Chromatographic purity
Buffer solution
,
Mobile phase, and
System suitability solutionProceed as directed in the
Assay.
Standard solution
Transfer 10.0 mL of the System suitability solution to a 100-mL volumetric flask, dilute with Mobile phase to volume, and mix. Transfer 10.0 mL of this solution to a 50 mL volumetric flask, dilute with Mobile phase to volume, and mix.
Test solution
Transfer 50 mg of Selegiline Hydrochloride to a 50-mL volumetric flask, dissolve in and dilute with Mobile phase to volume, and mix.
Chromatographic system
Proceed as directed in the
Assay. Inject about 20 µL of the
Standard solution, and record the peak responses as directed in the
Procedure: the resolution,
R, between the methamphetamine and selegiline peaks is not less than 3, and the relative standard deviation for replicate injections is not more than 5.0%.
Procedure
Separately inject equal volumes (about 20 µL) of the
Standard solution and the
Test solution into the chromatograph, and allow the
Test solution to elute for not less than three times the retention time of selegiline. Record the chromatograms, and measure the peak responses. Calculate the percentage of each impurity in the portion of Selegiline Hydrochloride taken by the formula:
5000(C / W)(ri / rS),
in which
C is the concentration, in mg per mL, of
USP Selegiline Hydrochloride RS in the
Standard solution,
W is the weight, in mg, of Selegiline Hydrochloride taken to prepare the
Test solution,
ri is the peak response for each impurity in the chromatogram of the
Test solution, and
rS is the peak response for selegiline in the chromatogram of the
Standard solution. Not more than 0.2% of any individual impurity is found, and the sum of all impurities is not more than 1.0%.
Assay
Buffer solution
Prepare a solution of 0.1 M monobasic ammonium phosphate, adjust with phosphoric acid to a pH of 3.1, and mix.
Mobile phase
Prepare a filtered and degassed mixture of
Buffer solution and acetonitrile (80:20). Make adjustments if necessary (see
System Suitability under
Chromatography 621).
Standard preparation
Dissolve an accurately weighed quantity of
USP Selegiline Hydrochloride RS, and dilute quantitatively, and stepwise if necessary, with
Mobile phase to obtain a solution having a known concentration of about 0.1 mg per mL.
Assay preparation
Transfer an accurately weighed quantity, about 50 mg of Selegiline Hydrochloride, to a 50-mL volumetric flask, dissolve in and dilute with Mobile phase to volume, and mix. Transfer 10.0 mL of this solution to a 100-mL volumetric flask, dilute with Mobile phase to volume, and mix.
Chromatographic system
(see
Chromatography 621)The liquid chromatograph is equipped with a 205-nm detector and a 3.9-mm × 30-cm column that contains packing L1. The flow rate is about 1 mL per minute. Chromatograph the
System suitability solution, and record the peak responses as directed for
Procedure: the resolution,
R, between the methamphetamine and selegiline peaks is not less than 3, and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure
Separately inject equal volumes (about 20 µL) of the
Standard preparation and the
Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the quantity, in mg, of C
13H
17N·HCl in the portion of Selegiline Hydrochloride taken by the formula:
500C(rU / rS),
in which
C is the concentration, in mg per mL, of
USP Selegiline Hydrochloride RS in the
Standard preparation, and
rU and
rS are the selegiline peak responses obtained from the
Assay preparation and the
Standard preparation, respectively.