Allner R, Eggstein M, Kujawski H
Klin Wochenschr. 1975 Sep 15;53(18):867-71. doi: 10.1007/BF01466960.
A method is described for the quantitative determination of 20-ketosteroids in urine during the application of a 3alpha, 20beta-hydroxysteroid: NAD oxidoreductase. In the presence of NADH H+ the enzyme catalyses in an "equimolar" developing reaction the reduction of 20-keto groups of C-21 steroids giving 20 beta-alcohols. The measurable NADH H+ consumption is a measure for the quantity of those 20-ketosteroids present in the reaction mixture. The 3alpha-hydroxysteroid: NAD oxidoreductase activity of the enzyme does not influence the specificity of the method to determine the 20-ketosteroids because,-under the present extraction and purification conditions-those 3alpha-hydroxy-5alpha-androstan derivatives are not reaching the reaction mixture of determination. The demands towards the criterions of reliability (correctness and exactitude) come to be fulfilled by this method.
描述了一种在应用3α,20β-羟基类固醇:NAD氧化还原酶过程中定量测定尿中20-酮类固醇的方法。在NADH H⁺存在下,该酶在“等摩尔”显色反应中催化C-21类固醇的20-酮基还原生成20β-醇。可测量的NADH H⁺消耗量是反应混合物中存在的那些20-酮类固醇数量的度量。该酶的3α-羟基类固醇:NAD氧化还原酶活性不影响测定20-酮类固醇方法的特异性,因为在目前的提取和纯化条件下,那些3α-羟基-5α-雄甾烷衍生物不会进入测定反应混合物。该方法满足了对可靠性标准(正确性和精确性)的要求。