Packaging and storage
Preserve in tight 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
A:
The retention time of the major peak for clavulanic acid in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay.
B:
A solution of it responds to the tests for
Potassium 191.
Bacterial endotoxins 85
Where the label states that Clavulanate Potassium is sterile or must be subjected to further processing during the preparation of injectable dosage forms, it contains not more than 0.03 USP Endotoxin Unit per mg.
Sterility 71
Where the label states that Clavulanate Potassium is sterile, it meets the requirements when tested as directed for
Membrane Filtration under
Test for Sterility of the Product to be Examined.
pH 791:
between 5.5 and 8.0, in a solution (1 in 100).
Limit of clavam-2-carboxylate potassium
Mobile phase
Prepare 0.1 M monobasic sodium phosphate, adjust with phosphoric acid to a pH of 4.0 ± 0.1, and pass through a membrane filter having a 0.5-µm or finer porosity. Make adjustments if necessary (see
System Suitability under
Chromatography 621).
Test solution
Transfer about 100 mg of Clavulanate Potassium, accurately weighed, to a 10-mL volumetric flask, dissolve in and dilute with water to volume, and mix.
Chromatographic system (see Chromatography 621)
The liquid chromatograph is equipped with a 210-nm detector and a 4-mm × 30-cm column that contains 3- to 10-µm packing L1. The flow rate is about 0.5 mL per minute. Chromatograph the
Standard solution, and record the peak responses as directed for
Procedure: the column efficiency is not less than 4000 theoretical plates; the tailing factor is not more than 1.5; and the relative standard deviation for replicate injections is not more than 5%.
Procedure
Separately inject equal volumes (about 20 µL) of the
Standard solution and the
Test solution into the chromatograph, record the chromatograms, and measure the responses for the major peaks. The relative retention times are about 0.7 for clavam-2-carboxylic acid and 1.0 for clavulanic acid. Calculate the percentage of clavam-2-carboxylate potassium in the specimen taken by the formula:
(C/W)(rU / rS),
in which
C is the concentration, in µg per mL, of clavam-2-carboxylate potassium in the
Standard solution; W is the quantity, in mg, of Clavulanate Potassium taken to prepare the
Test solution; and
rU and
rS are the clavam-2-carboxylic acid peak responses obtained from the
Test solution and the
Standard solution, respectively: not more than 0.01% is found.
Limit of aliphatic amines
Internal standard solution
Dissolve 50 µL of 3-methyl-2-pentanone in water, dilute with water to 100.0 mL, and mix.
Test solution
Transfer 1.0 g of Clavulanate Potassium to a centrifuge tube, add 5.0 mL of Internal standard solution, 5.0 mL of 2 N sodium hydroxide, 5.0 mL of isopropyl alcohol, and 5 g of sodium chloride. Shake for 1 minute, and centrifuge to separate the layers. Use the upper layer.
Reference solution
Dissolve 80.0 mg of each of the following amines in 2 N hydrochloric acid: 1,1-dimethylethylamine, diethylamine, tetramethylethylenediamine, 1,1,3,3-tetramethylbutylamine, and N,N¢-diisopropylethylenediamine. Dilute with 2 N hydrochloric acid to 200.0 mL, and mix. Transfer 5.0 mL of this solution to a centrifuge tube. Add 5.0 mL of Internal standard solution, 10.0 mL of 2 N sodium hydroxide, 5.0 mL of isopropyl alcohol, and 5 g of sodium chloride. Shake for 1 minute, and centrifuge to separate the layers. Use the upper layer.
Chromatographic system (see Chromatography 621)
The gas chromatograph is equipped with a flame-ionization detector and a 0.53-mm × 50-m capillary fused silica column that contains a 5-µm film coating of stationary phase G41. The temperatures of the column, the injection port, and the detector block are programmed as follows.
|
Time (minutes) |
Tempera- ture () |
Elution |
Column |
07 |
35 |
isothermal |
|
710.8 |
35150 |
linear gradient |
|
10.825.8 |
150 |
isothermal |
Injection port |
|
200 |
|
Detector block |
|
250 |
|
The carrier gas is helium flowing at a constant rate of about 8 mL per minute. The split ratio is 1:10. Chromatograph the
Reference solution, and record the peak responses as directed for
Procedure: the relative retention times are about 0.55 for 1,1-dimethylethylamine, 0.76 for diethylamine, 1.0 for isopropyl alcohol, 1.07 for tetramethylethylenediamine, 1.13 for 1,1,3,3-tetramethylbutylamine, 1.33 for
N,N¢-diisopropylethylenediamine, and 1.57 for bis(2-methylamino)ethyl ether.
Procedure
Separately inject equal volumes (about 1 µL) of the
Test solution and the
Reference solution into the chromatograph, record the chromatograms, and measure the peak area responses. Calculate the percentage of each impurity in the Clavulanate Potassium taken by the formula:
0.2(ri / rS),
in which
ri is the peak response for an individual impurity observed in the chromatogram obtained from the
Test solution; and
rS is the response for the peak corresponding to the relevant analyte in the chromatogram obtained from the
Reference solution. Calculate the percentage of any individual impurity for which no relevant reference compound is provided in the
Reference solution by the same formula, except for
rS use the peak response corresponding to the 1,1-dimethylethylamine peak. The total of all aliphatic amines is not more than 0.2%.
Limit of 2-ethylhexanoic acid
Internal standard solution
Prepare a solution of 3-cyclohexylpropionic acid in cyclohexane containing 1 mg per mL.
Standard solution
Transfer about 75 mg of 2-ethylhexanoic acid, accurately weighed, to a 50-mL volumetric flask, dilute with Internal standard solution to volume, and mix. Transfer 1.0 mL of this solution to a centrifuge tube, and add 4.0 mL of 4 N hydrochloric acid. Shake for 1 minute, and allow the phases to separate, centrifuging if necessary. Withdraw the lower phase, and reserve the upper phase. To the lower phase add 1.0 mL of Internal standard solution, and shake for 1 minute. Allow the phases to separate, centrifuging if necessary. Withdraw the upper phase, and combine with the reserved upper layer.
Test solution
Transfer about 300 mg of Clavulanate Potassium, accurately weighed, to a centrifuge tube. Add 4.0 mL of 4 N hydrochloric acid, and shake with two successive 1.0 mL portions of Internal standard solution. Allow the phases to separate, centrifuging if necessary. Use the combined upper phases.
Chromatographic system (see Chromatography 621)
The gas chromatograph is equipped with a flame-ionization detector and a 0.53-mm × 25-m capillary fused silica column that contains a 1-µm film coating of stationary phase G35. The temperatures of the column, the injection port, and the detector block are programmed as follows.
|
Time (minutes) |
Tempera- ture () |
Rate ( per minute) |
Elution |
Column |
02 |
40 |
|
isothermal |
|
27.3 |
40200 |
30 |
linear gradient |
|
7.310.3 |
200 |
|
isothermal |
Injection port |
|
200 |
|
|
Detector block |
|
300 |
|
|
The carrier gas is hydrogen flowing at a constant rate of about 100 mL per minute. Chromatograph the
Standard solution, and record the peak responses as directed for
Procedure: the resolution,
R, between the 2-ethylhexanoic acid peak and the 3-cyclohexylpropionic acid peak is not less than 2.0.
Procedure
Separately inject equal volumes (about 1 µL) of the
Standard solution and the
Test solution into the chromatograph, record the chromatograms, and measure the peak area responses. Calculate the percentage of 2-ethylhexanoic acid in the Clavulanate Potassium taken by the formula:
2(WS / WU)(RU / RS),
in which
WS is the weight, in mg, of 2-ethylhexanoic acid taken to prepare the
Standard solution; WU is the weight, in mg, of Clavulanate Potassium taken to prepare the
Test solution; and
RU and
RS are the ratios of the 2-ethylhexanoic acid peak to the 3-cyclohexylpropionic acid peak observed in the chromatograms obtained from the
Test solution and the
Standard solution, respectively. Not more than 0.8% is found.
Chromatographic purity
Solution A
Prepare a solution of 0.05 M monobasic sodium phosphate, adjust with phosphoric acid to a pH of 4.0 ± 0.1, and pass through a filter having a 0.5-µm or finer porosity.
Solution B
Prepare a mixture of Solution A and methanol (50:50).
Mobile phase
Use variable mixtures of
Solution A and
Solution B as directed for
Chromatographic system. Make adjustments to either
Solution as necessary (see
System Suitability under
Chromatography 621).
Standard solution
Prepare a solution of
USP Clavulanate Lithium RS in
Solution A having a known concentration of about 0.1 mg per mL.
Test solution
Prepare a solution of Clavulanate Potassium in Solution A containing 10.0 mg per mL.
Resolution solution
Prepare a solution of
USP Clavulanate Lithium RS and amoxicillin in
Solution A containing 0.1 mg of each per mL.
Chromatographic system (see Chromatography 621)
The liquid chromatograph is equipped with a 230-nm detector and a 4.6-mm × 10-cm column that contains 5-µm packing L1 and is maintained at a constant temperature of about 40
. The flow rate is about 1 mL per minute. The system is programmed to provide a mobile phase consisting of variable mixtures of
Solution A and
Solution B. The system is equilibrated for 15 minutes with 100%
Solution A, and held at that composition for 4 minutes after injection of the solution under test, after which the proportion of
Solution B is increased linearly from 0% to 50% over a period of 11 minutes. The system is held at that composition for 3 minutes, and is then changed to 100%
Solution A for 6 minutes. Chromatograph the
Resolution solution, and record the peak responses as directed for
Procedure: the relative retention times are about 1.0 for clavulanic acid and 2.5 for amoxicillin; the tailing factor for the clavulanic acid peak is not more than 2.0; the column efficiency determined from the clavulanic acid peak is not less than 2000 theoretical plates; and the resolution,
R, between the clavulanic acid peak and the amoxicillin peak is not less than 13. Chromatograph the
Standard solution, 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 20 µL) of the
Standard solution and the
Test solution into the chromatograph, record the chromatograms, and measure the peak responses. Calculate the percentage, in terms of clavulanate potassium equivalent, of each impurity in the Clavulanate Potassium taken by the formula:
10(237.3/205.1)(C)(ri / rS),
in which 237.3 is the molecular weight of clavulanate potassium; 205.1 is the molecular weight of clavulanate lithium;
C is the concentration, in mg per mL, of USP Clavulanate Potassium RS in the
Standard solution; ri is the peak response of an individual impurity peak in the chromatogram obtained from the
Test solution; and
rS is the clavulanic acid peak response in the chromatogram obtained from the
Standard solution. The sum of all the impurity peaks is not greater than 2%.
Assay
pH 4.4 sodium phosphate buffer
Dissolve 7.8 g of monobasic sodium phosphate in 900 mL of water, adjust with phosphoric acid or 10 N sodium hydroxide to a pH of 4.4 ± 0.1, dilute with water to make 1000 mL, and mix.
Mobile phase
Prepare a suitable mixture of
pH 4.4 sodium phosphate buffer and methanol (95:5), and pass through a membrane filter having a 0.5-µm or finer porosity. Make adjustments if necessary (see
System Suitability under
Chromatography 621).
Standard preparation
Dissolve an accurately weighed quantity of
USP Clavulanate Lithium RS in water to obtain a solution having a known concentration of about 0.25 mg per mL.
Assay preparation
Transfer about 50 mg of Clavulanate Potassium, accurately weighed, to a 200-mL volumetric flask, dissolve in and dilute with water to volume, and mix.
Resolution solution
Dissolve a suitable quantity of amoxicillin in Standard preparation to obtain a solution containing about 0.5 mg of amoxicillin and 0.25 mg of clavulanate lithium per mL.
Chromatographic system (see Chromatography 621)
The liquid chromatograph is equipped with a 220-nm detector and a 4-mm × 30-cm column that contains 3- to 10-µm packing L1. The flow rate is about 2 mL per minute. Chromatograph the
Standard preparation, and record the peak responses as directed for
Procedure: the column efficiency determined from the analyte peak is not less than 550 theoretical plates; the tailing factor for the analyte peak is not more than 1.5; and the relative standard deviation for replicate injections is not more than 2.0%. Chromatograph the
Resolution solution, and record the peak responses as directed for
Procedure: the relative retention times are about 0.5 for clavulanic acid and 1.0 for amoxicillin; and the resolution,
R, between the amoxicillin and clavulanic acid peaks is not less than 3.5.
Procedure
Separately inject equal volumes (about 20 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 quantity, in µg, of clavulanic acid (C
8H
9NO
5) in each mg of the Clavulanate Potassium taken by the formula:
(200CP/W)(rU / rS),
in which
C is the concentration, in mg per mL, of
USP Clavulanate Lithium RS in the
Standard preparation; P is the designated potency, in µg of clavulanic acid per mg, of
USP Clavulanate Lithium RS;
W is the quantity, in mg, of Clavulanate Potassium taken to prepare the
Assay preparation; and
rU and
rS are the peak responses obtained from the
Assay preparation and the
Standard preparation, respectively.