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The use of acid for liberating the steroids of urine is based on the observation that these c m p u n d s are excreted in conjugated form as evidenced by their inmlulbility in fat solvents and by the isolation of steroid glucuronidesl* and sulfates?-6 m e evidence that such treatment causes considerable l o~s~-~O bas led to a consideration of enzymes for the liberation of steroids frum their conjugates.Cvhea and Marrianll and Patterson12 have previously reported the liberation of estrogen of pregnancy urine by bacterial action and BaTker, Bruuksbank, and H a s l m d J B have found that pregnanediol glucuronide, and in some experiments glucuronic acid, which presumbly was liberated from the conjugate were destroyed by some strains of StaPkyZococcw albus. Preparations possessing glucuronidase activity which were obtained frm mammalian tissues hydrolyze the glucuronides of estrio1,l4J3 pregna.nedio17J6 and other co,mp 0 u n d~s~~1 '~J~ and Cohen and B a t e P have Ima., 1935, 54, 1025.

3 rate of albumin. The remaining protein sedimented 2.2 times more rapidly than the main component. Discussion.A rapidy migrating component is readily observed in the electrophoretic patterns of concentrated spinal fluids equilibrated with a phosphate buffer (pH 7.5, r/2 .05) or a veronal buffer (pH 8.6, r/2 0.1). Kabat et al. (1) using a phosphate-sodium chloride buffer mixture at pH 7.4 and an ionic strength of about 0.2, recorded the presence of a fast component in 3 of 40 spinal fluids. Scheid and Scheid(2) studied spinal fluids which were not sufficiently concentrated and observed the fast component in 1 of 2 2 cases. These studies emphasize the necessity of using concentrated fluids and suitable buffer mixtures if the rapidly migrating components of spinal fluid are to be observed or separated electrophoretically.Component X has a greater anodic mobility than albumin over a pH range varying from 7.9 to 4.0. In contrast, rapidly migrating components observed in plasma do not behave similarly over this pH range ( 7). Furthermore, plasma adjusted to the protein concentration of concentrated spinal fluid and equilibrated against dilute phosphate buffer does not reveal the presence of component X. Under these conditions the albumin .boundary of plasma spreads appreciably and may prevent the detection of small amounts of the fast component. From the available data, it appears justifiable to assume that components X and XI originate in the nervous system and are proteins typical for cerebrospinal fluid.Summary. Electrophoretic studies of 65 cerebrospinal fluids showed the presence of a rapidly migrating protein in each case. This component migrated about 1.3 times faster than albumin and sedimented at the same rate as albumin in the ultracentrifuge. In 9 cases, 2 rapidly migrating components were observed. Under the experimental conditions used, the fast components comprise about 9% of the total spinal fluid proteins.The study of the metabo1,ism of steroids is, in part at least, dependent upon the determination of the quantities and types of steroids excreted in the urine. Important information could also be obtained if it were possible to estimate the individual steroid conjugates so excreted.The usual method of hydrolyzing the steroid conjugates by refluxing strongly acidified urine has :been shown to cause alterations

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