Suzuki E, Saegusa K, Matsuki Y, Nambara T
Hatano Research Institute, Food and Drug Safety Center, Kanagawa, Japan.
J Chromatogr. 1993 Aug 11;617(2):221-5. doi: 10.1016/0378-4347(93)80491-l.
A simple and sensitive method for the determination of guaiaicol oestrogens enzymatically formed from 2- or 4-hydroxyoestradiol, by means of high-performance liquid chromatography with coulometric detection, has been developed. Catechol and guaiacol oestrogens were efficiently separated on a reversed-phase column, using 0.5% ammonium phosphate buffer (pH 3.0)-acetonitrile (59:41, v/v) as the mobile phase, and detected coulometrically in a screening-oxidation mode at +0.10 V and +0.35 V, respectively. The method was applied to the assay of in vitro enzymic O-methylation of catechol oestrogens. After 2- or 4-hydroxyoestradiol had been incubated with rat red blood cells in the presence of S-adenosylmethionine, the resulting guaiacols and unchanged substrate were percolated through an Extrelut-3 cartridge. The dried eluate was redissolved and directly injected. This simple procedure was as sensitive as the previously reported method using gas chromatography-mass spectrometry in a selected ion monitoring mode.
已开发出一种简单灵敏的方法，通过高效液相色谱 - 库仑检测法测定由2 - 或4 - 羟基雌二醇酶促形成的愈创木酚雌激素。儿茶酚雌激素和愈创木酚雌激素在反相柱上得到有效分离，以0.5%磷酸铵缓冲液（pH 3.0） - 乙腈（59:41，v/v）作为流动相，并分别在+0.10 V和+0.35 V的筛选氧化模式下进行库仑检测。该方法应用于儿茶酚雌激素体外酶促O - 甲基化的测定。在S - 腺苷甲硫氨酸存在下，将2 - 或4 - 羟基雌二醇与大鼠红细胞孵育后，将所得愈创木酚和未反应的底物通过Extrelut - 3柱进行渗滤。干燥后的洗脱液重新溶解并直接进样。这个简单的过程与先前报道的在选择离子监测模式下使用气相色谱 - 质谱联用的方法一样灵敏。
