Ball P, Reu G, Schwab J, Knuppen R
Steroids. 1979 May;33(5):563-76. doi: 10.1016/0039-128x(79)90036-9.
An assay for the quantitative determination of 2-hydroxyestrone and 2-methoxyestrone in human urine is described. The analytical procedure involves several purification steps: XAD-2 column chromatography of urine (1 ml), hot acid hydrolysis under reducing conditions, extraction with benzene/ethyl acetate, separation of monophenolic steroids from catecholestrogens by the formation of borate complexes, and partition between different solvents. Quantitation is achieved by radioimmunoassay using highly specific antibodies. For correction of procedural losses [6, 7-3H2] 2-hydroxyestrone and [6, 7-3H2] 2-methoxyestrone are used as internal standards. The method is highly specific as checked by comparison to a double-isotope-derivative method and a newly developed gas chromatography-mass spectrometry procedure. Using this method the urinary excretion of 2-hydroxyestrone and 2-methoxyestrone was studied in children, men, cycling, pregnant and postmenopausal women. Special interest was focussed on the molar ratios of 2-hydroxyestrone to 2-methoxyestrone and the so called "total estrogens" which vary markedly within the different groups investigated. The excretion of 2-hydroxyestrone is especially favoured in women during the menstrual cycle and pregnancy.
本文描述了一种用于定量测定人尿中2-羟基雌酮和2-甲氧基雌酮的分析方法。分析过程包括几个纯化步骤：尿液（1毫升）的XAD-2柱色谱法、还原条件下的热酸水解、苯/乙酸乙酯萃取、通过形成硼酸络合物将单酚类甾体与儿茶酚雌激素分离以及在不同溶剂之间分配。通过使用高度特异性抗体的放射免疫测定法进行定量。为校正操作损失，使用[6,7-³H₂] 2-羟基雌酮和[6,7-³H₂] 2-甲氧基雌酮作为内标。通过与双同位素衍生法和新开发的气相色谱-质谱法比较验证，该方法具有高度特异性。使用该方法研究了儿童、男性、处于月经周期的女性、孕妇和绝经后女性中2-羟基雌酮和2-甲氧基雌酮的尿排泄情况。特别关注的是2-羟基雌酮与2-甲氧基雌酮的摩尔比以及所谓的“总雌激素”，它们在不同研究组中差异显著。在月经周期和怀孕期间，女性中2-羟基雌酮的排泄尤其明显。
