Abstract
The demonstration that ovary-stimulating extracts prepared from human placenta (Collip et al. 1 ), or from urine of pregnant women (Evans, Meyer, and Simpson 2 ) differ from anterior pituitary gland implants in their influence on the weights of immature rat ovaries, is of considerable biological interest. It seemed of importance, therefore, to determine if extracts made from the blood of pregnant women were similar in this characteristic.
A crude estrin-free extract was prepared from blood by a method developed with the assistance of Dr. M. L. Tainter of the Department of Pharmacology, based on that of Wallen-Lawrence and Van Dyke 3 for the extraction of “hebin” from anterior pituitary glands. Forty to 50 cc., obtained by venapuncture, from each patient was placed in a receptacle containing a few crystals of sodium citrate, centrifuged, the red blood cells discarded, and the resultant clear blood plasma shaken up 5 times with ether. Ten volumes of 95% alcohol were then added to the blood plasma, the solution centrifuged, and the resultant precipitate washed twice with ether, dried, powdered, and extracted over night with 20 to 30 cc. of an isotonic sodium acetate-acetic acid buffer solution having a pH of about 4.4. The next morning it was centrifuged and to the clear supernatant solution was added 6 volumes of 95% alcohol. A white precipitate resulted which was recovered by centrifugalization, again washed with ether, dried and powdered. This powder contained the active element and was fairly readily dissolved in small amounts of the buffer solution for injection. It was generally so made up that 1 cc. of the extract was equivalent to 2.3 cc. of blood plasma and various dilutions were prepared from this stock solution.
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