Piperacillin and Tazobactam 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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1 DEFINITION

Piperacillin and Tazobactam for Injection contains amounts of Piperacillin Sodium and Tazobactam Sodium equivalent to NLT 90.0% and NMT 110.0% of the labeled amounts of piperacillin (C₂₃H₂₇N₅O₇S) and tazobactam (C₁₀H₁₂N₄O₅S), the labeled amounts representing proportions of piperacillin to tazobactam of 8:1. It may contain small amounts of a suitable buffer and stabilizer.

2 IDENTIFICATION

A. The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Assay.

3 ASSAY

3.1 PROCEDURE

Buffer: 27.6 g/L of monobasic sodium phosphate

Solution A: 80 mL of 40% aqueous tetrabutylammonium hydroxide diluted with water to 100 mL

Mobile phase: Methanol, water, Buffer, and Solution A (510:432:50:8). Adjust with phosphoric acid to a pH of 5.5.

Standard solution: 0.1 mg/mL of USP Tazobactam RS and 1 mg/mL of USP Piperacillin RS in Mobile phase. Refrigerate the Standard solution immediately after preparation and during analysis, using a refrigerated autosampler set at 5 ± 3°. Analyze within 24 h of preparation.

Sample solution: Nominally 0.125 mg/mL of tazobactam and 1 mg/mL of piperacillin from Piperacillin and Tazobactam for Injection in Mobile phase. Refrigerate the Sample solution immediately after preparation and during analysis, using a refrigerated autosampler set at 5 ± 3°. Analyze within 24 h of preparation.

3.2 Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 230 nm

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

Flow rate: 1 mL/min

Injection volume: 10 µL

Autosampler temperature: 5 ± 3°

3.3 System suitability

[NOTE-The relative retention times for tazobactam and piperacillin are 0.36 and 1.0, respectively.]

Sample: Standard solution

Suitability requirements

Tailing factor: NMT 2.0 for tazobactam and piperacillin

Relative standard deviation: NMT 2.0% for tazobactam and piperacillin

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of the labeled amount of piperacillin (C₂₃H₂₇N₅O₇S) in the portion of Piperacillin and Tazobactam for Injection taken:

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

r_U = peak response of piperacillin 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 = nominal concentration of piperacillin in the Sample solution (mg/mL)

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

F = conversion factor, 0.001 mg/µg

Calculate the percentage of the labeled amount of tazobactam (C₁₀H₁₂N₄O₅S) in the portion of Piperacillin and Tazobactam for Injection taken:

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

r_U = peak response of tazobactam from the Sample solution

r_S = peak response of tazobactam from the Standard solution

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

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

P = potency of tazobactam in USP Tazobactam RS (mg/mg)

Acceptance criteria: 90.0%-110.0%

4 PERFORMANCE TESTS

UNIFORMITY OF DOSAGE UNITS (905): Meets the requirements

5 IMPURITIES

5.1 ORGANIC IMPURITIES, PROCEDURE 1

Organic Impurities, Procedure 1 is recommended when the impurity profile includes piperacillin impurities 4, 5, and 6.

Buffer: Dilute the contents of one vial of tetrabutylammonium hydrogen sulfate ion pairing reagent with water to 1 L.

Solution A: Phosphoric acid and water (1:4)

Mobile phase: Acetonitrile and Buffer (25:75). Adjust with Solution A to a pH of 3.8.

Diluent: Acetonitrile and water (25:75)

Standard stock solution 1: 60 µg/mL of USP Tazobactam Related Compound A RS in Diluent

Standard stock solution 2: 0.5 mg/mL of USP Tazobactam RS in Diluent

Standard stock solution 3: 1.0 mg/mL of USP Piperacillin RS in acetonitrile and Diluent (1:24). Dissolve first in acetonitrile, using about 4% of the final volume, and dilute with Diluent to volume.

System suitability solution: 6 µg/mL of tazobactam related compound A from Standard stock solution 1 and 25 µg/mL of tazobactam from Standard stock solution 2 in Diluent

Standard solution: 25 µg/mL of tazobactam from Standard stock solution 2 and 0.2 mg/mL of piperacillin from Standard stock solution 3 in Mobile phase. Refrigerate the solution immediately after preparation and during analysis, using a refrigerated autosampler set at 5 ± 3°. Analyze within 24 h of preparation.

Sample solution: Nominally 25 µg/mL of tazobactam and 0.2 mg/mL of piperacillin from Piperacillin and Tazobactam for Injection in Mobile phase. Refrigerate the solution immediately after preparation and during analysis, using a refrigerated autosampler set at 5 ± 3°. Analyze within 24 h of preparation.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 210 nm

Column: 4.6-mm x 15-cm; 3-µm packing L11

Flow rate: 1 mL/min

Injection volume: 20 µL

Autosampler temperature: 5 ± 3°

System suitability

Samples: System suitability solution and Standard solution

Suitability requirements

Resolution: NLT 3 between tazobactam related compound A and tazobactam, System suitability solution

Tailing factor: NMT 1.8 for tazobactam and piperacillin, Standard solution

Relative standard deviation: NMT 2% for tazobactam and piperacillin, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of each impurity in the portion of Piperacillin and Tazobactam for Injection taken:

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

r_U = peak response of each impurity 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 = nominal concentration of piperacillin in the Sample solution (mg/mL)

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

F₁ = correction factor, 0.001 mg/µg

F₂ = relative response factor (see Table 1)

Acceptance criteria: See Table 1.

Table 1

^a Calculated relative to the peak area of piperacillin.

^b (2S,3S)-2-Amino-3-methyl-3-sulfino-4-(1H-1,2,3-triazol-1-yl)butyric acid.

^c Specified unidentified impurities.

^d This compound has two epimers that usually co-elute but that may be separated as a result of minor changes in the chromatographic conditions.

^e (4S)-2-{[2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido]methyl}-5,5-dimethylthiazolidine-4-carboxylic acid.

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

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

^h Total impurities does not include piperacillin penicilloic acid.

5.2 ORGANIC IMPURITIES, PROCEDURE 2

Organic Impurities, Procedure 2 is recommended when the impurity profile includes piperacillin dimer ethyl ester and piperacillin dimer thiazolamide derivative.

Solution A: 3.12 g/L of monobasic sodium phosphate. Adjust with phosphoric acid to a pH of 3.5.

Solution B: Methanol

Mobile phase: See Table 2.

Table 2

System suitability solution 1: 10 µg/mL of USP Amoxicillin Related Compound A RS and 6 µg/mL of USP Tazobactam Related Compound A RS in Solution A. Refrigerate System suitability solution 1 immediately after preparation and during analysis, using a refrigerated autosampler set at 5 ± 3°. Analyze within 24 h of preparation.

System suitability solution 2: 0.2 mg/mL each of USP Piperacillin RS and USP Tazobactam RS in a mixture of methanol and Solution A (30:70). Prepare the solution by dissolving the compounds in methanol and diluting with Solution A to volume. Refrigerate System suitability solution 2 immediately after preparation and during analysis, using a refrigerated autosampler set at 5 ± 3°. Analyze within 24 h of preparation.

Sample solution: Nominally 2 mg/mL of piperacillin and 0.25 mg/mL of tazobactam from Piperacillin and Tazobactam for Injection in Solution A. Refrigerate the Sample solution immediately after preparation and during analysis, using a refrigerated autosampler set at 5 ± 3°. Analyze within 24 h of preparation.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 220 nm

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

Temperatures

Column: 30°

Autosampler: 5 ± 3°

Flow rate: 1 mL/min

Injection volume: 20 µL

System suitability

Samples: System suitability solution 1 and System suitability solution 2

Suitability requirements

Resolution: NLT 1.5 between tazobactam related compound A and amoxicillin related compound A, System suitability solution 1

Tailing factor: NMT 2.0 for the piperacillin and tazobactam peaks, System suitability solution 2

Relative standard deviation: NMT 10.0% for the piperacillin and tazobactam peaks, System suitability solution 2

Analysis

Sample: Sample solution

Calculate the percentage of each impurity in the portion of Piperacillin and Tazobactam for Injection taken:

Result = (r_U/r_T) × 100

r_U = peak response of each impurity from the Sample solution

r_T = sum of the responses of all peaks from the Sample solution

Acceptance criteria: See Table 3.

Table 3

^a (2S,3S)-2-Amino-3-methyl-3-sulfino-4-(1H-1,2,3-triazol-1-yl)butyric acid.

^b 6-Aminopenicillanic acid; (2S,5R,6R)-6-Amino-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid.

^c 1-Ethylpiperazine-2,3-dione.

^d 2-Formamido-3-mercapto-3-methylbutanoic acid.

^e (R)-2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetic acid.

^f N-Acetyl piperacillin open ring; (2R,4S)-3-Acetyl-2-{(1R)-carboxy[2-(4-ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido]methyl}-5,5-dimethylthiazolidine-4-carboxylic acid.

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

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

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

^j Process impurities that are controlled in the drug substance are not to be reported. They are listed here for information only.

^k Piperacillin penilloic analog; (4S)-2-{[2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido]methyl}-5,5-dimethylthiazolidine-4-carboxylic acid.

^l (2S,5R,6R)-Methyl 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-carboxylate.

^m 2-(((3-Acetyl-4-(ethoxycarbonyl)-5,5-dimethylthiazolidin-2-yl)methyl)amino)-2-oxo-1-phenylethyl 6-(2-(4-ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylate.

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

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

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

^q Piperacillin amide dimer; (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-phenyl acetamido)-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carb oxylic acid.

5.3 ORGANIC IMPURITIES, PROCEDURE 3

Organic Impurities, Procedure 3 is recommended when the impurity profile includes piperacillinpenicillenic acid and piperazinedionecarbonyl D-phenylglycylglycine.

Buffer: 27.6 g/L of monobasic sodium phosphate dihydrate

Solution A: 0.4 M aqueous tetrabutylammonium hydroxide

Solution B: Methanol, Solution A, Buffer, and water (275:3:100:622). Adjust with phosphoric acid to a pH of 5.5.

Solution C: Methanol, Solution A, Buffer, and water (615:3:100:282). Adjust with phosphoric acid to a pH of 5.5.

Mobile phase: See Table 4.

Table 4

System suitability solution: 60 µg/mL of USP Piperacillin Related Compound ERS, 0.1 mg/mL of USP Tazobactam Related Compound A RS, and 0.76 mg/mL of USP Tazobactam RS in Solution C

Standard solution 1: 6 mg/mL of USP Piperacillin RS in Solution C

Standard solution 2: 0.06 mg/mL of USP Piperacillin RS in Solution C

Sample solution: Nominally 5.1 mg/mL of piperacillin and 0.64 mg/mL of tazobactam from Piperacillin and Tazobactam for Injection in water. Refrigerate the Sample solution immediately after preparation and during analysis, using a refrigerated autosampler set at 4°. Analyze within 10 h of preparation.

Chromatographic system

(See Chromatography (621), System Suitability.)

Mode: LC

Detector: UV 220 nm

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

Temperatures

Column: 40°

Autosampler: 4°

Flow rate: 1 mL/min

Injection volume: 10 µL

System suitability

Samples: System suitability solution and Standard solution 2

Suitability requirements

Resolution: NLT 1.5 between tazobactam related compound A and piperacillin related compound E, System suitability solution

Tailing factor: NMT 2.0 for piperacillin, Standard solution 2

Relative standard deviation: NMT 10.0% for the tazobactam peak, System suitability solution

Analysis

Samples: Standard solution 2 and Sample solution

Calculate the percentage of each impurity in the portion of Piperacillin and Tazobactam for Injection taken:

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

r_U = peak response of each impurity 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 = nominal concentration of the Sample solution (mg/mL)

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

F₁ = correction factor, 0.001 mg/µg

F₂ = relative response factor (see Table 5)

Acceptance criteria: See Table 5. Disregard peaks that are 0.05 times the response of the peak in Standard solution 2. Disregard peaks that elute after piperacillinylampicillin.

Table 5

^a 1-Ethylpiperazine-2,3-dione.

^b (2S,3S)-2-Amino-3-methyl-3-sulfino-4-(1H-1,2,3-triazol-1-yl)butyric acid.

^c 2-Formamido-3-mercapto-3-methylbutanoic acid.

^d (R)-2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetic acid.

^e (R)-2-[2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenyl acetamido]acetic acid.

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

^g 2-{[(E)-{2-[(R)-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)(phenyl)methyl]-5-oxooxazol-4(5H)-ylidene}methyl]amino}-3-mercapto-3-methylbutanoic acid.

^h (2S,5R,6R)-6-(2,5-Dioxo-4-phenylimidazolidin-1-yl)-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid.

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

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

^k The limit is for the sum of the two epimers of piperacillin open ring.

^l (2S,5R,6R)-6-(2-{3-[2-(1-Carboxy-N-ethylformamido)ethyl]ureido}-2-phenylacetamido)-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid.

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

ⁿ Process impurities that are controlled in the drug substance are not to be reported. They are listed here for information only.

^o (4S)-2-{[2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido]methyl}-5,5-dimethylthiazolidine-4-carboxylic acid.

^p The limit is for the sum of the two isomers of piperacillin penilloic analog.

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

^r (2S,5R,6R)-6-{(2R)-2-(2-[(4S)-4-Carboxy-5,5-dimethylthiazolidin-2-yl]-2-[(R)-2-(4-ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido] acetamido)-2-phenylacetamido}-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid.

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

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

^u (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.

5.4 ORGANIC IMPURITIES, PROCEDURE 4

Organic Impurities, Procedure 4 is recommended when the impurity profile includes piperacillin sulfoxide and piperacillin methyl penicilloate.

Buffer: 4 g/L of monobasic sodium phosphate dihydrate

Solution A: Acetonitrile and Buffer (2:98) adjusted with 1 M sodium hydroxide to a pH of 6.0 ± 0.05

Solution B: Acetonitrile

Mobile phase: See Table 6.

Table 6

System suitability solution: 10 µg/mL of USP Amoxicillin Related Compound A RS and 6 µg/mL of USP Tazobactam Related Compound A RS in Buffer

Standard stock solution: 1 mg/mL of USP Piperacillin RS and 36 µg/mL of USP Tazobactam RS, prepared as follows. Dissolve suitable amounts of USP Piperacillin RS and USP Tazobactam RS in a small amount of acetonitrile. Dilute with Buffer to volume.

Standard solution: 50 µg/mL of piperacillin and 1.8 µg/mL of tazobactam from Standard stock solution in Buffer

Sample solution: Nominally 5 mg/mL of piperacillin and 0.625 mg/mL of tazobactam from Piperacillin and Tazobactam for Injection in Buffer. Store the Sample solution at 2°-8°, and use 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

Temperatures

Column: 30°

Autosampler: 2°-8°

Flow rate: 1.5 mL/min

Injection volume: 20 µL

System suitability

Samples: System suitability solution and Standard solution

Suitability requirements

Resolution: NLT 1.5 between tazobactam related compound A and amoxicillin related compound A, System suitability solution

Tailing factor: NMT 2.0 for the piperacillin and tazobactam peaks, Standard solution

Relative standard deviation: NMT 5.0% for the piperacillin and tazobactam peaks, Standard solution

Analysis

Samples: Standard solution and Sample solution

Calculate the percentage of each impurity other than tazobactam related compound A in the portion of Piperacillin and Tazobactam for Injection taken:

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

r_U = peak response of each impurity other than tazobactam related compound A 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 = nominal concentration of piperacillin in the Sample solution (mg/mL)

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

F = conversion factor, 0.001 mg/µg

Calculate the percentage of tazobactam related compound A in the portion of Piperacillin and Tazobactam for Injection taken:

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

r_U = peak response of tazobactam related compound A from the Sample solution

r_S = peak response of tazobactam from the Standard solution s

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

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

P = potency of tazobactam in USP Tazobactam RS (mg/mg)

Acceptance criteria: See Table 7.

Table 7

^a (2S,3S)-2-Amino-3-methyl-3-sulfino-4-(1H-1,2,3-triazol-1-yl)butyric acid.

^b 6-Aminopenicillanic acid; (2S,5R,6R)-6-Amino-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid.

^c 1-Ethylpiperazine-2,3-dione.

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

^e (R)-2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetic acid.

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

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

^h (2S,5R,6R)-6-(2,5-Dioxo-4-phenylimidazolidin-1-yl)-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid.

ⁱ (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-carboxylic acid 4-oxide.

^j (4S)-2-{[2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido]methyl}-5,5-dimethylthiazolidine-4-carboxylic acid.

^k (2R,4S)-2-{(R)-1-[(R)-2-(4-Ethyl-2,3-dioxopiperazine-1-carboxamido)-2-phenylacetamido]-2-methoxy-2-oxoethyl}-5,5-dimethylthiazolidine-4-carboxylic acid.

^l Process impurities that are controlled in the drug substance are not to be reported. They are listed here for information only.

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

ⁿ (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.

6 SPECIFIC TESTS

6.1 BACTERIAL ENDOTOXINS TEST (85)

It contains NMT 0.08 USP Endotoxin Units in a portion equivalent to 1 mg of a mixture of piperacillin and tazobactam (0.89 and 0.11 mg, respectively).

6.2 STERILITY TESTS (71)

Meets the requirements

6.3 PARTICULATE MATTER IN INJECTIONS (788)

Meets the requirements

6.4 PH (791)

Sample solution: Nominally 40 mg/mL of piperacillin

Acceptance criteria: 5.0-7.0

6.5 WATER DETERMINATION (921), Method I

NMT 2.5%

6.6 OTHER REQUIREMENTS

It meets the requirements in Injections and Implanted Drug Products (1).

7 ADDITIONAL REQUIREMENTS

7.1 PACKAGING AND STORAGE

Preserve as described in Packaging and Storage Requirements (659), Injection Packaging, Packaging for constitution. Store at controlled room temperature.

7.2 LABELING

Label it to indicate its sodium content. If a test for Organic Impurities other than Procedure 1 is used, then the labeling states with which Organic Impurities test the article complies.

7.3 USP REFERENCE STANDARDS (11)

USP Amoxicillin Related Compound A RS

(2S,5R,6R)-6-Amino-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid.

C₈H₁₂N₂O₃S 216.26

USP Piperacillin RS

USP Piperacillin Related Compound E RS

1-Ethylpiperazine-2,3-dione.

C₆H₁₀N₂O₂ 142.16

USP Tazobactam RS

USP Tazobactam Related Compound A RS

(2S,3S)-2-Amino-3-methyl-3-sulfino-4-(1H-1,2,3-triazol-1-yl)butyric acid.

C₇H₁₂N₄O₄S 248.26