Piperacillin and Tazobactam for Injection USP – Specification & STP

Explore detailed specifications and analysis methods for Piperacillin and Tazobactam for Injection USP. Enhance your pharmaceutical knowledge and practices.

Specification – Piperacillin and Tazobactam for Injection USP

Sr. No. Test Parameter Specification
1. Description White to off-white, crystalline powder, filled in 20 ml colourless transparant moulded vial, Type – I with lavender colour transparent seal. (OR as per manufacturer specification)
2. Identification

(Piperacillin Sodium & Tazobactam Sodium)

 By HPLC; The retention times of the major peaks of the sample solution corresponds to those of the standard solution, as obtained in the assay.
3. pH 5.00  to  7.00
4. Average Filled Weight ± 7.5% of theoretical average weight.
5. Uniformity of Filled Weight ± 10.0% of theoretical average weight.
6. Constituted Solution

 

 The solid should dissolve completely leaving no visible residue. The constituted solution should not be significantly less clear than an equal volume of diluent or purified water.
7. Uniformity of Dosage Units 85.00  to  115.00%
8. Particulate Matter :
a)  ≥ 10 µm Not more than 6,000 particles/container.
b)  ≥ 25 µm Not more than 600 particles/container.
9. Water Determination Not more than 2.5%
10. Related Compounds :
Tazobactam Related Compound Ab Not more than 1.0%
Piperacillin Impurity 4 Not more than 1.0%
Piperacillin penilloic acid  d, e Not more than 1.0%
Piperacillin penicilloic acid d,f Not more than 5.0%
Acetylated penicilloic acid of piperacillin g Not more than 1.0%
Piperacillin Impurity 5c Not more than 1.0%
Piperacillin Impurity 6c Not more than 1.0%
Any Individual Unspecified Impurity Not more than 1.0%
Total impurities Not more than 5.0%
13. Assay  – By HPLC.
Each vial contains :

Piperacillin Sodium USP  eq. to Piperacillin  2 g.

 1.800   to   2.200 g.

(90.00   to   110.00% of labeled amount).
Tazobactam Sodium  eq. to  Tazobactam USP  0.25 g. 0.225   to   0.275 g.

(90.00   to   110.00% of labeled amount).

Download the Specification – Piperacillin and Tazobactam for Injection USP

STP – Piperacillin and Tazobactam for Injection USP

1.0     Description

2.0     Identification

    • The retention times of the major peaks of the sample solution corresponds to those of the standard solution, as obtained in the assay.

3.0     pH

    • To be reconstituted with WFI (In a solution containing the equivalent of 40 mg/ml of Piperacillin).  

4.0     Average Filled Weight

    • Average filled weight will vary batch to batch according to the active ingredient added after adjusting their potency. 

5.0     Uniformity of Filled Weight – Piperacillin Sodium and Tazobactam Sodium Injection

6.0     Constituted Solution

    • Carry out the test as per procedure.

7.0     Uniformity of Dosage Units

    • Carry out the test as per procedure.

8.0     Particulate Matter

    • Carry out the test as per procedure.

9.0     Water Determination

    • Weigh sample accurately 250 mg;

10.0    Sterility

    • Carry out the test as per procedure.

11.0    Bacterial Endotoxins

    • Carry out the test as per procedure.

12.0     Related Compounds – Piperacillin Sodium and Tazobactam Sodium Injection

    • Buffer  :  Dilute the contents of one vial of tetrabutylammonium hydrogen sulfate ion pairing reagent with water to 1 L.
    • Procedure for Buffer Preparation :
    • Gently pour all of the powder or solution from the TBAHS vial into a sterile, volumetric flask (ideally 1000 mL).
    • Fill the flask with a lower amount of filtered water (such as 500–800 mL), then gently shake or swirl to dissolve the material entirely.
    • Gently fill the flask with more water until the meniscus’s bottom lines up with the volumetric flask’s 1 L mark.
    • To make sure the solution is perfectly homogenous, stopper the flask and invert it multiple times
    • Solution A:  Phosphoric acid and water (1 : 4)

Dilution : Take 10 ml of orthophosphoric Acid (H₃PO₄) in 50 ml volumetric flask, Slowey add purified water to make up the volume of 50 ml volumetric flask.

    • 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
    • Dilution : Weigh accurately  6 mg of USP Tazobactam Related Compound A RS in 100 ml volumetric flask, Slowey add Diluent to make up the volume of 100 ml volumetric flask. (60ppm/60mcg.)
    • Standard Stock Solution 2  :  0.5 mg/mL of USP Tazobactam RS in Diluent.
    • Dilution : Weigh accurately  25 mg of USP Tazobactam Related Compound A RS in 50 ml volumetric flask, add Diluent to make up the volume of 50 ml volumetric flask. (500ppm/500mcg.)
    • 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  :

Mode                     :         LC

Detector                :         UV 210 nm

Column                 :         4.6-mm × 15-cm; 3-µm packing L11 (Inertsil Ph, InertSustain Phenyl)

Flow rate               :         1 mL/min.

Injection volume    :         20 µL

Autosampler temp. :         5 ± 3°

    • Suitability Requirements
    • Resolution  :  NLT 3 between tazobactam related compound A and tazobactam.
    • Tailing Factor  :  NMT 1.8 for tazobactam and piperacillin, Standard solution.
    • Relative Standard Deviation  :  NMT 2% for tazobactam and piperacillin, Standard solution.

SOP for Good Chromatographic Practices

  • HPLC 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) × (C S / C U) × P × (F 1 / F 2) × 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)
F 1 = correction factor, 0.001 mg/µg
F 2 = relative response factor (see Table 1)

Table 1

Name Relative
Retention
Time		 Relative
Response
Factor		 Acceptance
Criteria,
NMT (%)
    Tazobactam related compound Ab 0.12 0.75 1.0
  Tazobactam 0.25 — —
Piperacillin impurity 4c 0.31 1.0 1.0
  Piperacillin penilloic acid  d, e 0.36 1.0 1.0
Piperacillin penicilloic acid d, f 0.51 0.56 5.0
Acetylated penicilloic acid of piperacillin g 0.55 1.0 1.0
Piperacillin impurity 5c 0.62 1.0 1.0
Piperacillin impurity 6c 0.67 1.0 1.0
Piperacillin 1.0 — —
Any individual unspecified impurity — 1.0 1.0
Total impuritiesh — — 5.0

 

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.

13.0     Assay – Piperacillin Sodium and Tazobactam Sodium Injection USP

  • Procedure:  By HPLC.
    • Buffer  :   27.6 g/L of monobasic sodium phosphate
    • Solution A  :  80 mL of 40% aqueous tetrabutylammonium hydroxide diluted with water to 100mL.
    • 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.

Dilution:  About 10.5 mg Tazobactam & 105 mg Piperacillin ——–> 100 ml).

    • 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.
    • (Dilution: About 125mg sample ——>100 ml).
    • Chromatographic System:

Mode                     :         LC

Detector                :         UV 230 nm

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

Flow rate               :         1 mL/min.

Injection volume    :         10 µL

Autosampler temp. :         5 ± 3°

    • System Suitability

    • Note:-  The relative retention times for tazobactam and piperacillin are 0.36 and 1.0, respectively.]
    • Suitability Requirements
    • Tailing factor           :  NMT 2.0 for tazobactam and piperacillin
    • Relative Standard Deviation  :  NMT 2.0% for tazobactam and piperacillin
    • Calculate the percentage of the labeled amount of piperacillin (C23 H27 N5 O7 S) in the portion of Piperacillin and Tazobactam for Injection taken:

          Result         =        (r U / r S) × (C S / C U) × P × F × 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 (C10 H12 N4 O5 S) in the portion of Piperacillin and Tazobactam for Injection taken :

Result         =        (r U / r S) × (C S / C U) × P × 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).

 

Download the Method of Analysis – Piperacillin and Tazobactam Injection USP

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