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Biosynthetic preparation of the tritium labelled 3α‐hydroxy‐5α‐pregnan‐20‐one, an allosteric modulator of gabaergic neurotransmission
Author(s) -
Jussofie Astrid,
Jussofie Ibrahim,
Stoppok Martin,
Lax E. Rodney
Publication year - 1992
Publication title -
journal of labelled compounds and radiopharmaceuticals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.432
H-Index - 47
eISSN - 1099-1344
pISSN - 0362-4803
DOI - 10.1002/jlcr.2580310708
Subject(s) - chemistry , sephadex , tritium , metabolite , chromatography , yield (engineering) , high performance liquid chromatography , specific activity , silica gel , isotope dilution , allosteric regulation , biochemistry , receptor , mass spectrometry , enzyme , physics , materials science , nuclear physics , metallurgy
A procedure for the biosynthetic preparation of 3α‐hydroxy‐5α‐[1,2,6,7‐ 3 H]‐pregnan‐20‐one ( 3 H‐HPO) with high specific activity convenient for binding studies was developed. 3 H‐HPO was synthesized from [1,2,6,7‐ 3 H]progesterone using a microsomal preparation from rat liver and NADPH. The progesterone metabolite was purified by silica gel 60 and Sephadex LH 20 chromatography. The identity of the product was confirmed by reverse isotopic dilution of chemically synthesized and highly purified 3α‐hydroxy‐5α‐pregnan‐20‐one. The yield for radiolabelled HPO was 60% and its purity was > 98%.