Triamcinolone Acetonide Nasal Spray

This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition

Issued and maintained by the United States Pharmacopeial Convention (USP)

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1 DEFINITION

Triamcinolone Acetonide Nasal Spray is an aqueous suspension of Triamcinolone Acetonide. It is supplied in a form suitable for nasal administration. It contains NLT 90.0% and NMT 110.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆).

2 IDENTIFICATION

A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.

B. The UV spectrum of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.

3 ASSAY

PROCEDURE

Buffer A: 3.4 g/L of monobasic potassium phosphate prepared as follows. Dissolve 3.4 g of monobasic potassium phosphate in 900 mL of water, adjust with 5 M sodium hydroxide to a pH of 7.0, and dilute with water to 1000 mL.

Buffer B: 3.4 g/L of monobasic potassium phosphate prepared as follows. Dissolve 3.4 g of monobasic potassium phosphate in 900 mL of water, adjust with phosphoric acid to a pH of 3.0, and dilute with water to 1000 mL.

Solution A: Acetonitrile and Buffer A (27.5: 72.5)

Solution B: Acetonitrile and Buffer A (60:40)

Mobile phase: See Table 1.

Table 1
 

Diluent: Acetonitrile and Buffer B (27.5:72.5)

Standard stock solution: 0.4 mg/mL of USP Triamcinolone Acetonide RS in acetonitrile. Sonication for 15 min may be used to aid in dissolution.

Standard solution: 40 µg/mL of USP Triamcinolone Acetonide RS from Standard stock solution in Diluent

System suitability stock solution: 0.04 mg/mL of USP Triamcinolone Acetonide Related Compound B. RS and USP Triamcinolone Acetonide Related Compound C RS in Diluent

System suitability solution: 40 µg/mL of USP Triamcinolone Acetonide RS and 0.8 µg/mL each of USP Triamcinolone Acetonide Related Compound B RS and USP Triamcinolone Acetonide Related Compound C RS from suitable volumes of Standard stock solution and System suitability stock solution in Diluent

Sample solution: Nominally 40 µg/mL of triamcinolone acetonide prepared as follows. Discharge the contents of NLT 5 units of Nasal Spray to a suitable container. Transfer a portion of the Nasal Spray, equivalent to 4 mg of triamcinolone acetonide, to a 100-mL volumetric flask.

Dissolve in 28 mL of acetonitrile with the aid of sonication. Allow to cool to room temperature and dilute with Buffer B to volume. Sonicate, if necessary. Centrifuge and use the clear supernatant [NOTE-Centrifuging at 3600 rpm for 30 min may be suitable.]

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 239 nm. For Identification B, use a diode array detector in the range of 210-400 nm.

Column: 4.6-mm x 25-cm; 5-µm packing L1

Column temperature: 40°

Flow rate: 0.75 mL/min

Injection volume: 100 µL

System suitability

Samples: Standard solution and System suitability solution

[NOTE-See Table 4 for the relative retention times.]

Suitability requirements

Resolution: NLT 3.0 between triamcinolone acetonide related compound C and triamcinolone acetonide related compound B; NLT 3.0 between triamcinolone acetonide related compound B and triamcinolone acetonide, System suitability solution

Tailing factor: NMT 1.3 for triamcinolone acetonide, System suitability solution

Relative standard deviation: NMT 1.0%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆) in the portion of Nasal Spray taken:

                         Result = (r_U/r_S) × (C_S/C_U) × 100

r_U = peak response from the Sample solution

r_S = peak response from the Standard solution

C_S = concentration of USP Triamcinolone Acetonide RS in the Standard solution (µg/mL)

C_U = nominal concentration of triamcinolone acetonide in the Sample solution (µg/mL)

Acceptance criteria: 90.0%-110.0%

4 OTHER COMPONENTS

4.1 CONTENT OF EDETATE DISODIUM

Perform this test if edetate disodium is a known component in the Nasal Spray.

Buffer: Add 990 mL of water into a 1000-mL beaker, followed by 10.0 mL of 1.0 M tetrabutylammonium hydroxide in methanol. Adjust with phosphoric acid to a pH of 7.0.

Mobile phase: Acetonitrile and Buffer (15:85)

Solution A: 40 g/L of sodium chloride and 2 g/L of sodium acetate. Adjust with glacial acetic acid to a pH of 5.5.

Solution B: 1.0 g/L of cupric sulfate in water

Diluent: Acetonitrile and water (50:50)

Standard stock solution: 0.5 mg/mL of edetate disodium in water. Sonication may be used to aid in dissolution.

Standard solution: 0.05 mg/mL of edetate disodium. Transfer 5.0 mL of Standard stock solution to a 50-mL volumetric flask. Next add 10 mL of Solution A and then add 5.0 mL of acetonitrile. Mix the resulting solution and then add 20.0 mL of Solution B, dilute with Diluent to volume, and mix.

Sample solution: Combine the contents of NLT 5 units of Nasal Spray and mix the contents to obtain a composite suspension. Transfer a 5.0-g portion of the Nasal Spray to a 50-mL volumetric flask. Add 10 mL of Solution A and 5.0 ml. of acetonitrile. Mix and sonicate for 10 min and allow the solution to equilibrate to room temperature. Add 20.0 mL of Solution B, and sonicate for 10 min. Allow the sample to equilibrate to room temperature and dilute with Diluent to volume. Centrifuge a portion for 15 min, and use the supernatant. [NOTE-Centrifuging at 4000 rpm for 15 min may be suitable.]

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 265 nm

Column: 4.1-mm x 15-cm; 5-µm packing L21

Flow rate: 1 mL/min

Injection volume: 25 µL

System suitability

Sample: Standard solution

Suitability requirements

Tailing factor: NMT 2.0

Relative standard deviation: NMT 2.0%

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of edetate disodium (C₁₀H₁₄N₂Na₂O₈) in the portion of Nasal Spray taken:

                         Result = (r_U/r_S) × C_S x (V/W) x 100

r_U = peak response from the Sample solution

r_S = peak response from the Standard solution

C_S = concentration of edetate disodium in the Standard solution (mg/mL)

V = volume of the Sample solution (mL)

W = weight of Nasal Spray in the Sample solution (mg)

Acceptance criteria: 0.045%-0.055%

Change to read:

4.2 CONTENT OF BENZALKONIUM CHLORIDE

Perform this test if benzalkonium chloride is a known component in the Nasal Spray.

Buffer: Dissolve 10.8 g of monobasic sodium phosphate dihydrate in 90 mL of water, and adjust with phosphoric acid a pH of 2.5. Dilute with water to 100 mL.

Solution A: Mix 50 mL of Buffer, 750 mL of water, and 200 ml. of methanol. Add 5 ml. of triethylamine. Mix and adjust with phosphoric acid to a pH of 2.5.

Solution B: Mix 1 L of methanol with 50 mL of phosphoric acid.

Mobile phase: See Table 2.

Table 2

Diluent: 1% (v/v) hydrochloric acid in methanol

System suitability solution: 0.04 mg/mL of USP Benzalkonium Chloride RS ^▲_{▲(IRA 1-Mar-2021)} prepared as follows. Transfer a suitable volume of USP Benzalkonium Chloride RS to a suitable volumetric flask and ^▲add 30% of the flask volume of water. Dilute with Diluent to volume._{▲(IRA 1-Mar-2021)}

Standard stock solution: 0.2 mg/mL of USP Benzalkonium Bromide RS in water. [NOTE-A few drops of methanol may be used to resolve the formation of foam prior to dilution.]

Standard solution: 0.04 mg/ml. of USP Benzalkonium Bromide RS. Transfer an aliquot of Standard stock solution to a suitable volumetric flask, and add water equal to 30% of the flask volume. Dilute with Diluent to volume.

Sample solution: Combine the contents of NLT 5 units of Nasal Spray and mix the contents to obtain a composite suspension. Transfer a 5.0-g portion of the Nasal Spray to a 10-ml volumetric flask. Dilute with Diluent to volume. Centrifuge and use the supernatant. [NOTE-Centrifuging at 4000 rpm for 15 min may be suitable. The supernatant may be passed through a suitable filter of NMT 0.2-µm pore size.]

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 210 nm

Column: 4.6-mm x 3.0-cm; 2.6-µm packing L1

Column temperature: 50°

Flow rate: 2 mL/min

Injection volume: 100 µL

System suitability

Samples: System suitability solution and Standard solution

[NOTE-See Table 3 for the relative retention times. The peak due to the C10 analog may not be visible due to its low concentration in the System suitability solution. The Standard solution may contain only one peak as it is predominantly the C12 analog.]

Suitability requirements

Resolution: NLT 2.5 between the pairs of C12 and C14 homologs and C14 and C16 homologs of benzalkonium, System suitability solution

Tailing factor: NMT 2.0, Standard solution

Relative standard deviation: NMT 2%, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the sum of the corrected benzalkonium peak responses (r) in the portion of Sample solution taken:

                         Result = Σ[r_U × (1/F)]

r_U = peak response of each benzalkonium homolog from the Sample solution

F = relative response factor of the corresponding benzalkonium homolog relative to benzalkonium bromide (see Table 3)

Table 3

Calculate the percentage of benzalkonium chloride in the portion of Nasal Spray taken:

                         Result = (r_CU/r_S) × C_S × (V/W) × 100

r_CU = sum of the corrected peak responses of the benzalkonium homologs from the Sample solution 

r_S = peak response of benzalkonium from the Standard solution

C_S  = concentration of USP Benzalkonium Bromide RS in the Standard solution (mg/mL) 

V = volume of the Sample solution (mL)

W = weight of Nasal Spray in the Sample solution (mg)

Acceptance criteria: 0.0135%-0.0165%

5 PERFORMANCE TESTS

5.1 DELIVERED DOSE UNIFORMITY (BETWEEN UNITS)

Buffer: 7.0 g/L of sodium perchlorate prepared as follows. Dissolve 7.0 g of sodium perchlorate in 900 ml of water, adjust with perchloric acid to a pH of 3.0, and dilute with water to 1000 mL.

Mobile phase: Acetonitrile and Buffer (50:50)

Standard solution: 40 µg/mL of USP Triamcinolone Acetonide RS in Mobile phase

Sample solution: Nominally 40 µg/mL of triamcinolone acetonide prepared as follows. Transfer a portion of Nasal Spray, equivalent to 4.0 mg of triamcinolone acetonide, to a suitable volumetric flask. Add Mobile phase to 80% of the flask volume. Sonicate for 15 min. Allow to equilibrate to room temperature. Dilute with Mobile phase to volume. Centrifuge and pass the supernatant through a filter of 0.45-um pore size. [NOTE-Centrifuging at 4000 rpm for 45 min may be suitable.]

Repeat this procedure with 9 additional units.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 239 nm

Column: 4.6-mm x 15-cm; 5-µm packing L1

Flow rate: 1 mL/min

Injection volume: 40 µL

System suitability

Sample: Standard solution

Suitability requirements

Tailing factor: NMT 2.0

Relative standard deviation: NMT 1.0%

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆) in the portion of Nasal Spray taken: 24 31

                         Result = (r_U/r_S) × (C_S/C_U) × 100 

r_U = peak response from the Sample solution

r_S = peak response from the Standard solution 

C_S = concentration of USP Triamcinolone Acetonide RS in the Standard solution (µg/mL)

C_U = nominal concentration of triamcinolone acetonide in the Sample solution (µg/mL)

Acceptance criteria

Tier 1: The content of each of the 10 units is within 90.0%-110.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆). If the criterion in Tier 1 cannot be met, proceed to Tier 2.

Tier 2: If the content of 1 unit is outside 90.0%-110.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆) and the content of none of the units is outside 85.0%-115.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆), test an additional 20 units. All 30 of the results (including the results from Tier 1) meet the following acceptance criteria.

1. The content of each of 29 out of 30 units is within 90.0%-110.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆).

2. The content of each of the 30 units is within 85.0%-115.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆)

5.2 DELIVERED DOSE UNIFORMITY (WITHIN UNIT)

Buffer: 7.0 g/L of sodium perchlorate prepared as follows. Dissolve 7.0 g of sodium perchlorate in 900 mL of water, adjust with perchloric acid to a pH of 3.0, and dilute with water to 1000 mL.

Mobile phase: Acetonitrile and Buffer (50:50)

Standard solution: 4.8 µg/mL of USP Triamcinolone Acetonide RS in Mobile phase

Beginning sample solution (BOU): Hold the pump upright, actuate 5 times, and wipe the nosepiece dry. Hold a 25-mL volumetric flask in an inverted position and actuate the pump. Quickly turn the flask upright, wait 10 s, and repeat the process. Add 15 mL of Mobile phase while rinsing the neck of the flask and sonicate for 15 min. Allow the flask to equilibrate to room temperature, and dilute with Mobile phase to volume. Centrifuge and use the clear supernatant. This is the BOU sample. [NOTE-Centrifuging at 4000 rpm for 15 min may be suitable.]

Middle sample solution (MOU): Using the same pump as above, discharge an appropriate number of actuations to arrive at 50% of the labeled number of actuations to waste and wipe the nosepiece dry. Hold a 25-ml volumetric flask in an inverted position and actuate the pump to collect the next actuation. Quickly turn the flask upright, wait 10 s, and repeat the process to collect the next actuation. Add 15 mL of Mobile phase while rinsing the neck of the flask and sonicate for 15 min. Allow the flask to equilibrate to room temperature, and dilute with Mobile phase to volume. Centrifuge and use the clear supernatant. This is the MOU sample. [NOTE-Centrifuging at 4000 rpm for 15 min may be suitable.]

End sample solution (EOU): Using the same pump as above, discharge the next appropriate number of actuations to arrive at 100% of the labeled number of actuations to waste, and wipe the nosepiece dry. Hold a 25-mL volumetric flask in an inverted position and actuate the pump to collect the next actuation. Quickly turn the flask upright, wait 10 s, and repeat the process to collect the next actuation. Add 15 mL of Mobile phase while rinsing the neck of the flask and sonicate for 15 min. Allow the flask to equilibrate to room temperature, and dilute with Mobile phase to volume. Centrifuge and use the clear supernatant. This is the EOU sample. [NOTE-Centrifuging at 4000 rpm for 15 min may be suitable.]

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 239 nm

Column: 4.6-mm x 15-cm; 5-µm packing L1

Flow rate: 1 mL/min

Injection volume: 200 µL

System suitability

Sample: Standard solution

Suitability requirements

Tailing factor: NMT 2.0

Relative standard deviation: NMT 1.0%

Analysis

Samples: Standard solution, Beginning sample solution, Middle sample solution, and End sample solution Calculate the percentage of the labeled amount of the triamcinolone acetonide (C₂₄H₃₁FO₆) delivered dose:

                         Result = (r_U/r_S) × (C_S/C_U) × 100

r_U = peak response from the appropriate Sample solution

r_S = peak response from the Standard solution

C_S = concentration of USP Triamcinolone Acetonide RS in the Standard solution (µg/mL)

C_U = nominal concentration of triamcinolone acetonide in the appropriate Sample solution (µg/mL)

Acceptance criteria

Calculate the mean delivered dose from the BOU results from all 10 units.

Calculate the mean delivered dose from the MOU results from all 10units.

Calculate the mean delivered dose from the EOU results from all 10 units.

Calculate the unit delivery mean from the BOU, MOU, and EOU results from each of the 10 units.

Tier 1:

1. Mean delivered dose of BOU samples from all 10 units is within 85.0%-115.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆)

2. Mean delivered dose of MOU samples from all 10 units is within 85.0%-115.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆).

3. Mean delivered dose of EOU samples from all 10 units is within 85.0%-115.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆)

4. NMT 1 unit delivery mean from the 10 units is outside 80.0%-120.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆)

5. None of the unit delivery means from the 10 units is outside 75.0%-125.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆)

Tier 1 criteria 1-3 must be met. If criterion 4 or 5 cannot be met, proceed to Tier 2.

Tier 2: If NMT 3 unit delivery means are outside 80.0%-120.0% of the labeled amount of triamcinolone acetonide (C₂H, FO) and none of the unit delivery means is outside 75.0%-125.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆), test an additional units. All 30 unit delivery means (including the results from Tier 1) meet the following acceptance criteria.

1. NMT 3 of 30 unit delivery means are outside 80.0%-120.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆)

2. None of the 30 unit delivery means is outside 75.0%-125.0% of the labeled amount of triamcinolone acetonide (C₂₄H₃₁FO₆).

6 IMPURITIES

ORGANIC IMPURITIES

Buffer A, Buffer B, Solution A, Solution B, Mobile phase, Diluent, Standard solution, System suitability solution, Sample solution, Chromatographic system, and System suitability: Proceed as directed in the Assay.

Analysis

Sample: Sample solution

Calculate the percentage of each degradation product in the portion of Nasal Spray taken:

                         Result = (r_l/r_S) x 100

r_l = peak response of each degradation product from the Sample solution

r_S = peak response of triamcinolone acetonide from the Sample solution.

Acceptance criteria: See Table 4. Disregard any peak below 0.05%.

Table 4

^a 9-Fluoro-11-hydroxy-16,17-[(1-methylethylidene)bis(oxy)]-(118,16a)-3,20-dioxopregna-1,4-diene-21-oic acid,

7 SPECIFIC TESTS

pH (791): 4.5-6.0

MICROBIAL ENUMERATION TESTS (61) and TESTS FOR SPECIFIED MICROORGANISMS (62): It meets the requirements of the tests for absence of Staphylococcus aureus, Escherichia coli, Salmonella species, and Pseudomonas aeruginosa. The total aerobic microbial count is NMT 101 cfu/mL and the total combined molds and yeasts count is NMT 10¹ cfu/mL.

PARTICLE SIZE

Analysis: Shake the Nasal Spray and prime the pump by spraying 3-4 times. Actuate the spray and collect the sample on a glass microscope slide held above the nozzle, and repeat to prepare a second slide. Using light microscopy, determine the dimension of NLT 200 particles of triamcinolone acetonide by measuring NLT 100 particles from 20 random fields of view for each slide prepared. Repeat the procedure using a second unit of Nasal Spray.

Acceptance criteria: See Table 5.

Table 5

8 ADDITIONAL REQUIREMENTS

PACKAGING AND STORAGE: Preserve in a tight, light-resistant container, and store at controlled room temperature.

USP REFERENCE STANDARDS (11)

USP Benzalkonium Bromide RS

USP Benzalkonium Chloride RS

USP Triamcinolone Acetonide RS

USP Triamcinolone Acetonide Related Compound B.RS

9-Fluoro-11,21-dihydroxy-16,17-[(1-methylethylidene)bis(oxy))-(11ẞ,16a)-pregna-1,4,14-triene-3,20-dione.

C₂₄H₂₉FO₆         432.48

USP Triamcinolone Acetonide Related Compound C RS

9-Fluoro-11,21,21-trihydroxy-16,17-[(1-methylethylidene)bis(oxy)]-(113,164)-pregna-1,4-diene-3,20-dione.

C₂₄H₃₁FO₇           450.50