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11
Antibacterial Activity of Aluminum Potassium Sulfate and
Syzygium Aromaticum Extract Against Pathogenic
Microorganisms

Ilha( )*ass +n,an Ph-
1
' )*.ulsa(ie /assan )lta0ee Ph-
21
'Na.ia /assan 2a.hu( 3Sc.
3

1.3icro*iolog, -et.' 4ollege of 3e.icine' 5ni6ersit, of +a*,lon. /illa' P.%. +o7 483' Ira9
:-(ail; ilha(alsae.i<,ahoo.co(
2.+ioche(istr, -et.' 4ollege of 3e.icine' 5ni6ersit, of +a*,lon. /illa' P.%. +o7 483' Ira9
:-(ail; a*.ulsa(ie68<g(ail.co(
3.+ioche(istr, -et.' 4ollege of Phar(acolog,' 5ni6ersit, of +a*,lon' /illa' P.%. +o7 4' Ira9
:-(ail; na.iasafaa<,ahoo.co(

Abstract
Natural ro.ucts ha6e *een use. for centuries in treating hu(an .isease. =he resent stu., ai(e. to .eter(ine
the anti*acterial a*ilit, of alu(inu( otassiu( sulfate (alu(! an. Syzygium aromaticum (clo6e! e7tract against
.ifferent t,es of athogenic *acteria in scientific wa,. )nti(icro*ial acti6it, of cru.e e7tract fro( two
co((onl, use. (e.icinall, (aterials' alu( an. clo6e were e6aluate. against four (icroorganis(s o6er
.ifferent concentration (1#' 2#' 3#' 4# an. "#! w>6 ?. 3ini(u( inhi*itor, concentration (3I4! an. the
.ia(eter of inhi*ition @one were .eter(ine. *, in 6itro *ioassa, using agar well .iffusion (etho. against S.
aureus' S. epidermidis' E. coli' Klebsiella pneumonia. =he two (e.icinal (aterials e7hi*ite. anti*acterial
acti6it, an. inhi*ition growth of gra( ositi6e an. gra( negati6e *acteria isolate. fro( .ifferent sites of
infection. =he inhi*ition effect of these e7tracts co(are. with stan.ar. anti*iotics cefota7i(e. In conclusions'
the results of resent stu., re6eale. that the alu( an. clo6e coul. *e use. as natural anti(icro*ial in .ifferent
wa,s.
Keywords:)lu(' clo6e' S,@,giu( aro(aticu(' S. aureus' S. ei.er(i.is' :. coli' 2le*siella neu(onia'
cefota7i(e

Introduction
Natural ro.ucts ha6e *een use. for a long ti(e in treating hu(an .isease an. the, contain (an, constituents of
theraeutic 6alue. Natural ro.ucts are en6iron(entall, safer' easil, a6aila*le an. chea (%suala et al., 2##$!.
)lu( (alu(inu( otassiu( sulfate! the cr,stalli@e. .ou*le sulfates with the for(ula 2)l(S%
4
!.12/
2
% are
generall, o.orless' colorless cr,stalline soli.s that turn white in air' which is use. in foo. reser6ation an.
antisetic for (an, rearation rocess such as icAling an. fer(entation an. water urification as a flocculants
(+estoon' 2#12!. 3oreo6er' alu( has reco((en.e. as categor, I acti6e ingre.ient in (outhwashes *, the
4ounter ).6isor, Panel of 5.S. Boo. an. -rug ).(inistration (B-)s! (%l(e@ et al., 1$$8!.
4lo6es or Syzygium aromaticum are .rie. aro(atic *u.s of unoene. floral of an e6ergreen tree 1#-2#( in
height' *elonging to the fa(il, 3,rtacceae' .istri*ute. in In.ia' In.onesia' Can@i*ar' 3auritius an. 4e,lon
()neDa an. Joshi' 2#1#!. =he .rie.' clo6e *u. contains free eugenol' eugenol acetate an. car,ohllen. )lthough
these su*stances (a.e u to $$? of the oil e7tract fro( .rie. flower-*u.s of Syzygium aromaticum' *eta-
car,ohellene' is still one of the (ain constituents of this oil e7tract' an. it has local anesthetic acti6it,. =he
essential oil of clo6e is a colorless or light ,ellowish e7tract o*taine. fro( .rie. flower-*u.s *, stea(
.istillation (Ehelara.ini et al., 2##1!. Phen,lroene eugenol is the other (ost i(ortant co(oun. resent in
Syzygium aromaticum .ue to which it has strong characteristic aro(a. +acteria' 3ol.' an. ,east growth coul. *e
inhi*ite. *, the a..ition of clo6e essential oil (+urt' 2##4!. =he essential oils of clo6e ha6e *een reorte. to
ossess nu(erous antio7i.ant an. anti(icro*ial roerties (Bu et al., 2##8!.
Material and Methods
est organism; *oth gra( ositi6e an. gra( negati6e (icroorganis(s were use. for the test. =he gra( ositi6e
organis( inclu.e S. aureus' S. epidermidis (sAin!' an. gra( negati6e *acteria inclu.e E. coli' Klebsiella
pneumonia (otitis (e.ia!. )ll *acterial strains were (aintaine. on freshl, reare. *loo. agar. =he *acterial
strains were isolate. fro( sAin an. otitis (e.ia an. i.entifie. in 3icro*iolog, -eart(ent' 4ollege of 3e.icine'
5ni6ersit, of +a*,lon' Ira9.
Pre!aration of different concentration of a"ueous extracts:
Alum extract:
-ifferent concentration of alu( (1#' 2#' 3#' 4# an. "#! w>6 ? was reare.. )lu( (aterial was urchase. fro(
the local (arAet at /illa cit,' Ira9 2#14' an. was i.entifie. in the 4ollege of 3e.icine' -eart(ent of
+ioche(istr,' 5ni6ersit, of +a*,lon. 4r,stals of alu( 2)l (S%
4
!.12/
2
% .issol6e. co(letel, in hot (1## (l!
.istille. water at $2
F
4' to reare final concentration of (1#' 2#' 3#' 4# an. "#! w>6 ? at / 3.6.
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&ol.4' No.1"' 2#14

12
#loves extract:
-rie. lants use. in this stu., were o*taine. fro( the local (arAet at /illa cit,' Ira9 2#14. -istille. water was
*oile.' .rie. lants were a..e. to the (1## (l! an. left to cool' this content were (i7e. *, the *len.er an.
filtere. to re(o6e the large un ho(ogeni@e. articles to get clear a9ueous e7tract. -ifferent concentration were
reare. (1#' 2#' 3#' 4# an. "#! w>6 ?. =he e7tracts were Aet at 4
F
4 until *e use..
In vitro antibacterial activity testing using agar well diffusion assay $##%s:
=he agar well .iffusion (etho. was use. for the .eter(ination of anti*acterial acti6it, of alu(inu( otassiu(
sulfate (alu(! an. Syzygium aromaticum a9ueous e7tract *, using *acterial isolates taAen fro( (sAin an. otitis
(e.ia! to e6aluate its effects on the isolate. *acteria. Goo full growth fro( *acterial isolate was inoculate. into
nutrient *roth incu*ate. at 38
F
4 for 18 hrs. =he *acterial susension was .ilute. with nor(al saline. ).Dust the
tur*i.it, an. co(are with stan.ar. tu*e (3cBarlan. nu(*er #."! to ,iel. a unifor( susension containing
1."H1#8 4B5>(l. 3uller-/inton agar was inoculate. with #.1 (l of *acterial inoculu(. 5sing corA *orer' wells
were (a.e on the culture. (e.ia. =he a9ueous e7tract was consi.ere. as (1#' 2#' 3#' 4# an. "#! w>6 ?
concentration. =hen' #.1 (l of a9ueous e7tract were a..e. to wells' then the lates left for 3# (in in refrigerator
at 4
F
4' thereafter' the, were incu*ate. at 38
F
4 for 24 hrs. =he inhi*ition @one was .eter(ine. to e6aluate the
acti6it, of a9ueous e7tract *, (easuring the .ia(eter in (illi(eter (4reso et al., 1$$#!.
&etermination of minimal inhibitory concentration 'MI#( of a"ueous Alum and clove extracts:
=o .eter(ine the lowest concentration of anti(icro*ial agent' the (ini(u( inhi*itor, test was .one to fi7e. the
*est concentration that inhi*it 6isi*le *acterial growth of e7eri(ental (icroorganis(s for an o6ernight
incu*ation erio. (Pan.e, an. Singh' 2#11!. =he 3I4 of a9ueous e7tract of alu( an. clo6es were .eter(ine.
*, *oth .ilution (etho.s. ) two fol. serial .ilution of a9ueous e7tract of alu( an. clo6e were reare..
Mc)arland tube standard '*.+(:
) *ariu( sulfate tur*i.it, stan.ar. solution e9ui6alent to #." 3cBarlan. stan.ar.s was reare. as .escri*e. *,
(+n,an' 2#13!.
,esults
Antibacterial activity of a"ueous extract of alum and clove in different concentrations.
In the resent stu., in6estigation anti(icro*ial effects of a9ueous e7tract *, agar well .iffusion assa,' in the
resent .ifferent concentration (1#' 2#' 3#' 4# an. "#! w>6 ? of *oth alu( an. clo6e an. inhi*ition @one
increase. *, increases of concentration. =he (ini(u( inhi*ition @one was o*ser6e. in alu( an. clo6e a9ueous
e7tracts' (4#' an. 33! (( resecti6el, an. the (ini(u( were in 2#? for each e7tract 2# an.18 resecti6el,.
=a*le 1 an. =a*le 2.
able - )nti*acterial )cti6it, of )9ueous :7tract of )lu( in -ifferent 4oncentrations.
Pathogens .one of inhibition 'mm(
cefat7i(e )lu(
1#? 2#? 3#? 4#? "#?
S. aureus # Negati6e 2# 2" 3# 3"
S. epidermidis 18 Negati6e 22 28 3" 4#
E. coli 2# Negati6e 2# 23 28 36
Klebsiella pneumonia 16 Negati6e 23 28 3# 38

able / )nti*acterial acti6it, of a9ueous e7tract of clo6e
Pathogens .one of inhibition 'mm(
cefat7i(e clo6e
1#? 2#? 3#? 4#? "#?
S. aureus # Negati6e 16 23 28 33
S. epidermidis 18 Negati6e 18 22 28 32
E. coli 2# Negati6e 2# 23 2$ 33
Klebsiella pneumonia 16 Negati6e 1$ 21 2" 3#
=he anti(icro*ial stu., *, agar well .iffusion (etho.s shows that the alu( an. clo6e has anti(icro*ial acti6it,
co(ara*le to that of co((ercial anti*iotic cefota7i(e.
he antibacterial activity of a"ueous extract of alum with time of incubation.
=he anti*acterial acti6it, of a9ueous e7tract of alu( was teste. against .ifferent t,e of gra( ositi6e an. gra(
negati6e (icroorganis(s with ti(e of incu*ation an. was shown inhi*ition ti(e of 24 hours. Bigure 1.
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&ol.4' No.1"' 2#14

13

)igure - :ffect of )lu( on +acterial Erowth in -ifferent =i(es.
he antibacterial activity of a"ueous extract of clove with time of incubation.
)lso' the anti*acterial acti6it, of a9ueous e7tract of clo6e was teste. against .ifferent t,e of gra( ositi6e an.
gra( negati6e (icroorganis(s with ti(e of incu*ation an. was shown inhi*ition ti(e of 24 hours. Bigure 2.

)igure / :ffect of 4lo6e on +acterial Erowth in -ifferent =i(es.

&iscussion
)lu(inu( otassiu( sulfate an. Syzygium aromaticum are 6alua*le (aterials source of (e.icinall, useful
co(oun.s ha. *een tra.itionall, use. for se6eral alications. )9ueous e7tracts were *eing goo. sources for
the *ioacti6e co(oun.s that e7hi*ite. goo. anti(icro*ial roerties. -ifferent concentrations of alu( were
teste. against four *acterial isolates an. the results showe. that the 2#? of alu( consi.er as the 3I4. )lso'
fro( the results' it was o*ser6e. that the *acterial growth inhi*ition was increase when the alu( concentration
increase. Since the *o., .oesnIt a*sor* alu(inu(' alu( coul. *e consi.er as a har(less (aterial an. has a low
to7icit, in la*orator, ani(als. +ut it has *een .ocu(ente. that the ingestion of 3# gra(s coul. Aill an a.ult
hu(ans. /igh le6el of alu( solution coul. cause. .istraction of gu( tissue' Ai.ne, .a(age an. high (ortalit,
rate .ue to intestinal *lee.ing (3allincAreo. che(ical. J. +aAr. Inc.' 2##$!. =he *acterial resistance to .ifferent
anti-(icro*ial agents consi.er as a (aDor ro*le( in (e.icine an. u*lic health. Nowa.a,s stu.ies focuse. with
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&ol.4' No.1"' 2#14

14
new anti*acterial targets which attenuating 6irulence factor an. caacit, of certain (icroorganis( to eli(inate
the infection (%ne.a et al., 1$$4!. =he uses of alu( as co(*inations with chlorhe7i.ine or er,thro(,cin
increase its acti6it, as anti*acterial agents than uses as single salt (4heong et al., 2##8!. /owe6er (ore stu.ies
a*out alu( an. its h,sical an. antio7i.ant roerties if there' as well as to7icit, are necessar,. In the resent
stu., the a9ueous e7tract of clo6e ha6e *een stu.ie. against so(e gra( ositi6e an. gra( negati6e *acteria
na(el, S. aureus' S. epidermidis' E. coli an. Klebsiella pneumonia. =he screening was erfor(e. *, agar well
.iffusion (etho.. =he a9ueous e7tract of clo6e was acti6e against all t,es of *acteria isolate. fro( sAin an.
otitis (e.ia an. effecti6e against a large nu(*er of other *acteria (4ail' 1$$6! an. ( 4u(an' et al. 2##$!.
(4haie* et al.' 2##8! ha6e reorte. the inhi*itor, effect of clo6e e7tract against Staphylococcus aureus *acteria.
)nti*acterial co(oun. of clo6e effecti6e against gra( ositi6e an. gra( negati6e *acteria .

#onclusions
=he reli(inar, stu., of this roDect re6eale. that the alu( an. clo6e can *e use. as natural anti(icro*ial in
.ifferent wa,s liAe natural .eo.orants (alu(! since is a 6er, natural an. health, alternati6e to all the che(ical
*ase. .eo.orants. Sices (clo6e! can *e use. as natural anti*acterial in foo. ro.ucts .ue to their less lethal
effects as co(are. to s,nthetic che(ical a..iti6es.

Ac0nowledgements
)uthors woul. liAe to thanA the resi.enc, of 3icro*iolog, an. +ioche(istr, -eart(ents in the 4ollege of
3e.icine' 5ni6ersit, of +a*,lon' Ira9' .ue to their facilities to acco(lish the tasA of this stu.,.

,eferences
)neDa' 2.R. an. Joshi' R. 2#1#. )nti(icro*ial acti6it, of Syzygium aromaticum an. its *u. oil against .ental
caries causing (icroorganis(s. J. Microbiol. 1' 1-".
+estoon' 3.B. 2#12. :6i.ance for feasi*ilit, of alu(inu( otassiu( sulfate (alu(! solution as a root canal
irrigant. J. Bagh. Coll. Dent. 24' 1-".
+n,an' I.). 2#13. ) no6el *iological rote of J-G-fucose in (utans grou stretococci. J. Nat. Sci. es. 3' 1-8.
+urt' S. 2##4. :ssential oils; their anti*acterial roerties an. otential alications in foo.s-a re6iew. !nt. J.
"ood Microbiol. $4' 223-2"3.
4ail' K. 1$$6. 4o(oun.s fro( Syzygium aromaticum ossessing growth inhi*itor, acti6it, against oral
athogenes. J. Nature, #rod. "$' $88-$$#.
4haie*(' 2.' /aDlaoui' /.' C(antar' =.' NaA*i' 2.).+.' Roua*hia' 3.' an. 3ah.ouani' 2.+. 2##8. =he che(ical
co(osition an. *iological acti6it, of essential oil' Eugenia cryophyllata (Syzygium aromaticum G. 3,rtaceae!;
a short re6iew. #hytother es. 21' "#1-"#6.
4heong' /.E.' Paul' K.S./.' :sther' P.:./.' +enn,' 2./.G.' an. Gai K.4. 2##8. Interactions of anti(icro*ial
co(oun.s with cross-linAing agents of alginate .ressings. J. antimicrobial. Chemoth.' 62' 1#"-1#8.
4reso' 3.:.' Ji(ene@' J.' Eo(is' :.' an. Na6arro' 4.1$$#. )nti(icro*ial acti6it, of essential oil of th,(us
ser,lloi.es su* secies gar.orensis. Microbial. 61'181-184.
4u(an R' +ersani )' 4aarro@ S; )nti-infla((ator, an. anti-noceti6e acti6ities of eugenol essential oil in
e7eri(ental ani(al (o.els. Braz. J. #harm.,2##$' 1$' 212-218.
Bu' L.L.' 4hen' G.' Shi' M.' Kang' C.' Sun' S.' an. :fferth' =. 2##8. )nti(icro*ial acti6it, of clo6e an.
rose(ar, essential oils alone an. in co(*ination. #hytother, es.' 21' $8$-$$4.
Ehelara.ini' 4.' Ealeottti' N.' -icesanr' 3.G.' 3a@@anti' E.' an. +artolini' ). 2##1. Gocal anaesthetic acti6it, of
(*eta!-car,oh,llene 1. J.!.$. %armaco. "' 388-38$.
3allincAreo. che(ical. J. +aAr. Inc. 2##$. 3aterial an. safet, -ata Sheet (3S-S!. )lu(inu( otassiu(
sulfate. 5S).
%l(e@' ).' 4an' /.' ),han' /.' an. %lur' /. 1$$8. :ffect of alu(-containing (outhrins in chil.ren for la9ue
an. sali6ar, le6els of selecte. oral (icroflora. J. Clin. #ediat. Dent. 22' 33"-341.
%ne.a' S.' =aAasaAi' =.' 2uriwaAi' 2.' %hi' L.' an. Loshi.a' /. 1$$4. 4hronic to7icit, an. tu(origenicit, stu.,
of alu(inu( otassiu( sulfate in +643 B1 (ice. !n &itro' 8' 281-288.
%suala' B.=.' I*i.ao o*o' 3.=.' %Aoh' /.I.' )ina' %.%.' Ig*asi' 5.=.' an. Nchiogu 3.:. 2##$. :6aluation of the
effecinc, an. safet, of otassiu( alu(inu( tetra o7osuihate in the treat(ent of tu*erculosis. European. J. Biol.
Sci. 1'1#-14.
Pan.e,' ). an. Singh' P. 2#11. )nti*acterial acti6it, of S,@,giu( aro(aticu( (clo6e! with (etal ion effect
against foo. *orne athogens. 'sian. J. plant. Sci. es. 1' 6$-8#.
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