De Beer J O
Afdeling Geneesmiddelenanalyse, Instituut voor Hygiene en Epidemiologie, Brussels, Belgium.
J Chromatogr. 1989 Apr 7;489(1):139-55. doi: 10.1016/s0378-4347(00)82891-1.
An on-line high-performance liquid chromatographic diode array spectroscopic analytical method for the identification of more than 60 different steroidal compounds is described. For the chromatographic separation, a gradient elution that could distinguish the esterified and non-esterified steroids in the same run on a reversed-phase C18 column, using methanol as modifier, was developed. For both types of compound an internal standard was chosen to establish reproducible relative retention times that could be used as one element of the identification; the second element is the UV spectrum, which is recorded on-line during the separation. The combination of chromatographic and UV spectroscopic recordings selects only a few probable steroids, which could be the unknown. This approach has been applied to forensic analysis of illicit preparations used in cattle-breeding, some examples of which are shown. For those steroids that are very difficult to distinguish using this procedure, because of their chromatographic and spectroscopic similarity on this system, other solvent mixtures are used in place of methanol as modifier, namely acetonitrile or tetrahydrofuran, or both, with the same solvent strength, as proposed by Snyder. In this way totally different elution patterns and separations are obtained, providing complementary information for identification, as shown by the systematic analysis on two other isoeluotropic solvent systems.
描述了一种用于鉴定60多种不同甾体化合物的在线高效液相色谱二极管阵列光谱分析方法。对于色谱分离，开发了一种梯度洗脱方法，该方法可以在反相C18柱上使用甲醇作为改性剂，在同一次运行中区分酯化和未酯化的甾体。对于这两种类型的化合物，选择内标来建立可重现的相对保留时间，该时间可作为鉴定的一个要素；第二个要素是紫外光谱，它在分离过程中在线记录。色谱和紫外光谱记录的结合仅选择了少数可能是未知物的甾体。这种方法已应用于牛养殖中非法制剂的法医分析，并给出了一些示例。对于那些由于在此系统上的色谱和光谱相似性而很难用此方法区分的甾体，使用其他溶剂混合物代替甲醇作为改性剂，即乙腈或四氢呋喃，或两者都用，具有与斯奈德提出的相同的溶剂强度。通过这种方式，可以获得完全不同的洗脱模式和分离效果，为鉴定提供补充信息，这在对另外两种等度洗脱溶剂系统的系统分析中得到了体现。
