomshave also been reported as living organisms with antioxidantactivity which is correlated with their phenolic and polysac-charide compounds [13ndash15] Their global economic value isnow incredible and the reason for the rise in consumption is acombination of their value as food [16 17] and theirmedicinalor nutraceutical properties [18ndash21]

Harmful effects of disturbed antioxidant-prooxidant bal-ance can be largely prevented by intake of antioxidantsubstances [22 23] Antioxidants have already been found inmushroom materials and supplements [24 25] Due to theirnatural origin the antioxidants obtained from living organ-isms are of greater benefit in comparison to synthetic ones[26 27] The use of natural antioxidants from mushroomsdoes not induce side effects while synthetic antioxidantswerefound to have genotoxic effect [28 29]

Free radicals reactive nitrogen species (RNS) and otherreactive oxygen species (ROS) such as superoxide anionshydroxyl radicals and hydrogen peroxide are an entireclass of highly reactive species derived from the normalmetabolism of oxygen or from exogenous factors and agentssuch as pollutants and ultraviolet (UV) radiation Accordingto [30 31] oxidative damage to crucial cellular moleculesinduced by ROS has been implicated as a possible factorin the etiology of several human diseases such as cancercardiovascular diseases atherosclerosis Alzheimer diseaseand ageing [32] reported that irradiated flavonoid solutionsproduce chemical reactions between the solute and reactivespecies which are the result of the radiolyzed solvent Thesereactions according to [32] take place in radiolyzed alcoholsolutions a medium that is very rich in many differenthighly reactive species and that hosts specific reactionsThe reactivity of the solute (flavonoid) with the alkoxyradicals CH

3Olowast and CH

3CH2Olowast formed in methanol and

ethanol respectively and their carbon-centered isomersthe 1-hydroxy-methyl lowastCH

2OH and the 1-hydroxy-ethyl

CH3

lowastCHOH radicals operate to quickly repair the chemicalmodifications to biomolecules

Published studies by several researchers [33 34] showhow polyphenol compounds in our diets help to improveendothelial function which is a critical factor in preventingatherosclerosis They have also been shown to inhibit theabnormal blood platelet aggregation that cause most suddenheart attacks and strokes while fighting inflammation andsupporting healthy blood lipids They have the potential toreduce allergic conditions [35] by blocking the release ofhistamine (an irritating substance causing inflammation anditching) from the mast cells that mediate allergic reactionsLetenneur [36] reported that polyphenols from differentnatural sources may work synergistically when consumedtogether with benefits from the combination resulting inmore than the sum of the parts

Antimicrobial activity of polyphenols occurring in veg-etable foods medicinal plants and mushrooms has been

extensively investigated against a wide range of microor-ganisms Among polyphenols flavan-3-ols flavonols andtannins received most attention due to their wide spectrumand higher antimicrobial activity in comparison with otherpolyphenols and to the fact that most of them are able tosuppress a number of microbial virulence factors (such asinhibition of biofilm formation reduction of host ligandsadhesion and neutralization of bacterial toxins) and showsynergism with antibiotics [4] The antimicrobial propertiesof certain classes of polyphenols have been proposed eitherto develop new food preservatives [37] due to the increasingconsumer pressure on the food industry to avoid syntheticpreservatives or to develop innovative therapies for the treat-ment of various microbial infections [38 39] consideringthe increase in microbial resistance against conventionalantibiotic therapy

This study is to investigate the effect of gamma radiationon total phenolic content flavonoids and antioxidant activityin various extracts of the species Pleurotus ostreatus in differ-ent packaging materials using spectrophotometric methods

2 Materials and Methods

21 Chemicals Analytical ethanol methanol and sodiumhydrogen carbonate NAHCO

3were purchased from Sigma-

Aldrich USA Standards of phenolic acids (gallic acid[345-trihydroxybenzoic acid]) and of flavonoids potas-sium acetate quercetin [3310158404101584057-pentahydroxyflavone]and 22-dyphenyl-1-picrylhydrazyl (DPPH) were obtainedfrom Sigma Chemicals Co St Louis MO USA The Folin-Ciocalteursquos phenol reagent and Aluminium chloride (AlCl

3)

were from Fluka Chemie AG Buchs Switzerland

22 Mushroom Material Pleurotus ostreatus originally fromMauritius was cultivated on Triplochiton scleroxylon sawdustcomposted for 28 days supplemented with 1 CaCO

3and

10 wheat bran [40] at the Mushroom Unit of CSIR-FoodResearch Institute Accra Ghana Cultivation and harvestingwere from the period between September and December2012 Mushroom fruit bodies were solar-dried at temperaturerange of 20ndash40∘C to a moisture content of about 12 Driedmushroom parts were cut up to average size of 2 cm times 3 cmand stored in tight-sealed polythene (P) and polypropylene(Bx) containers at room temperature until needed

23 Irradiation of Mushroom Materials Forty (40) gramsof dried mushrooms were packed into various containersand irradiated at doses of 0 05 1 15 and 2 kGy at adose rate of 17 kGy per hour from a cobalt60 source (SLL515 Hungary) in air Absorbed doses were confirmed usingthe ethanol-chlorobenzene (ECB) dosimetry system at theRadiation Technology Centre of the Ghana Atomic EnergyCommission Accra Ghana

24 Preparation of Mushroom Extracts Mushroom extractswere prepared according to a standard protocol [41] Preparedmushroom material (10 g) was transferred to dark-coloredflasks and mixed with 200mL of solvents with different









3solution dissolved






times 100 (1)

IC50







TreatmentExtract

Aqueous(mgGAEg)

Ethanol(mgGAEg)

Methanol(mgGAEg)












0

005

01

015

02

025

03

0 005 01 015 02 025

Abso

rban

ce (120582

nm)


y = 12253x minus 00034

R2= 09888






TreatmentsExtracts


















0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30

y = 00058x + 00005

R2= 09863

Abso

rban

ce (120582

nm)




TreatmentsExtracts

Aqueous(mgGAEg)

Ethanol(mgGAEg)

Methanol(mgGAEg)













50lt



0

01

02

03

04

05

06

07

08

09

0 005 01 015 02 025

y = minus3309x + 07374

R2= 08507

Abso

rban

ce (120582

nm)




50)





50= 0064mgmLminus1


4 Conclusions




TreatmentsExtracts

Aqueous ()(mgGAEg)

Ethanol ()(mgGAEg)

Methanol ()(mgGAEg)












minus04 minus02 0 02 04 06 08 1 12 14

Methanol

Ethanol

Quercetin

Aqueous

Extr

acts

IC50 (mg mLminus1)

Figure 4 IC50




Acknowledgment


References








































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of









3solution dissolved






times 100 (1)

IC50







TreatmentExtract

Aqueous(mgGAEg)

Ethanol(mgGAEg)

Methanol(mgGAEg)












0

005

01

015

02

025

03

0 005 01 015 02 025

Abso

rban

ce (120582

nm)


y = 12253x minus 00034

R2= 09888






TreatmentsExtracts


















0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30

y = 00058x + 00005

R2= 09863

Abso

rban

ce (120582

nm)




TreatmentsExtracts

Aqueous(mgGAEg)

Ethanol(mgGAEg)

Methanol(mgGAEg)













50lt



0

01

02

03

04

05

06

07

08

09

0 005 01 015 02 025

y = minus3309x + 07374

R2= 08507

Abso

rban

ce (120582

nm)




50)





50= 0064mgmLminus1


4 Conclusions




TreatmentsExtracts

Aqueous ()(mgGAEg)

Ethanol ()(mgGAEg)

Methanol ()(mgGAEg)












minus04 minus02 0 02 04 06 08 1 12 14

Methanol

Ethanol

Quercetin

Aqueous

Extr

acts

IC50 (mg mLminus1)

Figure 4 IC50




Acknowledgment


References








































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of



TreatmentExtract

Aqueous(mgGAEg)

Ethanol(mgGAEg)

Methanol(mgGAEg)












0

005

01

015

02

025

03

0 005 01 015 02 025

Abso

rban

ce (120582

nm)


y = 12253x minus 00034

R2= 09888






TreatmentsExtracts


















0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30

y = 00058x + 00005

R2= 09863

Abso

rban

ce (120582

nm)




TreatmentsExtracts

Aqueous(mgGAEg)

Ethanol(mgGAEg)

Methanol(mgGAEg)













50lt



0

01

02

03

04

05

06

07

08

09

0 005 01 015 02 025

y = minus3309x + 07374

R2= 08507

Abso

rban

ce (120582

nm)




50)





50= 0064mgmLminus1


4 Conclusions




TreatmentsExtracts

Aqueous ()(mgGAEg)

Ethanol ()(mgGAEg)

Methanol ()(mgGAEg)












minus04 minus02 0 02 04 06 08 1 12 14

Methanol

Ethanol

Quercetin

Aqueous

Extr

acts

IC50 (mg mLminus1)

Figure 4 IC50




Acknowledgment


References








































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of


0

002

004

006

008

01

012

014

016

0 5 10 15 20 25 30

y = 00058x + 00005

R2= 09863

Abso

rban

ce (120582

nm)




TreatmentsExtracts

Aqueous(mgGAEg)

Ethanol(mgGAEg)

Methanol(mgGAEg)













50lt



0

01

02

03

04

05

06

07

08

09

0 005 01 015 02 025

y = minus3309x + 07374

R2= 08507

Abso

rban

ce (120582

nm)




50)





50= 0064mgmLminus1


4 Conclusions




TreatmentsExtracts

Aqueous ()(mgGAEg)

Ethanol ()(mgGAEg)

Methanol ()(mgGAEg)












minus04 minus02 0 02 04 06 08 1 12 14

Methanol

Ethanol

Quercetin

Aqueous

Extr

acts

IC50 (mg mLminus1)

Figure 4 IC50




Acknowledgment


References








































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of



TreatmentsExtracts

Aqueous ()(mgGAEg)

Ethanol ()(mgGAEg)

Methanol ()(mgGAEg)












minus04 minus02 0 02 04 06 08 1 12 14

Methanol

Ethanol

Quercetin

Aqueous

Extr

acts

IC50 (mg mLminus1)

Figure 4 IC50




Acknowledgment


References








































































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of






















































Volume 2014

ToxinsJournal of

VaccinesJournal of















AddictionJournal of






Autoimmune Diseases




Journal of


Pharmaceutics



of

nii korley kortei (Antioxidants)

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Transcript of nii korley kortei (Antioxidants)