Investigation of Steroid Sulfate Conjugates in Neonatal Serum-Elucidation of the Mechanism of the Biosynthesis of the Steroids and Its Application for Pathological Analysis.

本研究旨在研究本实验室新生儿血清中已鉴定的16-脱氢孕烯醇酮(16-DHP)的结合形式及其代谢部位。用甲醇提取可能含有结合类固醇的残留物，将甲醇抽提物用于高效液相色谱，收集洗脱出真正的16-DHP-3-硫酸盐的级分。这一组分用芳基硫酸酯酶处理。用GC/MS和GC/FT-IR对释放出的类固醇进行了分析，所有数据与正品16-DHP的结果一致。结果表明，16-DHP是与硫酸偶联的。为了探讨16-DHP 3-硫酸盐是否在胎儿肾上腺合成，观察了血清16-DHP 3-硫酸盐水平与胎龄的关系。结果表明，这两个参数之间存在显著的负相关。这些结果表明，16-DHP-3-硫酸盐是在胎儿肾上腺合成的。

The present study was designed to investigate the conjugation form of 16-dehydropregnenolone (16-DHP) which had been identified in the serum of neonates in our laboratory and its metabolic site.Free steroids were washed out from neonatal serum by CH_2Cl_2 treatment. The residue which may contain conjugated steroids was extracted with MeOH.The MeOH extracts were applied to HPLC.The fraction, in which authentic 16-DHP 3-sulfate was eluted, was collected. This fraction was treated with arylsulfatase. Liberated free steroids was analyzed by GC/MS and GC/FT-IR.All of the data obtained was identical with those of authentic 16-DHP.These results showed that 16-DHP is conjugated with sulfuric acid.To investigate if 16-DHP 3-sulfate is synthesized in fetal adrenal, correlation of serum levels of 16-DHP 3-sulfate and gestational age was observed. As a result, significant negative correlation was observed between these two parameters. These results suggest that 16-DHP 3-sulfate is synthesized in fetal adrenal.