Determination of Antioxidant and Antimicrobial Activities of Rumex

cri spus L. Extracts

Al i Y l d r m,*
,
Ahmet Mavi ,

and Ays e Aydan Kara

Kaz m Karabeki r Egi ti m Fakul tesi , Ki mya Egi ti mi Anabi l i m Dal , and Fen-Edebi yat Fakul tesi ,
Bi yol oji Bol umu, Ataturk U ni versi tesi , 25240 Erzurum, Turkey
The anti oxi dant acti vi ti es, reduci ng powers, 2,2-di phenyl -1-pi cryl hydrazyl (DPPH) scavengi ng
acti vi ti es, amount of total phenol i c compounds, and anti mi crobi al acti vi ti es of ether, ethanol , and
hot water extracts of the l eaves and seeds of RumexcrispusL. were studi ed. The anti oxi dant acti vi ti es
of extracts i ncrease wi th i ncreasi ng amount of extracts (50-150 g). However, the water extracts
of both the l eaves and seeds have shown the hi ghest anti oxi dant acti vi ti es. Thus, addi ti on of 75 g
of each of the above extracts to the l i nol ei c aci d emul si on caused the i nhi bi ti on of peroxi de formati on
by 96 and 94%, respecti vel y. Al though the anti oxi dant acti vi ty of the ethanol extract of seed was
l ower than the water extract, the di fference between these was not stati sti cal l y si gni fi cant, P >
0.05. Unl i ke the other extracts, 75 g of the ether extract of seeds was unabl e to show stati sti cal l y
si gni fi cant anti oxi dant acti vi ty, P > 0.05 (between thi s extract and control i n that there i s no extract
i n the test sampl e). Among al l of the extracts, the hi ghest amount of total phenol i c compound was
found i n the ethanol extract of seeds, whereas the l owest amount was found i n the ether extract of
seeds. Li ke phenol i c compounds, the hi ghest reduci ng power and the hi ghest DPPH scavengi ng
acti vi ty were found i n the ethanol extract of seeds. However, the reduci ng acti vi ty of the ethanol
extract of seeds was 40% that of ascorbi c aci d, whereas i n the presence of 400 g of water and
ethanol extracts of seeds scavengi ng acti vi ti es were about 85 and 90%, respecti vel y. There were
stati sti cal l y si gni fi cant correl ati ons between amount of phenol i c compounds and reduci ng power
and between amount of phenol i c compounds and percent DPPH scavengi ng acti vi ti es (r ) 0.99, P
< 0.01, and r ) 0.864, P < 0.05, respecti vel y) and al so between reduci ng powers and percent DPPH
scavengi ng acti vi ti es (r ) 0.892, P < 0.05). The ether extracts of both the l eaves and seeds and
ethanol extract of l eaves had shown anti mi crobi al acti vi ti es on Staphylococcus aureus and Bacillus
subtilis. However, none of the water extracts showed anti mi crobi al acti vi ty on the studi ed
mi croorgani sms.
Keywords: Antioxidant activity; reducingpower; antimicrobial activity; Rumexcrispus L.; DPPH
scavenging
I NTRODUCTI ON
Reacti ve oxygen speci es (ROS) and reacti ve ni trogen
speci es (RNS) are vari ous forms of acti vated oxygen and
ni trogen, whi ch i ncl ude free radi cal s such as superoxi de
i ons (O
2
-
), hydroxyl (OH

), and ni tri c oxi de radi cal s

(NO

) as wel l as non-free-radi cal speci es such as hydro-

gen peroxi de (H
2
O
2
) and ni trous aci d (HNO
2
) (1-3). I n
l i vi ng organi sms vari ous ROS and RNS can form by
di fferent ways. Normal aerobi c respi rati on, sti mul ated
pol ymorphonucl ear l eukocytes and macrophages, and
peroxi somes appear to the mai n endogenous sources of
most of the oxi dants produced by cel l s (4-6). Exogenous
sources of free radi cal s i ncl ude tobacco smoke, i oni zi ng
radi ati on, certai n pol l utants, organi c sol vents, and
pesti ci des (7-10). Free radi cal s can cause l i pi d peroxi -
dati on i n foods that l eads to the deteri orati on of them
(11, 12). Oxi dati on does not affect onl y l i pi ds. ROS and
RNS may cause DNA damage that coul d l ead to muta-
ti on (13, 14). I n addi ti on, ROS and RNS have been
i mpl i cated i n >100 di seases, i ncl udi ng mal ari a, ac-
quired immunodeficiency syndrome, heart disease, stroke,
arteri oscl erosi s, di abetes, and cancer (6, 15-17). When
produced i n excess, ROS can cause ti ssue i njury.
However, ti ssue i njury can i tsel f cause ROS generati on
(18). Neverthel ess, al l aerobi c organi sms, i ncl udi ng
human bei ngs, have anti oxi dant defenses that protect
agai nst oxi dati ve damages and numerous damage re-
moval and repai r enzymes to remove or repai r damaged
mol ecul es (8, 19-21). However, thi s natural anti oxi dant
mechani sm can be i neffi ci ent; hence, di etary i ntake of
anti oxi dant compounds wi l l become i mportant (5, 17,
22, 23). Al though there are some syntheti c anti oxi dant
compounds, such as butyl ated hydroxytol uene (BHT)
and butyl ated hydroxyani sol e (BHA), whi ch are com-
monl y used i n processed foods, i t has been reported that
these compounds have some si de effects (24, 25). I n
addi ti on, i t has been suggested that there i s an i nverse
rel ati onshi p between di etary i ntake of anti oxi dant-ri ch
foods and the i nci dence of human di seases (26, 27).
Therefore, research i nto the determi nati on of the natu-
ral anti oxi dant source i s i mportant.
* Author to whom correspondence shoul d be addressed (e-
mai l ayi l di ri m6l @yahoo.com; tel ephone +90-442 2314020; fax
+90-442 2184172).

Kazim Karabekir Egitim Fakultesi, Kimya Egitimi Anabilim

Dal i .

Fen-Edebi yat Fakul tesi , Bi yol oji Bol umu.

4083 J. Agric. Food Chem. 2001, 49, 40834089
10.1021/jf0103572 CCC: $20.00 2001 American Chemical Society
Published on Web 08/04/2001
Rumexcrispus L. i s a perenni al wi l d pl ant, and i t has
l ength of 30-150 cm. I ts basal l eaves are acute and
narrowl y l anceol ate to obl anceol ate. Peti ol es are canal i -
cul ate above, and thei r i nfl orescence i s dense. The
pedi cel s are l onger than the frui ti ng peri anth segments
and arti cul ate bel ow the mi ddl e. Frui ti ng peri anth
segments cordate to tri angul ar, at l east one tubercul ate,
4-5 3-4 mm. I t can grow on banks, marshes, and
waste pl aces and can be found at al ti tudes of up to 2300
m (28, 29). I ts young l eaves are cul ti vated i n spri ng and
used as a vegetabl e, whereas the seeds of thi s pl ant are
cul ti vated i n the summer and used i n Turki sh fol k
medi ci ne.
I n the present study, the anti oxi dant acti vi ti es of
ether, ethanol , and hot water extracts of R. crispus L.
were determi ned. Al so, i t was of i nterest to determi ne
the anti mi crobi al acti vi ti es of these extracts, and thi s
was carri ed out by the di sk di ffusi on method.
MATERI ALS AND METHODS
Preparation of Extracts. The l eaves of R. crispus L. were
cul ti vated i n May, and the seeds of thi s pl ant were cul ti vated
at the end of June, i n the central part of Turkey (Gol ova,
Si vas), and were l eft on a bench to dry. The dri ed sampl e was
chopped i nto smal l parts wi th a bl ender. Ether extracti on was
performed wi th a Soxhl et apparatus unti l extracti on sol vents
became col orl ess. Extracti on was fol l owed by fi l trati on and
evaporati on of the fi l trate to dryness by a rotary evaporator
at 30 C.
The resi due, after ether had been removed from i t, was
mi xed wi th ethanol i n an screw-capped Erl enmeyer fl ask and
was shaken i n a shaker to obtai n ethanol extract. Extracti on
was conti nued unti l extracti on sol vents became col orl ess. The
obtai ned extracts were fi l tered, the fi l trate was col l ected, and
then ethanol was removed by a rotary evaporator at 40 C to
obtai n dry extract.
The resi due obtai ned after fi l trati on was l eft i n a dark pl ace
at room temperature to become dry. Dri ed resi due was mi xed
wi th boi l i ng di sti l l ed water and then was sti rred on a hot pl ate
for 15 mi n and subsequentl y was fi l tered. Fi nal l y, the fi l trate
was freeze-dri ed i n a freeze-dryer at 5 mHg pressure at -50
C.
Determination of theAmount of Total Phenolic Com-
pounds. Thi s was carri ed out as descri bed previ ousl y (30).
Bri efl y, 0.1 mL of extract sol uti on (contai ns 500 g of extract)
was transferred to a 100 mL Erl enmeyer fl ask, and then the
fi nal vol ume was adjusted to 46 mL by the addi ti on of di sti l l ed
water. Afterward, 1 mL of Fol i n-Ci ocal teu reacti ve (FCR)
(Fl uka) was added i nto thi s mi xture, and after 3 mi n 3 mL of
Na
2CO3 (2%) was added. Subsequentl y, the mi xture was
shaken on a shaker for 2 h at room temperature, and then
absorbance was measured at 760 nm. Pyrocatechol (Si gma)
was used as the standard for the cal i brati on curve. The
phenol i c compound content was determi ned as pyrocatechol
equi val ents usi ng the fol l owi ng l i near equati on based on the
cal i brati on curve:
A i s the absorbance, and C i s pyrocatechol equi val ents (g).
Reducing Power. Thi s was carri ed out as descri bed
previ ousl y (31). Bri efl y, extracts (50-500 g) i n 1 mL of
appropri ate sol vents were mi xed wi th 2.5 mL of phosphate
buffer (0.2 M, pH 6.6) and 2.5 mL of potassi um ferri cyani de
[K
3Fe(CN)6] (1%), and then the mi xture was i ncubated at 50
C for 30 mi n. Afterward, 2.5 mL of tri chl oroaceti c aci d (10%)
was added to the mi xture, whi ch was then centri fuged at 3000
rpm for 10 mi n. Fi nal l y, 2.5 mL of the upper l ayer sol uti on
was mi xed wi th 2.5 mL of di sti l l ed water and 0.5 mL of FeCl
3
(0.1%), and the absorbance was measured at 700 nm. I ncreased
absorbance of the reacti on mi xture i ndi cates i ncreased reduc-
i ng power.
DPPH Radical Scavenging Activity. Thi s was carri ed
accordi ng to Bl oi s method wi th a sl i ght modi fi cati on (32).
Bri efl y, a 1 mM sol uti on of DPPH (Fl uka) radi cal sol uti on i n
ethanol was prepared, and then 1 mL of thi s sol uti on was
mi xed wi th 3 mL of extract sol uti on i n ethanol contai ni ng 50-
400 g of dri ed extract; fi nal l y, after 30 mi n, the absorbance
was measured at 517 nm. Thi s acti vi ty i s gi ven as percent
DPPH scavengi ng that i s cal cul ated as
Antioxidant Activity. Anti oxi dant acti vi ty was determi ned
accordi ng to the thi ocyanate method. Bri efl y, each sampl e
(contai ni ng 50-150 g of extract) i n 0.5 mL of di sti l l ed water
was mi xed wi th 2.5 mL of l i nol ei c aci d (Fl uka) emul si on (0.02
M, i n 0.02 M, pH 7.0, phosphate-buffered sal i ne, Si gma) and
2 mL of phosphate-buffered sal i ne (0.02 M, pH 7.0) i n a test
tube and i ncubated i n darkness at 37 C. The amount of
peroxi de was determi ned by readi ng the absorbance at 500
nm after col ori ng wi th FeCl 2 and thi ocyanate at i nterval s
duri ng i ncubati on (33).
Antimicrobial Activity. To be abl e to determi ne anti mi -
crobi al acti vi ti es, Staphylococcus aureus ATCC 25923, Can-
dida albicans ATCC 60193, Escherichiacoli ATCC 25922,
Bacillus subtilis ATCC 6633, and Pseudomonas aeruginosa
ATCC 10145 were used. These were obtai ned from Karadeni z
Techni cal Uni versi ty, Medi cal School , Department of Mi cro-
bi ol ogy and Cl i ni cal Mi crobi ol ogy, Trabzon, Turkey.
Anti mi crobi al acti vi ti es were determi ned by usi ng the di sk
di ffusi on method (34). Bri efl y, 50 mg of dri ed extract sampl e
was di ssol ved i n 50 mL of a conveni ent sol vent, and then 500
L of thi s sol uti on were transferred i nto 6 mm di ameter
anti mi crobi al suscepti bi l i ty bl ank di sks (Oxoi d). Afterward,
di sks were l eft at room temperature to dry. Sol vent absorbed
di sks were used as control . As a standard peni ci l l i n G (Oxoi d)
anti bi oti c absorbed di sks were used.
Test mi croorgani sms grown on nutri ent agar (Oxoi d) (for
bacteri a) or on potato dextrose agar (Oxoi d) (for fungus) for
24 h were transferred i nto 12 cm di ameter Petri di shes
contai ni ng sol i d medi a by a steri l e cotton wool covered wand.
Subsequentl y, these mi croorgani sms were spread over the
surface of the sol i d medi a as a thi n fi l m. Fi nal l y, Petri di shes
were i ncubated at 37 C for 48 and 72 h for bacteri a and
fungus, respecti vel y, and then i nhi bi ti on zones were observed.
Statistical Analysis. Each datum represents the mean of
three di fferent experi ments i n each of whi ch two measure-
ments were made. SPSS 9.0 software was used for stati sti cal
cal cul ati ons. Val ues of P < 0.05 were consi dered to be
si gni fi cant and val ues of P < 0.01 very si gni fi cant.
RESULTS AND DI SCUSSI ON
Amount of Total Phenolic Compounds. As can be
seen i n Tabl e 1, the amounts of phenol i c compounds are
hi gher i n the ethanol and water extracts of seeds than
i n the l eaves extracts wi th the same sol vents. Among
al l of the extracts, the hi ghest amount was found i n the
ethanol extract of seeds and the l owest amount was
measured i n the ether extract of seeds.
A ) 0.0034C - 0.058, R
2
) 0.9996
Table 1. Amount of Total Phenolic Compounds in R.
cri spus L. Extracts
500 g of extract
av absorbance
(760 nm)
pyrocatechol
equi v (g)
control 0.000
ether extract of seeds 0.020 9.30
ethanol extract of seeds 0.736 220
water extract of seeds 0.252 77.6
ether extract of l eaves 0.028 11.6
ethanol extract of l eaves 0.028 11.6
water extract of l eaves 0.040 15.2
% DPPH scavengi ng )
[(control absorbance - extract absorbance)/
(control absorbance)] 100
4084 J. Agric. Food Chem., Vol. 49, No. 8, 2001 Yldrm et al.
Reducing Power. Li ke the amount of phenol i c
compounds, the reduci ng power of ethanol extract of
seeds of R. crispus L. was the hi ghest among al l of the
extracts, and i t i s i ncreased as the amount of extract
i ncreased (Fi gure 1). Al though the water extract of seeds
has shown a hi gher reduci ng power than the other
extracts, i t was l ower than that of the ethanol extract
of seeds, and thi s di fference was stati sti cal l y si gni fi cant,
P < 0.05. There was no si gni fi cant di fference among
l eaves extracts, whi ch were shown to have a l ower
reduci ng power compared wi th seeds extract.
The amount of phenol i c compound was the hi ghest
i n ethanol extract of seeds, and water extract of seeds
was the second hi ghest among al l of the extracts; si mi l ar
resul ts were obtai ned i n reduci ng power acti vi ti es.
Hence, combi ni ng these two resul ts, we can suggest that
there may be a rel ati onshi p between the amount of total
phenol i c compounds and reduci ng powers. I n fact, there
i s a stati sti cal l y si gni fi cant correl ati on between these
two, r ) 0.99, P < 0.01.
To be abl e to compare the reduci ng powers of these
extracts wi th a known reduci ng reagent, the reduci ng
powers of 500 g of each of extract and ascorbi c aci d
(35) were measured. As the absorbance val ues were l ow
i n the presence of 150 g of extracts, 500 g was chosen
i n compari son. As can be seen i n Tabl e 2, the reduci ng
powers of al l of these extracts were markedl y l ower than
that of ascorbi c aci d. However, among al l of the extracts
the ethanol extract of seeds has shown the hi ghest
reduci ng power, whi ch was 40% that of ascorbi c aci d.
Neverthel ess, al l of the extracts have shown hi gher
acti vi ti es than control , and these di fferences were
stati sti cal l y si gni fi cant (P < 0.01).
DPPH Radical ScavengingActivity. Percent DPPH
scavengi ng acti vi ti es of ethanol and water extracts of
seeds were concentrati on dependent. Li ke the amount
of phenol i c compounds and reduci ng power, the hi ghest
percent DPPH scavengi ng acti vi ty was shown by the
ethanol extract of seeds, and the second hi ghest acti vi ty
was determi ned i n the water extract of seeds. Al though
some acti vi ti es were determi ned i n the water and
ethanol extracts of l eaves, i t was too l i ttl e compared
wi th those of the ethanol or water extracts of seeds.
Thus, i n the presence of 400 g of water or ethanol
extracts of l eaves scavengi ng acti vi ti es were 12 and 4%,
respecti vel y, whereas i n the presence of 400 g of each
of the above sol vent extracts of seeds, scavengi ng
acti vi ti es were about 85 and 90%, respecti vel y (Fi gure
2). Unl i ke water and ethanol extracts, there were no
acti vi ti es i n ether extracts at the studi ed (50-400 g)
concentrati ons. There was a stati sti cal l y si gni fi cant
correl ati on between the amount of phenol i c compounds
and percent DPPH scavengi ng acti vi ti es, r ) 0.864, P
< 0.05, and al so between reduci ng powers and percent
DPPH scavengi ng acti vi ti es, r ) 0.892, P < 0.05.
Antioxidant Activity. I n the present study, the
anti oxi dant acti vi ti es of ether, ethanol , and hot water
extracts of l eaves and seeds of R. crispus L. determi ned
by usi ng the thi ocyanate method i n that amount of
peroxi des formed i n emul si on duri ng i ncubati on i s
determi ned spectrophotometri cal l y by measuri ng the
absorbance at 500 nm. Hi gh absorbance i s an i ndi cati on
of hi gh concentrati on of formed peroxi des. Therefore,
l ow absorbance i ndi cates hi gh anti oxi dant acti vi ty.
The anti oxi dant acti vi ti es of ethanol extracts of l eaves
i ncreased wi th i ncreasi ng amount of extract (Fi gure 3).
As si mi l ar resul ts were obtai ned wi th the other extracts,
onl y a si ngl e datum i s gi ven to prevent repeti ti on. As
can be seen i n thi s fi gure, even the addi ti on of 50 g of
dri ed ethanol extract i n the l i nol ei c aci d emul si on was
Figure 1. Reduci ng powers of ether, ethanol , and water extracts of seeds and l eaves of R. crispus L.
Table 2. Comparison of Reducing Powers of Ascorbic
Acid and R. cri spus L. Extracts
a
sampl e absorbance (700 nm)
control 0.045
ascorbi c aci d 3.540
ether extract of seeds 0.182
ethanol extract of seeds 1.52
water extract of seeds 0.724
ether extract of l eaves 0.294
ethanol extract of l eaves 0.158
water extract of l eaves 0.130
a
Fi ve hundred mi crograms of extract or ascorbi c aci d was used;
i n the control there was no extract. Hi gh absorbance i ndi cates hi gh
reduci ng power.
Antioxidant and Antimicrobial Activities of Rumex crispus L. J. Agric. Food Chem., Vol. 49, No. 8, 2001 4085
abl e to reduce the formati on of peroxi des. However,
there was no stati sti cal l y si gni fi cant di fference between
50 g of extract-contai ni ng sampl e and control , i n whi ch
there i s no extract, P > 0.05. However, there were
stati sti cal l y si gni fi cant di fferences between 75 g of
extract-contai ni ng sampl e and control , P < 0.05.
Al though the anti oxi dant acti vi ti es of extracts i n-
creased wi th i ncreasi ng amount of extracts, there was
no stati sti cal l y si gni fi cant di fference between 100 g of
extract-contai ni ng sampl es and 150 g of extract-
contai ni ng sampl es i n both l eaves and seeds extracts,
P > 0.05. Therefore, >150 g of extract was not studi ed.
To be abl e to compare the anti oxi dant acti vi ti es of
water, ethanol , and ether extracts of both of the l eaves
and seeds, peroxi de formati on was measured i n the
presence of 75 g of each of the above extracts. As can
be seen i n Fi gure 4, al l of the l eaves extracts showed
stati sti cal l y si gni fi cant acti vi ty. Among al l of the l eaves
extracts the hi ghest anti oxi dant acti vi ty was measured
i n the water extract. However, there were no stati sti -
cal l y si gni fi cant di fferences between l eaves extracts, P
> 0.05.
Al l of the seed extracts showed an anti oxi dati ve effect
(Fi gure 5). However, unl i ke the others, the anti oxi dant
acti vi ty of the ether extract of seed was not stati sti cal l y
si gni fi cant. Thus, the di fference between 75 g of ether
extract contai ni ng sampl e and control was not si gni fi -
cant, P > 0.05. The most effecti ve anti oxi dant acti vi ty
was seen i n the water extract (Fi gure 5). Al though the
di fference between thi s extract and ethanol was not
si gni fi cant, P > 0.05, the di fference between the water
and ether extracts was stati sti cal l y si gni fi cant, P <0.05.
Figure 2. Percent DPPH scavengi ng acti vi ti es of ether, ethanol , and water extracts of seeds and l eaves of R. crispus L.
Figure3. Anti oxi dant acti vi ty of ethanol extract of l eaves of R. crispus L. I ndi cated amounts of dri ed extracts were added to the
l i nol ei c aci d emul si on.
4086 J. Agric. Food Chem., Vol. 49, No. 8, 2001 Yldrm et al.
Water extracts of both l eaves and seeds have shown
the hi ghest anti oxi dant acti vi ti es, and there was no
stati sti cal l y si gni fi cant di fference between these, P >
0.05.
To i ndi cate the anti oxi dant acti vi ti es of these extracts
more cl earl y, absorbance val ues at 98 h of i ncubati on
and the equati on bel ow were used to cal cul ate the
i nhi bi ti on percentages of peroxi de formati on by 75 g
of these extracts:
As can be seen i n Tabl e 3, the hi ghest acti vi ti es were
i n both of the water extracts, whereas the l owest
acti vi ti es were shown by the ether extract of seeds.
The l eaves of R. crispus L. are cooked i n water.
Therefore, i t was i nteresti ng to fi nd that the hi ghest
anti oxi dant acti vi ty was shown by the water extract of
l eaves and seeds. However, there were no stati sti cal l y
si gni fi cant di fferences among the anti oxi dant acti vi ti es
of l eaves extract.
Al though the hi ghest anti oxi dant acti vi ti es were
shown by water extracts of l eaves and seeds, the hi ghest
Figure 4. Compari son of anti oxi dant acti vi ti es of ether, ethanol , and water extracts of l eaves of R. crispus L. Seventy-fi ve
mi crograms of each of the extracts was added to the l i nol ei c aci d emul si on.
Figure 5. Compari son of anti oxi dant acti vi ti es of ether, ethanol , and water extracts of seeds of R. crispus L. Seventy-fi ve
mi crograms of each of the extracts was added to the l i nol ei c aci d emul si on.
% i nhi bi ti on)
[(control absorbance -
sampl e absorbance)/control absorbance] 100
Table 3. Percent Inhibition of Peroxide Formation by 75
g of Extract
sampl e % i nhi bi ti on
ether extract of seeds 8.8
ethanol extract of seeds 61
water extract of seeds 96
ether extract of l eaves 90
ethanol extract of l eaves 81
water extract of l eaves 94
Antioxidant and Antimicrobial Activities of Rumex crispus L. J. Agric. Food Chem., Vol. 49, No. 8, 2001 4087
reduci ng power was found i n the ethanol extract of
seeds. I n addi ti on, the DPPH scavengi ng acti vi ty and
the amount of phenol i c compound were hi ghest i n the
ethanol extract of seeds. Phenol i c compounds coul d
easi l y donate a hydroxyl hydrogen due to resonance
stabi l i zati on (36). Combi ni ng thi s fact wi th the obtai ned
resul ts we coul d suggest that as the amount of phenol i c
compounds i ncreases, reduci ng power i ncreases as wel l .
Reduci ng power of a compound i s rel ated to el ectron
transfer abi l i ty of that compound. Therefore, the reduc-
i ng capaci ty of a compound may serve as a si gni fi cant
i ndi cator of i ts potenti al anti oxi dant acti vi ty (35).
However, we have previ ousl y found that thi s may not
al ways be the case (37).
The hi gh potenti al of phenol i c compounds to scavenge
radi cal s may be expl ai ned by thei r abi l i ty to donate a
hydrogen atom from thei r phenol i c hydroxyl groups (14).
The accepted way to i nhi bi t l i pi d oxi dati on by anti oxi -
dants i s through thei r anti radi cal acti vi ty
The new radi cal formed (A

) can mai nl y fol l ow radi cal -

radi cal i nteracti on to render stabl e mol ecul es vi a radi cal
di sproporti onati on wi th abstracti on of an atom by one
radi cal to another (23):
However, the anti oxi dant acti vi ti es of putati ve anti -
oxi dants have been attri buted to vari ous mechani sms;
among these are preventi on of chai n i ni ti ati on, bi ndi ng
of transi ti on metal i on catal ysts, decomposi ti on of
peroxi des, preventi on of conti nued hydrogen abstrac-
ti on, and radi cal scavengi ng (38, 39).
I n the present study i t was found that the hot water
extracts of both the l eaves and seeds of R. crispus L.
have shown the hi ghest anti oxi dant acti vi ty. Thi s i s a
wi del y grown pl ant and i s used as a vegetabl e, especi al l y
i n eastern Turkey. As i t has been reported that there
i s an i nverse rel ati onshi p between the di etary i ntake
of anti oxi dant-ri ch foods and the i nci dence of a number
of human di seases (26, 40), these resul ts are i nteresti ng.
I n addi ti on, anti oxi dant compounds, whi ch are respon-
si bl e for thi s acti vi ty, coul d be i sol ated and then can be
used as food addi ti ves to del ay the deteri orati on of food
due to oxi dati on. Therefore, research i nto the determi -
nati on of anti oxi dant-ri ch foods i s i mportant.
Antimicrobial Activity. The ether extracts of both
the l eaves and seeds and ethanol extracts of l eaves had
shown anti mi crobi al acti vi ti es on S. aureusATCC 25923
and B. subtilis ATCC 6633, whi ch are Gram-posi ti ve
bacteri a (Tabl e 4). However, these extracts di d not show
detectabl e anti mi crobi al acti vi ty on E. coli ATCC 25922
and C. albicans ATCC 60193, whi ch are Gram-negati ve
bacteri a. Al so, none of the extracts showed anti mi crobi al
acti vi ty on P. aeruginosa ATCC 10145, whi ch i s a
fungus. These resul ts coul d suggest that some of the
studi ed extracts are effecti ve agai nst the Gram-posi ti ve
bacteri a, whereas none of the extracts i s effecti ve on the
Gram-negati ve bacteri a and fungi . The reason for thi s
coul d be the subject of further studi es.
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a
extracts of l eaves extracts of seeds
mi croorgani sm ether EtOH H2O ether EtOH H2O
S. aureus ATCC 25923 1 0.8 - 1.1 - -
B. subtilis ATCC 6633 1.1 0.8 - 1.1 - -
C. albicans ATCC 60193 - - - - - -
E. coli ATCC 25922 - - - - - -
P. aeruginosa ATCC 10145 - - - - - -
a
(-) i ndi cates no i nhi bi ti on zone; the numbers i n the tabl e
i ndi cate the di ameters of i nhi bi ti on zones (cm).
DPPH

+ (AH)
n
f DPPH-H + (A

)
n
DPPH

+ A

f DPPH-A
A

+ A

f A-A
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Recei ved for revi ew March 15, 2001. Revi sed manuscri pt
recei ved June 11, 2001. Accepted June 12, 2001. We express
si ncere thanks to the Ataturk Uni versi ty Rectorate for fi nan-
ci al support (Grant 1999/49 Uni versi ty Research Fund).
JF0103572
Antioxidant and Antimicrobial Activities of Rumex crispus L. J. Agric. Food Chem., Vol. 49, No. 8, 2001 4089