Moxifloxacin Ophthalmic Solution
» Moxifloxacin Ophthalmic Solution is a sterile, self-preserved aqueous solution of Moxifloxacin Hydrochloride. It contains not less than 90.0 percent and not more than 110.0 percent of the labeled amount of moxifloxacin (C21H24FN3O4).
Packaging and storage—
Preserve in tight containers. Store between 2
and 25.
and 25.
USP Reference standards 
11
—
11—
USP Moxifloxacin Hydrochloride RS 
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USP Moxifloxacin Related Compound A RS
1-Cyclopropyl-6,8-difluoro-1,4-dihydro-7-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]-4-oxo-3-quinolinecarboxylic acid.
C20H21F2N3O3 389.40
1-Cyclopropyl-6,8-difluoro-1,4-dihydro-7-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]-4-oxo-3-quinolinecarboxylic acid.
C20H21F2N3O3 389.40
Identification—
The retention time of the major peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay.
Sterility 71—
It meets the requirements when tested as directed for Membrane Filtration under Test for Sterility of the Product to be Examined.
71—
It meets the requirements when tested as directed for Membrane Filtration under Test for Sterility of the Product to be Examined.
pH 791:
between 6.3 and 7.3.
791:
between 6.3 and 7.3.
Osmolality 785:
between 260 and 320 mOsmol per kg.
785:
between 260 and 320 mOsmol per kg.
Related compounds—
[note—Protect solutions from light. Analyze the Test solution immediately after preparation. ]
test 1—
Early-eluting related compounds (relative retention time less than 1.8).
Buffer solution—
Use the Buffer solution prepared as directed in the Assay.
Mobile phase—
Use variable mixtures of Buffer solution and methanol as directed in Table 1. Make adjustments if necessary (see System Suitability under Chromatography 621).
621).
Blank solution—
Use the Buffer solution.
Resolution solution—
Use the Resolution solution prepared as directed in the Assay.
Standard solution—
Dissolve an accurately weighed quantity of USP Moxifloxacin Hydrochloride RS in water to obtain a solution containing about 6 mg per mL. Dilute quantitatively, and stepwise if necessary, with the Buffer solution to obtain a solution having a known concentration of about 0.002 mg per mL.
Sensitivity solution—
Dilute an accurately measured volume of the Standard solution with Buffer solution to obtain a solution having a known concentration of about 0.05 µg per mL. [note—Store the Sensitivity solution under refrigeration and protected from light. ]
Test solution—
Use the Assay preparation.
Chromatographic system (see Chromatography 621)—
The liquid chromatograph is equipped with a 293-nm detector and 4.0-mm × 25-cm column that contains 5-µm packing L11. The chromatograph is programmed as shown in Table 1.
The flow rate is about 0.5 mL per minute. The column temperature is maintained at 45. Chromatograph the Resolution solution, and record the peak responses as directed for Procedure: the resolution, R, between moxifloxacin and moxifloxacin related compound A is not less than 2.0. Chromatograph the Standard solution, and record peak responses as directed for Procedure: the column efficiency is not less than 4000 theoretical plates; the tailing factor is not more than 2.0; and the relative standard deviation for replicate injections is not more than 2.0%. In addition, chromatograph the Sensitivity solution, and record the peak response as directed for Procedure. Confirm that the signal-to-noise ratio of moxifloxacin is not less than 10.
621)—
The liquid chromatograph is equipped with a 293-nm detector and 4.0-mm × 25-cm column that contains 5-µm packing L11. The chromatograph is programmed as shown in Table 1.
Table 1
| Time (minutes) |
Flow Rate (mL/minute) |
Buffer solution (%) |
Methanol (%) |
Elution |
|---|---|---|---|---|
| 0–30 | 0.5 | 69 | 31 | isocratic |
| 30–31 | 0.5 | 69®60 | 31®40 | linear gradient |
| 31–36 | 0.9 | 60 | 40 | isocratic |
| 36–37 | 0.9 | 60®69 | 40®31 | linear gradient |
| 37–42 | 0.5 | 69 | 31 | re-equilibration |
. Chromatograph the Resolution solution, and record the peak responses as directed for Procedure: the resolution, R, between moxifloxacin and moxifloxacin related compound A is not less than 2.0. Chromatograph the Standard solution, and record peak responses as directed for Procedure: the column efficiency is not less than 4000 theoretical plates; the tailing factor is not more than 2.0; and the relative standard deviation for replicate injections is not more than 2.0%. In addition, chromatograph the Sensitivity solution, and record the peak response as directed for Procedure. Confirm that the signal-to-noise ratio of moxifloxacin is not less than 10.
Procedure—
Separately inject equal volumes (about 25 µL) of the Blank solution, the Standard solution, and the Test solution into the chromatograph, record the chromatograms, and measure the peak responses, disregarding any peaks corresponding to those obtained from the Blank solution. Calculate the percentage of each related compound in the portion of Ophthalmic Solution taken by the formula:
(401.43/437.89)( CS /CU)(1/F)(ri / rS)(100)
in which 401.43 and 437.89 are the molecular weights of moxifloxacin free base and moxifloxacin hydrochoride, respectively; CS is the concentration, in mg per mL, of USP Moxifloxacin Hydrochloride RS in the Standard solution; CU is the concentration, in mg per mL, of Test solution based on the label claim; F is the relative response factor for the individual related compound; ri is the peak response of each individual impurity in the Test solution; rS is the peak response of moxifloxacin in the Standard solution; and 100 is the percent factor. The following limits as shown in Table 2 are met.
Table 2
| Related Compound | F | Relative Retention Time vs. Moxifloxacin |
Limit (%) |
|---|---|---|---|
| Specified unknown impurity #1 | 1.0 | 0.3 | 0.2 |
| Decarboxy1 | 0.13 | 0.4 | 0.3 |
| Specified unknown impurity #2 | 1.0 | 0.9 | 0.3 |
| Any specified and identified impurity | 1.0 | — | 1.0 |
| Other single impurities | 1.0 | — | 0.1 |
| *Moxifloxacin related compound A | — | 1.1 | — |
| **8-Hydroxy2 | — | 1.8 | — |
|
1
1-Cyclopropyl-6-fluoro-8-methoxy-7-[(4aS,7aS)-octahydro-pyrrolo[3,4-b]pyridin-6-yl]-1H-quinolin-4-one.
2
1-Cyclopropyl-6-fluoro-8-hydroxy-7-[(4aS,7aS)-octahydro-6H-pyrrolo[3,4-b]pyridin-6-yl]-4-oxo-1,4-dihydroquinoline-3-carboxylic acid.
*
Disregard this peak because this is a process impurity controlled for the drug substance.
**
Disregard this peak because it is quantitated using Test 2.
|
|||
test 2—
Late-eluting related compounds (relative retention time equal to more than 1.8).
Buffer solution—
Use the Buffer solution prepared as directed in the Assay.
Mobile phase—
Prepare a filtered and degassed mixture of Buffer solution and methanol (60:40). Make adjustments if necessary (see System Suitability under Chromatography 621).
621).
Blank solution—
Use the Buffer solution.
Standard solution—
Dissolve an accurately weighed quantity of USP Moxifloxacin Hydrochloride RS in water to obtain a solution containing about 6 mg per mL. Dilute quantitatively, and stepwise if necessary, with the Buffer solution to obtain a solution having a known concentration of about 0.002 mg per mL.
Sensitivity solution—
Dilute an accurately measured volume of the Standard solution with Buffer solution to obtain a solution having a known concentration of about 0.05 µg per mL. [note—Store the Sensitivity solution under refrigeration and protected from light. ]
Test solution—
Use the Assay preparation. [note—The 8-Hydroxy compound is unstable in dilute aqueous solutions. Analyze the Test solution immediately after preparation. ]
Chromatographic system (see Chromatography 621)—
The liquid chromatograph is equipped with a 293-nm detector and 4.0-mm × 25-cm column that contains 5-µm packing L11. The flow rate is about 0.9 mL per minute. The column temperature is maintained at 45. Chromatograph the Standard solution, and record the peak responses as directed for Procedure: the column efficiency is not less than 2000 theoretical plates; the tailing factor is not more than 2.0; and the relative standard deviation for replicate injections is not more than 2.0%. In addition, chromatograph the Sensitivity solution, and record the peak response as directed for Procedure. Confirm that the signal-to-noise ratio of the moxifloxacin is not less than 10.
621)—
The liquid chromatograph is equipped with a 293-nm detector and 4.0-mm × 25-cm column that contains 5-µm packing L11. The flow rate is about 0.9 mL per minute. The column temperature is maintained at 45. Chromatograph the Standard solution, and record the peak responses as directed for Procedure: the column efficiency is not less than 2000 theoretical plates; the tailing factor is not more than 2.0; and the relative standard deviation for replicate injections is not more than 2.0%. In addition, chromatograph the Sensitivity solution, and record the peak response as directed for Procedure. Confirm that the signal-to-noise ratio of the moxifloxacin is not less than 10.
Procedure—
Separately inject equal volumes (about 25 µL) of the Blank solution, the Standard solution, and the Test solution into the chromatograph, record the chromatograms, and measure the peak responses, disregarding any peaks corresponding to those obtained from the Blank solution. Calculate the percentage of each related compound in the portion of Ophthalmic Solution taken by the formula:
(401.43/437.89)(CS /CU)(1/F))(ri / rS)(100)
in which 401.43 and 437.89 are the molecular weights of moxifloxacin free base and moxifloxacin hydrochoride, respectively; CS is the concentration, in mg per mL, of USP Moxifloxacin Hydrochloride RS in the Standard solution; CU is the concentration, in mg per mL, of Test solution based on the label claim; F is the relative response factor for the individual related compound; ri is the peak response of each individual impurity in the Test solution; rS is the peak response of moxifloxacin in the Standard solution; and 100 is the percent factor. The following limits as shown in Table 3 are met.
Table 3
| Related Compound | F | Relative Retention Time vs. Moxifloxacin |
Limit (%) |
|---|---|---|---|
| 8-Hydroxy | 0.29 | 1.8 | 0.2 |
| Specified unknown impurity #3 | 1.0 | 3.4 | 0.2 |
| Specified impurity #4* | 0.42 | 3.9 | 0.2 |
| Other single impurities | 1.0 | — | 0.1 |
| Total impurities (sum from both Test 1 and Test 2) | — | — | 1.5 |
|
*
7-Amino-1-cyclopropyl-6-fluoro-8-methoxy-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid.
|
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Assay—
Buffer solution—
Dissolve 0.5 g of tetrabutylammonium hydrogen sulfate and 1.0 g of monobasic potassium phosphate in 1000 mL of water, add 2 mL of phosphoric acid, filter, and degas.
Mobile phase—
Use variable mixtures of Buffer solution and methanol as directed in Table 1. Make adjustments if necessary (see System Suitability under Chromatography 621.
621.
Resolution solution—
Dissolve suitable quantities of USP Moxifloxacin Hydrochloride RS and USP Moxifloxacin Related Compound A RS in Buffer solution to obtain a solution containing about 0.1 mg per mg and 0.001 mg per mg, respectively.
Standard preparation—
Dissolve an accurately weighed quantity of USP Moxifloxacin Hydrochloride RS in water to obtain a solution containing about 6 mg per mL. Dilute quantitatively, and stepwise if necessary, with the Buffer solution to obtain a solution having a known concentration of about 0.1 mg per mL.
Assay preparation—
Transfer an accurately measured volume of Ophthalmic Solution, equivalent to about 5 mg of moxifloxacin, to a 50-mL volumetric flask, dilute with Buffer solution to volume, and mix.
Chromatographic system (see Chromatography 621)—
The liquid chromatograph is equipped with a 293-nm detector and 4.0-mm × 25-cm column that contains 5-µm packing L11. The chromatograph is programmed as shown in Table 1. The column temperature is maintained at 45. Chromatograph the Resolution solution, and record the peak responses as directed for Procedure: the resolution, R, between moxifloxacin and moxifloxacin related compound A is not less than 2.0. Chromatograph the Standard preparation, and record peak responses as directed for Procedure: the column efficiency is not less than 4000 theoretical plates; the tailing factor is not more than 2.0; and the relative standard deviation for replicate injections is not more than 2.0%.
621)—
The liquid chromatograph is equipped with a 293-nm detector and 4.0-mm × 25-cm column that contains 5-µm packing L11. The chromatograph is programmed as shown in Table 1. The column temperature is maintained at 45. Chromatograph the Resolution solution, and record the peak responses as directed for Procedure: the resolution, R, between moxifloxacin and moxifloxacin related compound A is not less than 2.0. Chromatograph the Standard preparation, and record peak responses as directed for Procedure: the column efficiency is not less than 4000 theoretical plates; the tailing factor is not more than 2.0; and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure—
Separately inject equal volumes (about 25 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the peak responses for the major peaks. Calculate the percent of labeled amount of C21H24FN3O4 in the portion of the Ophthalmic Solution taken by the formula:
(401.43/437.89)(CS /CU)/(rU /rS)(100)
in which 401.43 and 437.89 are the molecular weights of moxifloxacin free base and moxifloxacin hydrochoride, respectively; CS is the concentration, in mg per mL, of USP Moxifloxacin Hydrochloride RS in the Standard preparation; CU is the concentration, in mg per mL, of Assay preparation based on the label claim; rU and rS are the peak responses obtained from the Assay preparation and Standard preparation, respectively; and 100 is the percent factor.
Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Leonel M. Santos, Ph.D.
Senior Scientific Liaison 1-301-816-8168 |
(SM12010) Monographs - Small Molecules 1 |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
|
| 71 |
Radhakrishna S Tirumalai, Ph.D.
Principal Scientific Liaison 1-301-816-8339 |
(GCM2010) General Chapters - Microbiology |
USP35–NF30 Page 3960
Pharmacopeial Forum: Volume No. 34(5) Page 1173