Detection of Mammotropin ∗ in the Urine of Lactating Women

Abstract

In the course of studies on the hormonal control of the mammary gland, it has been of interest to ascertain by urinalysis to what extent a lactating woman is under the influence of estrin and the hypophyseal mammotropic hormone. A crude estrin may be prepared from the urine and tested by smearing it in the vaginae of ovariectomized rats. The urines of 8 lactating women (4–13 days postpartum) have been tested and all found to contain mammotropin in amounts that make it appear that at least as much of this hormone is excreted daily as is extractable from a bovine anterior lobe. The urine may be treated as follows: (1) to 100 cc. add 200 cc. acetone and 3 cc. HCl (concentrated); centrifuge and discard insoluble material; (2) add acetone to 90%; discard supernatant; (3) extract precipitate with mixture of 10 cc. stronger ammonia water, 20 cc. water, and 60 cc. acetone; discard insoluble; (4) add one volume of acetone; discard supernatant; (5) wash precipitate with 85% acetone, absolute acetone and ethyl ether (2 × 25 cc. in each case); (6) dry precipitate in warm desiccator; dissolve in 5.0 cc. water; adjust to pH 7.6; discard any insoluble.† Inject in doses of 0.1 and 1.0 cc. intradermally over the right and left crop sacs of squabs one month from hatching‡ for presumptive testing. Sacrifice birds at 48 hours, and retest in accordance with results until the minimal effective dose is determined. Until the hormone is purified it may suffice to consider the minimal effective dose in a statistically adequate number of birds as the temporary unit of urinary mammotropin. Sufficient amounts of untreated urine or blood plasma may be injected into the crop areas to allow for detection of mammotropin, but this procedure sometimes prevents accurate reading of the reactions because of inflammatory processes.