A: Pass hydrogen sulfide through 50 mL of it: no black discoloration or black precipitate is formed. To 50 mL of Topical Solution add 3 or 4 drops of bromine, mix, and warm on a steam bath to expel the excess bromine. Add 5 mL of 3 N hydrochloric acid, filter, and pass hydrogen sulfide through the filtrate: a black precipitate is formed.

B: To 1 mL of Topical Solution add 9 mL of water, mix, and add 1 mL of cupric sulfate TS: a green color is produced immediately and is followed by the gradual precipitation of flocculent, greenish brown particles.

(Official January 1, 2007)

Stannous chloride solution— Dissolve 50 g of stannous chloride in 100 mL of hydrochloric acid on a steam bath, cool, dilute with water to 500 mL, and mix. Use within 3 months.

Standard solutions— Prepare aqueous solutions of USP Thimerosal RS of known concentrations of about 1.8, 2.0, and 2.2 µg per mL.

Standard preparations— Pipet 20 mL of each Standard solution into separate 100-mL volumetric flasks, and treat each flask as follows. Add 5 mL of sulfuric acid, cool, add 3 mL of nitric acid, and mix. Add potassium permanganate crystals, while mixing, until the purple color persists for not less than 15 minutes. Add about 200 mg of potassium persulfate, mix, and heat on a steam bath for 2 hours. Cool, dilute with water to volume, and mix.

Assay solution— Pipet 2 mL of Topical Solution into a 1000-mL volumetric flask, dilute with water to volume, and mix.

Assay preparation— Pipet 20 mL of Assay solution into a 100-mL volumetric flask, and proceed as directed for Standard preparations, beginning with “Add 5 mL of sulfuric acid.”

Blank preparation— Pipet 20 mL of water into a 100-mL volumetric flask, and proceed as directed for Standard preparation, beginning with “Add 5 mL of sulfuric acid.”

Procedure— Proceed with each of the Standard preparations, the Assay preparation, and the Blank preparation as follows: Pipet 3 mL into the scrubbing chamber of a suitable system designed for determination of mercury by flameless atomic absorption, using a mercury hollow-cathode lamp, dilute with water to about 150 mL, and add hydroxylamine hydrochloride solution (1 in 10) just to reduce the excess permanganate. Add 5 mL of Stannous chloride solution, and immediately attach the scrubbing chamber to the system. Concomitantly determine the absorbance of each solution at an integration time of 15 seconds. Use the absorbance of the Blank preparation to correct the absorbances of the Standard preparations and the Assay preparation. Plot the corrected absorbances of the standards versus the respective concentrations of the Standard solutions, in µg per mL, and from the curve so obtained determine the concentration, C, in µg per mL, of the Assay solution. Calculate the quantity, in mg, of C9H9HgNaO2S in each 100 mL of Topical Solution taken by the formula: in which the terms are as defined therein.