Articaine Hydrochloride and Epinephrine Injection

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

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

DOWNLOAD PDF HERE

1 DEFINITION

Articaine Hydrochloride and Epinephrine Injection is a sterile solution of Articaine Hydrochloride and Epinephrine, in Water for Injection, and contains NLT 95.0% and NMT 105.0% of the labeled amount of articaine hydrochloride (C₁₃H₂₀N₂O₃S.HCI) and NLT 90.0% and NMT 115.0% of the labeled amount of epinephrine (C₉H₁₃NO₃).

2 IDENTIFICATION

A. The retention times of the articaine and epinephrine peaks from the Sample solution correspond to those of the Standard solution, as obtained in the Assays for Articaine Hydrochloride and Epinephrine, respectively.

3 ASSAY

3.1 ARTICAINE HYDROCHLORIDE

Buffer: Glacial acetic acid and water (50:930). Adjust with 2 N sodium hydroxide to a pH of 3.4.

Mobile phase: Acetonitrile and Buffer (22:78)

Standard stock solution: 40 mg/mL of USP Articaine Hydrochloride RS in water

Standard solution: 0.8 mg/mL of USP Articaine Hydrochloride RS in Mobile phase from Standard stock solution

Sample solution: Equivalent to 0.8 mg/mL of articaine hydrochloride in Mobile phase from a portion of Injection

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 270 nm

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

Flow rate: 1 mL/min

Injection size: 10 µL

Run time: 2.5 times the retention time of articaine

System suitability

Sample: Standard solution

Suitability requirements

Tailing factor: NMT 2.2

Relative standard deviation: NMT 1.0%, from six injections

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of articaine hydrochloride (C₁₃H₂₀N₂O₃S.HCI) in the portion of Injection 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 Articaine Hydrochloride RS in the Standard solution (mg/mL)

C_U = nominal concentration of articaine hydrochloride in the Sample solution (mg/mL)

Acceptance criteria: 95.0%-105.0%

3.2 EPINEPHRINE

Mobile phase: Mix 50 mL of glacial acetic acid and 930 mL of water. Adjust with 2 N sodium hydroxide to a pH of 3.4. In this solution, dissolve 1.2 g of sodium 1-heptanesulfonate, and add 1.0 mL of 0.1 M edetate disodium and 0.298 g of potassium chloride. Add 150 mL of methanol.

Diluent: 0.5 mg/mL potassium metabisulfite in water

System suitability solution: 22 µg/mL of epinephrine from USP Epinephrine Bitartrate RS and 20 µg/mL of norepinephrine from USP

Norepinephrine Bitartrate RS in Diluent

Standard stock solution: 0.55 mg/mL of epinephrine from USP Epinephrine Bitartrate RS in Diluent

Standard solution: Dilute a suitable volume of the Standard stock solution with Diluent to obtain a final concentration of L mg/mL of epinephrine, where L is the label claim of epinephrine in the Injection.

Sample solution: Use the Injection directly.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: Amperometric electrochemical

Reference electrode: Silver/silver chloride

Working electrode: Glassy carbon

Potential: +650 mV

Detector temperature: 28 ± 2°

Column: 4.0-mm x 25-cm; 5-µm packing L7

Flow rate: 1 mL/min

Injection size: 2 µL

Run time: 1.7 times the retention time of epinephrine

System suitability

Sample: System suitability solution

[NOTE-The relative retention times for norepinephrine and epinephrine are 0.90 and 1.0, respectively.]

Suitability requirements

Resolution: NLT 1.5 between the norepinephrine and epinephrine peaks

Tailing factor: NMT 2.0 for the epinephrine peak

Relative standard deviation: NMT 1.0% for the epinephrine peak, from six injections

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of epinephrine (C₉H₁₃NO₃) in the portion of Injection 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 epinephrine in the Standard solution (mg/mL)

C_U = nominal concentration of epinephrine in the Sample solution (mg/mL)

Acceptance criteria: 90.0%-115.0%

4 PERFORMANCE TESTS

4.1 DELIVERABLE VOLUME (698)

For Articaine Hydrochloride and Epinephrine Injection packaged in single-dose containers: Meets the requirements

5 IMPURITIES

5.1 ORGANIC IMPURITIES, LIMIT OF ARTICAINE RELATED COMPOUNDS

Mobile phase, Standard stock solution, Sample solution, and Chromatographic system: Proceed as directed in the Assay for Articaine Hydrochloride.

Standard solution: 0.8 mg/mL of USP Articaine Hydrochloride RS from Standard stock solution and 40 µg/mL of USP Articaine Related Compound B RS in Mobile phase

System suitability

Sample: Standard solution

[NOTE-See Table 1 for relative retention times.]

Suitability requirements

Tailing factor: NMT 2.2 for the articaine peak

Resolution: NLT 1.25 between the articaine related compound B and articaine peaks

Relative standard deviation: NMT 1.0% for the articaine peak

Analysis

Samples: Standard solution and Sample solution

[NOTE-Articaine related compounds elute at relative retention times of NMT 2.0 with respect to the articaine peak.]

Calculate the percentage of articaine related compounds and any other individual impurity in the portion of Injection taken:

Result = (r_U/r_S) × (C_S/C_U) × 100

r_U = response of each individual impurity from the Sample solution

r_S = response of articaine from the Standard solution

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

C_U = nominal concentration of articaine in the Sample solution (mg/mL)

[NOTE-Disregard any peak below 0.05%.]

Acceptance criteria: See Table 1.

Table 1

5.2 ORGANIC IMPURITIES, LIMIT OF EPINEPHRINE RELATED COMPOUNDS

Mobile phase, System suitability solution, Standard solution, Sample solution, Chromatographic system, and System suitability: Proceed as directed in the Assay for Epinephrine.

Analysis

Samples: Standard solution and Sample solution

[NOTE-Epinephrine related compounds elute between relative retention times of 0.35 and 1.0, with respect to the epinephrine peak.]

Calculate the percentage of epinephrine related compounds and any other individual impurity in the portion of Injection taken:

Result = (r_U/r_S) × (C_S/C_U) × 100

r_U = response of each individual impurity from the Sample solution

r_S = response of epinephrine from the Standard solution 

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

C_U = nominal concentration of epinephrine in the Sample solution (mg/mL)

Acceptance criteria: See Table 2.

Table 2

^a 1-(3,4-Dihydroxyphenyl)-2-(methylamino) ethanesulfonic acid.

6 SPECIFIC TESTS

PH (791): 2.7-5.2

BACTERIAL ENDOTOXINS TEST (85): NMT 0.7 USP Endotoxin Unit/mg of articaine hydrochloride

STERILITY TESTS (71): It meets the requirements when tested as directed for Test for Sterility of the Product to Be Examined, Membrane Filtration.

PARTICULATE MATTER IN INJECTIONS (788): Meets the requirements

OTHER REQUIREMENTS: It meets the requirements under Injections and Implanted Drug Products (1).

7 ADDITIONAL REQUIREMENTS

7.1 PACKAGING AND STORAGE

Preserve in single-dose containers, preferably of Type I glass. Store at controlled room temperature.

7.2 USP REFERENCE STANDARDS (11)

USP Articaine Hydrochloride RS

USP Articaine Related Compound B RS

4-Methyl-3-{[2-(propylamino) propanoyl)amino}thiophene-2-carboxylic acid.

C₁₂H₁₈N₂O₃S  270.35

USP Epinephrine Bitartrate RS

USP Norepinephrine Bitartrate RS