I
Anne\ rr
((lause 3.3.1.1)
I)ETERNII\ATION OT MIITALAXYL _M CONTtrNT
Crntenl is dctcrmincd Li! '(lus (lhfomatograph)' (GC), Lrsing an intemrl slandaf,l
Icllo\\cd b) Clhjr.rl HIll C external standard mclhod lo detemine the fatios 01R anil
isonrers. Final assa)' fcprrrtcd .rs tolal of R and S isontcrs.
.\nncxurcA-Ilartl
Dctrrmination of Chlorolhilonil and Nlctnlaxll content
c PIiINCIPLE
\- L | \lelala\\ I contcnt is dctcnnined b\ Gas chronatosraphic rncthod using intemel
sLiudrrd rnelhod.
\-2 .\PPAIIATT S
\-l.j Oes Chrornarogfafh (Cjl-(l tequipFed\\ith l-lD an,:l couplcd 1o I pfinter plotler curir
irlcgralor or an\ operrti\e data handlin-q s)stclr. Thc opcrati\'c corditions slr!!en:d
bclo$ arc npical *hich can be chaneed pror idcd standardizalion is done.
aoluDrn: OV l?01 lrused silica capillary colurxn
I:iIn thiakncss LllmI
lcnqrlr = l :mi lntcnrdl I)iarnctcr (r i: nrnr
Tempefalufes: c!.lunr rl
Ritc irr'C rninutc l lol,l in IlinLrles
150
210
2:r.) .l
In.lcci(n' 250'C
Detectof 280 "C
(larrier gas I IeliLrn
Carficr gas Iiorr : l0 nrl per ninure.
Split Italio - Splitlcss
.-\-l.l ln ection \olume , lpl
\ :.r Strndxrd glass$are
,\.3 IiT,AGLN'I'S
.1'l 1 A:ctune AR gradc or eq i!alent
\-l I I|lcnral standxrd Slrcuric acid \{clh}l estef.
\-r..r llclala\\l Nl reli'rcnce stalrdard ofkno\\n content of Nietalax\l_ R and S isLrtnc15.
\--r.I ( rlorLrlhalonil rel'erancc standaftl ol kno\n prrit\.
A ] PIIOCEI)LRE
\ II Preparation of internal \tandurd stock solution
\\ ei.qh (1. rhc n.arest 0 lrr! ) oul accLlratel\ it.li to 0.l6 llnts ,il- Stcafi. rLcld nt.rlr\ I
clter ill l. a il](l ml \oLrm€trc fl.rsk. Dissolre in l00lnl .\.ernnc ll,J rtrrke tp r l iii
\\ l1h r:etonc. Sh:rkc s cll to ltonrogcnize and d ii\ol\ c
\ .1 I Prel)arition of ]IetRtr\t I fl jirock sotution
\,eilh rccLrralcl\ l7 r. llam of rcl!:fcnce sub\tancc to tlte ncaresl 0 ()l llra iir 100|rl
\.lumcrfi. flask. Dissol\e in i(l{)ntl ol itcetonc and make up to mark \r r1h rcelonc. Shakc
\\ cll 11, honr(]senlze and d is(i l\ e.
,\-J.l ['rcparAtion ol sinndard -\olulion ., |f/
\ieigh accuratcl\ l7 to.l7 gnt ol chlorolhalonil fcfefence -slandaid t{r the Icatc\t al.0l ng
ir:i.)inl \olumctfic flask. Add l0 ml of\lelala\\l NI slock solulion. \dJ irl t)rl iri
.. e'r. .l,ld.rJ.or i,, .1..t)-,.!e zc., r- Ji.. lrr
\ l.l h-eprr.rtio of srrnplr soluti,,n
-l \\eigh 1'r LhL'ncarest 0.1 nrg clrough sallple tc conLarn about l0 cnr ..1chlorc,thelor l
r' rnd l nrr ol'nrerala\tl \1 ..\dcl lU rnl ofintcrnal stanctard. .{dtltorliii l0 ml ol icet!-.nc.
liaficartc and dissol\ e.
//
\-.1.3 ESTIItA t'ION
\ I 1.1 In.ielt I irl offefcrence standarcl solutjon unlil thc inlernal standarcl , feferencr
\ubstance afea !uoticnt\ oft\\o successivc chrotnalogralns do nol de\ iatc tfont
eacll ollref b! lnore lhLrn 2oi.
L,-r. r...le' | .i
- irr:r..inr.<rlr...c
------4.\' \ ( \.\r \ \ ------
l. - Standard !olUtion
S = Sanplc solLrrioD (1.2.:]._.|)
,^j:I""ii;;itill:'j:,,,:Tl,il,il;iHllfilxli:ili*Jiili;:ii::::;,i::::::
jt::
.\-.1..1.2 Rclention tinre (Guidc
\ alucs)
Chlorolhalonil
10.95 nin
[Ieta]:r\\ I \.1
| 1.30 nrin.
llcth\ Slerratc
i2.90 nin
.\ J.J,J Tolal nrr tiDrc
22 min.
.\5 C.rLCUI,AI'IONS
ChlorothrloIiI r.nteIt. pefrcnl ,\lrxA:\AtXp
tr\ ntas\ =
i!l: x Arx r\i
\'1
\'1: .l l; j,l. ,
i,l .';1,,,""1.,;,,r,,!-.,r "
.r.r
rc.r[ .rrrr ot ( il, /1,r]r.rlunrlrrr
.\l
, rhe.hr,,,.
,rlron,r,"iu,,r ;' ,;;'.],;, j" ''' I r
I'r' i ' , ..
1, ',.' ;'r"t ' t. '-
1..:, " rr.rerr'.r .r.rnu,.rd ;,,; ];,li
\ f" J r',,: r
l.
r
p. r.,
P ",,
,c..,,.ucr,,,ir\",,.,,,,i"ti.,,iil:,li:ll
"rr:r,i"ili,;,',i','Jll,.;li:].;1,r,
.",
\lcrrla\\ j con1cn1. pcrccnl Lrr rnasr =
0I \ \J \ \ \ -\. \ |
M: x Ar r ,\t
\1 = rllass itl rnt ol reference strndard ofNictale\)l
n1a\s in nr; oflhe sanple takcn lor
tcst'
\tl l(' 1
f.'^.'|= o' \lclrlJ\\i;rlInc(hr"ln'rl'urnll' l'r'nJrrl
.\: P..,r..,rcr'|\lcrJl.r)'\l In ll_c ihr'rnr" ?r'rrn 1'rrlfrccrrt,"t] ,...
so rlllrn
.
- f stanclafd in thc chromalogram ofslandard
""f. "r.".ffnternal solrlle
- f'.,tf. or"".ff"tcmrl stalldard in the chrolndlogran oisample
j - p"r."ntrg. purlry of\lelala\]l refcrence standrrd
hpi.ai chrolnalogfanl is enc|rscd pagc l7
\NNtiX A Prrt 2
(clauselll)
Detenninrtion ofllctallx)l contcnt R: S r:!tios'
0 Cicnrral
0-l lJelermiraliott ol lUclala\)llll s ratios is carfied oul llsing iligh Perfonnance i iquid
Cf.\ratogriphi (i IPI C)
B.I PRINCIPLI
I|,1 Corrlenl is dcteniilred bJ chiral cQloumn usinS High Pcrfirnnance Liquid CronatL'grlfh)
(l lPl.C) and c\tclnal standard lechniquc
B-2 .\I'PAIiATT]S
cquipfed \\ith isocratic ptrnrpcnd torrplcdtr'1 prirtcr.llnfier!rrrn Integfalnr
" - - 'llPl-C
B-2.1
.tf,. pr'l i'1cd
."nditiorls strggcsted bcio\\ 'rre t\ pirnl \ hicll !'rrr bt r lr'ln3'-Ll
"p.rotit.
slandardiTalicrn ls d{'llc
C olLrnrn: (lhiral Ol'
Parlicle sizc 5lnl
lcngth=250 nrml I D =+6mln'
Deleclol LrV Detcctor 2i0 nnl
carlier gas; nlle)iane: Isop|op)l' Alcohol 90:!0
Flo\\falcI I0nllmln
B-1.2 lnicction !olume - l0 Pl
B-l I Ultrasonic bath
B-l l Slanda|1] lass\\ afe
g
Il-3 R[A(;L\-] s
li 3.1 n He\anc IIPI C gfaclc or cqlrl\alenl
B-1.2 lsrifr,rp! flPLC gradc or eqrrr!aLcnl
I alcohol
B-J.3 \lc1alar\ I Nl lcfercnce s{andard of l(llo\!n p'lritr'
B-.I PROCED(IItE
B-1 | I'repar:rtion of sti dard sol{tion
\\cigh (t() th. nearest 0.0lIng) 16111 l8 mg ofNl{ala\)l \t relercrcc llrndard inro rhc
:11 mL \olLlrclric 0ask. ,\dd cluent l0 trl and !onicate ibr l0 nrinutes. Cool Lo r.rrrrr
lenrpcralufc irfd ntalic up Lo \olLrnrc \\ith cluenr.
B l.: Ilt-eparation {,f srmple (tcst) solution
\\crgh !o thc t)cdr.\t 0.lmg cnNgh sanrplc t{) cortain about 16 l. lS r g !.1 .,i
rrrclela\\l NI into thr i0lrll volu|netr.ic flask add 5 nl ofisopfopafol. SLrnicltte fL]r ll.i
ninulcs. Cool 1C] rcotn lemperaturc and ntake up io lolume \\ illt rHe\ane
B-{.3 ES'llX{,\TtON
Iniecl I Lrl ofrcfefence standerd solution until the peak arcas obtrined ibr oft\!o
successlve chromrtograms do not de\iate lioln e.rrh othef b\ more than li,0
Thcr ' . r'eIll^.\:r:rreIr..Ltrcr ..
------cr. s src sr s:c S: Sr -----
(l Slandrrd solulion
S - Sumllc so[r1ion ( 1.2.].-.n)
L-sc l|c drndaKl chftrmatogfams to idcntrl\ the pcaks ol l{ rnd S ircmers \fearut;
lhc p.ak areri ofN{ctalaxrl I{ and S isomef and coulputc rhe fercetlagc as gi\el
8.1.3.1 lletc lion li c (Guide valucs)
\lctalar\ I (S cnanliomer) 9.80 lnin
\letala\) I M (ll enalrtionrcr) l-1.0 nrin
r]-.1.1.2 'fotal run tine qpnfo\ :5 nr in.
Annc\ rc I (Part 3)
c-1.tI Arr(
1'o R F,lranlonlel
c l.l As\C
Flnrnliomer =
?,/o S
( i\r ' '\s)
t
il,r*:ir ii r*]'#: il *n I l; :r' l'
iii ilii*$J,il";lt
ol, \Ictala{-\'l Coltent t'- r' L I
(C'l 0)' o
n S Etanlir'lner
Conlcnl = % I{ Enarlti$nef
q\ "; l\'lctalc\\l
,\ l\'pilirl chfomarcgfall
-B
APPcndix
Do ItrR\IlNArloN
o[ lfl
xl'l
\\.iqh 1 0 gnr (lo the ,l".ii:::l,ll-':,ii$':lT';,,i'ir"l i' :"';l''-'':':'i:'"T:';'i'i''
{leir;isecl rrrtcr tlorl
medium sPecd bc L'cli
conbincd clcclr"cle th' lliaFhrsqm shr'old
ln $e cese ol
llnlnefse thc elcctrodc(s)
lc\c\ ofthe ll(lLlro sPeed'
ar llrc s1o*est
stirrel fof lilrthef l0 'scconds
Prrrcess {llir')
on thc nlagnctio
,/ | 0 l pli unrt:
\ulue \\'ith !n accrrroc] of
It.ild (]1f thc |lt
