Iron-Phenol TS (Iron-Kober Reagent)
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This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition
Issued and maintained by the United States Pharmacopeial Convention (USP)
Dissolve 1.054 g of ferrous ammonium sulfate in 20 mL of water, and add 1 mL of sulfuric acid and 1 mL of 30 percent Hydrogen peroxide. Mix, heat until effervescence ceases, and dilute with water to 50 mL. To 3 volumes of this solution contained in a volumetric flask add sulfuric acid, with cooling, to make 100 volumes. Purify phenol by distillation, discarding the first 10% and the last 5%, collecting the distillate, with exclusion of moisture, in a dry, tared glass-stoppered flask of about twice the volume of the phenol. Solidify the phenol in an ice bath, breaking the top crust with a glass rod to ensure complete crystallization. Weigh the flask and its contents, add to the phenol 1.13 times its weight of the iron-sulfuric acid solution prepared as directed, insert the stopper in the flask, and allow to stand, without cooling but with occasional mixing, until the phenol is liquefied. Shake the mixture vigorously until mixed, allow to stand in the dark for 16 to 24 hours, and again weigh the flask and its contents. To the mixture add 23.5% of its weight of a solution of 100 volumes of sulfuric acid in 110 volumes of water, mix, transfer to dry glass-stoppered bottles, and store in the dark, protected from atmospheric moisture. Use within 6 months. Dispense the reagent from a small-bore buret, arranged to exclude moisture, capable of delivering 1 mL in 30 seconds or less, and having no lubricant, other than reagent, on its stopcock. Wipe the buret tip with tissue before each addition.
