Determination of 19-Hydroxytestosterone in Human Ovarian Venous Blood by Selected Ion Monitoring

Abstract

A highly specific method, selected ion monitoring (SIM), using [7,7-2H2] 19-hydroxytestosterone (19-OHT) as an internal standard, was newly developed to determine 19-OHT in serum. Steroids in serum obtained from the ovarian vein were extracted with ethyl acetate. Following separation by thin layer chromatography, the 19-OHT fraction was subjected to SIM after formation of the methoxime-trimethylsilyl (MOTMS) derivative. Mass spectra of the 19-OHT-MOTMS derivative was dominated by the molecular cation [M] + (m/e 477). Serum concentration of 19-OHT was 280 pg/ml in ovarian vein. This is the first evidence of 19-OHT in human blood.

Biochemical Significance of 19-Hydroxytestosterone in the Process of Aromatization in Human Corpus Luteum.

“…19-Hydroxyandrogens are known to be an intermediary metabolite in the aromatizing reaction. Previously, we measured the concentration of 19-OHT in the ovarian vein by selected ion monitoring (SIM) [4], and suggested that 19-OHT is an endogenous steroid in human ovary.…”