Printed on: Thu Jul 29 2021, 06:54:43 PM Official Status: Currently Official on 29-Jul-2021 DocId: 1_GUID-E4F23E9B-3A4D-4730-A017-FCF421A250AF_4_en-US

(EST)
Printed by: Le Tran Official Date: Official as of 01-May-2020 Document Type: USP @2021 USPC
1

Analysis
Piperacillin Samples: Standard solution and Sample solution
Calculate the potency of piperacillin (C23H27N5O7S) in the
portion of Piperacillin taken:

Result = (r U/r S) × (C S/C U) × P

rU = peak response from the Sample solution

rS = peak response from the Standard solution
CS = concentration of USP Piperacillin RS in the
Standard solution (mg/mL)
CU = concentration of the Sample solution (mg/mL)
P = potency of piperacillin in USP Piperacillin RS
C23H27N5O7S · H2O 535.57 (µg/mg)
C23H27N5O7S 517.56
4-Thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid, Acceptance criteria: 960–1030 µg/mg on the anhydrous
6-[[[[(4-ethyl-2,3-dioxo-1-piperazinyl)carbonyl] amino] basis
phenylacetyl]amino]-3,3-dimethyl-7-oxo-, monohydrate, IMPURITIES
[2S-2α,5α,6β(S*)]]; • AMPICILLIN, PIPERACILLIN PENICILLOIC ACID, PIPERACILLIN
(2S,5R,6R)-6-[(R)-2-(4-Ethyl-2,3-dioxo-1- RELATED COMPOUND E, AND ACETYLATED PENICILLOIC
piperazinecarboxamido)-2-phenylacetamido]-3,3-dimethyl- ACID OF PIPERACILLIN
7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid Mobile phase, Sample solution, Chromatographic

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monohydrate [66258-76-2]. system, and System suitability: Proceed as directed in the
Anhydrous [61477-96-1]. Assay.
DEFINITION Standard solution 1: 0.08 mg/mL of USP Ampicillin RS in
Piperacillin contains NLT 960 µg/mg and NMT 1030 µg/mg Mobile phase
of piperacillin (C23H27N5O7S), calculated on the anhydrous Standard solution 2: 0.04 mg/mL of USP Piperacillin RS in
basis.
IDENTIFICATION
ci Mobile phase. Dissolve in a few drops of methanol, and
dilute with Mobile phase to volume. Use this solution within
1 h.
Analysis
Change to read: Samples: Standard solution 1, Standard solution 2, and
ffi
Sample solution
• A. ▲SPECTROSCOPIC IDENTIFICATION TESTS á197ñ, Infrared
Calculate the percentage of ampicillin in the portion of
Spectroscopy: 197K▲ (CN 1-May-2020)
Piperacillin taken:
ASSAY
• PROCEDURE Result = (r U/r S) × (C S/C U) × P × F × 100
Mobile phase: Methanol, water, 0.2 M monobasic sodium
O

phosphate, and 0.4 M tetrabutylammonium hydroxide rU = peak response of ampicillin from the Sample
(450:447:100:3). Adjust with phosphoric acid to a pH of solution
5.50. rS = peak response of ampicillin from Standard solution
System suitability solution: 0.1 mg/mL of USP 1
Ampicillin RS and 0.2 mg/mL of USP Piperacillin RS in CS = concentration of USP Ampicillin RS in Standard
Mobile phase solution 1 (mg/mL)
Standard solution: 0.4 mg/mL of USP Piperacillin RS in CU = concentration of the Sample solution (mg/mL)
Mobile phase. Dissolve in a few drops of methanol, and P = potency of ampicillin in USP Ampicillin RS
dilute with Mobile phase to volume. Use this solution within (µg/mg)
1 h. F = conversion factor, 0.001 mg/µg
Sample solution: 0.4 mg/mL of Piperacillin in Mobile phase.
Dissolve in a few drops of methanol, and dilute with Calculate the percentages of specified impurities other than
Mobile phase to volume. Use this solution within 1 h. ampicillin in the portion of Piperacillin taken:
Chromatographic system
(See Chromatography á621ñ, System Suitability.) Result = (r U/r S) × (C S/C U) × P × (1/F 1) × F 2 × 100
Mode: LC
Detector: UV 220 nm rU = peak response of each specified impurity other
Column: 4.6-mm × 25-cm; 5-µm packing L1 than ampicillin from the Sample solution
Flow rate: 1 mL/min rS = peak response of piperacillin from Standard
Injection volume: 10 µL solution 2
System suitability CS = concentration of USP Piperacillin RS in Standard
Samples: System suitability solution and Standard solution solution 2 (mg/mL)
[NOTE—See Table 1 for the relative retention times.] CU = concentration of the Sample solution (mg/mL)
Suitability requirements P = potency of piperacillin in USP Piperacillin RS
Resolution: NLT 16 between ampicillin and piperacillin, (µg/mg)
System suitability solution F1 = relative response factor (see Table 1)
Tailing factor: NMT 1.2 for the piperacillin peak, System F2 = conversion factor, 0.001 mg/µg
suitability solution
Relative standard deviation: NMT 2%, Standard Acceptance criteria: See Table 1.
solution

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Printed on: Thu Jul 29 2021, 06:54:43 PM Official Status: Currently Official on 29-Jul-2021 DocId: 1_GUID-E4F23E9B-3A4D-4730-A017-FCF421A250AF_4_en-US
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Printed by: Le Tran Official Date: Official as of 01-May-2020 Document Type: USP @2021 USPC
2

Table 1 CS = concentration of USP Piperacillin RS in the

Relative Relative Acceptance Standard solution (mg/mL)
Retention Response Criteria, CU = concentration of the Sample solution (mg/mL)
Name Time Factor NMT (%) P = potency of piperacillin in USP Piperacillin RS
Piperacillin related (µg/mg)
compound Ea 0.24 2.4 0.2 F1 = relative response factor (see Table 2)
Ampicillin 0.31 1.0 0.2 F2 = conversion factor, 0.001 mg/µg
Acetylated penicilloic acid Acceptance criteria: See Table 2.
of piperacillinb 0.37 1.1 0.4
Piperacillin penicillo- Table 2
ic acidc 0.62 0.7 1.0 Relative Relative Acceptance
Piperacillin 1.0 — — Retention Response Criteria,
Name Time Factor NMT (%)
a 1-Ethyl-2,3-piperazinedione.
Piperacillin 1.0 — —
b (2R,4S)-3-Acetyl-2-{(1R)-carboxy[2-(4-ethyl-2,3-dioxopiperazine-1-
carboxamido)-2-phenylacetamido]methyl}-5,5-dimethylthiazolidine-4- Piperacillinylampicillina 2.55 0.7 2.0
carboxylic acid.
c (2R,4S)-2-{(1R)-Carboxy[2-(4-ethyl-2,3-dioxopiperazine-1-carboxamido)-2- Total impuritiesb — — 3.8
phenylacetamido]methyl}-5,5-dimethylthiazolidine-4-carboxylic acid.
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-
• PIPERACILLINYLAMPICILLIN azabicyclo[3.2.0]heptane-2-carboxamido}-2-phenylacetamido)-3,3-dimethyl-
Mobile phase: Methanol, water, 0.2 M monobasic sodium 7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid.

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phosphate, and 0.4 M tetrabutylammonium hydroxide b Total impurities is the sum of all impurities reported in the tests for Ampicillin,
(615:282:100:3). Adjust with phosphoric acid to a pH of Piperacillin Penicilloic Acid, Piperacillin Related Compound E, and Acetylated
Penicilloic Acid of Piperacillin, and Piperacillinylampicillin.
5.50.
Standard solution: 0.04 mg/mL of USP Piperacillin RS in
Mobile phase. Dissolve in a few drops of methanol, and SPECIFIC TESTS
• WATER DETERMINATION, Method I á921ñ: 2.0%–4.0%
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
ci • OPTICAL ROTATION, Specific Rotation á781Sñ
Sample solution: 40 mg/mL in methanol
Acceptance criteria: +155° to +175°
Mobile phase to volume. Use this solution within 1 h. • BACTERIAL ENDOTOXINS TEST á85ñ: Where the label states
that Piperacillin is sterile or that it must be subjected to
ffi
Chromatographic system
(See Chromatography á621ñ, System Suitability.) further processing during the preparation of injectable
Mode: LC dosage forms, it contains NMT 0.07 USP Endotoxin
Detector: UV 220 nm Unit/mg of piperacillin.
Column: 4.6-mm × 25-cm; 5-µm packing L1 • STERILITY TESTS á71ñ: Where the label states that Piperacillin
Flow rate: 1 mL/min is sterile or that it must be subjected to further processing
during the preparation of injectable dosage forms, it meets
O

Injection volume: 10 µL
System suitability the requirements when tested as directed in Test for Sterility
Sample: Standard solution of the Product to Be Examined, Membrane Filtration.
[NOTE—See Table 2 for the relative retention times.] ADDITIONAL REQUIREMENTS
Suitability requirements • PACKAGING AND STORAGE: Preserve in well-closed
Relative standard deviation: NMT 2% containers.
Analysis • LABELING: Where it is intended for use in preparing
Samples: Standard solution and Sample solution injectable dosage forms, the label states that it is sterile or
Calculate the percentage of piperacillinylampicillin in the must be subjected to further processing during the
portion of Piperacillin taken: preparation of injectable dosage forms.
• USP REFERENCE STANDARDS á11ñ
Result = (r U/r S) × (C S/C U) × P × (1/F 1) × F 2 × 100 USP Ampicillin RS
USP Piperacillin RS
rU = peak response of piperacillinylampicillin from the
Sample solution
rS = peak response of piperacillin from the Standard
solution

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