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Evaluation of Antioxidant and Iron Chelating Activities of A Wild

Edible Oyster MushroomPleurotus Cystidiosus from Tanzania

Masoud *adi Muru+e

,e!artment o Molecular iology and iotec/nology ollege o Natural and 1!!lied Sciences 3umu5i

oad Mwalimu 7..Nyerere Mlimani am!us ni3ersity o ,ar es Salaam .%.o9 :#)$' ,ar es Salaam

;an5ania

udsm.ac.t5? /smuru+e>gmail.com

Abstract

;/is study was conducted in order to assess t/e le3els acti3ities and estalis/ t/e inluence o !ost/ar3est

storage met/ods on antio9idants o t/e giant wild edile mus/room Pleurotus cystidiosus.Standard analytical

met/ods were em!loyed to determine t/e le3el and acti3ities o antio9idants. It was ollowed y com!aring t/e

results in order to estalis/ t/e inluence o !ost/ar3est storage met/ods namely /eat drying ree5ing sun

drying rerigeration salt drenc/ing and de/ydration using silica gel eads. ;/e mus/room ca!s and sti!es were

treated se!arately. esults s/owed t/at irres!ecti3e o t/e !reser3ation met/od and !ortion o t/e mus/room

used P. cystidiosus e9/iited 3ery /ig/ 3alues o t/e numer o antiradical acti3ity units in ) mg o e9tract

(

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Figure ). Pleurotus cystidiosus

ni3ersity o ,ar es S

$#% Materials and Methods

2.1 ample collection

Fres/ sam!les o mus/room identi

tree at t/e Mwalimu 7..Nyerere

20)4. Field oser3ation and mor!/was a!!ortioned and suGected to di

200

sun drying or se3en days remus/room ca!s and sti!es were trea

2.2 Crude e!tract preparation

Met/anolic e9tractions were carriesti!e rom eac/ !reser3ation treatm

o met/anol.

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2.% &easurement of antiradical acti'ity

Measurement o antiradical acti3ity is ased on t/e !rinci!le t/at ,* ())-dienly-2-!icryl/dra5yl" radical in

its orm /as a c/aracteristic asorance at #)# nm w/ic/ disa!!ears ater its reduction y an antiradical

com!ound. ;/us t/e reduction o ,* can e monitored using s!ectro!/otometer y measuring decrease in its

asorance at #)# nm during t/e reaction met/od as descried y rand-Cilliams et al. ()''#".

In t/is study 40 Jl o met/anolic e9tract were added to )460 Jl o 0.00:$B ,*. 1sorance was measuredat #)# nm at time 0 and ater ) minute. ;/e antiradical acti3ity unit (1#)#" o eac/ e9tract was calculated

according to t/e eEuation=

1#)# (10@ 1)" @ (10@ 1)"D w/ere

1#)# 1ntiradical acti3ity o t/e e9tract

10 1sorance o t/e sam!le at t/e eginning o t/e reaction (0 minute"

1) 1sorance o t/e sam!le ater ) minute o t/e reaction

10 1sorance o t/e control sam!le at t/e eginning o t/e reaction

1) 1sorance o t/e control sam!le ater ) minute o t/e reaction.;/e numer o antiradical acti3ity units (

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al.20)2".

2.- Total $la'onoid Content;otal la3onoid content was determined according to al et al.(20)0" using Euercetin as standard. %ne () ml" o

eac/ mus/room e9tract was diluted wit/ 4.: ml o 80B aEueous et/anol containing 0.) ml o )0B aluminum

nitrate and 0.) ml o ) M aEueous !otassium acetate and let to stand or 40 minutes at room tem!erature (gi3e

t/e tem!erature range since room tem!erature is relati3e". ;/e asorance was determineds!ectro!/otometrically at 4.# nm using 3-3is model 6:0# 7enway . ;otal concentration was calculated

using Euercetin standard cur3e and e9!ressed as utin eEui3alent?)00 g o mus/room e9tract.

2. )etermination of /itamin C

;/e 3itamin content was determined y titration using 26 ,ic/loro!/eno indo!/enols met/ods descried y

lummer ()'8$".nown weig/t oground sam!le was mi9ed wit/ 2# ml o #B meta!/os!/oric acid solution

and s/a+en or :0 min. ;/e mi9ture was t/en iltered wit/ C/atman no. 42 ilter !a!er using suction !um!. )0

ml was !i!!eted rom t/e e9tract into 2#0 ml conical las+ and titrated against 0.02#B o 26 ,ic/loro!/enol

Indo!/enol reagents. ;/e amount o 3itamin in eac/ e9tract was calculated rom ollowing eEuation=

( )( )CH&

)00H&HIH1g)00!eracidascoricomg

2

)=

C/ere= 1 Euantity o ascoric acid (mg" reacting wit/ ) ml o 2 6 Indo!/enol

I 3olume o Indo!/enol (ml" reEuired or com!letion o t/e e9tract titration&) total 3olume o e9tract

&2 &olume o aliEuotC weig/t o ground mus/room

2.0 determination of Carotene and Lycopene Contents

K-Carotene and Ayco!ene contents were measured using t/e met/od o Nagata and Lamas/ita ()''2". %ne

/undred ()00" mg o a mus/room e9tract was s/a+en wit/ )0 ml o 1cetone-/e9ane mi9ture ('2=:" or ) minute

and iltered t/roug/ C/atman numer 4 ilter !a!er. ;/e asorance o t/e iltrate was measured at 4#: #0#and 66: nm. ;/e K @ carotene and lyco!ene content were calculated as=

Ayco!ene Jg?mg 0.04#8166: 0.:$21#0#@ 0.080614#:

K@carotene Jg?mg 0.2)6166: @ 0.:041#0# 0.4#214#:

% 'esults and (iscussion

%.1 Crude e!tract yieldesults s/owed signiicant 3ariations in crude e9tract yield among t/e dierent !reser3ation met/ods and

etween t/e ca! and t/e sti!e !arts o t/e same mus/room (Figure 2".

Figure 2.ercentage yield o e9tracts rom mus/room sam!les.

&alues are means o t/ree dierent measurements (MeanS, n:"

In t/e o3erall /ig/est yields were otained in t/e ca! !ortions !reser3ed y o3en drying ()).#'B" ollowed y

rerigeration (res/" and silica gel drying. Aowest yields in t/e ca! !ortions were recorded in ro5en sam!les

().40B" sun dried (#.#)B" and salt drenc/ed (#.68B" sam!les. Cit/ t/e e9ce!tion o ro5en and sun dried

sam!les crude e9tract yields were consistently /ig/er in t/e ca! t/an in t/e sti!e !ortions. &ery low yield

oser3ed in ro5en sam!les could e associated wit/ signiicant losses o mus/room content e9!erienced during

derosting !rocess o t/e ca! sam!les.%.2 Antiradical acti'ity

0.00

2.00

4.00

6.00

8.00

10.00

12.00

14.00

%Y

ield

Treatment

CAP

STIPE

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;/e results o antio9idati3e !ro!erties o t/e mus/room PleurotuscyctidiosussuGected to dierent !reser3ation

met/ods are !resented in ;ale ). emar+ale dierences in t/e antiradical acti3ity units (1#)#" numer o

antiradical acti3ity units in ) mg o e9tract (

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Figure :.,* o mus/room ca! and sti!e stored under dierent !reser3ation conditions

re3ious studies y im et al.(200'" /a3e s/own radical sca3enging acti3ities o oyster mus/rooms at 2 mg?ml

concentration to e 84.4 #4.# and 2'B or yellow !in+ and dar+-grey mus/room strains res!ecti3ely. Mau et al.

(2002" and Ao (200#" re!orted similar 3alues at 6.4 mg?ml (8).8B" and # mg?ml (68.4B" res!ecti3ely. Studies

y ;iu/wa (20)2" re!orted sca3enging acti3ity 3alues at concentration o ) mg?ml or Termitomyces

microcarpus ('8.6#B" T. letestui ('#.'#B" and T. titanicus (62.)4B". esults rom t/is study /a3e s/own

/ig/er radical sca3enging aility o Pleurotus cystidiosus at lower concentrations o 0.2 to ).0 mg?ml. ;/e totalantio9idants reEuired to decrease t/e initial ,* radical concentration y #0B (

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metal ion ound to t/e c/elator is c/emically inert. ;/us one o t/eir im!ortant roles is to !ro3ide !rotection o

our odies y remo3ing e9cess iron rom t/e lood /ence !re3ent !oisoning. ;/e !rinci!le and trend in t/e iron

c/elating aility was similar to t/at o t/e ,* radical sca3enging. Ferro5ine Euantitati3ely orms com!le9es

wit/ Fe2

. In t/e !resence o c/elating agent t/e com!le9 ormation is disru!ted t/us resulting in t/e reduction

o t/e red colour (al et al.20)0". eduction t/ereore allows estimation o c/elating aility o t/e mus/room

e9tract.Iron c/elating aility o P. cystidiosuse9tracts increased wit/ t/e concentration o e9tracts rom 0.02# to 0.40

mg?ml ut also 3aried wit/ t/e met/od o !reser3ation and t/e !art o mus/room ca! or sti!e (Figure 4". ;/e

/ig/est iron c/elating acti3ities at a concentration o 0.40 mg?ml were oser3ed in e9tracts rom o3en dried

sam!le ('8.:B" ollowed y silica gel dried sam!le ('6.'B" and rerigerated sam!le ('4.$B". ;/e least

c/elating ca!ailities were s/own y salt drenc/ed ca! and sti!e sam!les. enerally t/e sti!e /ad lower iron

c/elating aility t/an t/e ca!. ;/is trend 3ery muc/ mimics t/e ,* radical sca3enging acti3ities. /elating

eects on errous ion rom met/anolic e9tracts rom ot/er mus/rooms /a3e een re!orted. For e9am!le

Pleurotus citrinopileatus/a3e een re!orted y Aee et al. (200$" to c/elate errous ions y 82.)B at #mg?ml. Ao(200#" re!orted c/elating ailities oPleurotus eryngii, P. ferulaeandP. ostreatus at a concentration o # mg?ml

to e 4).4 to 64B. Similarly im et al.(200'" re!orted /ig/er c/elating ailities ($8-80B" o coloured oyster

mus/rooms at e3en lower concentration o ).# mg?ml. In t/is re!ort t/e studied mus/room Pleurotus

cystidiosus e9/iited /ig/er c/elating ailities y all t/e crude e9tracts (6$.) @ '8.:B" at a 3ery low

concentration o 0.4 mg?ml t/an so ar re!orted. ;/is is an indication t/at t/e mus/room studied is a 3eryeneicial ood source wit/ !owerul antio9idant ailities.

Figure 4. Ferrous iron c/elating aility o e9tracts romPleurotus cystidiosusca! and sti!e

;/e

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/ad est acti3ity 3alues salt drenc/ing and ro5en sam!les !erormed least. It is also e3ident rom t/e results t/at

/ig/er antio9idant and iron c/elating acti3ities were ound in t/e ca! !ortions o t/e mus/room t/an in t/e sti!e.

;/ese molecules may naturally e ewer in t/e irous sti!e t/an in t/e ca! !ortions or t/e met/anolic e9traction

met/od was not suitale or remo3ing out t/e molecules rom t/e irous sti!e material.

%.+ Phenolic and fla'onoid content

Mus/rooms contain 3arious !oly!/enolic com!ounds recogni5ed as e9cellent antio9idants not only ecause ot/eir aility to sca3enge ree radicals y single electron transer (Mau et al.2004 /eung et al. 200:" ut also

ecause o t/eir stale radical intermediates (S/a/idi and Canasundra )''2". In t/e !resent study !/enolic

content e9!ressed in mg gallic acid eEui3alent !er)00g mus/room e9tract diered among t/e dierent

!reser3ation met/ods and etween t/e mus/room ca! and sti!e (Figure #". &ery /ig/ !/enolic content (''.26

mg 1

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mus/room ca!s t/an in t/e sti!es. ;/e results suggest t/at t/e two mus/room !arts (ca! and sti!e" contain

dierent le3els o !/enolic com!ounds and la3onoids.

%. 9Carotene and lycopene

K-carotene and lyco!ene are !owerul antio9idants and !lay im!ortant roles o en/ancing 3itamin 1 in mammals

(al et al. 20)0" lowering ris+ o cancer and /eart diseases (ao and 1garwal 2000". In t/is re!ort t/e le3els o

K-carotene in t/e mus/room e9tracts were inluenced y t/e !ost-/ar3est storage in t/e same !attern.

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esults s/ow t/at e9ce!t or t/e sundried sam!les 3itamin 3alues did not 3ary signiicantly among t/e

dierent !reser3ation met/ods and etween t/e ca! and sti!e !art o t/e ruiting ody. &itamin 3alues ranged

etween #4.42 and 8:.6# mg?)00g eing lowest in sundried sam!les. ;/e 3alues were wit/in t/e range wit/

t/ose re!orted y Ms/andete and u (200$" on : mus/room genera (::-## mg?)00g". *owe3er muc/ /ig/er

3itamin contents /a3e een re!orted recently on mus/rooms. For e9am!le ;iu/wa (20)2" re!orted 200-480

mg?)00g in termitarian mus/rooms and ;iu/wa et al. (20)2" re!orted 60-200 mg?)00g in Coprinus cinereus. Ita!!ears drying y e9!osing mus/room sam!les on t/e sun radiation decreases 3itamin content.

1# Conclusion

;/is study re!orts or t/e irst time t/e e3aluation o t/e le3els and acti3ities o antio9idants o t/e wild edile

mus/room Pleurotus cystidiosus. P. cystidiosus /as s/own uniEuely /ig/ antiradical sca3enging and iron

c/elating ailities ma+ing it one o t/e mus/rooms wit/ t/e /ig/est antio9idants !ro!erties. Its enormous si5e

deliciousness and now /ig/ antio9idant ailities not only ma+e it a ood o t/e c/oice ut also en3isage its

domestication and !ossile u!-scaling and commerciali5ation.

Ac2noledgement

;/e aut/or is grateul to t/e ,e!artment o Molecular iology and iotec/nology (M" ollege o Natural

and 1!!lied Sciences (oN1S" ni3ersity o ,ar es Salaam (,SM" ;an5ania or !ro3iding laoratory

acilitiesD ,r. ,.,. ;iu/wa o M ,e!artment or identiying t/e mus/room,Mr. /arles weyunga o t/e,e!artment o otany or sam!le analysis w/ile Mr. ros!er aymond o t/e M ,e!artment is a!!reciatedor /is tec/nical ad3ice on data analysis. I am indeted to t/e anonymous re3iewer or constructi3e and

insig/tul comments and suggestions t/at /a3e im!ro3ed t/e manuscri!t.

'eferences

). 1c/arya . (200$". Medical !ro!erties o mus/room. 7n Ad'ances in &edicinal Plant :esearch, edited y

1c/arya SN and ;/omas ;

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)8.im M.L. Seguin . 1/n 7.. im 7.7. /un S.. im

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