Polymyxin B Sulfate and Trimethoprim Ophthalmic Solution
» Polymyxin B Sulfate and Trimethoprim Ophthalmic Solution is a sterile, isotonic, aqueous solution of Polymyxin B Sulfate and Trimethoprim Sulfate or of Polymyxin B Sulfate and Trimethoprim that has been solubilized with Sulfuric Acid. It contains not less than 90.0 percent and not more than 130.0 percent of the labeled amount of polymyxin B and the equivalent of not less than 90.0 percent and not more than 110.0 percent of the labeled amount of trimethoprim (C14H18N4O3). It contains one or more preservatives.
Packaging and storage Preserve in tight, light-resistant containers, and store at controlled room temperature.
Labeling Label it to indicate that it is to be stored at 15 to 25, protected from light.
USP Reference standards 11
USP Polymyxin B Sulfate RS
A: It meets the requirements for polymyxin B under Thin-Layer Chromatographic Identification Test 201BNP.
B: The retention time of the trimethoprim peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay for trimethoprim.
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 4.0 and 6.2.
Assay for polymyxin B Proceed as directed for polymyxin B under AntibioticsMicrobial Assays 81, using an accurately measured volume of Ophthalmic Solution, diluted quantitatively and stepwise with Buffer No. 6, to obtain a Test Dilution having a concentration of polymyxin B assumed to be equal to the median dose level of the Standard.
Assay for trimethoprim
Diluent Prepare a mixture of 0.01 N hydrochloric acid and acetonitrile (870:130).
Mobile phase Dissolve 1.65 g of ethanesulfonic acid in 1000 mL of a mixture of water and acetonitrile (870:130). Adjust with 10 N sodium hydroxide or 0.1 N hydrochloric acid to a pH of 3.5. Pass this solution through a filter having a 0.5-µm or finer porosity, and degas. Make adjustments if necessary (see System Suitability under Chromatography 621).
Standard preparation Dissolve an accurately weighed quantity of USP Trimethoprim RS in Diluent to obtain a solution having a known concentration of about 0.04 mg per mL.
Assay preparation Transfer an accurately measured volume of Ophthalmic Solution, equivalent to about 1 mg of trimethoprim, to a 25-mL volumetric flask, dilute with Diluent to volume, and mix.
Chromatographic system (see Chromatography 621) The liquid chromatograph is equipped with a 254-nm detector and a 3.9-mm × 30-cm column that contains packing L11. The flow rate is about 1.5 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the tailing factor is not more than 1.5, when calculated at 10% height of the peak; and the relative standard deviation for replicate injections is not more than 2.0%.
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 quantity, in mg, of trimethoprim (C14H18N4O3) in each mL of the Ophthalmic Solution taken by the formula:
25(C/V)(rU / rS)in which C is the concentration, in mg per mL, of USP Trimethoprim 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 trimethoprim peak area responses obtained from the Assay preparation and the Standard preparation, respectively.
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USP35NF30 Page 4351Pharmacopeial Forum: Volume No. 28(4) Page 1185