ROUTINE DETERMINATIONS OF 17-KETOSTEROIDS AND PORTER-SILBER CHROMOGENS COMPARED WITH THE CHROMATOGRAPHIC MEASUREMENT OF GROUPED AND INDIVIDUAL STEROIDS

1964 ◽  
Vol 46 (4) ◽  
pp. 552-562 ◽  
Author(s):  
Ingrid Ernest ◽  
Britt Håkansson ◽  
Jörgen Lehmann ◽  
Björn Sjögren

ABSTRACT The accuracy of two routine methods for the determination of urinary steroids – 17-ketosteroids (17-KS) and Porter-Silber chromogens – has been investigated by chromatographic separation and quantitative determination of individual 17-KS and Porter-Silber reacting steroids in 141 urine samples. For this purpose urine was submitted to enzyme hydrolysis and subsequent solvolysis. The pertinent steroids were separated by column chromatography into three groups, 11-deoxy-17-KS, 11-oxy-17-KS and Porter-Silber reacting steroids. The final separation was accomplished by paper chromatography. As a mean, the urinary excretion of true 17-KS corresponded to about 40–50 per cent of the routine figures. Due to non specific chromogens, individual routine figures were completely unreliable and probably had no more significance than showing a difference between a high and a low urinary content of 17-KS A true figure, however, was never higher than that indicated by the routine method. The routine method for determination of Porter-Silber chromogens also overestimated the urinary content of steroids, the true excretion of Porter-Silber steroids being, on an average, about 25 % lower. Again, the significance of individual determinations was low. The determination of 17-KS and Porter-Silber steroids by column chromatography was found to be rather simple and reliable as only minor amounts of unspecific chromogens were included in the results. Moreover with this method, the 17-KS were separated into 11-deoxy-17-KS and 11-oxy-17-KS.

1962 ◽  
Vol 41 (2) ◽  
pp. 234-246 ◽  
Author(s):  
H. J. van der Molen

ABSTRACT A procedure for the quantitative determination of 5β-pregnan-3α-ol-20-one in urine is described. After acid hydrolysis of the pregnanolone-conjugates in urine, the free steroids are extracted with toluene. Pregnanolone is isolated in a pure form as its acetate; after chromatographic separation of the free steroids on alumina, the fraction containing pregnanolone is acetylated and rechromatographed on alumina. Quantitative determination of the isolated pregnanolone-acetate is carried out with the aid of the infrared spectrum recorded by a micro KBr-wafermethod. The reliability of the method under various conditions is discussed under the headings, specificity, accuracy, precision and sensitivity. It is possible to determine 30–40 μg pregnanolone in a 24-hours urine portion with a precision of 25%.


1968 ◽  
Vol 109 (1) ◽  
pp. 13-18 ◽  
Author(s):  
D. E. S. Stewart-Tull

A colorimetric method is described whereby the direct quantitative determination of glucosamine, galactosamine and muramic acid can be achieved without previous treatment of the cell-wall hydrolysate, for example by column chromatography. Molar ratios of hexosamines in cell-wall preparations, from a number of bacterial species, determined by this method were found to be in general agreement with previously published results.


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