United States Pharmacopeia online
Select Pharmacopoeia
Piperacillin
Click to View Image
C23H27N5O7S·H2O 535.57

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-2,5,6(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]heptalic acid monohydrate [66258-76-2].

Anhydrous 517.56 [61477-96-1].
» Piperacillin contains not less than 960 µg and not more than 1030 µg of piperacillin (C23H27N5O7S) per mg, calculated on the anhydrous basis.
Packaging and storage— Preserve in well-closed containers.
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.
Identification, Infrared Absorption 197K.
Water, Method I 921: between 2.0% and 4.0%.
Specific rotation 781S: between +155 and +175.
Test solution: 40 mg per mL, in methanol.
Related compounds—
test 1
Mobile phase and Chromatographic system— Prepare as directed in the Assay.
Standard piperacillin solution— Transfer about 40 mg of USP Piperacillin RS, accurately weighed, to a 100-mL volumetric flask, add a few drops of methanol to dissolve it, dilute with Mobile phase to volume, and mix. Transfer 5.0 mL of this solution to a 50-mL volumetric flask, dilute with Mobile phase to volume, and mix. [note—Use these solutions within 1 hour.]
Standard ampicillin solution— Prepare a solution of USP Ampicillin RS in Mobile phase having a known concentration of about 0.08 mg per mL.
Test solution— Use the Assay preparation.
Procedure— Separately inject equal volumes (about 10 µL) of the Test solution, the Standard ampicillin solution, and the Standard piperacillin solution, and proceed as directed in the Assay. Calculate the percentage of ampicillin in the portion of Piperacillin taken by the formula:
10C(P / W)(rU / rSa)
in which C is the concentration, in mg per mL, of USP Ampicillin RS in the Standard ampicillin solution; P is the designated potency, in µg of ampicillin per mg, of USP Ampicillin RS; W is the weight, in mg, of Piperacillin taken to prepare the Test solution; and rU and rSa are the peak responses of ampicillin obtained from the Test solution and the Standard ampicillin solution, respectively: not more than 0.2% of ampicillin is found.
Calculate the percentage of piperacillin related compound A (4-carboxy--[2-(4-ethyl-2,3-dioxo-1-piperazinecarboxamido-2-phenylacetamido]-5,5-dimethyl-2-thiazolidinacetic acid), piperacillin related compound B (1-ethyl-2,3-piperazinedione), and piperacillin related compound C (2-(3-acetyl-4-carboxy-5,5-dimethyl-2-thiazolidinyl)-N-[N-[(4-ethyl-2,3-dioxo-1-piperazinyl)-carbonyl]-2-phenylglycyl]glycine) in the portion of Piperacillin taken by the formula:
10C(P / W)(RRFi)(ri / rSp)
in which C is the concentration, in mg per mL, of USP Piperacillin RS in the Standard piperacillin solution; P is the designated potency, in µg of piperacillin per mg, of USP Piperacillin RS; W is the weight, in mg, of Piperacillin taken to prepare the Test solution; RRFi is the response factor of an individual piperacillin related compound relative to the response of piperacillin, specifically 1.4 for piperacillin related compound A, 0.41 for piperacillin related compound B, and 0.93 for piperacillin related compound C; ri is the response of each impurity peak obtained from the Test solution; and rSp is the peak response of piperacillin obtained from the Standard piperacillin solution: not more than 1.0% of piperacillin related compound A, not more than 0.2% of piperacillin related compound B, and not more than 0.4% of piperacillin related compound C is found.
test 2
Mobile phase— Prepare a mixture of 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 ± 0.02, and degas. Make adjustments if necessary (see System Suitability under Chromatography 621).
Standard stock solution— Transfer about 20 mg of USP Piperacillin RS, accurately weighed, to a 50-mL volumetric flask, add a few drops of methanol to dissolve it, dilute with Mobile phase to volume, and mix. [note—Use this solution within 1 hour.]
Standard solution— Transfer 5.0 mL of the Standard stock solution to a 50-mL volumetric flask, dilute with Mobile phase to volume, and mix. [note—Use this solution within 1 hour.]
Test solution— Transfer about 40 mg of Piperacillin, accurately weighed, to a 100-mL volumetric flask, add a few drops of methanol to dissolve it, dilute with Mobile phase to volume, and mix. [note—Use this solution within 1 hour.]
Chromatographic system (see Chromatography 621) The liquid chromatograph is equipped with a 220-nm detector and a 4.6-mm × 25-cm column that contains packing L1. The flow rate is about 1 mL per minute. Chromatograph the Standard solution, and record the peak responses as directed for Procedure: the relative retention times are about 1.0 for piperacillin and 2.55 for 6-[2-[6-[2-(4-ethyl-2,3-dioxo-1-piperazinecarboxamido)-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 (piperacillin related compound D); and the relative standard deviation for replicate injections is not more than 2%.
Procedure— Separately inject equal volumes (about 10 µL) of the Standard solution and the Test solution into the chromatograph, record the chromatograms, and measure the peak responses for piperacillin and piperacillin related compound D. Calculate the percentage of piperacillin related compound D in the portion of Piperacillin taken by the formula:
10C(1.47)(P / W)(ri / rS)
in which C is the concentration, in mg per mL, of USP Piperacillin RS in the Standard solution; 1.47 is the relative response factor for piperacillin related compound D; P is the designated potency, in µg of piperacillin per mg, of USP Piperacillin RS; W is the weight, in mg, of Piperacillin taken to prepare the Test solution; ri is the impurity peak response obtained from the Test solution; and rS is the peak response of piperacillin obtained from the Standard solution: not more than 2.0% of piperacillin related compound D is found. The sum of all impurities found in Test 1 and Test 2 is not more than 3.8%.
Other requirements— Where the label states that Piperacillin is sterile, it meets the requirements for Sterility and Bacterial endotoxins under Piperacillin for Injection. Where the label states that Piperacillin must be subjected to further processing during the preparation of injectable dosage forms, it meets the requirements for Bacterial endotoxins under Piperacillin for Injection.
Assay—
Mobile phase— Prepare a mixture of 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 ± 0.02. Make adjustments if necessary (see System Suitability under Chromatography 621).
Standard preparation— Transfer about 40 mg of USP Piperacillin RS, accurately weighed, to a 100-mL volumetric flask, add a few drops of methanol to dissolve it, dilute with Mobile phase to volume, and mix. [note—Use this solution within 1 hour.]
Resolution solution— Prepare a solution in Mobile phase containing in each mL about 0.1 mg of USP Ampicillin RS and 0.2 mg of USP Piperacillin RS.
Assay preparation— Transfer about 40 mg of Piperacillin, accurately weighed, to a 100-mL volumetric flask, add a few drops of methanol to dissolve it, dilute with Mobile phase to volume, and mix. [note—Use this solution within 1 hour.]
Chromatographic system (see Chromatography 621) The liquid chromatograph is equipped with a 220-nm detector and a 4.6-mm × 25-cm column that contains packing L1. The flow rate is about 1 mL per minute. Chromatograph the Resolution solution, and record the peak responses as directed for Procedure: the relative retention times are about 0.24 for piperacillin related compound B, 0.31 for ampicillin, 0.37 for piperacillin related compound C, 0.62 for related compound A, and 1.0 for piperacillin, the resolution, R, between ampicillin and piperacillin is not less than 16, and the tailing factor for the piperacillin peak is not more than 1.2. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the relative standard deviation for replicate injections is not more than 2%.
Procedure— Separately inject equal volumes (about 10 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the major peaks. Calculate the potency, in µg of piperacillin (C23H27N5O7S) per mg, of the portion of Piperacillin taken by the formula:
100(CP / W)(rU / rS)
in which C is the concentration, in mg per mL, of USP Piperacillin RS in the Standard preparation; P is the designated potency, in µg of piperacillin per mg, of USP Piperacillin RS; W is the weight, in mg, of Piperacillin taken to prepare the Assay preparation; and rU and rS are the piperacillin peak responses obtained from the Assay preparation and the Standard preparation, respectively.
Auxiliary Information— Staff Liaison : Ahalya Wise, M.S. , Scientist
Expert Committee : (MDANT05) Monograph Development-Antibiotics
USP31–NF26 Page 3011
Pharmacopeial Forum : Volume No. 28(4) Page 1182
Phone Number : 1-301-816-8161