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

本研究旨在探讨16-脱氢孕烯醇酮（16-DHP）在实验室新生儿血清中的结合形式及其代谢位点。CH_2Cl_2处理可清除新生儿血清中的游离甾体。用甲醇提取可能含有偶联甾体的残留物。甲醇提取物采用高效液相色谱法。收集正品16- dhp3 -硫酸盐洗脱后的馏分。该馏分用芳基磺化酶处理。采用GC/MS和GC/FT-IR分析游离甾体。所得数据与正品16-DHP完全一致。结果表明，16-DHP与硫酸偶联。为了研究16-DHP 3-硫酸盐是否在胎儿肾上腺中合成，观察了血清16-DHP 3-硫酸盐水平与胎龄的相关性。因此，这两个参数之间存在显著的负相关。这些结果表明，16-DHP 3-硫酸盐在胎儿肾上腺中合成。

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.