Reid J J, Stitzel R E, Head R J
J Pharmacol Methods. 1985 Aug;14(1):25-39. doi: 10.1016/0160-5402(85)90040-3.
A precolumn extraction procedure combined with high-performance liquid chromatography with electrochemical detection (HPLC-ECD) has been developed for the identification and quantitation of catechol estrogens, their immediate precursors (estrogens), and their 0-methylated metabolites. These compounds were isolated from Krebs solutions and from perchloric acid tissue extracts or whole tissues by simple solvent extraction prior to quantitation. The optimal conditions for detection by HPLC-ECD were those employing a radial compression module, a C18 Bondapak cartridge, and an isocratic mobile phase of ammonium phosphate (pH 3.0) combined with 30-70% acetonitrile. A biological application of this procedure includes the investigation of the 0-methylation of catechol estrogens by vascular tissue. In the present study, the 0-methylation of 2-hydroxyestradiol by rabbit aorta has been examined. The 0-methylation in segments of thoracic aorta was significantly less than that in abdominal aortic segments, but was reproducible along the length of the thoracic aorta.
已开发出一种柱前萃取程序，结合高效液相色谱 - 电化学检测（HPLC - ECD）用于儿茶酚雌激素、其直接前体（雌激素）及其O - 甲基化代谢物的鉴定和定量。在定量之前，通过简单的溶剂萃取从克雷布斯溶液、高氯酸组织提取物或全组织中分离出这些化合物。HPLC - ECD检测的最佳条件是采用径向压缩模块、C18 Bondapak柱以及磷酸铵（pH 3.0）与30 - 70%乙腈组成的等度流动相。该程序的一个生物学应用包括研究血管组织对儿茶酚雌激素的O - 甲基化作用。在本研究中，已检测了兔主动脉对2 - 羟基雌二醇的O - 甲基化作用。胸主动脉段的O - 甲基化作用明显低于腹主动脉段，但在胸主动脉全长范围内具有可重复性。
