Ofloxacin Ophthalmic Solution
» Ofloxacin Ophthalmic Solution is a sterile aqueous solution of Ofloxacin. It contains not less than 90.0 percent and not more than 110.0 percent of the labeled amount of ofloxacin (C18H20FN3O4).
Packaging and storage Preserve in tight containers at controlled room temperature.
A: Thin-Layer Chromatographic Identification Test 201
Test solution Dilute a portion of Ophthalmic Solution with a mixture of chloroform and methanol (1:1) to obtain a solution having a concentration of about 0.3 mg of ofloxacin per mL.
Standard solution Dissolve an accurately weighed quantity of USP Ofloxacin RS in a mixture of chloroform and methanol (1:1) to obtain a solution having a concentration of 3.0 mg per mL. Transfer 5.0 mL of this solution to a 50-mL volumetric flask, add 5 mL of water, dilute with a mixture of chloroform and methanol (1:1) to volume, and mix.
Application volume: 2 µL.
Developing solvent system: a mixture of chloroform, methanol, and a solution (1 in 30) of ammonium hydroxide (150:75:15). Saturate a paper-lined chromatographic chamber with this mixture.
B: The retention time of the ofloxacin 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.
pH 791: between 6.0 and 6.8.
Mobile phase Prepare a filtered and degassed mixture of sodium dodecyl sulfate (0.24% aqueous solution), acetonitrile, and glacial acetic acid (580:400:20). Make adjustments if necessary (see System Suitability under Chromatography 621).
0.05 N Hydrochloric acid Add 4.0 mL of hydrochloric acid to 500 mL of water, dilute with water to 1000 mL, and mix.
Resolution solution Prepare a solution of about 0.1 mg of USP Ofloxacin RS and about 2.4 mg of propylparaben in each mL of acetonitrile.
Standard preparation Quantitatively dissolve an accurately weighed quantity of USP Ofloxacin RS in 0.05 N Hydrochloric acid to obtain a solution having a known concentration of about 0.06 mg per mL.
Assay preparation Transfer an accurately measured volume of Ophthalmic Solution, equivalent to about 3 mg of ofloxacin, to a 50-mL volumetric flask, dilute with 0.05 N Hydrochloric acid to volume, and mix.
Chromatographic system (see Chromatography 621) The liquid chromatograph is equipped with a 294-nm detector and a 4.6-mm × 25-cm column that contains 5-µm packing L1. The flow rate is about 1.5 mL per minute. The column temperature is maintained at a constant temperature of about 35. Chromatograph the Resolution solution, and record the peak responses as directed for Procedure: the resolution, R, between propylparaben and ofloxacin is not less than 2. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the tailing factor is not more than 3; and the relative standard deviation for replicate injections is not more than 2.0%.
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 areas for the major peaks. Calculate the quantity, in mg, of ofloxacin (C18H20FN3O4) in each mL of the Ophthalmic Solution taken by the formula:
50(C / V)(rU / rS)in which C is the concentration, in mg per mL, of USP Ofloxacin RS in the Standard preparation; V is the volume, in mL, of Ophthalmic Solution taken to prepare the Assay preparation; and rU and rS are the ofloxacin peak areas obtained from the Assay preparation and the Standard preparation, respectively.
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USP32NF27 Page 3129Pharmacopeial Forum: Volume No. 28(4) Page 1170