Numazawa M, Osada R, Tsuji M, Osawa Y
Anal Biochem. 1985 Apr;146(1):75-81. doi: 10.1016/0003-2697(85)90398-7.
A sensitive nonradiometric assay of aromatization of 16 alpha-hydroxylated androgens, 16 alpha-hydroxy-4-androstene-3,17-dione (16 alpha-OHA), and 16 alpha-hydroxytestosterone (16 alpha-OHT), has been developed using reverse-phase high-performance liquid chromatography with voltametric detector. The estrogens produced by human placental microsomes, estriol (E3) and 16 alpha-hydroxyestrone (16 alpha-OHE1), were simultaneously detected in quantities as low as 1-2 ng using 3-methoxy-1,3,5(10)-estratriene-2, 16 alpha,17 beta-triol as an internal standard. E3 was the only estrogen detected from the incubate of 16 alpha-OHT with the microsomes and NADPH, while 16 alpha-OHA gave 16 alpha-OHE1 and E3 under the same conditions. Apparent Km and Vmax of the microsomal aromatase for 16 alpha-OHA and 16 alpha-OHT were 2.56 microM and 71.4 pmol/min/mg and 13.33 microM and 15.4 pmol/min/mg, respectively.
利用带有伏安检测器的反相高效液相色谱法，已开发出一种灵敏的非放射性测定方法，用于检测16α-羟基化雄激素、16α-羟基-4-雄烯-3,17-二酮（16α-OHA）和16α-羟基睾酮（16α-OHT）的芳香化反应。使用3-甲氧基-1,3,5(10)-雌三烯-2,16α,17β-三醇作为内标，可同时检测出人胎盘微粒体产生的雌激素，即雌三醇（E3）和16α-羟基雌酮（16α-OHE1），检测量低至1-2纳克。E3是16α-OHT与微粒体和NADPH孵育后唯一检测到的雌激素，而在相同条件下，16α-OHA产生16α-OHE1和E3。微粒体芳香化酶对16α-OHA和16α-OHT的表观Km和Vmax分别为2.56微摩尔和71.4皮摩尔/分钟/毫克以及13.33微摩尔和15.4皮摩尔/分钟/毫克。
