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(EST)
Printed by: Fabio Camargo-Trujillo Official Date: Official as of 01-May-2020 Document Type: USP @2021 USPC
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IMPURITIES
Benzocaine • RESIDUE ON IGNITION á281ñ: NMT 0.1%
• CHLORIDE
Analysis: To a solution of 200 mg in 5 mL of alcohol,
previously acidified with a few drops of diluted nitric acid,
add a few drops of silver nitrate TS.
Acceptance criteria: No turbidity is produced immediately.
• ORGANIC IMPURITIES
Solution A: Add 1.0 mL of trifluoroacetic acid in 1 L of water.
Solution B: Acetonitrile
Mobile phase: See Table 1.
C9H11NO2 165.19
Benzoic acid, 4-amino-, ethyl ester; Table 1
Ethyl p-aminobenzoate [94-09-7]. Time Solution A Solution B
(min) (%) (%)
DEFINITION
0 90 10
Benzocaine, dried over phosphorus pentoxide for 3 h, contains
NLT 98.0% and NMT 102.0% of benzocaine (C9H11NO2). 34 50 50

IDENTIFICATION 35 90 10

38 90 10
Change to read:

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• A. ▲SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared Diluent: Solution A and Solution B (1:1)
Spectroscopy: 197K▲ (CN 1-May-2020) Standard stock solution: 0.1 mg/mL each of USP
Sample: Previously dried over phosphorus pentoxide for 3 h Benzocaine RS, USP Aminobenzoic Acid RS, and USP Ethyl
Acceptance criteria: Meets the requirements 4-nitrobenzoate RS in Diluent. Sonicate for 2–5 min to
• B. The retention time of the major peak of the Sample dissolve before diluting to final volume.
solution corresponds to that of the Standard solution, as
obtained in the Assay.
ASSAY
ci Standard solution: 1 µg/mL each of USP Benzocaine RS,
USP Aminobenzoic Acid RS, and USP Ethyl
4-nitrobenzoate RS in Diluent from the Standard stock
solution
• PROCEDURE
Solution A: Acetic acid, triethylamine, and water Sample solution: 1 mg/mL of Benzocaine in Diluent.
Sonicate for 2–5 min to assist in dissolution as needed
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(20:1:980). The pH should be between 2.95 and 3.0 (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 in (See Chromatography á621ñ, System Suitability.)
Mobile phase Mode: LC
Sample solution: 0.024 mg/mL of Benzocaine in Mobile Detector: UV 280 nm
Column: 4.6-mm × 25-cm; 5-µm packing L7
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phase
Chromatographic system Flow rate: 1.5 mL/min
(See Chromatography á621ñ, 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 peak
System suitability corresponding to benzocaine, aminobenzoic acid, and
Sample: Standard solution 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 ethyl
Analysis 4-nitrobenzoate in the portion of Benzocaine taken:
Samples: Standard solution and Sample solution Result = (r U/r S) × (C S/C U) × 100
Calculate the percentage of benzocaine (C9H11NO2) in the
portion of Benzocaine taken: rU = peak response of aminobenzoic acid or ethyl
4-nitrobenzoate from the Sample solution
Result = (r U/r S) × (C S/C U) × 100 rS = peak response of corresponding reference
standard from the Standard solution
rU = peak response from the Sample solution CS = concentration of USP Aminobenzoic Acid RS or
rS = peak response from the Standard solution USP Ethyl 4-nitrobenzoate RS in the Standard
CS = concentration of USP Benzocaine RS in the solution (mg/mL)
Standard solution (mg/mL) CU = concentration of Benzocaine in the Sample
CU = concentration of Benzocaine in the Sample solution (mg/mL)
solution (mg/mL)
Calculate the percentage of any other individual
Acceptance criteria: 98.0%–102.0% on the previously unspecified impurity in the portion of Benzocaine taken:
dried basis
Result = (r U/r S) × (C S/C U) × 100

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Printed by: Fabio Camargo-Trujillo Official Date: Official as of 01-May-2020 Document Type: USP @2021 USPC
2

rU = peak response of any other individual impurity Table 2 (continued)

from the Sample solution Relative Acceptance
rS = peak response of benzocaine from the Standard Retention Criteria,
solution Name Time NMT (%)
CS = concentration of USP Benzocaine RS in the Total impurities — 1.0
Standard solution (mg/mL)
CU = concentration of Benzocaine in the Sample
solution (mg/mL) SPECIFIC TESTS
• LOSS ON DRYING á731ñ
Acceptance criteria: See Table 2. Disregard peaks less than Analysis: Dry over phosphorus pentoxide for 3 h.
0.05%. Acceptance criteria: NMT 1.0%
ADDITIONAL REQUIREMENTS
Table 2 • PACKAGING AND STORAGE: Preserve in well-closed
Relative Acceptance containers.
Retention Criteria, • USP REFERENCE STANDARDS á11ñ
Name Time NMT (%)
USP Aminobenzoic Acid RS
Aminobenzoic acid 0.29 0.10 Benzoic acid, 4-amino.
C7H7NO2 137.14
Benzocaine 1.0 —
USP Benzocaine RS
Ethyl 4-nitrobenzoate 2.1 0.10 USP Ethyl 4-nitrobenzoate RS
Any other unspecified impur-
Benzoic acid, 4-nitro-, ethyl ester.
— C9H9NO4 195.17
ity 0.10

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