A: Infrared Absorption á197Kñ.

B: Ultraviolet Absorption á197Uñ—

C: Prepare a test solution in methanol containing 20mg of Moricizine Hydrochloride per mL.Similarly prepare a Standard solution in methanol containing 20mg ofUSP Moricizine Hydrochloride RS.Separately apply 5µLof each solution on a thin-layer chromatographic plate (seeChromatography á621ñ)coated with a 0.25-mm layer of chromatographic silica gel mixture.Place the plate in a chromatographic chamber lined with filter paper saturated with a solvent system consisting of a mixture of chloroform,methanol,and diethylamine (91:7:2).Develop the chromatogram,protected from light,until the solvent front has moved about three-fourths of the length of the plate.Remove the plate from the chamber.Spray the plate with a freshly prepared ferric chloride solution prepared by adding 20mLof 10%ferric chloride solution to 200mg of potassium ferricyanide dissolved in 20mLof water:the RFvalue of the blue spot in the chromatogram obtained from the test solution corresponds to that in the chromatogram obtained from the Standard solution.

Mobile phase— Prepare a mixture of water,acetonitrile,and triethylamine (580:420:1)containing 0.005Msodium 1-octane sulfonate,and adjust with glacial acetic acid to a pHof 4.2.Make adjustments if necessary (seeSystem SuitabilityunderChromatography á621ñ).

Diluent— Prepare a mixture of 0.02Nhydrochloric acid and acetonitrile (58:42).

Internal standard solution— Prepare a solution of butamben inDiluentcontaining about 0.1mg per mL.

Standard solution— Prepare a solution ofUSP Moricizine Hydrochloride RSinDiluenthaving a known concentration of about 0.10mg per mL.Transfer 10.0mLof this solution to a 500-mLvolumetric flask,add 25.0mLofInternal standard solution,dilute withDiluentto volume,and mix to obtain a solution containing about 0.0020mg ofUSP Moricizine Hydrochloride RSper mL.[NOTE—Protect this solution from light.]

Test solution— Transfer about 100mg of Moricizine Hydrochloride,accurately weighed,to a 100-mLlow-actinic volumetric flask,add 5.0mLofInternal standard solution,dilute withDiluentto volume,and mix.[NOTE—Protect this solution from light.]

Chromatographic system(seeChromatography á621ñ)— The liquid chromatograph is equipped with a 254-nm detector and a 4.6-mm ×25-cm column that contains packing L7and is maintained at a constant temperature of about 35.The flow rate is about 2.5mLper minute.Chromatograph theStandard solution,and record the peak responses as directed forProcedure:the relative retention times are about 0.6for moricizine and 1.0for butamben;the resolution,R,between the moricizine peak and the butamben peak is not less than 2;and the relative standard deviation for replicate injections is not more than 5%.

Procedure— Separately inject equal volumes (about 20µL)of theStandard solutionand theTest solutioninto the chromatograph,record the chromatograms for a period of time that is five times the elution time of moricizine,and measure the responses for the peaks,except for the solvent peak.Calculate the percentage of each impurity peak in the portion of Moricizine Hydrochloride taken by the formula: in which Cis the concentration,in mg per mL,ofUSP Moricizine Hydrochloride RSin theStandard solution,Riis the ratio of the peak areas of an individual impurity peak to the butamben peak obtained from theTest solution,and RSis the ratio of the peak areas of the moricizine peak to the butamben peak obtained from theStandard solution.Any impurity eluting before the moricizine peak is not more than 0.25%,any impurity eluting after the moricizine peak is not more than 0.20%,and the total of all impurities is not more than 1.5%,any impurity of less than 0.1%being disregarded.

Standard solution— Transfer 6.0mLof dehydrated alcohol to a 100-mLvolumetric flask,dilute with water to volume,and mix.Transfer 5.0mLof this solution to a second 100-mLvolumetric flask,dilute with water to volume,and mix.Transfer 5.0mLof this solution to a third 100-mLvolumetric flask,dilute with water to volume,and mix.This solution contains 0.1184mg of C2H5OHper mL.

Test solution— Transfer about 1g of Moricizine Hydrochloride,accurately weighed,to a 50-mLglass-stoppered centrifuge tube,add 19.0mLof water,and sonicate to dissolve.Transfer 1.0mLof 3Nammonium hydroxide to the tube,insert the stopper,and shake the tube by mechanical means for 30minutes.Centrifuge,draw off a portion of the clear supernatant and filter through a filter having a porosity of 0.5µm or finer.

Chromatographic system(seeChromatography á621ñ)— The gas chromatograph is equipped with a flame-ionization detector and a 4-mm ×1.8-m glass column that contains support S2.The column is maintained at 150,and the injection port and detector block are maintained at 170.Helium is used as the carrier gas at a flow rate of about 50mLper minute.Chromatograph theStandard solution,and record the peak responses as directed forProcedure:the relative standard deviation for replicate injections is not more than 3%.

Procedure— Separately inject equal volumes (about 5µL)of theStandard solutionand theTest solutioninto the chromatograph,record the chromatograms,and measure the peak responses.Calculate the percentage of C2H5OHin the portion of Moricizine Hydrochloride taken by the formula: in which Cis the concentration,in mg per mL,of C2H5OHin the Standard solution;Wis the weight,in g,of Moricizine Hydrochloride taken to prepare theTest solution;and rUand rSare the alcohol peak responses obtained from theTest solutionand theStandard solution,respectively.Not more than 0.25%is found.

Mobile phase— Prepare a mixture of water,acetonitrile,glacial acetic acid,and triethylamine (580:420:20:1)containing 0.005Msodium 1-octane sulfonate.Make adjustments if necessary (seeSystem SuitabilityunderChromatography á621ñ).

Diluent— Prepare a mixture of 0.02Nhydrochloric acid and acetonitrile (58:42).

Internal standard solution— Prepare a solution of butamben inDiluentcontaining about 5mg per mL.

Standard preparation— Transfer about 25mg ofUSP Moricizine Hydrochloride RS,accurately weighed,to a 25-mLlow-actinic volumetric flask,add 5.0mLofInternal standard solution,dilute withDiluentto volume,and mix.[NOTE—Protect this solution from light.]

Assay preparation— Transfer about 1mg per mLof Moricizine Hydrochloride,accurately weighed,to a 50-mLlow-actinic volumetric flask,add 10.0mLofInternal standard solution,dilute withDiluentto volume,and mix.[NOTE—Protect this solution from light.]

Chromatographic system(seeChromatography á621ñ)— The liquid chromatograph is equipped with a 254-nm detector and a 4.6-mm ×25-cm column that contains packing L7and is maintained at a constant temperature of about 35.The flow rate is about 2.5mLper minute.Chromatograph theStandard preparation,and record the peak responses as directed forProcedure:the relative retention times are about 0.6for moricizine,1.7for the reverse Mannich product,2.0for the amide hydrolysis product,and 1.0for butamben;the resolution,R,between the moricizine peak and the butamben peak is not less than 2;and the relative standard deviation for replicate injections is not more than 2%.

Procedure— Separately inject equal volumes (about 10µL)of theStandard preparationand theAssay preparationinto the chromatograph,record the chromatograms,and measure the responses for the major peaks.Calculate the quantity,in mg,of C22H25N3O4S·HCl in the portion of Moricizine Hydrochloride taken by the formula: in which Cis the concentration,in mg per mL,ofUSP Moricizine Hydrochloride RSin theStandard preparation;and RUand RSare the ratios of the peak area responses of moricizine and butamben obtained from theAssay preparationand theStandard preparation,respectively.