Synthesis of Isotope-labeled Cholesterol-3,4-13C2

Abstract

The 13C-labeled synthesis of cholesterol-3,4-13C2 was based on phenyl acetate-1,2-13C2 and 4-cholesten-3-one. 4-Cholesten-3-one was reacted in a certain proportion solvent mixed by isopropanol and water to obtain the carboxylic acid intermediate through ring opening oxidation reaction using potassium permanganate and sodium periodate as oxidant，and then reacted with acetate under heat and reflux in nitrogen protection to produce the ring-closure carbonyl intermediate. The acetylation labeled intermediate was synthesized with the ring-closure carbonyl intermediate and phenyl acetate-1,2-13C2 in tetrahydrofuran as solvent，using sodium hydride as catalyzer through acetylation reaction. Finally the cholesterol-3,4-13C2 was obtained sequentially by carbon ring reconstruction reaction，esterification and reduction reaction with the acetylation labeled intermediate as raw material. The reduction reaction showed a certain degree of asymmetric synthesis phenomenon. The yield based on the amount of substance of acetate-1,2-13C2 was 16.2%. The structure of target product was confirmed by MS，NMR and HPLC. Its chemical purity (HPLC) was higher than 96%，and isotopic abundance was higher than 98% (atom 13C). This product can be used as the stable isotope internal standard reagent for detection in the field of clinical mass spectrometry and biomedical. It can also be used as a raw material of the test kit for clinical mass spectrometry disease detection.

Keywords

• cholesterol-3
• 4-13c2
• stable isotope labeled compound
• ssteroid hormone
• internal standard reagent
• test kit