Lidia Matei1, Cristian Postolache1, Eduard Condac2, Marieta Costache2, Mihai Ionica3

SYNTHESIS of testosteron-1,2-D
Lidia Matei¹, Cristian Postolache¹, Eduard Condac², Marieta Costache², Mihai Ionica^3
1Natinal Institute of Research and Development for Physics and Nuclear Energy “Horia Hulubei”, Măgurele, Atomistilor Str. nr. 407, CP- MG-6.

²University of Bucharest, Faculty of Biology, Splaiul Independentei, nr. 91-95, Bucharest.

³Deffence Medical Center, Bucharest

Labelling of testosterone with deuterium was necessary for carring out the molecular biology studies concerning androgen dependent diseases.

Testosterone was labelled with deuterium by selective hydrogenation of 1,2 dehydrotestosteron acetate. The precursor was synthesized in two steps: 1) esterification of testosterone using acetic anhydride, and 2) selective dehydrogenation with 2,6-dichloro-3,5-dicyan-1,4 quinone (DDQ) of the ester, formed in the first step. Testosterone acetate was synthesized with yields of 73%. Ester was purified by recristalisation in acetone:n hexane (15:1) with a 80% yield. The dehydrogenation process was characterized by yields of 82% for synthesis. Product was purified by two steps recristalisation in acetone : n-hexane (15:1) and in ethanol. Obtained products were analysed by melting points determination, thin layer chromatography, and GC MS. In GC MS analysis was used N-phenilsuccinimide-D2 as internal standard.

The GC MS analysis of ¹- testosterone acetate, purified by recristalisation, conduced to a multitude of picks from which we identified: testosterone acetate, ¹- testosterone acetate, 1,6 Δ- testosterone acetate and costebole acetate-Hal. The separation of ¹- testosterone acetate was realised by preparative thin layer chromatography, using silicagel GF 254 as substrate and chloroform : acetone (9:1) mixture, as eluent. Purified product was caracterised by GC MS.

The deuterium labelled hormone was obtained by: (a) selective hydrogenation of ¹- testosterone acetate in dioxane, in presence of Pd/C, 10%, at low pressure D₂ gas, (b) separation of reaction mixture by filtration and evaporation of solvent and (c) hydrolysis of the ester at basic pH. The obtained raw product was purified by preparative thin layer cromatography. The characterization of labelled testosterone by GC MS reveals a purity higher than 98%.