Related
substances:
Solution preparation:
Mobile phase: 10 ml of trifluoroacetic acid, 300 ml of acetonitrile and 690
ml of water.
Standard
impurity Solution : Weigh accurately about 3 mg of Tramadol
impurity A CRS in 100 ml volumetric flask dilute to the volume with mobile
phase.
System
Suitability Solution (SST):
Weigh accurately about 3 mg of tramadol hydrochloride CRS and 3 mg of Tramadol
impurity A CRS in 100 ml volumetric flask dilute to the volume with mobile
phase.
Test
Solution : Weigh and
finely powder 20 tablets. Weigh and transfer ___ mg of the powder, equivalent
to 500 mg of Tramadol Hydrochloride Mix with the aid of ultrasound, with 75 ml
of the mobile phase, cool, add sufficient mobile phase to produce 100 ml and
filter through a glass- fibre filter.
Reference
Solution : Dilute 2 ml of
test solution to 100 ml with the mobile
phase and further dilute 1 ml to 10 ml with the mobile phase.
Chromatographic
condition:
Column : Use a stainless steel column
(25 cm × 4.6 mm) C18(5 µl)
Flow rate : 1.0 ml/min.
Injection volume : 20 µl
Column temperature: Ambient
Wavelength
: 271 nm.
For test solution allow the chromatography to
process for four times of Tramadol HCl.
Procedure: Inject blank (one time) SST solution (one
time) standard impurity solution ( six time)reference (six time )blank (one
time) Test solution ( one time ) into
the chromatograph.
In chromatogram obtained with the test solution
disregard any peak with area than 0.5 times of the principle peak in
chromatogram obtained with reference solution.
System suitability: In the chromatogram obtained with SST Solution
, the resolution factor between the two principal peaks is at least 3.0.
Calculation:
|
Impurity
A
|
=
|
Area
of impurity A in test solution
|
x
|
Conc. of std
impurity solution
|
x
|
100
|
|
Mean
area of principle peak of std impurity solution
|
Conc. of
test solution
|
|
Any other impurity
|
=
|
Area
of any other impurity test in solution
|
x
|
Conc. of reference
solution
|
x
|
100
|
|
Mean area of Principle peak in
reference solution
|
Conc. of
test solution
|
|
Total impurities
|
=
|
Sum
of area of any other impurity in test solution
|
x
|
Conc. of reference solution
|
x
|
100
|
|
Mean area of principle peak in reference solution
|
Conc. of
test solution
|
Acceptance criteria:
Impurity A : Not
more than 0.3%
Any unspecified impurity:
Not more than 0.2%
Total
impurities : Not more than 1.0 %
