Piperacillin

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

C₂₃H₂₇N₅O₇S·H₂O 535.57

C₂₃H₂₇N₅O₇S 517.56

4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid,

6-[[[[(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl]amino]phenylacetyl]amino]-3,3-dimethyl-7-oxo-, monohydrate, [2S-2α, 5α,6β(S*)]];

(2S,5R,6R)-6-[(R)-2-(4-Ethyl-2,3-dioxo-1-piperazinecarboxamido)-2-phenylacetamido]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid monohydrate CAS RN^®: 66258-76-2; UNII: X00B0D5O0E.

Anhydrous CAS RN®: 61477-96-1; UNII: 91628532GX.

1 DEFINITION

Piperacillin contains NLT 960 µg/mg and NMT 1030 µg/mg of piperacillin (C₂₃H₂₇N₅O₇S), calculated on the anhydrous basis.

2 IDENTIFICATION

A. SPECTROSCOPIC IDENTIFICATION TESTS (197), Infrared Spectroscopy: 197K

3 ASSAY

3.1 PROCEDURE

Mobile phase: Methanol, water, 0.2 M monobasic sodium phosphate, and 0.4 M tetrabutylammonium hydroxide (450:447:100:3). Adjust with phosphoric acid to a pH of 5.50.

System suitability solution: 0.1 mg/mL of USP Ampicillin RS and 0.2 mg/mL of USP Piperacillin RS in Mobile phase

Standard solution: 0.4 mg/mL of USP Piperacillin RS in Mobile phase. Dissolve in a few drops of methanol, and dilute with Mobile phase to volume. Use this solution within 1 h.

Sample solution: 0.4 mg/mL of Piperacillin in Mobile phase. Dissolve in a few drops of methanol, and dilute with Mobile phase to volume. Use this solution within 1 h.

3.2 Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 220 nm

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

Flow rate: 1 mL/min

Injection volume: 10 µL

3.3 System suitability

Samples: System suitability solution and Standard solution

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

Suitability requirements

Resolution: NLT 16 between ampicillin and piperacillin, System suitability solution

Tailing factor: NMT 1.2 for the piperacillin peak, System suitability solution

Relative standard deviation: NMT 2%, Standard solution

3.4 Analysis

Samples: Standard solution and Sample solution

Calculate the potency of piperacillin (C₂₃H₂₇N₅O₇S) in the portion of Piperacillin taken:

Result = (r_U/r_S) x (C_S/C_U) x P

r_U = peak response from the Sample solution

r_S = peak response from the Standard solution

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

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

P = potency of piperacillin in USP Piperacillin RS (µg/mg)

Acceptance criteria: 960-1030 µg/mg on the anhydrous basis

4 IMPURITIES

4.1 AMPICILLIN, PIPERACILLIN PENICILLOIC ACID, PIPERACILLIN RELATED COMPOUND E, AND ACETYLATED PENICILLOIC ACID OF PIPERACILLIN

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

Standard solution 1: 0.08 mg/mL of USP Ampicillin RS in Mobile phase

Standard solution 2: 0.04 mg/mL of USP Piperacillin RS in Mobile phase. Dissolve in a few drops of methanol, and dilute with Mobile phase to volume. Use this solution within 1 h.

Analysis

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

Calculate the percentage of ampicillin in the portion of Piperacillin taken:

Result = (r_U/r_S) x (C_S/C_U) x P x F x 100

r_U = peak response of ampicillin from the Sample solution

r_S = peak response of ampicillin from Standard solution 1

C_S = concentration of USP Ampicillin RS in Standard solution 1 (mg/mL)

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

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

F = conversion factor, 0.001 mg/µg

Calculate the percentages of specified impurities other than ampicillin in the portion of Piperacillin taken:

Result = (r_U/r_S) x (C_S/C_U) x P x (1/F₁) x F₂ x 100

r_U = peak response of each specified impurity other than ampicillin from the Sample solution

r_S = peak response of piperacillin from Standard solution 2

C_S = concentration of USP Piperacillin RS in Standard solution 2 (mg/mL)

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

P = potency of piperacillin in USP Piperacillin RS (µg/mg)

F₁ = relative response factor (see Table 1)

F₂ = conversion factor, 0.001 mg/µg

Acceptance criteria: See Table 1.

Table 1

^a 1-Ethyl-2,3-piperazinedione.

^b (2R,4S)-3-Acetyl-2-{(1R)-carboxy[2-(4-ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido]methyl}-5,5-dimethylthiazolidine-4-carboxylic acid.

^c (2R,4S)-2-{(1R)-Carboxy[2-(4-ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido]methyl}-5,5-dimethylthiazolidine-4-carboxylic acid.

4.2 PIPERACILLINYLAMPICILLIN

Mobile phase: Methanol, water, 0.2 M monobasic sodium phosphate, and 0.4 M tetrabutylammonium hydroxide (615:282:100:3). Adjust with phosphoric acid to a pH of 5.50.

Standard solution: 0.04 mg/mL of USP Piperacillin RS in Mobile phase. Dissolve in a few drops of methanol, and dilute with Mobile phase to volume. Use this solution within 1 h.

Sample solution: 0.4 mg/mL of Piperacillin in Mobile phase. Dissolve in a few drops of methanol, and dilute with Mobile phase to volume. Use this solution within 1 h.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 220 nm

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

Flow rate: 1 mL/min

Injection volume: 10 µL

System suitability

Sample: Standard solution

[NOTE-See Table 2 for the relative retention times.]

Suitability requirements

Relative standard deviation: NMT 2%

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of piperacillinylampicillin in the portion of Piperacillin taken:

Result = (r_U/r_S) x (C_S/C_U) x P x (1/F₁) x F₂ x 100

r_U = peak response of piperacillinylampicillin from the Sample solution

r_S = peak response of piperacillin from the Standard solution

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

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

P = potency of piperacillin in USP Piperacillin RS (µg/mg)

F₁ = relative response factor (see Table 2)

F₂ = conversion factor, 0.001 mg/µg

Acceptance criteria: See Table 2.

Table 2

^a (2S,5R,6R)-6-((R)-2-{(2S,5R,6R)-6-[(R)-2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxamido}-2-phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid.

^b Total impurities is the sum of all impurities reported in the tests for Ampicillin, Piperacillin Penicilloic Acid, Piperacillin Related Compound E, and Acetylated Penicilloic Acid of Piperacillin, and Piperacillinylampicillin.

5 SPECIFIC TESTS

5.1 WATER DETERMINATION, Method I (921)

2.0%-4.0%

5.2 OPTICAL ROTATION, Specific Rotation (781S)

Sample solution: 40 mg/mL in methanol

Acceptance criteria: +155° to +175°

5.3 BACTERIAL ENDOTOXINS TEST (85)

Where the label states that Piperacillin is sterile or that it must be subjected to further processing during the preparation of injectable dosage forms, it contains NMT 0.07 USP Endotoxin Unit/mg of piperacillin.

Change to read:

5.4 STERILITY TESTS (71)

Where the label states that Piperacillin is sterile, it meets the requirements when tested as directed in Test for Sterility of the Product to Be Examined, Membrane Filtration. (ERR 1-Aug-2023)

6 ADDITIONAL REQUIREMENTS

6.1 PACKAGING AND STORAGE

Preserve in well-closed containers.

6.2 LABELING

Where it is intended for use in preparing injectable dosage forms, the label states that it is sterile or must be subjected to further processing during the preparation of injectable dosage forms.

6.3 USP REFERENCE STANDARDS (11)

USP Ampicillin RS

USP Piperacillin RS