A: Transfer about 100 mg to a suitable container, and dissolve in about 10 mL of water. In rapid succession, add 1 mL of 3 N hydrochloric acid, shake briefly, immediately decant the aqueous layer, and wash the precipitate with two additional 10-mL portions of water, each time removing the water by decanting. Remove as much of the water as possible, spread the precipitate in a suitable container, and dry in vacuum at about 60 for 3 hours: the IR absorption spectrum of a mineral oil dispersion of the precipitate so obtained exhibits maxima only at the same wavelengths as that of a similar preparation of USP Hydrocortisone Hemisuccinate RS.

B: Ultraviolet Absorption 197U—

Solution : 20 µg per mL.

Medium : methanol.

C: It responds to the flame test for Sodium 191.

Test solution: 10 mg per mL, in alcohol.

(Official January 1, 2007)

Standard preparation— Prepare as directed for Standard Preparation under Assay for Steroids 351, using USP Hydrocortisone Hemisuccinate RS, but dilute the solution with alcohol to a concentration of about 12.5 µg per mL.

Assay preparation— Weigh accurately about 100 mg of Hydrocortisone Sodium Succinate, dissolve in sufficient alcohol to make 200.0 mL, and mix. Pipet 5 mL of this solution into a 200-mL volumetric flask, add alcohol to volume, and mix. Pipet 20 mL of the resulting solution into a glass-stoppered, 50-mL conical flask.

Procedure— To each of the two flasks containing the Assay preparation and the Standard preparation, respectively, and to a similar flask containing 20.0 mL of alcohol to provide the blank, add 2.0 mL of a solution prepared by dissolving 50 mg of blue tetrazolium in 10 mL of alcohol, and mix. Then to each flask add 4.0 mL of a 1 in 10 solution of tetramethylammonium hydroxide TS in alcohol, mix, allow to stand in the dark for 90 minutes, add 1.0 mL of glacial acetic acid, mix, and proceed as directed for Procedure under Assay for Steroids 351, beginning with “Concomitantly determine the absorbances.” Calculate the quantity, in mg, of C25H33NaO8 in the Hydrocortisone Sodium Succinate taken by the formula: