Ampicillin for Injection

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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Ampicillin for Injection

1 DEFINITION

Ampicillin for Injection contains an amount of Ampicillin Sodium equivalent to NLT 90.0% and NMT 115.0% of the labeled amount of ampicillin (C₁₆H₁₉N₃O₄S).

2 ASSAY

Procedure

Mobile phase: Acetonitrile, water, 1 M monobasic potassium phosphate, and 1 N acetic acid (80:909:10:1)

Diluent: Water, 1 M monobasic potassium phosphate, and 1 N acetic acid (989:10:1)

Standard solution: 1 mg/mL of USP Ampicillin RS in Diluent. Shake and sonicate, if necessary, to dissolve. Use this solution promptly after preparation.

System suitability solution: 0.12 mg/mL of caffeine in the Standard solution

Sample solution 1 (where it is represented as being in a single-dose container): 1 mg/mL of ampicillin in Diluent. Constitute Ampicillin for Injection in a volume of Diluent, corresponding to the volume of solvent specified in the labeling. Withdraw all of the withdrawable contents, using a suitable hypodermic needle and syringe, and dilute with Diluent. Use this solution promptly after preparation.

Sample solution 2 (where the label states the quantity of ampicillin in a given volume of constituted solution): 1 mg/mL of ampicillin in Diluent. Constitute 1 container of Ampicillin for Injection in a volume of Diluent, corresponding to the volume of solvent specified in the labeling. Dilute a suitable aliquot of the constituted solution with Diluent. Use this solution promptly after preparation.

Chromatographic system

(See Chromatography 〈621〉, System Suitability.)

Mode: LC

Detector: UV 254 nm

Columns

Precolumn: 4-mm × 5-cm; 5- to 10-µm packing L1

Analytical: 4-mm × 30-cm; 5- to 10-µm packing L1

Flow rate: 2 mL/min

Injection volume: 20 µL

System suitability

Samples: Standard solution and System suitability solution

[Note—The relative retention times for ampicillin and caffeine are 0.5 and 1.0, respectively, System suitability solution.]

Suitability requirements

Resolution: NLT 2.0 between caffeine and ampicillin, System suitability solution

Tailing factor: NMT 1.4, Standard solution

Capacity factor: NMT 2.5, Standard solution

Relative standard deviation: NMT 2.0%, Standard solution

Analysis

Samples: Standard solution and Sample solution 1 or Sample solution 2

Calculate the percentage of the labeled amount of ampicillin (C₁₆H₁₉N₃O₄S) in the container or in the volume of constituted solution taken:

Result = (rᵤ/rₛ) × (Cₛ/Cₓ) × P × (1/F) × 100

rᵤ = peak response from the Sample solution

rₛ = peak response from the Standard solution

Cₛ = concentration of USP Ampicillin RS in the Standard solution (mg/mL)

Cₓ = concentration of Sample solution 1 or Sample solution 2 (mg/mL)

P = potency of ampicillin in USP Ampicillin RS (µg/mg)

F = conversion factor, 0.001 mg/µg

Where the test for Uniformity of Dosage Units has been performed using the Procedure for content uniformity, use the average of these determinations as the Assay value.

Acceptance criteria: 90.0%–115.0%

3 PERFORMANCE TESTS

Uniformity of Dosage Units 〈905〉: ▲Meets the requirements▲ (CN 1-Aug-2023)

Procedure for content uniformity

Analysis: Perform the Assay on individual containers using Sample solution 1 or Sample solution 2, or both, as appropriate. ▲ ▲ (CN 1-Aug-2023)

4 SPECIFIC TESTS

Crystallinity 〈695〉: Meets the requirements. Freeze-dried products are exempt from this requirement.

pH 〈791〉

Sample solution: 10.0 mg/mL of ampicillin

Acceptance criteria: 8.0–10.0

Water Determination, Method I〈921〉: NMT 2.0%

Particulate Matter in Injections 〈788〉: Meets the requirements for small-volume injections

Sterility Tests 〈71〉: Meets the requirements

Bacterial Endotoxins Test 〈85〉: NMT 0.15 USP Endotoxin Units/mg of ampicillin

Constituted Solution: At the time of use, it meets the requirements for Injections and Implanted Drug Products 〈1〉, Specific Tests, Completeness and clarity of solutions.

Other Requirements: It meets the requirements of the tests for Identification in Ampicillin Sodium. It also meets the requirements in Labeling 〈7〉, Labels and Labeling for Injectable Products.

5 ADDITIONAL REQUIREMENTS

Packaging and Storage: Preserve as described in Packaging and Storage Requirements 〈659〉, Injection Packaging, Packaging for constitution. Protect the constituted solution from freezing.

USP Reference Standards 〈11〉

USP Ampicillin RS

USP Ampicillin Sodium RS