Abstract
To determine 170S sulfates and glucuronides, which separately extracted by ion pair extraction as described in the previous report, by high-performance liquid chromatography using a reversed phase Capcell-Pak C8 column, we conducted experiments on labeling condition of the steroids with dansylhydrazine and on chromatographic condition for the separation of the labeled steroids.
As a result, we found a method that obtained much more labeled steroids under lower temperature than already reported methods, and obtained three mobile phase solutions that can separate individual steroid. The labeled 170S sulfates were perfectly separated by using methanol-0.5% (w/v) sodium acetate-50% (v/v) acetic acid (57: 42: 1, v/v) at 30°C. The labeled 170S glucuronides were also clearly separated by using methanol-0.03%acetic acid (v/v)(46: 54, v/v) and methanol-0.01% acetic acid (v/v)(56: 44, v/v) at 55°C.
