Harris Rosenkrantz

An antimony trichloride reagent suitable for the detection and estimation of nonketonic steroids

Although a number of excellent paper chromatographic technics are available for resolution of steroid mixtures (l-3), the separation of nouketonic hydroxy steroids has not been satisfactorily investigated. The difficulty in determining the chromatographic behavior of the free nonketonic st)eroids has been the lack of a suitable color reaction for locating the compounds on a chromatogram. Estrogens have been chromatographed on paper, but their detection required coupling with a dye (3). Antimony trichloride appeared to have the necessary characteristicas for reacting with steroids to give colored products. Earlier investigators had utilized solutions of this reagent in glacial acetic acid or chloroform for estimation of urinary steroid metabolites (5) and for spectrophotomet,ric comparison of steroidSbC4 product,s (6). More recently atItimony trichloride (20 % in chloroform) (7, 8), antimony pentachloride (8, IO), and antimony trichloride in an atmosphere of chlorine (I I ) have been used for the detection of steroids on paper chromatograms. The use of another antimony trichloride preparation whose application has proven to be very simple, versatile, and effective in the detection of steroids is to be discussed here. Characteristic colors were observed on paper when various steroids were treat’ed lvith antimon! trichloride in nitrobenzene (13), hydroxylated compounds being partic+ularly reactive. This proredure will permit. the examination of non-

Studies on the biosynthesis of cholesterol. II. Isolation of 3 beta, 5 alpha-dihydroxy-6-ketocholestane from cholesterol

It has been found that C4-cholesterol as synthesized in the perfusion of surviving livers (1) from carboxyl-labeled CL4-sodium acetate always contains impurities which show a higher radioactivity than the Cl”cholesterol itself. Part of these companions are precipitated by digitonin and therefore may be steroids. Obviously such higher radioactive companions could be precursors in the biosynthesis of cholesterol from acetate, and their identification would be of great significance. Experiments to this end have been started at this laboratory, in the course of which a substance has been isolated from cholesterol which may be derived from a hitherto not discovered companion of cholesterol, the 3/3,5qGp-trihydroxycholestane. This substance was isolated not only from C!l4-cholesterol but also from pure commercial cholesterol and from a crude liver extract. Its analysis suggests a ,formula C27H4603 or C27H4403. The melting point is 233-6” which is also that of the known 3/3,5Lu,G&trihydroxycholestane of the formula C&H4603 recently investigated by Fieser and Rajagopalan (2), and there was no melting-point depression with a sample of the latter, obtained through the kindness of Dr. Fieser. However, an infrared spectrum disclosed the presence of a carbonyl group. This was confirmed by reaction with dinitrophenylhydrazine which gave a yellow

The reactivity of catechol amines, 5-hydroxyindoles and phenols with blue tetrazolium

Abstract A comparison of the intensity and rates of reaction of catechol amines, 5-hydroxyindoles, and phenols with blue tetrazolium and ferric chloridedipyridyl was carried out. It was demonstrated that catechol amines reacted approximately four times more intensely with blue tetrazolium than the 5-hydroxyindoles or α-ketolic steroids. The ferric chloride-dipyridyl test was useful in distinguishing the catechol amines from the corresponding N -methylated derivatives.

A comparison of reducing systems in vitamin E-deprived rabbits and mice with dystrophia muscularis.

Abstract 1. 1. It has been found that the ability to produce formazan from triphenyltetrazolium chloride (TPTZ) in the presence of isocitric acid and α-ketoglutaric acid is reduced in the muscle of vitamin E-deficient rabbits. 2. 2. Some differences in the levels of reducing enzyme systems were observed in the liver and kidney of the rabbits. 3. 3. The muscle content of enzymes capable of reducing TPTZ in the presence of glutamic acid was elevated in mice with inherited dystrophy. 4. 4. An increase in oxygen uptake was also obtained for the dystrophic muscle from the mice. 5. 5. It is possible that the histopathological similarities of nutritional muscular dystrophy, the inherited myopathy of the mouse, and human muscular dystrophy result from a biochemical lesion at a different site in the same biochemical pathway or cycle.

The C-20α reduction of steroids by hog liver preparations

Abstract The presence of a steroid C-20α-reductase has been demonstrated in hog liver brei by the isolation and identification of Δ 4 -pregnene-20α, 21-diol-3-one and Δ 4 -pregnene-17α, 20α, 21-triol-3-one after the incubation of 11-deoxycorticosterone and 11-deoxycortisol, respectively. Some properties of the 20α-reductase are described.

Nucleic acid content of rat neoplasms

Abstract Determinations of nucleic acids in spleen and neoplastic tissue were carried out in several rat strains. Two mammary adenocarcinomas designated R-36 and MT/W9A, and a mammary fibroadenoma, referred to as the C-tumor, in addition to a lymphocytic leukemia, WR-6, were studied. The RNA/DNA ratio of normal spleen was similar in 5 strains of rat. No appreciable influence on host splenic nucleic acid content could be discerned due to the presence of the neoplasms studied. The RNA/DNA ratios were capable of differentiating the various tumor systems, the ratio appearing to increase with survival time. Cytoxan influenced the RNA/DNA ratio by producing reduction in DNA and RNA levels in both splenic and tumor tissue from R35 tumor-bearing rats. Chlorambucil did not affect spleen nucleic acid content but may have depressed the RNA/DNA ratio of tumor tissue from WR-6 tumor-bearing animals.

A Comparison of Infrared Frequencies of 19-Nor Steroids and their 19-Methyl Analogues

The infrared absorption spectra of thirteen 19-nor steroids in the C-19 series were recorded by the potassium bromide dispersal technique It was found that a shift to higher frequencies occurred for the bands that have been correlated with structures in saturated 3-hydroxysteroids, 3α, 5α shifted from 998 to 1017 cm−1, 3β, 5α from 1042 to 1060 cm−1, 3α, 5β from 1035 to 1060 cm−1, and 3β 5β from 1033 to 1055 cm−1. A band near 881 cm−1 helped identify Δ4-19-nor structures, while a similar band near 885 and/or 825 cm−1 seemed characteristic for the Δ1-19-nor compound

Purification of canine prostatic acid phosphatase by gel filtration

Abstract A colony of dogs with cystopreputiostomy prostatic fistulas has provided a continuous supply of prostatic secretion rich in acid phosphatase and proteolytic activities. Purification of acid phosphatase enzymes was accomplished by dialysis alone, dialysis followed by DEAE-cellulose chromatography, and by gel filtration on A50 Sephadex followed by dialysis. The latter procedure was simple, rapid, and provided a 45-fold purification. Large amounts of prostatic fluid could be processed and separation of acid phosphatase from proteolytic activities was readily achieved.