(Official January 1, 2007)

Mobile phase and Chromatographic system—Proceed as directed in the Assay under Chloramphenicol.

Standard preparation— Transfer about 25 mg of USP Chloramphenicol RS, accurately weighed, to a 100-mL volumetric flask, dissolve in methanol, dilute with methanol to volume, and mix. Transfer 10.0 mL of the resulting solution to a 25-mL volumetric flask, dilute with Mobile phase to volume, and mix. Filter a portion of this solution through a 0.5-µm or finer porosity filter, and use the clear filtrate as the Standard preparation.

Assay preparation— Transfer an accurately weighed quantity of Ophthalmic Ointment, equivalent to about 25 mg of chloramphenicol, to a suitable conical flask, add 20 mL of cyclohexane, mix, and sonicate for about 2 minutes. Add 60 mL of methanol, and mix. Filter this mixture, collecting the filtrate in a 100-mL volumetric flask. Wash the filter with methanol, collecting the washings in the volumetric flask. Dilute with methanol to volume, and mix. Transfer 50.0 mL of the resulting solution to a suitable round-bottom flask, and evaporate to dryness by rotating the flask under vacuum in a water bath at 35. Dissolve the residue in 50.0 mL of methanol. Transfer 10.0 mL of the resulting solution to a 25-mL volumetric flask, dilute with Mobile phase to volume, and mix. Filter a portion of this solution through a 0.5-µm or finer porosity filter, and use the clear filtrate as the Assay preparation.

Procedure— Proceed as directed for Procedure in the Assay under Chloramphenicol. Calculate the quantity, in mg, of C11H12Cl2N2O5 in the portion of Ophthalmic Ointment taken by the formula: in which the terms are as defined therein.