Fluticasone Propionate Nasal Spray
DEFINITION
Fluticasone Propionate Nasal Spray is a white, opaque suspension of Fluticasone Propionate in water. It is supplied in a form suitable for nasal administration. It contains NLT 95.0% and NMT 115.0% of the labeled amount of fluticasone propionate (C25H31F3O5S).
IDENTIFICATION
•  A. Infrared Absorption 197M
Sample:  Transfer 30 g of Nasal Spray equally into two 50-mL centrifuge tubes. Add 10 mL of water to each tube, insert the stopper, and shake vigorously for 2 min. Centri-fuge at 3500 rpm for 10 min, and discard the supernatant. Add 10 mL of water to each tube, insert the stopper, and shake vigorously for 2 min. Centrifuge at 3500 rpm for 10 min, and discard the supernatant. Add 10 mL of water to each tube, insert the stopper, and shake vigorously for 2 min. Centrifuge at 3500 rpm for 10 min, and discard the supernatant. To one tube add 10 mL of methanol. Shake to disperse the residue, and transfer to the other tube. Shake the other tube for 1 min. Centrifuge at 3500 rpm for 10 min. Decant the supernatant into an agate mortar. Evaporate the methanol either by carefully blowing dry with compressed air or nitrogen, or by allowing the methanol to evaporate naturally. If using an air or nitrogen line, use a suitable in-line filter to avoid contamination. Allow the residue to dry overnight in a desiccator over silica gel.
Acceptance criteria:  Meets the requirements
•  B. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
ASSAY
•  Procedure
Diluent:  Acetonitrile and 0.001 M hydrochloric acid (60:40)
Buffer:  1.2 g/L of monobasic ammonium phosphate. Adjust with phosphoric acid to a pH of 3.5 ± 0.05.
Mobile phase:  Methanol, acetonitrile, and Buffer (50:15:35)
System suitability solution:  50 µg/mL of USP Phenylethyl Alcohol RS and 10 µg/mL of USP Fluticasone Propionate Nasal Spray Resolution Mixture RS in Diluent
Standard solution:  10 µg/mL of USP Fluticasone Propionate RS in Diluent
Sample solution:  Nominally 10 µg/mL prepared as follows: Transfer an amount of the Nasal Spray containing 0.5 mg of fluticasone propionate to a 50-mL volumetric flask. Add about 40 mL of Diluent, and sonicate the flask for 10 min. Dilute with Diluent to volume, and shake. Allow to stand for 10 min until the supernatant is a clear solution. Inject the clear supernatant into the chromatograph.
Chromatographic system 
Mode:  LC
Detector:  UV 210 and 239 nm
Column:  4.6-mm × 25-cm; 5-µm packing L1
Column temperature:  40
Flow rate:  1.5 mL/min
Injection size:  20 µL
System suitability 
[Note—Record the peak areas at 210 nm for 5 min, then change the wavelength to 239 nm and record the peak areas. ]
Samples:  System suitability solution and Standard solution
Record the chromatogram at 210 nm for 5 min and then change the wavelength to 239 nm.
[Note—The relative retention times for phenylethyl alcohol, fluticasone propionate, and fluticasone propionate related compound D are about 0.42, 1.0, and 1.10, respectively. ]
Suitability requirements 
Resolution:  NLT 1.5 between fluticasone propionate and fluticasone propionate related compound D, System suitability solution
Relative standard deviation:  NMT 2.0%, Standard solution
Analysis 
Samples:  Standard solution and Sample solution
Calculate the percentage of the label claim of fluticasone propionate (C25H31F3O5S) in the portion of Nasal Spray taken:
Result = (rU/rS) × (CS/CU) × 100
rU== peak response from the Sample solution
rS== peak response from the Standard solution
CS== concentration of USP Fluticasone Propionate RS in the Standard solution (µg/mL)
CU== nominal concentration of Fluticasone Propionate in the Sample solution (µg/mL)
Acceptance criteria:  95.0%–115.0%
OTHER COMPONENTS
•  Content of Phenylethyl Alcohol
Diluent, Mobile phase, System suitability solution, Chromatographic system, and System suitability:  Proceed as directed in the Assay.
Standard solution:  0.05 mg/mL of USP Phenylethyl Alcohol RS in Diluent
Sample solution:  Transfer 1.0 g of the Nasal Spray to a 50-mL volumetric flask. Add about 40 mL of Diluent, and sonicate for 10 min until supernatant is clear. Use the clear supernatant for analysis.
Analysis 
Samples:  Standard solution and Sample solution
Calculate the quantity, in mg/g, of phenylethyl alcohol in the portion of Nasal Spray taken:
Result = (rU/rS) × CS × V × (1/W)
rU== peak response of phenylethyl alcohol from the Sample solution
rS== peak response of phenylethyl alcohol from the Standard solution
CS== concentration of USP Phenylethyl Alcohol RS in the Standard solution (mg/mL)
V== volume of the Sample solution, 50 mL
W== weight of the Nasal Spray in the Sample solution (g)
Acceptance criteria 

For 50 sprays: 1.75–2.63 mg/g
For 120 sprays: 1.88–2.63 mg/g
•  Content of Benzalkonium Chloride
Buffer:  250 mg/mL of citric acid. Adjust the solution with 2 N sodium hydroxide to a pH of 3.5 ± 0.05.
Standard solution:  200 µg/mL [0.02% (w/w)] of USP Benzalkonium Chloride RS in water
Docusate sodium titrant:  Dissolve 0.22 g of USP Docusate Sodium RS in 100 mL of warm water. Dilute with water to make 1000 mL.
Eosin Y indicator:  Dissolve 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.
Titer value of docusate sodium:  Pipet 10 mL of the Standard solution into a 250-mL glass-stoppered flask containing 40 mL of water, 5 mL of Eosin Y indicator, and 2 mL of Buffer. 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 Standard solution, and make any necessary correction (see Titrimetry 541).
Calculate the titer value (T) of the Docusate sodium titrant, in µg/mL, of benzalkonium chloride:
Result = CS × (VS/VD)
CS== concentration of USP Benzalkonium Chloride RS in the Standard solution (µg/mL)
VS== volume of the Standard solution, 10 mL
VD== volume of the Docusate sodium titrant used in the titration of the Standard solution (mL)
Analysis 
Sample:  10 g of Nasal Spray
Transfer the Sample into a 250-mL glass-stoppered flask containing 40 mL of water, 5 mL of Eosin Y indicator, and 2 mL of Buffer. Repeat the procedure as given above for the Standard solution. To clarify the endpoint, place the flask in an ultrasonic bath for 1–2 min to separate the chloroform layer from the aqueous phase. Perform a blank determination.
Calculate the concentration of benzalkonium chloride, in µg/g, in the portion of Nasal Spray taken:
Result = TV/W
T== titer value of the Docusate sodium titrant
V== volume of the Docusate sodium titrant used in the titration of the Nasal Spray (mL)
W== weight of the portion of Nasal Spray taken (g)
Acceptance criteria:  140–220 µg/g
PERFORMANCE TESTS
•  Delivered Dose Uniformity (within container)
Diluent:  Acetonitrile and 0.001 M hydrochloric acid (60:40)
Buffer:  1.2 g/L of monobasic ammonium phosphate. Adjust with phosphoric acid to a pH of 3.5 ± 0.05.
Mobile phase:  Methanol, acetonitrile, and Buffer (50:15:35)
System suitability solution:  5 µg/mL of USP Fluticasone Propionate Nasal Spray Resolution Mixture RS in Diluent
Standard solution:  4 µg/mL of USP Fluticasone Propionate RS in Diluent
Sample solution:  Wipe the pump clean. Shake the bottle for 30 s, 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 3–48 (50-spray pack) or 3–118 (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 Diluent to each flask, and shake well for 10 min to disperse the suspension. Dilute with Diluent to volume, and mix thoroughly. Allow the flask to stand until the excipients have settled. Inject the clear supernatant. Repeat this procedure with 4 additional bottles.
Chromatographic system 
Mode:  LC
Detector:  UV 239 nm
Column:  4.6-mm × 25-cm; 5-µm packing L1
Column temperature:  40
Flow rate:  1.5 mL/min
Injection size:  50 µL
System suitability 
Samples:  System suitability solution and Standard solution
[Note—The relative retention times for fluticasone propionate and fluticasone propionate related compound D are about 1.0 and 1.10, respectively. ]
Suitability requirements 
Resolution:  NLT 1.5 between fluticasone propionate and fluticasone propionate related compound D, System suitability solution
Relative standard deviation:  NMT 2.0%, Standard solution
Analysis 
Samples:  Standard solution and Sample solution
Calculate the percentage of the label claim of fluticasone propionate (C25H31F3O5S) in each dose:
Result = (rU/rS) × (CS/CU) × 100
rU== peak response from the Sample solution
rS== peak response from the Standard solution
CS== concentration of USP Fluticasone Propionate RS in the Standard solution (µg/mL)
CU== nominal concentration of the Sample solution (µg/mL)
Acceptance criteria:  The mean dose delivered from 10 doses is within 85%–115% of the label claim. NMT 1 dose is outside 80%–120% of the label claim. No doses are outside 75%–125% of the label claim. If 2 or 3 doses are outside outside 80%–120% of the label claim, test an additional 10 bottles. The mean dose delivered from 30 doses is within 85%–115% of the label claim. NMT 3 doses are outside 80%–120% of the label claim. No doses are outside 75%–125% of the label claim.
•  Delivered Dose Uniformity (within batch)
Diluent, Buffer, Mobile phase, System suitability solution, Standard solution, Chromatographic system, and System suitability:  Prepare as directed in the test for Delivered Dose Uniformity (within container).
Sample solution:  Wipe the pump clean. Shake the bottle for 30 s, 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.
Analysis 
Samples:  Standard solution and Sample solution
Calculate the percentage of the label claim of fluticasone propionate (C25H31F3O5S) in each dose:
Result = (rU/rS) × (CS/CU) × 100
rU== peak response from the Sample solution
rS== peak response from the Standard solution
CS== concentration of USP Fluticasone Propionate RS in the Standard solution (µg/mL)
CU== nominal concentration of the Sample solution (µg/mL)
Acceptance criteria:  The mean dose delivered from 10 doses is within 85%–115% of the label claim. NMT 1 dose is outside 80%–120% of the label claim. No doses are outside 75%–125% of the label claim. If 2 or 3 doses are outside 80%–120% of the label claim, test an additional 20 bottles. The mean dose delivered from 2 actuations in the beginning of the 30 bottles (30 doses) is within 85%–115% of the label claim. NMT 3 doses are outside 80%–120% of the label claim. No doses are outside 75%–125% of the label claim.
IMPURITIES
•  Organic Impurities
Diluent:  Acetonitrile and 0.001 M hydrochloric acid (60:40)
Solution A:  Methanol and acetonitrile (77:23)
Buffer:  1.2 g/L of monobasic ammonium phosphate. Adjust with phosphoric acid to a pH of 3.4 ± 0.1.
Mobile phase:  Solution A and Buffer (60:40)
System suitability solution:  0.1 mg/mL of USP Fluticasone Propionate Related Compounds Mixture RS and 0.5 mg/mL of USP Phenylethyl Alcohol RS in Diluent
Control solution:  0.5 mg/mL of USP Phenylethyl Alcohol RS and 0.08 mg/mL of USP Benzalkonium Chloride RS in a mixture of Diluent and water (4:1)
Sample solution:  0.2 g/mL of Nasal Spray in Diluent. Shake the flask vigorously to dissolve. Pass through a filter of 0.5-µm pore size.
Chromatographic system 
Mode:  LC
Detector:  UV 239 nm
Column:  4.6-mm × 25-cm; 5-µm packing L1
Column temperature:  40
Flow rate:  1.5 mL/min
Injection size:  50 µL
System suitability 
Sample:  System suitability solution
[Note—See Table 1 for the relative retention times. ]
Suitability requirements 
Resolution:  NLT 1.5 between fluticasone propionate related compound F and phenylethyl alcohol. NLT 2 between fluticasone propionate related compound D and fluticasone propionate
Analysis 
Samples:  System suitability solution, Control solution, and Sample solution
Calculate the percentage of each impurity in the portion of Nasal Spray taken:
Result = (rU/rT) × 100
rU== peak response of each impurity from the Sample solution
rT== sum of all the peak responses from the Sample solution, excluding the peaks from the Control solution
Acceptance criteria:  See Table 1.
Table 1
Name Relative
Retention
Time
Acceptance
Criteria,
NMT (%)
S-Fluoromethyl 17-acetyloxy-
6,9-difluoro-11-hydroxy-16-methyl-3-oxoandrosta-1,4-diene-17-carbothioate;
S-Fluoromethyl 9-fluoro-11-hydroxy-16-methyl-3,6-dioxo-17-propionyloxyandrosta-1,4-diene-17-carbothioate
0.7 0.3
Fluticasone propionate related compound D 1.1 0.3
6,9-Difluoro-11,17-dihydroxy-16-methyl-3-oxoandrosta-1,4-diene-17-carboxylic acid-6,9-difluoro-17-(fluoromethylthio)carbonyl-11-hydroxy-16-methyl-3-oxoandrosta-1,4-dien-17-yl ester 2.1 0.3
Any unspecified related impurities 0.2
Total impurities 1.5
SPECIFIC TESTS
•  Microbial Enumeration Tests 61 and Tests for Specified Microorganisms 62: Meets the requirements of the tests for absence of Staphylococcus aureus, Escherichia coli, Salmonella species, and Pseudomonas aeruginosa. The total aerobic microbial count does not exceed 25 cfu/mL, and the total combined molds and yeasts count does not exceed 25 cfu/mL.
•  pH 791: 5.0–7.0
•  Particle Size
Analysis:  Remove the pump system after shaking the test bottle to ensure product uniformity. Transfer 1 drop of the Nasal Spray onto a clean microscope slide. Examine 10 random fields of view on the slide using 400× magnification. Drug substance particles are irregular in shape, whereas the excipient particles are elongated and angular. Record the number of individual drug substance particles that are less than 5 µm in diameter, greater than 5 µm but less than 15 µm in diameter, and greater than 15 µm in diameter. Calculate the percentage of each category by number.
Acceptance criteria:  See Table 2.
Table 2
Particle Size Acceptance Criteria
<5 µm NLT 98%
>5 µm – <15 µm NMT 1.8%
>15 µm NMT 0.2%
•  Foreign Particulates
Analysis:  Shake the required number of bottles to ensure uniformity. Remove the pump system carefully to minimize contamination of the sample. Collect about 100 g of Nasal Spray, and pass it through a wetted 250-µm screen. Rinse each bottle with a portion of water equal to twice the volume of each bottle. Pass the rinse through the 250-µm screen. Visually observe the screen and filtrate for any foreign particulates. Also examine the screen under a microscope using transmitted light.
Acceptance criteria:  No foreign particulates greater than 250 µm are visible.
•  Droplet Size Distribution: Determine using a validated laser diffraction technique and method that measures the volume diameters of droplets. In preparation shake the bottle to ensure product uniformity. Prime the pump by discharging a predetermined (refer to the product label) number of actuations to waste, at which time a fine mist should appear. Measure and record the average of three sprays per bottle, and report the mean diameter defining the population of particles, by volume, below 10% (D10), 50% (D50), and 90% (D90) of five bottles.
•  Spray Pattern: Determine the spray pattern using a validated method that measures the size of the pattern. Gently shake the bottle to ensure product uniformity. Prime the pump by discharging a predetermined number (refer to the product label) of actuations to waste, at which time a fine mist should appear. Measure and record the average of two sprays per bottle and report the longest axis (x axis), and the ratio of longest to shortest axes (x/y ratio) of two bottles.
ADDITIONAL REQUIREMENTS
•  Packaging and Storage: Preserve in tight, light-resistant containers. Store between 4 and 30.
•  USP Reference Standards 11
USP Benzalkonium Chloride RS Click to View Structure
USP Docusate Sodium RS Click to View Structure
USP Fluticasone Propionate RS Click to View Structure
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.
USP Fluticasone Propionate Related Compounds Mixture RS
This Reference Standard is a mixture of fluticasone propionate and fluticasone propionate related compounds D and F, and the chemical names for all 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.
Fluticasone propionate related compound F: 6,9-Difluoro-11,17-dihydroxy-16-methyl-3-oxoandrosta-1,4-diene-17-carboxylic acid.
USP Phenylethyl Alcohol RS Click to View Structure
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