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In the preceding paper (1) a method was described which permits the simple calorimetric estimation of microgram amounts of estrogens in the presence of tissues. The results obtained by the use of this method in a study of the in vitro metabolic action of rat liver on 17-p8 estradiol were also reported.In the present paper we are reporting a similar study on human liver. The original motive for this study was to evaluate whether a deficiency of the metabolic function of the liver exists in patients with cancer of the uterus. Patients with this type of cancer and other diseases were studied. METHODSThe procedures described in the preceding paper for the incubation of liver slices, the extraction of the estrogens and the calorimetric method for the measurement of estrogens were used in the present work and found to be applicable without modification to the study of human liver.Human liver specimens were obtained at laparotomy from patients with various surgical diseases. The patients were unselected. The liver specimen was immersed into ice-cold Krebs solution in the operating room, immediately transferred to the laboratory and the incubation experiment begun. The time interval between surgical removal of the specimen and the beginning of the incubation period did not exceed 10 minutes and was comparable to the time interval obtaining in the experiments with rat liver.The liver specimen consisted of a wedge of tissue resected from the anterior border and weighing approximately 2 gms. Sufficient tissue was available in each case to run the metabolic, the control and the blank experiments with slices from the same bloc of tissue.

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Since the work of Zondek (1) the in vitro action of liver slices and liver pulp on natural estrogens is usually called inactivation because the transformation undergone by these compounds results in loss of estrogenic activity as measured by bioassay methods (2). Using cc. of toluene were added to the mixture. All these operations were carried out in an ice bath.5 The colored material was extracted into toluene by shaking for 10 minutes in a shaking machine. After centrifuging for two minutes at 2,500 r.p.m., the toluene layer was pipetted off and read in a Beckman quartz spectrophotometer (Model DU) at 425 m#* in a 1 cm. cell against the blank.The blank was treated as the experimental sample except that no estrogen was present.Quantitative measurements of 17-j estradiol, estrone and estriol were obtained by this procedure between 10 and 150 micrograms, as indicated on Figure 1. The relation between optical densities and concentrations followed Beer's law between these two values. The slope of the curve was found to be similar for estrone and estradiol but different for estriol (Figure 1).The absorption spectrum of the colored compound showed a plateau-like peak between 400 mAi' and 450 miA with the maximum at 425 my approximately (Figure 2). 4 Kindly supplied by Drs. E. Schwenk and E. B. Hirshbirg, Schering Corporation, Bloomfield, New Jersey. 5 The tetrazotized dianisidine solution was prepared as follows: 25 mg. of dianisidine dihydrochloride (recrystallized from ethanol) were dissolved in 3 cc. of water. Three tenths cc. of concentrated hydrochloric acid, followed by 0.6 cc. of freshly prepared 5% sodium nitrite solution were added. After thorough mixing and a five minute waiting period, 0.66 cc. of a 5% urea solution was added. The solution was prepared in an ice bath, and made freshly for each experiment.341