FIELD OF INVEN'I'ION:
The present invention in general relates to s),nthesis of silver nanoparticles from
pltlnt source. Tlie present invention in particular relates to synthesis ol' silver
na~~oparticlefrso m . ~ I . I I L J ~,~I o~ h ilr\ and an herbal forriii~latio~M ih icli can be easilj
applied for local andlor systemic effect 011 nound and skin burn.
L~ISSCIIIPI'ION 01: 1'H E RE1,A'l'ED AR'I':
Skin burns and \\ounds are colnnion accidents in life cai~sed by thermal.
chemical, or electrical contact and physical trauma. All i~!juries to tlie skin
compron~ise tlie skin's ~neclianical integrity. the degree of compromise var),ing
froni superlicial disruption 01' the outer~iiost layer of the skin to complete
destruction of all layers of sliin. Because skin is tlie bod).'s tirst deltnse against
environ~iiental microorganisms as \veil as against fluid loss. a burn that dcst~.o!s
this barrier can cause life-threatening complications for the victini.
At the site ot' a wound, tlie victim can be affected by. for example. pain. local
inl;'ctlon, \found edema. loss ot' intravascular tluid \olumc di~c tu i~l~re,~\c.cI
\~ascular permeability. or a combination thereof. Sjstemic ef'lkct4 incluclc. 101
e\ample. liypovolemic sliocl\, s)steriiic inkction, respirator) tract i~i.jur!. 01 a
combination tliereof Tlie s)stemic effects ol'ten pose n greater threat to the l~lc01
the victim than do tlic localized effects.
Ei'fective burn and cliro~iic ~vound treatment must prevent infect~on as \\ell a4
prepare the ~\ounds ite for healing and closul-e. Wounds that do not readil! heal
can cause the suk~ject considerable physical. emotional. and social di5tres4 a\ \\ell
a5 great financial e\pense. Indecci. wounds that fail to heal propcrl! and beconic
infected may recluire excision of the affected tissue. A number of treat~ncnt
modalities have been developed as scientists' basic understanding of \\oi~nd\a nd
\ V O L I I I ~Il ealing ~iieclianisnis has progressed. Generally. the tlicrap! use5 dsugi
suc.11 as antiplilogi5tic<. antiseptics or analgesics. to mitigate inllam~nation.
prevcnt infection or relieve pain or disco~nfort of' patients. A biolog~cal drc\\ing
(such as pipkin) or non-biological dreising (sucli as 13iohranc1c) i \ al\o
emplo! ed to facilitate the ~vound healing.
7
-3-
Silver conipounds such as silver sulfadiazine in cream f'ormi~lations (c.g..
FlammazineE. silvadeneO) have also been used for L L O L I I It~re atment. I lo\\c\cr.
such formi~lationsa lso liave li~iiitedr csidual activity and have to be applied ti\ ice
a cia!. Bacterial resistance does develop lo these fi)rmulations, ancl. impaired reepitlielialization
lias also been observed. Bone marrow toxicity lias been obscrvcd
\vitIi silver sulfadiazine. primarily due to its propylene gljcol component.
Additionall), in sonie methods. silker itself is incorporated into tlie drc\sing
instcad of being applied as a separate t'ormi~lation. Co~itrolled and prolonged
release of silver to tlie woi~nd allows dressings to be changed less frequentl!.
Ho\vever, dressings have to be impregnated with large amount ol silver. \\li~cli
resi~lts in cytoto\icit\ to ~nammalian cells. Silver released ti.oni a comme~~ciall!
available wound-dressing (ActicoatTb') conta~ning~ ianocrystallincs il\cr ( I > l ~ n ne t
al. 2004. Burns 30 (supplement I):SI; herein i~icorporated b) rcfcrencr: in itentirety) is toxic to in vitro monolayer cell cultures of keratinoc?tes and
fibroblast\ (Poon et al. 2004, Burns 39: 140: ?'sop et al.. 2006. .I. 1 raunia 00:04X:
each herein incorporated by reference in its entirety).
I'ublication No. 624/L)EL/2006 provides an ayurvedic composition usel'i~l ill thc
trei~t~nenotsf bur11 i~i.jiiries~ IiicIic omprises: ( i ) oil ol'seeds ol'.\'c.srntlr~ni~~~ tlic.ri~t~
sesame, dried leave powder of Art~~hic1r1 ohi1i.s. was mised liomogenously ill a
ratio of 0.8 to 1.2: 0.15 to 0.3 to 0.5 respectivel~.
fada Khaloon. et a1 (Natural 1'1 o d ~ ~ c l
Sciences. pp. 286-290, 2003) that tall\s about ratari.jot which is regarded as one uf'
tlie irnporla~ith erbal d~.ugso f' indigenous \!stems of medicine. ?I lie root and root
stocl,. \vliicli f'or~ii Ilie acti~al drug, are considered to be all ~inthelni~ntic.
;tntip!retic and antiseptic. T h e are also claimed to be i~seti~inl burn. ecrcrila.
\vounds and eruptions. and are used for treating the diseases of eyes. broiicliitis.
abdomiiial pains. itch. etc.
-4-
I lo\\ever. the comple\ nature of patliologic \vounds. and thc lacl, of' signilicir~it
clinical progress based on current tlierapies. indicates the urgent need for ne\\ and
uncon\/c~itionap proaches.
Accordingll, tliere remains a need in tlie art for an effective and non-invasivc
regimen for nianaging i~i.jurie\t o skin caused by burn. acute or chronic \\ouilcIs.
~l'lieretore. the present invention provides a topical herbal formulation {'or skin
burns and wounds comprising silver nanoparticles extracted from Arnebia /iohlli.which lias improved efticacy over tlie alread~ marl prop! I mctli! lcellulosc)
(Metliocel 1<4) 3% \ilv slowlq in colloidal solution of silver nanoparticles. ~ v i t l i
-6-
continuous stirring till gel is Ibrmed. The gel is kept overnight to s\\ell. then lilled
in tubes and stored at 4°C. 0.2% \+/v Methyl parabe11 is used as prc\crvati\e.
In jet iinother en~botlin~cnotf the prcscnt invention. for thc pl.oduct~ono l
AI.IIO~I(IIIO /II/I\ root e\tract, 5 g AI.IICJ/)I~III ~ / I I / I \r oot po~zdero f 2 0 - ~ i i ~ sisl ia dcie~l
to ,I 500 mL Erlenme! e l flash \\ itli 100 1iiL sterile distilled \\atel allti then
eipobed to micro\\a~cf o ~3 miniites. l'lien tlie e\tract obtained I\ lil~ereclI n
hot condition \\/it11 lilter p'ipcl. to rcmokc tibroi~si~ iipurities.
111 still ;inother en~l)otli~ncoft the present invention. tlie s ! ~ i t l i c ~ i ~\ci l~~lc r
nanop:~rticlcs alone or in combin:itio~i \i it11 other s j ntlietic and/or herbal drug\.
Ins! be fol-mulated as an ointment \vIiicli can be easily applied li)r local anclior
s j stemic effect on \vound and skin burn.
l3lt~.ateXsli ll, \l?ectri~~ofi'ib io-s! ntliesized silver nanoparticle\:
I:ig. 3 il lusrrates 'FliM image of bio-syntlicsizcd silver nanoparticles:
Fig. 4 illi~stratesF 'I'II( absorption spectra of'biosyntliesized silver nanoparticlcs:
t ig. 5 illustrates comparison in healing activit) of control. 5tandal.cl and
11) clrogel of bios! ntliesi/ed \il\ecr ~la~ioparticles.
I)I and does not proiluce an). ud\er.se el'l'cc~.
'I'llc in\.eotion is tlcscril)ed in tletuil ~ i t hre t'ercnce to the e\i~rnl)les gi\c~l
bclo\\. T ~ I Cex iitlll)les i1t.e ~)ro\~itletjlu st to illustriite the ill\cntioll ;inti
thcrcfore, slioultl not I)c construct1 to li~nitt lie scol)e of tlie in\ cntion.
I'sainple 1 :
S! ntliesis ol'silver nanoparticles i~singci queous extract o f. -lruehiti 11o11ili.sr oo~s
-8-
,A~~/c>IhI Ot/r) I~/ I \ root\ \LCI.Cpu rchased from the local ~nnrl\eta ncl \\ashccl \\rtll
deioni~ed \\ater to rclnove an! impurities, dried in dad\ to completel) remove the
rno~\turea nd po\vdcrcd in a mixer grinder and then sieved usi~iga 20-mesh \le\c.
to get unilbr~ns ize range. For the production of extract. 5 g po\vde~\.\ as added to
;r 500 mL Erlenmejcr flask \\it11 100 mL sterile distilled \Later a11cl then e\poiccl
to micro~ave for 3 miniites. Then the sa\\ extract obtained \+a\ tiltered in hot
co~lilit~o\n\i t11 filter paper to rcliiove tjbroi~s i~iipurities. 'l'lie ~c\i~ltan~t l c , ~ r
e\tr.nct \ \ a \ i.~\ed ti)r tlic s! ntlicsis of \iI\/cr nanoparticles.
For reductio~io f Agf ions. I0 ml nclueous . lr17chtct t/ohtlt\ root c\tlact I\ a\ adclcd
to 50 1111 01' 10-' M ac,ucous AgNO; w l ~ ~ t iaon~d it he solution nli\turc \\;I\ c'\l~o\c~I
to niicro\\ave radiation at a li\ed ti-ecluencj of 2450 MH/ and po\\er ot 450
\vat[\. I'er~ndicallj. aliquots of tlic reaction solirtion \\ere renio~ecl crnd siibicctcd
to IJV-Vis spectroscopy ~neasurcments. The synthesized nalioparticlec \\ere
ccn~rili~gedat 8000 rpm ti)r 10 ~ninutc\ and subsequentl!/ re-dispcrsccl 111
deivn~zed\ \ater t\vice to get r~do l'any unbound biological molecules.
Exutnl~le2 :
I'~cpn~'atio0n1' tll'l\/lC hydrogel
-1 lle lijdrogcls \\ere prepared bj cold \i~spension method. tll'kl(' (Vlcthocel I<4)
3% \\I\ \\a\ \lo\\l! di\pcr5ed in colloidal \elution of'\ilvcr nanop:~rticlci \\ltri
continuou\ \tirring till gel \\as tbrmed. It \\as kept o\;crniglit to \\\ell. tIic11 lillcL1
in ti~besa nd stored at 3°C' until f'i~rtheru se. 0.2% ui/v Methyl parabcn \\as ii\cd
press1 \ a ~ ie\.
k:~i~t~ll3):l e
Cliaracterization of the synthesized silver nanopal-ticles
A. [I\'-vis at~alysiso f's?.r itllesizetl sil\~clt-i ;~noparticles
Ilic pr.ogl'e\s ol' iil\er ~ianopart~cleIib ~~iiatio\n$ a\ monitorccl tr\lng I I' \ I spectra by cmplojing a UV-visible Shimadzu double bean1 s p e c ~ ~ ~ ) p l l o t ~ ~ ~ ~ e t ~ r
1800 ope~.ated at u rc\olution of' 1 11111 u ill1 optical path length 01' I0 Inm O1>ticCil
dc~isit! \\YIS meahurcd b! diluting the colloidal solution t\\o t~lncs1 1\i11dg cio~i~/cd
\\ atel
-9-
The color change ol' the colloidal solution into reddi4i brou 11 indicatecl tlie
formation of silver ~ianoparticles. The intetisitj of tlie color of' reaction mi\ti~rc
increased evenl~\\ it11 tinie o f ' m i c r ~ ~ a vc\ep osilre. tlii~sc ontir~ii~ntlige l i j ~ ~ i i ~ ~ t i o n
of \il\/er nanoparticles. It \\as due to [lie exitation of tlie surliice pl;~\rnon
0 \ibration in metal nnnoparticles. I'he reduction of pure Ag' Ions to Ag I\;Imonitored b j measuring IJV-\is spectril~n of the reaction media at regular
intc~.\,~l\.1 lie metal ions reduction occurred \cry rapidly and reduction of ~iiosot l
the Ag' ions was completed in 210 seconds. Absorption spectra of' \il\cr
nnnoparticles Ibrmed in the rcaction mi\ture \vas obtained at diffkrent \vavelcrlgtIi
(11111)1 .e. 250 to 550 nm. fhe particle slio\\ed sharp absorption ma\imu~iip eal, at
4 I0 nlil and gi.aduallj dec~.eased\ \ liile uc~vclengtilni creased (F'ig. I )
13. 'I'ri~nsrnission I ;~llo\vcdto dr! prior to ~neasureme~it.
rl'lic. p;trticles \{:ere almost spherical in shape \\lit11 diameters in tlie r;llige i ~ i 1' 0~0
Iim and were \veil dispersed (Fig 3).
C. 1iouric.1. tr:lnsfor~n infraretl spectroscopy (F'I'IK) analysis of ~)~ntllcsizctl
sil\.cr nanol)articles
The F'I'IR spectl.urn \\,as recorded using Dri~ker- I-TIR Alpha instru~iicnta [ ;I
~.ewli~tioonf 4 c~n-II.t \\as obsci.ved that colloidal silver nnnopr\~.ticlesw li~ticili
was cstremcl!, stable l'o~. more than four months. It has been reported that proteins
car1 bind silver nanoparticles t l i r ~ ~ ~frgelei aniine groups nnci pro\ ide a goocl
protecting environment for ~netalij,drosols during the gro\\itli processes. I:ig 4
slio\\< some pru110~11iceadb sorbance halids at around 3400-3430 c r n - ' ( ~ -I- l
I st~.ctcliing) and 1620- 1650 cm- (aromatic rings), suggesti~ig tlic pl.esclicc of'
~xoteiriso n [he surfhcc of' Ag-core particles. I'he peal; around .3400-3430 ~111.'0 1'
-10-
p1,11it proleins in the nanoparticles shell \+as rni~ch nal.ro\\cr I lii\ ~ ~ 1 1be1
c\pl,~~ncda s I-I-bonds can be i'ormed bct\cee~i tlic amicle g~.oi~psA. s plant
~iiolcculesa dsorbed onto the silver nanoparticles surface, tlie amide groi~piin tend
to Io1.111 slr.origer bond\ \\ill? Ag atonis. \\hicIi \+ill breal, most ol' tlie I I-bondbct\\c.cn the W - l i groi~p\a nd Icad lo the narroning and b l u e - i l i ~ l t \01 ' tlic ' l ~ i i ~ c i c .
bond. It bas concli~clccl tliat the At.richto trohilt\ proleins hacl adsol bccl a i n I,i!c~
occ~ ~ l i c s~lcer nnno particles. \\hicIi in turn stabili~ed the sjnrhcs~re~l
~ i : ~ ~ ~ ~ ~ ~ ) ~ i r t i c l c s .
I). X-ray tliffri~ctio(~X~It 1)) itn;rlysis of synthesized silver nanopiirticles
1-01 XI<[) measurements, tlie 5) nthesi/ed nanoparticles \\ere ccntr.iftlged at 8000
rpm 1;)r I0 ~ninutesL inil subsequentlq re-dispersed in deionized \\alcr t\\ ice to gct
r~col I':11ij ~111bounbdio logical ~nolecules.I 'hc solid residi~cla !cr. containing \ i l \ c r
nanop;wticles \\/as re-dispersed in sterile dcionised water for three timcb to I . C I ~ - I O \ ~
i~~iattac~liebciol logical componenl\ to the 4ilrface of' ~ i a ~ i o l ? i ~ r t itclial ct ~.i i.c not
re\po~lsible for bio-Iu~i~tio~iiili~atoiro tc~;r pping. 'I he pure r-e\iduc \\:I\ LIriccl
pc1.12ctl! in an oven overnight at 60°C'.
tor the crystallinit!, studies, powcler was irsed for XRD stud!. A l31.ul,er S-ra!
clil't'ractomerer (11-2 I)ha\e) \\it11 ('11 I<(tr) radiation \\as uscd 10 a\\c\\ the
I- .~ g .7 \ho\vj tlie XRL) patterns o f silver nanoparticles sjntlies~/cdt luin root
e\tl;icl o f .lr-rrs/~~rulo l?i/i\. A nirmber o f Braggs reflection \ \ i l l 1 20 ~ : ~ l u colj
38 30. 44 50, 64.60. ,tnd 77 50 corrcsponcling to I I I. 200. 230. ;inel 3 l I 4ct ol
lattice plalies \\ere obser\~ecl (JCI'IIS. sil\ier file no. 04-0783) 7Tlic. X-la!
cliflr;~c~tion (XRI)) anal) \is sIio\? ecl dil'l'r'~ction pcahs corrcspond~ng to
structure anci cr~stallinit) o f siI\ier nanoparticles.
.Zg~\r\ ell d~t'l'i~siomne thod \\as ubcd to evaluate the bactericidal activit! o1'\iI\ei
llano-colloid \elution. Sterile nutrient agar medium \\as pourecl into \tcr~lc 1'ctl.i
pli~ti.\ ;111d; 1110\\ed to \oIicIilj . TIic I'ctsi l?lc~te~\ c r ie11 ciiI1att'il ;it 37"( 1'01- 24
- I I -
hours to cliecl, for bterility. The medium \\/as seeded \\ it11 the olganivn c u l ~ ~(~ I r c
~iil)b! pour plate ~iietliod.B ores were made 011 tlie medium using 4terilc horcr.
10. 20. 40 and 80 pgiml of silver rianoparticles was added to tlie re\pcctive bore\.
I'hc I'etri plates \\c~.cI, ept in refrigerator at 4°C' lhr 30 rnini~tc\l or d~l'lus~on.
Alicr clil'fi~sion the I'ctri plates were i~icubutcd at .37"(' ti)r 34 hour.\ and /one ol
inhibition \\ere ob$er~cda nd mea\i~rcd.
Nit110 silver slio\ved activit!, to\vnrds Cir;~m p0sitiL.e ah \\ell as (ir;\ln negatilc
b;~c~c~.hiatrla in. A goocl zone ot' inlii1)ition \\as observetl for Grnrn ~~eg:~li(\I::c.
caoli) bacterial strains than Cram positive (.5'. c~llr.eo.vb) acterial strains. '['he ionc ol'
iilllihition i~~creahcc\\l'i t11 incre:\sing concc~itratio~91i ' silvcr nanoparticles ( 1 ;~blc
I ) . I.'rom [lie data it \\,as inl'errcd [hat tlie s!~tithesizrd silver nanop:\~.ticle>\ \ c I . ~ '
el'l'ecti\e 1;11. both tlpe of' microbes. Moreover, the zone of' inhibition obsc.r\~cd
;iroi111cGI ram i~egati\.eb acterial strains uas found to be reproducibl~o f the biggel.
si/c 111311 the (;rani positive bacterial strains.
E\ii~lll)le5 :
I:\,~lu,~tioonl 'wound lieali~igp otential - 1-\ci\ion Woi~ndM oclel
lie:~ltli!/ Albino M'istar rats ( 150-200 g) 01' either sex Lvere obtainecl l'rom anillla1
lioi~hc.Y I L': 'I' C'oIIegc. (;~.tfi\t~N'~oi.c i;l. 'I'he~w ere lioi~scdi nili\~idirall! uilclcr
sta~l~lartlla borator! conclitions and ti.d \\it11 pellet rodent dict aid \\;t[cr ncl
libiti~~nC.' I'C'SEA g~~idelinc\sy ere aclliered to during the mainlenancc and
csl>criiiic~l(tA pp~'o~ln;~o.l 1 I 2 1 lac! CI'CSEAl07).
/\~iiliial!,\ \crc di\ idetl into three ~ ~ O L(coI11Itr0~1. SIIII~CIa;nIc~l Jr est). 411i1nalh\\ itl.c.
a~i;res~lietize\d\i t11 anaesthetic ether and placed on operation tablc in its I I ~ I L L I I . ~ I I
position. An impression \\as madc on the tkwsal region i~si~ai gr 0i11ic1s ~ ; I I 01'3.5
c11i diameter. .T'he skin of tlie impressed area was excised to li~ll thiclincs.; to
o1x1i11:I \~OLIIK iIre ;^ ol.i113oi1t4 00 scl 1111ii. I llc \ V O L I I I L\\~c~r c t~.i~cc(c1l1 1 ~ ~ ; I I ~ ~ ~ : I I . C I I ~
sheet on thc day 01' \\'oi~ndinga rid subsccluc~i~loyn claj 0. 7. 14 ;\ncl 2 I . 'l'he
I X I . C ~ I I ~ ; I0~1'C W OLIIIII cli)st~~a.necl period ol'cpithelialisation \\ere rcco~ded.
- 13-
l'lic control gl-ot~p\ \a> not sub.jected to an! medication and \+as lcti open. the
stanclirrd g~.oup\ \a\ trc;rtcd topically wit11 Sil\cr sulfrrdi;~zine& the tc\t groi~p\ \ a trc'rlctl top~cally \\ it11 li! d~.ogcI.
I is. 5 \lio\\s the potcntinl of' \\)ound hcal~ng acti\lit! of contrul. \tanclal.d alicl
li>drogel of bios~ntliecired silver nanoparticlcs.
'I';lble 1: sho\zs ~lntimicrobiala ctivity of'sil~eln-a noparticles against 1; c,oli and .\'
~ / l / l ' ( ' / / \ .
Conccn tration .Avcruge zone of' inllibition Averilgc zonc ot' inhil~itio~~
(~lg/lllI) (111111) (111 111)
I<. ~ 0 l i s. itllrell~
I 0 I 0 I 5
20 17 I 0
40 I8 17
XO 2 0 I9
Nulllcrous nlotlilicwtions i~ntl atli~ptiltio~osf ' the s!stem of' the prcscrit
in\.cntion nil1 be apl)arcot to those skilled in the art, antl thus it is inteotlctl
by tllc al)l)endetl clainls to cover all such modilic;~tioas alltl ;itlal)tittionwhich fill1 within tllc true spirit and scol)c of this inbeation.
-13-
lIIplic;itions ill
biotcclinologj . .I Dispersion Sci7feclinol 26: 389-3 14
3. Chan WCM' (2006) 13ionanotecl1nology progress and advirnccs. 13iologi)
13loocl Mnrrou Transpla~ltatio~1l2 : 87-9 I.
3. 13iossrlier L. Astr-i~c L) (2000) (;old nanoparticles in nirno meclici~ic:
I'rc17ar;rtion. imaging, diagnostics. tlierapies and toxicity. C'lie~n Soc I~.i osy~i~Ii0~1s'i~si lvern a11op:irticlcs sing the l1111guh
I ; ' l i . s c l r i r ~ ~ l.r~ . ~ ~ I I I ~ I O C I I IMI I Iu. ter Iici
synthesis ol' silverna~ioparticles using culture supernatants 01'
I:ntet-obactesi;~a: novel biological upp~.oacli.I 'r.c)cc.>s I3i~)clicrn4 3:c11 0 2.;.
I I . ~ia1~cle:~-'1'or1~csJcL1e. yf 'arsons JCi. 1)okken I<. I'eraltu-Vide; .IK. I'roiani
I I . Santiago 1.' Josc-Yaca~na~Ml (3002) For~nation ant1 g~.o\~t0h1' Au
nanoparticlcs i~isidol ive all':llli~ plants. Nano L.ett 2 :-307-40 1 .

-15-
23.Arora A. Il:lstogi I). Gupta I). Culra-jani ML (2012) 1;inctic.s 1111cl
tIiesniod!~namics of djje extracted from Arnebia nobilis llech. 1'. on \\ ool.
lSl~~'17-3R7 : 9 1-97
34. Morton SJP. hlalone HI-1. Evalualion of vulne~ary aclivit) by an open
\\lound psoceclusc in rats. Arch. 1111. pharmacoclyn. 'l'hcr. 1072: 100: 117-
126.
2 5 , Mitra IiN. Illiis 1'1; (2008) In sit11 preparation of'goltl ~iarioparticlc..; 0 1
\/ar\,ing shape in moleci~li~hry drogel of peptide a~npliiphilcs.. I l'l~!'\.
C'licm. C' 1 12:8 159--8 166.
20. (;ole A, I>asli C'. lianiak~~i~linVa. nS ainkas S. Mandale A. llao M. Sns~r!
21 (200 1 ) I'cpsin-golil colloid co~i.iugares: prcl->nratic)~~l. I l i ~ ~ . ; ~ ~ l ~ r i / . i ~ t i r ) ~ i .
ant1 e n ~ ~ ~ ~ nacatiivi cil~ ,L. angniuir 17: 1674--I6 79.
- 1 GWE
CLAIM:
1. An herbal formulation comprising ~ilvern anopar-ticles and a niethod for
synthesis of same using 31"17(1/71(1 noh111\ root eltract comprising:
a. adding 10 nil aqueous root eltract to 50 ml of 10-' M aqucou,
AgN03 solutio~l:
b. exposing the abo\e solution ~niltut-eto micro~zaver ~d~ntioJI ~Li~
fixed frequencq:
c. centrifuging the synthesized nanoparticles at 8000 rpm for about
10 minutes:
d. redispersing the nanoparticles in deionized water tmice to get rid OF
any unbound biological molecules:
e. loading the synthesized silver nanoparticles in hydrogel.
2. The herbal formulation colnprising silver nanoparticles and a method
synthesis of same using ..llnehitr nohi1i.v root extract as claimed in claim I .
wherein Arnebia nobili.~ro ot extract is exposed to niicrowave radiation at
a fixed frequency of 2450MHz and power of 450 \vatts.
3. The herbal formulation comprising silver nanoparticles ancl a niethod f\)r
synthesis of same using .-ll-~rc>ht~~tro hil~ro\o t extract as claimed in clailn I.
wherein the preparation of hydrogel-silver nanoparticles co~npriscb the
steps of:
a. dispersing HPMC (hydroxypropyl methylcellulose) (hlcthoccl ti41
3% W/V slowly in colloidal solution of silver nanoparticles. Ivith
continuoi~ss tirring r i l l gel is formed:
b. keeping the gel o\erniglit to s~vell, then filling i t in tubes artci
storing at 4 degree C.
4. The herbal formulation comprising silver nanoparticles and a nierhod fi)r
synthesis of same using .4meAicr 1tohili.5 root extract as claimed in clai~n I .
wherein 0.2% wlv MethcI paraberi is used as preservative.
5. The herbal formulation comprising silver nanoparticles and a method f(31.
synthesis of same using .4r11cl/?itlrl ohili~ro ot extract as claimed in claim I .
-17-
wherein the method for the production of .-lr.nehin noh111\ root c\t~-dct
comprises the steps of:
a. adding 5 g Ar.nchrir i~ohili, root powder of 20-rnesli to a 500 nil
Erlennie~erfl ash \\it11 100 mL sterile tiistilled \barer:
b. exposing the above solittion to microwave for 3 minutes;
c. filtering the ran eitroct in hot condition nit11 filter paper to remo\c
fibrous impurities.
6. The herbal formulation comprising silver nanoparticles and a method for
synthesis of same using Arnrbia nobilis root extract as claimed in claim 1 .
wherein the silver nanoparticles are spherical shaped. fcc structured wirh
diameter range of 20 to 80 nm.
7. The herbal formulation comprising silver nanoparticles and a niethod fbr
synthesis of same using Arnrbia nobilis root extract as claimed in claim 1. wherein the silver nanoparticles shows antibacterial activity against both
gram negative particularly, E. coli and gram positive particularly S. aureus strains.
8. The herbal formulation comprising silver. nanoparticles and a method for synthesis of same using Arnrbia nobilis root extract as clairned in claim 1 .
wherein the colloidal silver nanoparticles solution is stable fix more than
four months.