Ogawa H
Nihon Naibunpi Gakkai Zasshi. 1983 May 20;59(5):801-14. doi: 10.1507/endocrine1927.59.5_801.
A simple method for the estimation of a urinary steroids profile with glass capillary gas chromatography is described. A 10ml portion of a urinary specimen was hydrolysed by Helix pomatia juice. Additional hydrolysis with ox-liver beta glucuronidase was necessary for the complete liberation of the urinary steroids from their glucuronides conjugates. Liberated free steroids were extracted with ethyl acetate, and the extract was washed and evaporated under reduced pressure. Finally extracted steroids were derivatised as MO-TMS (methoxime-trimethylsilyl) ethers. For the full-silylation of the steroids, the Sakauchi-Hornig method was employed and good results were obtained. An OV-101-WCOT column (30m length) was connected to the "solventless injection system" (a type of glass falling needle injection) and temperature-programmed gas chromatography was performed. By this method, eight fractions of 17-KS, Pregnanediol, Pregnanetriol, Pregnanetriolone, 5-Pregnentriol, tetrahydro-metabolites of 11-Deoxy-Cortisol and of Cortisol, and hexahydrometabolites of Cortisol were separated and quantitatively determined. Data processing was performed by an on-line microcomputer. The value in normal male and female controls was compatible with the results of glass capillary gas chromatography measured by Shackleton. Urine specimens from children with adrenocortical cancer and 21-hydroxylase deficiency were analyzed. Characteristic metabolic profiles of each patient were easily demonstrated. This method seems suitable for the routine clinical elucidation of abnormal steroid metabolism.
本文描述了一种用玻璃毛细管气相色谱法估算尿类固醇谱的简单方法。取10ml尿样，用蜗牛汁进行水解。为了使尿类固醇从其葡糖醛酸结合物中完全释放出来，还需要用牛肝β-葡糖醛酸酶进行额外水解。游离类固醇用乙酸乙酯萃取，萃取液经洗涤并减压蒸发。最后，将萃取的类固醇衍生化为甲氧肟-三甲基硅烷基（MO-TMS）醚。对于类固醇的全硅烷化，采用坂内-霍尼希法，获得了良好的结果。将一根OV-101壁涂开管柱（30m长）连接到“无溶剂进样系统”（一种玻璃落针进样器）上，并进行程序升温气相色谱分析。通过这种方法，分离并定量测定了17-酮类固醇、孕二醇、孕三醇、孕三酮、5-孕烯三醇、11-脱氧皮质醇和皮质醇的四氢代谢物以及皮质醇的六氢代谢物的八个组分。数据处理由在线微型计算机完成。正常男性和女性对照的值与沙克尔顿用玻璃毛细管气相色谱法测得的结果相符。对患有肾上腺皮质癌和21-羟化酶缺乏症儿童的尿样进行了分析。每位患者的特征性代谢谱很容易得到显示。该方法似乎适用于常规临床分析异常的类固醇代谢。
