Fluticasone Propionate Cream
Fluticasone Propionate Cream contains NLT 90.0% and NMT 110.0% of the labeled amount of fluticasone propionate (C25H31F3O5S).
• A. Thin-Layer Chromatographic Identification Test 201
Standard solution: 0.4 mg/mL of USP Fluticasone Propionate RS in acetonitrile
Test solution: Transfer a quantity of Cream, equivalent to 1000 µg of fluticasone propionate, to a 125-mL separatory funnel. Add 25 mL of acetonitrile and 25 mL of hexane to the separatory funnel. Stopper and shake the funnel until the Cream is completely dispersed. Shake the separatory funnel for an additional 3 min, and allow the phases to separate. Filter the lower layer through a 20-mL syringe containing a cotton plug into a 50-mL volumetric flask. Repeat the extraction with one 7-mL aliquot of acetonitrile, and filter the lower layer into the volumetric flask. Wash the cotton plug with 2 mL of acetonitrile, and collect the washings into the volumetric flask. Dilute the sample extract with acetonitrile to volume. Transfer 12 mL of the sample extract to a glass tube suitable for evaporation, and evaporate to dryness at about 40. Dissolve the residue in 0.6 mL of acetonitrile. [NoteThe Test solution may be cloudy because of the presence of undissolved excipients. ]
Adsorbent: 0.2-mm layer of chromatographic silica gel mixture on a high-performance thin-layer chromatographic plate, 5-µm particle size
Application volume: 40 µL
Developing solvent system: Dichloromethane, ethyl acetate, and glacial acetic acid (30:8:1)
Samples: Standard solution and Test solution
Separately apply the Standard solution and the Test solution to the plate. On the same plate, apply 20 µL of the Standard solution, allow the application to dry, and apply 20 µL of the Test solution on top of the dried 20-µL Standard solution spot. Allow each of the applications to dry thoroughly. Place the plate in a tank equilibrated with the developing solvent, and allow the developing solvent to travel about 8 cm from the point of application. Remove the plate and allow to air-dry. Examine the plate under ultraviolet light at 254 nm.
Acceptance criteria: The RF value of the principal spot from the Test solution corresponds to that of the Standard solution. [NoteIf the excipients in the Cream interfere with the appearance of the principal spot obtained for the Test solution, use the Standard solution and the Test solution overspot to confirm identity. ]
• B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
[NoteProtect the Standard solution and the Sample solution from direct light by using a light-protective volumetric flask and autosampler vials. ]
Buffer: 1.2 g/L of monobasic ammonium phosphate. Adjust with phosphoric acid to a pH of 3.50 ± 0.03.
Mobile phase: Methanol, acetonitrile, and Buffer (46:14:40)
Diluent: Alcohol and water (65:35)
System suitability stock solution: 0.5 mg/mL of USP Fluticasone Propionate Nasal Spray Resolution Mixture RS in methanol. [NoteUSP Fluticasone Propionate Nasal Spray Resolution Mixture RS is a mixture of fluticasone propionate and fluticasone propionate related compound D. ]
System suitability solution: 10 µg/mL of USP Fluticasone Propionate Nasal Spray Resolution Mixture RS in Diluent from System suitability stock solution
Standard stock solution: 0.5 mg/mL USP Fluticasone Propionate RS. Dissolve the standard first in a volume of methanol, equivalent to 80% of the final volume, and dilute with water to volume.
Standard solution: 20 µg/mL USP Fluticasone Propionate RS in Diluent, from Standard stock solution
Sample solution: Equivalent to 20 µg/mL of fluticasone propionate. Transfer a quantity of Cream, equivalent to 1000 µg of fluticasone propionate, to a 125-mL separatory funnel. Add to the separatory funnel 25 mL of Diluent. Stopper and shake vigorously until the Cream is completely dispersed. Add 25 mL of hexane, shake for an additional 3 min, and allow the phases to separate. Filter the lower layer through a 20-mL syringe containing a cotton plug into a 50-mL volumetric flask. Repeat the extraction with one 5-mL and one 2-mL aliquot of Diluent, filtering the lower layers into the volumetric flask. Wash the cotton plug with 1 mL of Diluent, and collect the washings into the volumetric flask. Dilute the sample with Diluent to volume.
Detector: 240 nm
Column: 4.6-mm × 25-cm; 5-µm packing L1
Column temperature: 50
Flow rate: 1.5 mL/min
Injection size: 20 µL
Samples: System suitability solution and Standard solution
Resolution: NLT 1.4 between fluticasone propionate and fluticasone propionate related compound D, System suitability solution
Tailing factor: NMT 1.4, Standard solution (calculated using the width of the peak at 10% of the height)
Relative standard deviation: NMT 2.0%, Standard solution
Samples: Standard solution and Sample solution
Calculate the percentage of Fluticasone Propionate (C25H31F3O5S) in the portion of Cream taken:
Result = (rU/rS) × (CS/CU) × 100
Acceptance criteria: 90.0%110.0%
• Minimum Fill 755: Meets the requirements
• pH 791: 4.56.5
• Microbial Enumeration Tests 61 and Tests For Specified Microorganisms 62: Meets the requirements of the tests for absence of Staphylococcus aureus and Pseudomonas aeruginosa. The total aerobic microbial count is NMT 100 cfu/g, and the total combined molds and yeasts count is NMT 10 cfu/g.
• Packaging and Storage: Preserve in collapsible tubes or tight containers, protected from light. Store between 2 and 30.
• USP Reference Standards 11
USP Fluticasone Propionate Nasal Spray Resolution Mixture RS
This Reference Standard is a mixture of fluticasone propionate and fluticasone propionate related compound D, and the chemical names for both are given below:
Fluticasone propionate: S-Fluoromethyl 6,9-difluoro-11-hydroxy-16-methyl-3-oxo-17-propionyloxyandrosta-1,4-diene-17-carbothioate.
Fluticasone propionate related compound D: S-Methyl-6,9-difluoro-11-hydroxy-16-methyl-3-oxo-17-propionyloxy-androsta-1,4-diene-17-carbothioate.
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USP35NF30 Page 3263Pharmacopeial Forum: Volume No. 34(3) Page 609