A: Infrared Absorption 197M—

B: The retention time of the major peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay.

0.01 M Monobasic ammonium phosphate buffer, Mobile phase, and Chromatographic system— Prepare as directed in the Assay under Fluticasone Propionate.

Diluent— Prepare a mixture of acetonitrile and 0.001 M hydrochloric acid (60:40).

Resolution solution— Dissolve an accurately weighed quantity of USP Fluticasone Propionate Nasal Spray Resolution Mixture RS in Diluent, and dilute quantitatively, and stepwise if necessary, with Diluent to obtain a solution containing 5 µg per mL of USP Fluticasone Propionate Nasal Spray Resolution Mixture RS.

Standard solution— Dissolve an accurately weighed quantity of USP Fluticasone Propionate RS in Diluent to obtain a solution having a known concentration of about 4 µg per mL.

Test solution— Wipe the pump clean. Shake the bottle for 30 seconds, and mechanically prime the bottle. Hold a 25-mL volumetric flask in an inverted position, and discharge the first two actuations (1 dose) into the flask. Turn the flask to the upright position immediately after each actuation. Insert the stopper into the flask after collecting two actuations. Discharge actuations 3 to 48 (50-spray pack) or 3 to 118 (120-spray pack) to waste. Wipe the bottle clean, and collect the last two actuations (49 and 50 or 119 and 120) in a second 25-mL volumetric flask. Turn the flask to the upright position immediately after each actuation, and insert the stopper into the flask. Add 20 mL of the Diluent to each flask, and shake well for 10 minutes to disperse the suspension. Dilute with Diluent to volume, and mix thoroughly. Allow the flask to stand until the excipients have settled. Transfer a portion of the clear supernatant to an HPLC vial, and inject. Repeat this procedure with 4 additional bottles.

Procedure— Separately inject equal volumes (about 50 µL) of the Standard solution and the Test solution into the chromatograph, record the chromatograms, and measure the areas for the major peaks. Calculate the quantity, in µg, of C25H31F3O5S per dose taken by the formula: in which C is the concentration of USP Fluticasone Propionate RS, in µg per mL, in the Standard solution; V is the total volume of Test solution, in mL; and rU and rS are the peak responses obtained from the Test solution and the Standard solution, respectively. The mean dose delivered from 10 doses is within 85% to 115% of the label claim. Not more than 1 dose is outside 80% to 120% of the label claim. No doses are outside 75% to 125% of the label claim. Test an additional 10 bottles if 2 or 3 doses are outside ±20% of the label claim. The mean dose delivered from 30 doses should be within 85% to 115% of the label claim. Not more than 3 doses are outside 80% to 120% of the label claim. No doses are outside 75% to 125% of the label claim.

Diluent, 0.01 M Monobasic ammonium phosphate buffer, Mobile phase, Resolution solution, Standard solution, and Chromatographic system— Prepare as directed in the test for Delivered dose uniformity (within container).

Test solution— Wipe the pump clean. Shake the bottle for 30 seconds, and mechanically prime the bottle. Hold a 25-mL volumetric flask in an inverted position, and discharge the first two actuations into the flask. Turn the flask to the upright position immediately after each actuation. Insert the stopper into the flask after collecting two actuations (1 dose). Repeat this procedure with 9 additional bottles.

Procedure— Separately inject equal volumes (about 50 µL) of the Standard solution and the Test solution into the chromatograph, record the chromatograms, and measure the areas for the major peaks. Calculate the quantity, in µg, of C25H31F3O5S per dose taken by the formula: in which C is the concentration of USP Fluticasone Propionate RS, in µg per mL, in the Standard solution; V is the total volume of Test solution, in mL; and rU and rS are the peak responses obtained from the Test solution and the Standard solution, respectively. The mean dose delivered from 10 doses is within 85% to 115% of the label claim. Not more than 1 dose is outside 80% to 120% of the label claim. No doses are outside 75% to 125% of the label claim. Test an additional 20 bottles if 2 or 3 doses are outside ±20% of the label claim. The mean dose delivered from the two actuations at the beginning of the 30 bottles (30 doses) is within 85% to 115% of the label claim. Not more than 3 doses are outside 80% to 120% of the label claim. No doses are outside 75% to 125% of the label claim.

Solution A— Prepare a mixture of methanol and acetonitrile (77:23).

0.01 M Monobasic ammonium phosphate buffer— Dissolve 1.15 g of monobasic ammonium phosphate in 1000 mL of water, adjust with phosphoric acid to a pH of 3.4 ± 0.1, and mix.

Mobile phase— Prepare a filtered and degassed mixture of Solution A and 0.01 M Monobasic ammonium phosphate buffer (60:40). Make adjustments if necessary (see System Suitability under Chromatography 621).

Diluent— Prepare a mixture of acetonitrile and 0.001 M hydrochloric acid (60:40).

Control solution— Prepare a solution in a mixture of Diluent and water (4:1) containing about 0.5 mg per mL of USP Phenylethyl Alcohol RS and 0.08 mg per mL of USP Benzalkonium Chloride RS.

System suitability solution— Transfer accurately about 10 mg of USP Fluticasone Propionate Related Compounds Mixture RS to a 100-mL volumetric flask and add 50 mg of USP Phenylethyl Alcohol RS. Dilute with Diluent to volume to obtain a solution having a known concentration of about 100 µg per mL of USP Fluticasone Propionate Related Compounds Mixture RS and 500 µg per mL of USP Phenylethyl Alcohol RS.

Test solution— Transfer accurately about 1.0 g of the Nasal Spray to a 5-mL volumetric flask, dissolve in and dilute with Diluent to volume. Shake the flask vigorously to dissolve. Pass through a 0.5-µm porosity filter.

Chromatographic system (see Chromatography 621)— The liquid chromatograph is equipped with a 239-nm detector and a 4.6-mm × 25-cm column that contains 5-µm packing L1. The flow rate is about 1.5 mL per minute. The column is maintained at a temperature of 40. Chromatograph the System suitability solution, and measure the peak responses as directed for Procedure: the resolution, R, between fluticasone propionate related compound F and phenylethyl alcohol is not less than 1.5; the resolution, R, between fluticasone propionate related compound D and fluticasone propionate is not less than 2; and the relative retention times and limits are as provided in Table 1.

Procedure— Separately inject a volume (about 50 µL) of the Control solution, the System suitability solution, and the Test solution into the chromatograph, record the chromatograms, and measure all of the peak responses. Calculate the percentage of each impurity in the portion of Nasal Spray taken by the formula: in which ri is the peak response for each impurity, and rs is the sum of the responses of all the peaks, excluding the peaks obtained from the Control solution.

Diluent, Mobile phase, Resolution solution, and Chromatographic system— Proceed as directed in the Assay.

Standard solution— Dissolve an accurately weighed quantity of USP Phenylethyl Alcohol RS in Diluent to obtain a solution having a known concentration of about 50 µg per mL.

Test solution— Use the Assay preparation.

Procedure— Separately inject equal volumes (about 20 µL) of the Standard solution and the Test solution into the chromatograph, record the chromatograms, and measure the areas for the major peaks. Calculate the quantity, in mg per g, of phenylethyl alcohol in the portion of Nasal Spray taken by the formula: in which C is the concentration, in mg per mL, of the USP Phenylethyl Alcohol RS in the Standard solution; WU is the weight, in g, of the Nasal Spray taken to prepare the Test solution; and rU and rS are the peak responses obtained from the Test solution and the Standard solution, respectively. For 50 sprays: not less than 1.75 mg per g and not more than 2.63 mg per g. For 120 sprays: not less than 1.88 mg per g and not more than 2.63 mg per g.

Citric acid buffer solution— Dissolve 50 g of citric acid in 200 mL of water. Adjust the solution with 2 N sodium hydroxide to a pH of 3.5 ± 0.05.

Docusate sodium titrant— Dissolve 0.22 g of USP Docusate Sodium RS in 100 mL of warm water, and dilute with water to make 1000 mL.

Eosin Y indicator— Dissolve about 25 mg of eosin Y in 50 mL of acetone. Add 450 mL of chloroform and 5.0 ± 0.5 g of citric acid. Shake thoroughly until no discoloration occurs. Filter the mixture to remove any undissolved citric acid. Store in an amber bottle.

Benzalkonium chloride standard solution— Dissolve an accurately weighed quantity of USP Benzalkonium Chloride RS in water, and dilute quantitatively, and stepwise if necessary, with water to obtain a solution having a known concentration of about 0.02 % (w/w).

Procedure— Pipet 10 mL of Benzalkonium chloride standard solution into a 250-mL glass-stoppered flask containing 40 mL of water, 5 mL of Eosin Y indicator, and 2 mL of Citric acid buffer solution. Insert the stopper into the flask, and shake, releasing any build-up of pressure. Titrate with Docusate sodium titrant with vigorous shaking to a point when pink coloration is discharged from the chloroform layer. Perform a blank determination, substituting 10 mL of water for the Benzalkonium chloride standard solution, and make any necessary correction (see Titrimetry 541). Calculate the titer value of the Docusate sodium titrant, in µg of benzalkonium chloride per mL of Docusate sodium titrant, by the formula: in which WB is the weight, in µg, of benzalkonium chloride titrated; and VD is the volume, in mL, of Ducosate sodium titrant. Transfer accurately about 10 g of Nasal Spray into a 250-mL glass-stoppered flask containing 40 mL of water, 5 mL of Eosin Y indicator, and 2 mL of Citric acid buffer solution. Repeat the procedure as given above for Benzalkonium chloride standard solution. To clarify the endpoint, place the flask in an ultrasonic bath for 1 to 2 minutes to separate the chloroform layer from the aqueous phase. Perform a blank determination. Calculate the concentration of benzalkonium chloride, in µg per g, in the portion of Nasal Spray taken by the formula: in which T is the titer value of Ducosate sodium titrant; V is the volume, in mL, of the Docusate sodium titrant used in the titration of the Nasal Spray; and W is the weight, in g, of the portion of Nasal Spray taken: not less than 160 µg per g and not more than 210 µg per g.

Diluent— Prepare a mixture of acetonitrile and 0.001 M hydrochloric acid (60:40).

0.01 M Monobasic ammonium phosphate buffer— Dissolve 1.15 g of monobasic ammonium phosphate in 1000 mL of water, adjust with phosphoric acid to a pH of 3.5 ± 0.05, and mix.

Mobile phase— Prepare a filtered and degassed mixture of methanol, 0.01 M Monobasic ammonium phosphate buffer, and acetonitrile ( 50:35:15). Make adjustments if necessary (see System Suitability under Chromatography 621).

Resolution solution— Dissolve an accurately weighed quantity of USP Phenylethyl Alcohol RS and of USP Fluticasone Propionate Nasal Spray Resolution Mixture RS in Diluent, and dilute quantitatively, and stepwise if necessary, with Diluent to obtain a solution having known concentrations of about 50 µg per mL, and 10 µg per mL, respectively.

Standard preparation— Dissolve an accurately weighed quantity of USP Fluticasone Propionate RS in Diluent to obtain a solution having a known concentration of about 10 µg per mL.

Assay preparation— Transfer accurately about 1.0 g of the Nasal Spray to a 50-mL volumetric flask, add about 40 mL of Diluent, and sonicate the flask for 10 minutes. Dilute with Diluent to volume, and shake. Allow to stand for about 10 minutes until the supernatant is a clear solution. Inject the clear supernatant into the chromatograph.

Chromatographic system (see Chromatography 621)— The liquid chromatograph is equipped with a 4.6-mm × 25-cm column that contains 5-µm packing L1, and a programmable variable wavelength detector capable of monitoring at 210 nm and 239 nm. The flow rate is about 1.5 mL per minute. The column is maintained at a temperature of 40. Chromatograph the Resolution solution and the Standard preparation, record the peak areas at 210 nm for 5 minutes, then change the wavelength to 239 nm and record the peak areas: the relative retention times are about 0.42 for phenylethyl alcohol, 1.0 for fluticasone propionate, and 1.10 for fluticasone propionate related compound D; the resolution, R, between fluticasone propionate and fluticasone propionate related compound D is not less than 1.5; 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 areas for the major peaks. Calculate the quantity, in mg, of fluticasone propionate (C25H31F3O5S) in the portion of Nasal Spray taken by the formula: in which C is the concentration of USP Fluticasone Propionate RS, in mg per mL, in the Standard preparation; and rU and rS are the peak responses obtained from the Assay preparation and the Standard preparation, respectively.