Abstract
By the following method it has been possible to demonstrate the presence of luteinizing (in 62.5% of 16 women) as well as follicle-stimulating principle in the urine of both castrates and spontaneous menopause.
Two hundred cc. of urine is taken from a fresh 24 hours specimen and acidified with concentrated acetic acid to pH 3.5 (Congo Red). Four volumes of cold acetone are added to the urine, shaken vigorously and allowed to stand overnight in the refrigerator. The supernatant fluid is poured off and the precipitate extracted with weak NaOH. The pH of the mixture is adjusted to between 8 and 8.5, the residual precipitate stirred thoroughly, the mixture centrifuged, and the final precipitate discarded as the supernatant fluid contains the gonadotropic principle. The fluid is then adjusted to pH 7 with dilute acetic acid. The equivalent of 100, 50, and 25 cc. is injected into immature rats weighing 22 to 24 gm. The injections in each rat are divided into 5 doses, given over 3 days. The animals are autopsied at the end of 96 hours. The ovaries are examined in serial section.
The same procedure can be performed using alcohol instead of acetone. Acetone is preferable to alcohol as no deterioration or destruction of the hormone occurs on standing, whereas deterioration of the gonadotropic principle occurs when alcohol is used.
The opinion is gaining ground that “the principle in castrate or menopause urine seems to be identical with the follicle-stimulating fraction prepared from the pituitary glands.” 1 However, it should be emphasized that Lassen and Brandstrup 2 reported a luteinizing reaction in the urine (diagnosis by gross inspection of the mouse ovaries), and Fluhman 3 in the blood of such patients.
Katzman and Doisy's benzoic acid precipitation method 4 does not extract the factor found in the menopause or castrate urine. By using our new method, 16 cases, partly surgical castrates, X-ray castrations, and spontaneous menopause were studied.
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