Xu S H, Norton R A, Crumley F G, Nes W D
Plant Physiology Research Unit, Russell Research Center, U.S. Department of Agriculture, Athens, GA 30605.
J Chromatogr. 1988 Oct 28;452:377-98. doi: 10.1016/s0021-9673(01)81462-x.
The chromatographic properties of approximately 100 sterols, select steroids of plant origin (sapogenins and steroidal alkaloids) and triterpenoids has been evaluated in this laboratory by monitoring their elution characteristics in adsorption (gravity column and thin-layer methods with and without the addition of silver nitrates), gas and reversed-phase high-performance liquid chromatography. The utility of each methodology to act in one or another chromatographic mode-separation, radio-chemical purification, quantitation and structural elucidation, is discussed. The importance of the tilt of the -OH group at C-3 as well as the polarity, size, an shape of the rest of the molecule as it effects the hydrogen-bonding ability of the -OH group is demonstrated through changes in chromatographic behavior that result from the step-wise introduction of double bonds, methyl, bromo, oxygen, nitrogen and cyclopropyl groups into 5 alpha-cholestanol. An independent aid in the structure identification and quantitation of the compounds was use of a multiple-wavelength diode array detector in which different wavelengths of the UV spectrum (200-400 nm) were simultaneously monitored following passage of the sample through a reversed-phase C18 column.
本实验室通过监测约100种甾醇、精选的植物源甾体（皂甙元及甾体生物碱）和三萜类化合物在吸附色谱法（重力柱色谱法以及添加和不添加硝酸银的薄层色谱法）、气相色谱法和反相高效液相色谱法中的洗脱特性，对它们的色谱性质进行了评估。文中讨论了每种方法在一种或另一种色谱模式（分离、放射化学纯化、定量和结构解析）中的应用。通过在5α-胆甾烷醇中逐步引入双键、甲基、溴、氧、氮和环丙基导致的色谱行为变化，证明了C-3位-OH基团的取向以及分子其余部分的极性、大小和形状对-OH基团氢键结合能力的影响。在化合物的结构鉴定和定量分析中，一种独立的辅助手段是使用多波长二极管阵列检测器，在样品通过反相C18柱后，同时监测紫外光谱（200 - 400 nm）的不同波长。
