Santos-Montes A, Gasco-Lopez A I, Izquierdo-Hornillos R
Departamento de Química Analítica, Facultad de Química, Universidad Complutense de Madrid, Spain.
J Chromatogr. 1993 Oct 22;620(1):15-23. doi: 10.1016/0378-4347(93)80046-7.
Systematic optimization of the HPLC separation of a mixture of natural and synthetic corticosteroids was carried out for screening purposes. The method involves binary, ternary or quaternary mixtures containing water, methanol, acetonitrile and tetrahydrofuran. It was possible to separate thirteen out of fourteen corticosteroids contained in a sample in about 26 min, with a 5-microns Hypersil-C18 (250 mm x 4.6 mm I.D.) column thermostated at 30 degrees C, using a mobile phase composed of water-tetrahydrofuran (72:28, v/v). This separation was not improved using other C8 or C18 columns. The effect of temperature on the separation of these compounds was also studied. Calibration graphs were established for each corticosteroid up to 8 micrograms/ml using indapamide as internal standard. The detection limits were in the range 0.02-0.14 ng. The optimized method was applied to urine samples spiked with corticosteroids and showed potential for future applications.
为了筛选目的，对天然和合成皮质类固醇混合物的高效液相色谱（HPLC）分离进行了系统优化。该方法涉及含有水、甲醇、乙腈和四氢呋喃的二元、三元或四元混合物。使用内径为4.6 mm、长度为250 mm、粒径为5微米的Hypersil-C18柱，在30℃恒温条件下，以水 - 四氢呋喃（72:28，v/v）为流动相，在约26分钟内可以分离样品中所含的十四种皮质类固醇中的十三种。使用其他C8或C18柱并不能改善这种分离效果。还研究了温度对这些化合物分离的影响。以吲达帕胺为内标，为每种皮质类固醇建立了浓度高达8微克/毫升的校准曲线。检测限在0.02 - 0.14纳克范围内。优化后的方法应用于添加了皮质类固醇的尿液样本，并显示出未来应用的潜力。
