Neomycin Sulfate and Hydrocortisone Acetate Ophthalmic Suspension
»Neomycin Sulfate and Hydrocortisone Acetate Ophthalmic Suspension is a sterile,aqueous suspension containing the equivalent of not less than 90.0percent and not more than 130.0percent of the labeled amount of neomycin,and not less than 90.0percent and not more than 110.0percent of the labeled amount of hydrocortisone acetate (C23H32O6).
Packaging and storage— Preserve in tight containers.The containers or individual cartons are sealed and tamper-proof so that sterility is assured at time of first use.
Identification— The chromatogram of the Assay preparationobtained as directed in the Assay for hydrocortisone acetateexhibits a major peak for hydrocortisone acetate,the retention time of which corresponds with that exhibited in the chromatogram of the Standard preparationobtained as directed in the Assay for hydrocortisone acetate.
Sterility á71ñ: meets the requirements.
pHá791ñ: between 5.5and 7.5.
Assay for neomycin— Proceed as directed for neomycin under Antibiotics—Microbial Assays á81ñ,using an accurately measured volume of Ophthalmic Suspension,freshly mixed and free from air bubbles,diluted quantitatively and stepwise with Buffer No.3to yield a Test Dilutionhaving a concentration assumed to be equal to the median dose level of the Standard.
Assay for hydrocortisone acetate—
Mobile phase— Prepare a solution containing n-butyl chloride,water-saturated n-butyl chloride,tetrahydrofuran,methanol,and glacial acetic acid (95:95:14:7:6).
Internal standard solution— Prepare a solution of fluoxymesterone in chloroform containing 0.8mg per mL.
Standard preparation— Dissolve about 10mg of USP Hydrocortisone Acetate RS,accurately weighed,in 10.0mLof Internal standard solution,dilute with about 40mLof chloroform,and mix.
Assay preparation— Transfer an accurately measured volume of Ophthalmic Suspension,freshly mixed and free from air bubbles,equivalent to about 10mg of hydrocortisone acetate,to a suitable container.Add 10.0mLof Internal standard solutionand about 40mLof chloroform,shake vigorously for about 5minutes,and allow the phases to separate.Use the clear chloroform layer as the Assay preparation.
Chromatographic system (see Chromatography á621ñ)—The liquid chromatograph is equipped with a 254-nm detector and a 4-mm ×30-cm column that contains packing L3.The flow rate is about 1mLper minute.Chromatograph the Standard preparation,and record the peak responses as directed for Procedure:the resolution,R,between the analyte and internal standard peaks is not less than 3.0,and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure— Separately inject equal volumes (about 15µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the responses for the major peaks.The relative retention times are about 0.7and 1.0for hydrocortisone acetate and fluoxymesterone,respectively.Calculate the quantity,in mg,of hydrocortisone acetate (C23H32O6)in each mLof the Ophthalmic Suspension taken by the formula:
(W/V)(RU/RS),
in which Wis the quantity,in mg,of USP Hydrocortisone Acetate RStaken to prepare the Standard preparation,Vis the volume,in mL,of Ophthalmic Suspension taken,and RUand RSare the peak response ratios of the hydrocortisone acetate peak to the internal standard peak obtained from the Assay preparationand the Standard preparation,respectively.
Auxiliary Information— Staff Liaison:William W.Wright,Ph.D.,Scientific Fellow
Expert Committee:(PA7)Pharmaceutical Analysis 7
USP28–NF23Page 1349
Phone Number:1-301-816-8335