Abstract
In the course of investigations on the excretion of gonad stimulating hormones in the urine of mental patients 1 , 2 the following method was developed for the preparation of concentrates of the follicle stimulating hormone (F.S.H.).
The method is based on the observation that the stable aluminum hydroxide C (γ), prepared according to Willstaetter and Kraut 3 is a satisfactory absorbent for F.S.H. from urine. The effectiveness of this adsorption depends markedly on the pH, a definite maximum occurring at pH 4.5. Evans, Gustus and Simpson 4 have reported that solutions of the gonadotropic hormone from acetone-dried pregnant mare's serum are more satisfactorily adsorbed at pH 3.5 by aluminum hydroxides of Types A and B, prepared according to an earlier method of Willstaetter and Kraut, 5 while Type C was considerably less efficient.
The urine is collected in glass jars containing 5 cc. of a saturated alcoholic solution of thymol per liter capacity. Insoluble material is removed by filtration and the filtrate is adjusted to exactly pH 4.5 with 5 N HCl using bromcresol green as external indicator. A suspension containing 20 gm. per liter of aluminum hydroxide C (γ) is slowly poured into the urine until 1 gm. of aluminum hydroxide per liter of urine has been added. For 24-hour specimens of 500-1000 cc. a full 1.0 gm. of aluminum hydroxide is added; for samples of less than 500 cc. only 0.5 gm. is used. The resulting mixture is occasionally stirred by hand (mechanical stirring results in poor yields) during 20 minutes so as to maintain a homogeneous suspension; the solids are allowed to settle during 45 minutes. The supernatant fluid is removed by siphoning and centrifugation, readjusted to pH 4.5 and again treated with aluminum hydroxide by the same procedure.
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