Neostigmine Injection

Neostigmine Methylsulphate Injection

Neostigmine Injection is a sterile solution of NeostigmineMethylsulphate in Water for
Injections.Neostigmine Injection contains not less than 90.0 per centand not more
than 110.0 per cent of the stated amount of neostigmine methylsulphate, C
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Identification
%. &ilute, if necessary, a 'olume of the injection containing!.( mg of Neostigmine
Methylsulphate to ( ml )ith
)ater
,sha*e )ith three +uantities, each of 10 ml, of
ether
and discardthe ether e,tracts.When e,amined in the range !30 nm to 3#0 nm -!.../0, a
! cmlayer of the resulting solution sho)s a1sorption ma,ima ata1out !#0 nm and
!#/ nm.
N2"$3I4MIN2 M2356$768%32

919
I8 !00/
:. &etermine 1y thin;layer chromatography -!...1/0, coatingthe plate )ith
silica gel 4. Mo1ile phase
. % mi,ture of (0 'olumes of
chloroform
,3( 'olumes of
methanol
, 10 'olumes of
formic acid
and( 'olumes of
)ater
.
3est solution
. &ilute the injection under e,amination, if necessary, )ith
)ater
to produce a solution containing0.0( per cent )<' of Neostigmine Methylsulphate.
=eference solution -a0
. % 0.0( per cent )<' solution of
neostigmine methylsulphate =$
in
)ater
.
=eference solution -10
. % mi,ture of e+ual 'olumes of the testsolution and reference solution -a0.%pply to the
plate 10 >l of each solution. %fter de'elopment,dry the plate in air, spray )ith
dilute potassiumiodo1ismuthate solution
. 3he principal spot in thechromatogram o1tained )ith the test solution corresponds
tothat in the chromatogram o1tained )ith reference solution -a0.3he principal spot in
the chromatogram o1tained )ithreference solution -10 appears as a single, compact
spot.C. 3o 1 ml add 0.( ml of
sodium hydro,ide solution
ande'aporate to dryness on a )ater;1ath. eat +uic*ly in an oil;1ath to a1out !(0? and
maintain at this temperature for a1out30 seconds. Cool, dissol'e the residue in 1 ml
of
)ater
, cool inice
)ater
and add 1 ml of
dia@o1en@enesulphonic acid solution
A an orange;red colour is produced.
3ests
p
. -!...!.0 ..( to #.(.
3;ydro,y trimethylanilinium methyl sulphate
.

&etermine1y li+uid chromatography -!...1.0.
3est solution
. &ilute the injection if necessary, )ith
)ater
tocontain a 0.0( per cent )<' solution of NeostigmineMethylsulphate.
=eference solution -a0.
&ilute 1 'olume of the test solution to100 'olumes )ith
)ater
.
=eference solution -10
. %dd 0.0( ml of
( M sodium hydro,ide
to 1 ml of the test solution and allo) to stand for ( minutes.%dd 0.1 ml of
( M hydrochloric acid
and use immediately.Chromatographic systemBa stainless steel column !( cm C ..# mm
pac*ed )ithoctadecylsilane chemically 1onded to porous silicaparticles -( >m0 -such
as 6ichrosphere #0 =8;select :0,Bmo1ile phaseD
0.001( M
solution of
sodiumheptanesulphonate
in a mi,ture of 1( 'olumes of
acetonitrile
and 9( 'olumes of
0.0( M

potassiumdihydrogen orthophosphate
adjusted to p 3.0 )ith
orthophosphoric acid
,Bflo) rate. of 1.1 ml per minute,Bspectrophotometer set at !1( nm,Ba 10 >l loop
injector.In the chromatogram o1tained )ith reference solution -10 theprincipal pea*
has a retention time of a1out #.9 minutes-neostigmine methylsulphate0 and there is a
pea* )ith arelati'e retention time of a1out 0.( --3;hydro,y0trimethylanilinium
methylsulphate0. In the chromatogramo1tained )ith the test solution, the area of any
secondarypea* )ith a retention time corresponding to that of the pea* due to -3;
hydro,y0trimethylanilinium methylsulphate in thechromatogram o1tained )ith
reference solution -10 is notgreater than the area of the principal pea* in the
chromatogramo1tained )ith reference solution -a0 -1 per cent0.
"ther 3ests
. Complies )ith the tests stated under 8arenteral8reparations -Injections0.
%ssay
. &ilute an accurately measured 'olume containing a1out!( mg of Neostigmine
Methylsulphate to (0.0 ml )ith
)ater
.Measure the a1sor1ance of the resulting solution at thema,imum at a1out !#0 nm
-!.../0. Calculate the content of C
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!
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$ ta*ing 1..3( as the specific a1sor1ance at!#0 nm.
$torage
. $tore protected from light.