Emons G, Hoppen H O, Ball P, Knuppen R
Steroids. 1980 Jul;36(1):73-9. doi: 10.1016/0039-128x(80)90069-0.
Portions of pregnancy and midcycle urines were submitted to hot acid hydrolysis, extracted with benzene/ethyl acetate and the extracts washed with ascorbic acid buffer. From the remaining organic phase the catecholestrogens were removed with borate buffer and further purified on Sephadex LH-20 columns. After derivatisation 4-hydroxyestrone was separated from the isomeric 2-hydroxyestrone peak were identical with that of authentic 4-hydroxyestrone. After treatment of the extracts with sodium borohydride 4-hydroxyestradiol-17 beta was identified by GC-MS. By the addition of trace amounts of tritiated 4-hydroxyestrone a recovery of 40% was calculated. On the basis of this recovery and the peak heights of the gas chromatograms an excretion of 4 microgram (midcycle) and 40 microgram (pregnancy) of 4-hydroxyestrone/24 h was estimated.
取部分孕期尿液和月经周期中期尿液进行热酸水解，用苯/乙酸乙酯萃取，萃取液用抗坏血酸缓冲液洗涤。从剩余的有机相中用硼酸盐缓冲液除去儿茶酚雌激素，并在葡聚糖凝胶LH - 20柱上进一步纯化。衍生化后，从异构体2 - 羟基雌酮峰中分离出4 - 羟基雌酮，其峰与 authentic 4 - 羟基雌酮的峰相同。用硼氢化钠处理萃取液后，通过气相色谱 - 质谱联用仪鉴定出4 - 羟基雌二醇 - 17β。加入微量氚标记的4 - 羟基雌酮后，计算回收率为40%。根据该回收率和气相色谱图的峰高，估计4 - 羟基雌酮的排泄量为4微克（月经周期中期）和40微克（孕期）/24小时。
