Schöneshöfer M, Dulce H J
J Chromatogr. 1979 Sep 11;164(1):17-28. doi: 10.1016/s0378-4347(00)81567-4.
The high-performance liquid chromatography of nineteen hormonal steroids with special respect to its suitability for routine purification of these steroids from crude, organic extracts of biological fluids prior to final quantitation by immunoassay has been studied. In all systems the gradient elution technique was applied. Separation of steroids has been investigated using different stationary phases chemically coated with non-polar, hydroxyl, NO2 and CN groups. Reproducibility of retention times was studied on a stationary phase coated with hydroxyl groups (DIOL column) using different organic eluents. Coefficients of variation range from 0.76 to 8.16%. Reproducibility was shown to be unequivocally better in the gradient part than in the isocratic part of the chromatographic run. In contrast to the other steroids, 18-hydroxylated steroids were more or less unstable in certain systems studied. As to resolution and reproducibility, the DIOL column run with an n-hexane-dioxane gradient has been shown to be superior to the other systems studied.
研究了19种激素甾体的高效液相色谱法，特别关注其在通过免疫测定进行最终定量之前，从生物流体的粗有机提取物中常规纯化这些甾体的适用性。在所有系统中均采用梯度洗脱技术。使用化学涂覆有非极性、羟基、NO2和CN基团的不同固定相研究了甾体的分离。在涂覆有羟基的固定相（二醇柱）上使用不同的有机洗脱剂研究了保留时间的重现性。变异系数范围为0.76%至8.16%。结果表明，在色谱运行的梯度部分，重现性明显优于等度部分。与其他甾体不同，在所研究的某些系统中，18-羟基化甾体或多或少不稳定。就分离度和重现性而言，使用正己烷-二氧六环梯度的二醇柱已被证明优于所研究的其他系统。
