A:
Hydroxylamine reagent—Mix 5 mL of a solution of hydroxylamine hydrochloride in methanol (3.5 in 100) with 15 mL of a solution of potassium hydroxide in methanol (5.6 in 100), and use this reagent within 2 hours after mixing.
[NOTE—Store the solution of hydroxylamine hydrochloride in methanol and the solution of potassium hydroxide in methanol in a refrigerator, and discard both after 1 month.
]
Iron reagent—
Dissolve 1.5 g of ferric chloride in 30 mL of water, add 3 mL of perchloric acid and 15 mL of nitric acid, and heat the solution until dense, white fumes are produced. Cool, add 40 mL of water and 10 mL of nitric acid, and dilute with perchloric acid to 100 mL. [NOTE—This concentrate may be stored and used for several months.] Transfer 4.0 mL of this concentrate to a 100-mL volumetric flask, add 36 mL of alcohol, dilute with water to volume, and mix.
Procedure—
Place a portion of finely powdered Tablets, equivalent to about 250 mg of testolactone, in a 100-mL volumetric flask, add 85 mL of chloroform, and shake by mechanical means for 30 minutes. Add chloroform to volume, mix, and centrifuge the solution. Transfer 2 mL of the supernatant to a small test tube, and evaporate with the aid of a gentle current of air to dryness. Add 0.1 mL of
Hydroxylamine reagent, heat in a water bath at about 60
for 1 minute, then add 1 mL of
Iron reagent: a rose-violet color is produced.
Infrared Absorption 
197K
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