Iohan F, Vincze I, Monder C, Cohen S
Population Council, New York, NY 10021.
J Chromatogr. 1991 Mar 8;564(1):27-41. doi: 10.1016/0378-4347(91)80067-m.
A new procedure is described for the detection of the acidic metabolites of cortisol (cortoic acids) as the pyrenylmethyl-21-oic esters. The derivatizing reagent, diazomethylpyrene, was prepared by an improved procedure. The reagent was used at room temperature, required no catalyst, and was not restricted by stoichiometric requirements. The steroid esters were separated by reversed-phase high-performance liquid chromatography and analyzed simultaneously by their ultraviolet absorbance and fluorescence characteristics. Identities of the products were confirmed using the photodiode array detector to determine spectral profiles, absorbance maxima, and absorbance ratios. Further confirmation of identity of the cortoic acid esters used mass spectrometry under normal and collision-activated dissociation conditions. With the method described, a linear spectral response was obtained between 8 and 1680 fmol. Application of the technique to the analysis of steroid acids in human urine indicated the presence of cortoic acids.
描述了一种用于检测皮质醇酸性代谢物（皮质酸）的新方法，即检测其芘甲基-21-酸酯。衍生试剂重氮甲基芘通过改进的方法制备。该试剂在室温下使用，无需催化剂，不受化学计量要求的限制。甾体酯通过反相高效液相色谱分离，并通过其紫外吸收和荧光特性同时进行分析。使用光电二极管阵列检测器确定光谱图、最大吸收波长和吸收比，以确认产物的身份。在正常和碰撞激活解离条件下，通过质谱进一步确认皮质酸酯的身份。使用所描述的方法，在8至1680飞摩尔之间获得了线性光谱响应。将该技术应用于分析人尿中的甾体酸，结果表明存在皮质酸。
