A Direct Determination of Sulfate Conjugates of 17-Oxosteroids in Urine by Use of Benzyltributylammonium Chloride

Without Solvolysis and Enzymatic Hydrolysis

抄録

A method for the determination of urinary sulfate conjugates of 17-oxosteroids (17OS-sulfates), which can directly determine 17 OS-sulfates without hydrolysis of the sulfate conjugates, is described. The four major urinary 17OS-sulfates are effectively extracted (about 100% recovery) with benzene in the presence of benzyltributylammonium chloride (BTBA). The four chromogens produced by the reaction of 17OS-sulfates with m-dinitrobenzene reagent with BTBA added are quantitatively extracted with diethylether without occurrence of cloudy appearance. The chromogens can then be measured at 515 nm without interferences of contaminants. The proposed method shows a sufficient recovery and a good reproducibility. The values of 17OS-sulfates in urine samples that were determined by Kanbegawa's method after the solvolysis by Barstein and Lieberman's method showed good correlation with the values of 17OS-sulfates obtained by the proposed method.