Naphazoline Hydrochloride and Pheniramine Maleate Ophthalmic Solution
»Naphazoline Hydrochloride and Pheniramine Maleate Ophthalmic Solution is a sterile,buffered solution of Naphazoline Hydrochloride and Pheniramine Maleate in water adjusted to a suitable tonicity.It contains not less than 90.0percent and not more than 110.0percent of the labeled amount of naphazoline hydrochloride (C14H14N2·HCl)and pheniramine maleate (C16H20N2·C4H4O4).It contains a suitable preservative.
Packaging and storage— Preserve in tight containers,and store at a temperature between 20and 25,protected from light.
Identification—
A: Proceed as directed in the following thin-layer chromatographic procedure.
Naphazoline hydrochloride standard solution— Dissolve a quantity of USP Naphazoline Hydrochloride RSin water to obtain a solution containing about 1.5mg per mL.
Pheniramine maleate standard solution— Dissolve a quantity of USP Pheniramine Maleate RSin water to obtain a solution containing about 6.0mg per mL.
Test solution— Dilute,if necessary,a volume of Ophthalmic Solution with water to obtain a solution containing about 0.25mg of naphazoline hydrochloride per mLand 3mg of pheniramine maleate per mL.
Procedure— Separately apply 5µLof Naphazoline hydrochloride standard solution,10µLof Pheniramine maleate standard solution,and 30µLof the Test solutionto a 20-cm ×20-cm thin-layer chromatographic plate (see Chromatography á621ñ)coated with a 0.25-mm layer of silica gel.Allow the spots to dry,then place the plate in a saturated chromatographic chamber,and develop in a solvent system consisting of methanol,water,and acetic acid (8:1:1)until the solvent front has moved to about 1.5cm from the top of the plate.Remove the plate from the developing chamber,mark the solvent front,and allow to air-dry.Spray with ninhydrin TS,and place in an oven at 105to visualize the spots.Both the naphazoline and pheniramine spots are purplish grey in color.The RFvalues of the spots obtained from the Test solutioncorrespond to those obtained from theNaphazoline hydrochloride standard solution and thePheniramine maleate standard solution.
B: The retention times of the major peaks in the chromatogram of the Assay preparationcorrespond to those of the Standard preparation,as obtained in the Assay.
Sterility á71ñ— It meets the requirements when tested as directed for Membrane Filtrationunder Test for Sterility of the Product to be Examined.
pHá791ñ: between 5.7and 6.3.
Assay—
Buffer solution— Dissolve 14.2g of anhydrous dibasic sodium phosphate and 20mLof triethylamine in 1900mLof water,adjust with phosphoric acid to a pHof 5.6±0.1,dilute with water to make 2000mLof solution,and mix.
Mobile phase— Prepared a filtered and degassed mixture of Bufferand acetonitrile (80:20).Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).
Naphazoline hydrochloride stock standard solution— Dissolve an accurately weighed quantity of USP Naphazoline Hydrochloride RSin Mobile phaseto obtain a solution having a known concentration of about 0.75mg per mL.
Pheniramine maleate stock standard solution— Dissolve an accurately weighed quantity of USP Pheniramine Maleate RSin Mobile phaseto obtain a known concentration of about 3.00mg per mL.
Standard preparation— Transfer 1.0mLof Naphazoline hydrochloride stock standard solutionand 3.0mLof Pheniramine maleate stock standard solutionto a 25-mLvolumetric flask,dilute with Mobile phaseto volume,and mix to obtain a solution having known concentrations of naphazoline hydrochloride and pheniramine maleate of 0.03and 0.36mg per mL,respectively.
Assay preparation— Transfer an accurately measured volume of Ophthalmic Solution,equivalent to about 0.75mg of naphazoline hydrochloride and 9.0mg of pheniramine maleate,to a 25-mLvolumetric flask,dilute with Mobile phaseto volume,and mix.
Chromatographic system (see Chromatography á621ñ)— The liquid chromatograph is equipped with a 270-nm detector and a 4.6-mm ×15-cm column that contains packing L7.The flow rate is about 1.5mLper minute.Chromatograph the Standard preparation,and record the peak responses as directed for Procedure:the resolution,R,between the naphazoline peak and the pheniramine peak is not less than 2;the column efficiency,determined from the naphazoline and pheniramine peaks,is not less than 750theoretical plates;the tailing factor is not greater than 2.5for pheniramine;and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure— Separately inject equal volumes (about 25µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the responses for the peaks.Calculate the quantity,in mg,of naphazoline hydrochloride (C14H14N2·HCl)in each mLof the Ophthalmic Solution taken by the formula:
25(C/V)(rU/rS),
in which Cis the concentration in mg per mLof USP Naphazoline Hydrochloride RSin the Standard preparation;Vis the volume,in mL,of Ophthalmic solution taken;and rUand rSare the naphazoline peak responses obtained from the Assay preparation and the Standard preparation,respectively.Calculate the quantity,in mg,of pheniramine maleate (C16H20N2·C4H4O4)in each mLof the Ophthalmic Solution taken by the same formula,changing the terms to refer to pheniramine maleate.
Auxiliary Information— Staff Liaison:Lawrence Evans,III,Ph.D.,Scientist
Expert Committee:(PA6)Pharmaceutical Analysis 6
USP28–NF23Page 1334
Pharmacopeial Forum:Volume No.28(6)Page 1825
Phone Number:1-301-816-8389