Accessed from 201.105.98.

86 by sptng3zyg on Fri Oct 06 12:47:02 EDT 2017

2968 Benzethonium / Official Monographs USP 40

Prepare Benzethonium Chloride Tincture 2 mg/mL as Carrier gas: Dry helium

follows. Temperature
Injector port: 240°
Benzethonium Chloride 2g Detector block: 240°
Column: 120°
Alcohol 685 mL
Flow rate: 90 mL/min
Acetone 100 mL Injection size: 0.8 µL
Purified Water, a sufficient quantity to make 1000 mL Analysis
Samples: Standard solutions A, B, C, and D, and Sam-
Dissolve the Benzethonium Chloride in a mixture of Alcohol ple solution
and Acetone. Add sufficient Purified Water to make From the respective chromatograms obtained as de-
1000 mL. [NOTE—Benzethonium Chloride Tincture may be scribed previously, calculate the ratios of peak areas for
colored by the addition of any suitable color or combina- alcohol to internal standard and for acetone to internal
tion of colors certified by the FDA for use in drugs.] standard.
IDENTIFICATION Calculate the percentage of alcohol and of acetone in
• A. PROCEDURE the portion of Tincture taken:
Sample: 50 mL Result = [A(Y − Z) + B(Z − X)]/(Y − X)
Analysis: To the residue obtained by evaporating the
Sample on a steam bath, add 2 mL of alcohol, 0.5 mL A = percentage of alcohol, or of acetone, in
of 2 N nitric acid, and 1 mL of silver nitrate TS. Standard solution A and Standard solution C,
Acceptance criteria: A white precipitate, which is insol- respectively
uble in 2 N nitric acid but soluble in 6 N ammonium Y = ratio of the alcohol peak areas, or the acetone
hydroxide, is formed. peak areas, to the internal standard peak
• B. PROCEDURE areas for Standard solution B and Standard
Sample: 50 mL solution D, respectively
Analysis: Evaporate the Sample on a steam bath. Z = ratio of the alcohol peak areas, or the acetone
Acceptance criteria: The residue obtained forms preci- peak areas, to the internal standard peak
pitates with 2 N nitric acid and with mercuric chloride areas for the Sample solution
TS, both of which dissolve upon the addition of B = percentage of alcohol, or of acetone, in
alcohol. Standard solution B and Standard solution D,
ASSAY respectively
• PROCEDURE X = ratio of the alcohol peak areas, or the acetone
Sample: 50 mL peak areas, to the internal standard peak
Analysis: Transfer the Sample to a 150-mL beaker, and areas for Standard solution A and Standard
add, with continuous stirring, 10 mL of 25 mg/mL of solution C, respectively
sodium tetraphenylboron solution. Cover, and allow to Acceptance criteria
Alcohol (C2H5OH): 62.0%–68.0%
USP Monographs

stand for 16 h. Decant the supernatant into a tared

sintered-glass crucible, applying vacuum filtration. Sus- Acetone (C3H6O): 9.0%–11.0%
pend the precipitate in 20 mL of water. Transfer the SPECIFIC TESTS
precipitate to the crucible, washing well with water. • SPECIFIC GRAVITY 〈841〉: 0.868–0.876
Dry the precipitate and the crucible at 105° for 1 h,
cool, and weigh. The weight of the precipitate so ob- ADDITIONAL REQUIREMENTS
tained, multiplied by 0.6122, represents its equivalent • PACKAGING AND STORAGE: Package in tight, light-resistant
of benzethonium chloride (C27H42ClNO2). containers.
Acceptance criteria: 190–210 mg
OTHER COMPONENTS
• ALCOHOL AND ACETONE CONTENT
Standard solution A (alcohol low standard solution):
.

Add 5.0 mL of methanol and 11.0 mL of dehydrated Benzocaine

alcohol to a 100-mL volumetric flask, and dilute with
water to volume.
Standard solution B (alcohol high standard solution):
Add 5.0 mL of methanol and 14.0 mL of dehydrated
alcohol to a 100-mL volumetric flask, and dilute with
water to volume.
Standard solution C (acetone low standard solution): C9H11NO2 165.19
Add 5.0 mL of methanol and 1.7 mL of acetone to a Benzoic acid, 4-amino-, ethyl ester;
100-mL volumetric flask, and dilute with water to Ethyl p-aminobenzoate [94-09-7].
volume.
Standard solution D (acetone high standard solu- DEFINITION
tion): Add 5.0 mL of methanol and 2.2 mL of acetone Benzocaine, dried over phosphorus pentoxide for 3 h, con-
to a 100-mL volumetric flask, and dilute with water to tains NLT 98.0% and NMT 102.0% of benzocaine
volume. (C9H11NO2).
Sample solution: Transfer 20 mL of Tincture to a
100-mL volumetric flask, add 5.0 mL of methanol as the IDENTIFICATION
internal standard, and dilute with water to volume. • A. INFRARED ABSORPTION 〈197K〉
Chromatographic system Sample: Previously dried over phosphorus pentoxide for
(See Chromatography 〈621〉, System Suitability.) 3h
Mode: GC Acceptance criteria: Meets the requirements
Detector: Flame ionization • B. The retention time of the major peak of the Sample
Column: 120-cm × 4-mm packed with a suitable type solution corresponds to that of the Standard solution, as
of support, such as 80- to 100-mesh S3 obtained in the Assay.

Official from August 1, 2017

Copyright (c) 2017 The United States Pharmacopeial Convention. All rights reserved.
Accessed from 201.105.98.86 by sptng3zyg on Fri Oct 06 12:47:02 EDT 2017

USP 40 Official Monographs / Benzocaine 2969

ASSAY trobenzoate RS in Diluent from the Standard stock

• PROCEDURE solution
Solution A: Acetic acid, triethylamine, and water Sample solution: 1 mg/mL of Benzocaine in Diluent.
(20:1:980). The pH should be between 2.95 and 3.0 Sonicate for 2–5 min to assist in dissolution as needed
(adjust as needed). before diluting to final volume.
Mobile phase: Methanol and Solution A (40:60) Chromatographic system
Standard solution: 0.024 mg/mL of USP Benzocaine RS (See Chromatography 〈621〉, System Suitability.)
in Mobile phase Mode: LC
Sample solution: 0.024 mg/mL of Benzocaine in Mobile Detector: UV 280 nm
phase Column: 4.6-mm × 25-cm; 5-µm packing L7
Chromatographic system Flow rate: 1.5 mL/min
(See Chromatography 〈621〉, System Suitability.) Injection volume: 20 µL
Mode: LC System suitability
Detector: UV 285 nm Sample: Standard solution
Column: 2.0-mm × 15-cm; 5-µm packing L11 Suitability requirements
Flow rate: 0.2 mL/min Resolution: NLT 10 from any two peaks
Injection volume: 10 µL Relative standard deviation: NMT 2.0% for each
System suitability peak corresponding to benzocaine, aminobenzoic
Sample: Standard solution acid, and ethyl 4-nitrobenzoate
Suitability requirements Analysis
Tailing factor: NMT 2.0 Samples: Standard solution and Sample solution
Relative standard deviation: NMT 2.0% Calculate the percentage of aminobenzoic acid and
Analysis ethyl 4-nitrobenzoate in the portion of Benzocaine
Samples: Standard solution and Sample solution taken:
Calculate the percentage of benzocaine (C9H11NO2) in
the portion of Benzocaine taken: Result = (rU/rS) × (CS/CU) × 100
Result = (rU/rS) × (CS/CU) × 100 rU = peak response of aminobenzoic acid or ethyl
4-nitrobenzoate from the Sample solution
rU = peak response from the Sample solution rS = peak response of corresponding reference
rS = peak response from the Standard solution standard from the Standard solution
CS = concentration of USP Benzocaine RS in the CS = concentration of USP Aminobenzoic Acid RS
Standard solution (mg/mL) or USP Ethyl 4-nitrobenzoate RS in the
CU = concentration of Benzocaine in the Sample Standard solution (mg/mL)
solution (mg/mL) CU = concentration of Benzocaine in the Sample
Acceptance criteria: 98.0%–102.0% on the previously solution (mg/mL)
dried basis Calculate the percentage of any other individual
unspecified impurity in the portion of Benzocaine

USP Monographs
IMPURITIES taken:
• RESIDUE ON IGNITION 〈281〉: NMT 0.1%
Result = (rU/rS) × (CS/CU) × 100
Delete the following: rU = peak response of any other individual impurity
•• HEAVY METALS, Method II 〈231〉: NMT 10 ppm• (Official 1- from the Sample solution
.

rS = peak response of benzocaine from the

Jan-2018) Standard solution
• CHLORIDE CS = concentration of USP Benzocaine RS in the
Analysis: To a solution of 200 mg in 5 mL of alcohol, Standard solution (mg/mL)
previously acidified with a few drops of diluted nitric CU = concentration of Benzocaine in the Sample
acid, add a few drops of silver nitrate TS. solution (mg/mL)
Acceptance criteria: No turbidity is produced Acceptance criteria: See Table 2. Disregard peaks less
immediately. than 0.05%.
• ORGANIC IMPURITIES
Solution A: Add 1.0 mL of trifluoroacetic acid in 1 L of
water. Table 2
Solution B: Acetonitrile Relative Acceptance
Mobile phase: See Table 1. Retention Criteria,
Name Time NMT (%)
Table 1 Aminobenzoic acid 0.29 0.10
Time Solution A Solution B Benzocaine 1.0 —
(min) (%) (%) Ethyl 4-nitrobenzoate 2.1 0.10
0 90 10 Any other unspecified
—
34 50 50 impurity 0.10
35 90 10 Total impurities — 1.0
38 90 10
SPECIFIC TESTS
Diluent: Solution A and Solution B (1:1) • LOSS ON DRYING 〈731〉
Standard stock solution: 0.1 mg/mL each of USP Ben- Analysis: Dry over phosphorus pentoxide for 3 h.
zocaine RS, USP Aminobenzoic Acid RS, and USP Ethyl Acceptance criteria: NMT 1.0%
4-nitrobenzoate RS in Diluent. Sonicate for 2–5 min to
dissolve before diluting to final volume. ADDITIONAL REQUIREMENTS
Standard solution: 1 µg/mL each of USP Benzocaine • PACKAGING AND STORAGE: Preserve in well-closed
RS, USP Aminobenzoic Acid RS, and USP Ethyl 4-ni- containers.

Official from August 1, 2017

Copyright (c) 2017 The United States Pharmacopeial Convention. All rights reserved.
 Accessed from 201.105.98.86 by sptng3zyg on Fri Oct 06 12:47:02 EDT 2017

2970 Benzocaine / Official Monographs USP 40

• USP REFERENCE STANDARDS 〈11〉 [NOTE—See Table 2 for relative retention times.]
USP Aminobenzoic Acid RS Suitability requirements
Benzoic acid, 4-amino. Resolution: NLT 6 between aminobenzoic acid and
C7H7NO2 137.14 benzocaine; NLT 6 between benzocaine and ethyl
USP Benzocaine RS 4-nitrobenzoate, System suitability solution
USP Ethyl 4-nitrobenzoate RS Tailing factor: NMT 1.5, Standard solution
Benzoic acid, 4-nitro-, ethyl ester. Relative standard deviation: NMT 1.0%, Standard
C9H9NO4 195.17 solution
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of ben-
.

zocaine (C9H11NO2) in the portion of Topical Aerosol

Benzocaine Topical Aerosol taken:

DEFINITION Result = (rU/rS) × (CS/CU) × 100

Benzocaine Topical Aerosol is a solution of Benzocaine in a rU = peak response of benzocaine from the Sample
pressurized container. It contains NLT 90.0% and NMT solution
110.0% of the labeled amount of benzocaine (C9H11NO2). rS = peak response of benzocaine from the
IDENTIFICATION Standard solution
• A. The UV spectrum of the major peak of the Sample CS = concentration of USP Benzocaine RS in the
solution corresponds to that of the Standard solution, as Standard solution (mg/mL)
obtained in the Assay. CU = nominal concentration of benzocaine in the
• B. The retention time of the major peak of the Sample Sample solution (mg/mL)
solution corresponds to that of the Standard solution, as Acceptance criteria: 90.0%–110.0%
obtained in the Assay. IMPURITIES
ASSAY • ORGANIC IMPURITIES
• PROCEDURE Solution A: 0.1% Trifluoroacetic acid, prepared by di-
Solution A: 0.1% Trifluoroacetic acid, prepared by di- luting 1.0 mL of trifluoroacetic acid with water to 1 L
luting 1.0 mL of trifluoroacetic acid with water to 1 L Solution B: Acetonitrile
Solution B: Acetonitrile Mobile phase: See Table 1.
Mobile phase: See Table 1. Diluent: Solution A and Solution B (1:1)
Standard solution: 1 µg/mL each of USP Benzocaine
RS, USP Aminobenzoic Acid RS, and USP Ethyl 4-Ni-
Table 1 trobenzoate RS in Diluent
Time Solution A Solution B Sample composite: Spray a portion of the Topical Aer-
(min) (%) (%) osol into a beaker or a glass tube, and heat on a steam
USP Monographs

0 90 10 bath or a heating module at 100° for a few min to

34 50 50
expel residual propellant. Use the resulting benzocaine
solution.
35 90 10 Sample solution: Nominally 0.5 mg/mL of benzocaine
38 90 10 in Diluent prepared as follows. Transfer 50 mg of benzo-
caine from a portion of Sample composite to a 100-mL
Diluent: Solution A and Solution B (1:1) volumetric flask, dissolve, and dilute with Diluent to
System suitability solution: 1 µg/mL each of USP Ben- volume.
zocaine RS, USP Aminobenzoic Acid RS, and USP Ethyl Chromatographic system
4-Nitrobenzoate RS in Diluent (See Chromatography 〈621〉, System Suitability.)
Standard solution: 0.1 mg/mL of USP Benzocaine RS in Mode: LC
Diluent Detector: UV 280 nm
Sample composite: Spray a portion of Topical Aerosol Column: 4.6-mm × 25-cm; 5-µm packing L7
into a beaker or glass tube, and heat on a steam bath Flow rate: 1.5 mL/min
or a heating module at 100° for a few min to expel Injection volume: 20 µL
residual propellant. Use the resulting benzocaine System suitability
solution. Sample: Standard solution
Sample solution: Nominally 0.1 mg/mL of benzocaine [NOTE—See Table 2 for relative retention times.]
in Diluent prepared as follows. Transfer a known quan- Suitability requirements
tity of benzocaine solution from a portion of Sample Resolution: NLT 6 between aminobenzoic acid and
composite to an appropriate volumetric flask, dissolve, benzocaine; NLT 6 between benzocaine and ethyl
and dilute with Diluent to volume. 4-nitrobenzoate
Chromatographic system Relative standard deviation: NMT 3% for each peak
(See Chromatography 〈621〉, System Suitability.) corresponding to benzocaine, aminobenzoic acid,
Mode: LC and ethyl 4-nitrobenzoate
Detector: UV 280 nm. For Identification test A, use a Analysis
diode array detector in the range of 200–400 nm. Samples: Standard solution and Sample solution
Column: 4.6-mm × 25-cm; 5-µm packing L7 Calculate the percentage of aminobenzoic acid and
Flow rate: 1.5 mL/min ethyl 4-nitrobenzoate in the portion of Topical Aerosol
Injection volume: 20 µL taken:
System suitability
Samples: System suitability solution and Standard Result = (rU/rS) × (CS/CU) × 100
solution
rU = peak response of aminobenzoic acid or ethyl
4-nitrobenzoate from the Sample solution
rS = peak response of aminobenzoic acid or ethyl
4-nitrobenzoate from the Standard solution

Official from August 1, 2017

Copyright (c) 2017 The United States Pharmacopeial Convention. All rights reserved.