Zhang Xin, Cambrai Amandine, Miesch Michel, Roussi Stamatiki, Raul Francis, Aoude-Werner Dalal, Marchioni Eric
Laboratoire de Chimie Analytique et Sciences de l'Aliment (LC4, UMR 7178), Faculté de Pharmacie, Université Louis Pasteur, 74 route du Rhin, 67400 Illkirch, France.
J Agric Food Chem. 2006 Feb 22;54(4):1196-202. doi: 10.1021/jf052761x.
A method for the separation, isolation, and identification of phytosterols was developed. A commercial phytosterols mixture, Generol 95S, was fractionated first by adsorption silica gel column chromatography and then separated by means of a semipreparative reverse phase high-performance liquid chromatography fitted with a Polaris C8-A column (250 mm x 10 mm i.d., 5 microm) using isocratic acetonitrile:2-propanol:water (2:1:1, v/v/v) as the mobile phase. Milligram scales of six individual phytosterols, including citrostadienol, campesterol, beta-sitosterol, Delta7-avenasterol, Delta7-campesterol, and Delta7-sitosterol, were obtained. Purities of these isolated sterols were 85-98%. Relative response factors (RRF) of these phytosterols were calculated against cholestanol as an authentic commercial standard. These RRF values were used to quantify by gas chromatography-mass spectrometry (GC-MS) the phytosterols content in a reference material, oils, and chocolates.
开发了一种分离、提纯和鉴定植物甾醇的方法。首先通过吸附硅胶柱色谱法对市售植物甾醇混合物Generol 95S进行分馏，然后采用配备Polaris C8-A柱（250 mm×10 mm内径，5微米）的半制备反相高效液相色谱法进行分离，以等度乙腈：2-丙醇：水（2:1:1，v/v/v）作为流动相。获得了毫克级的六种单一植物甾醇，包括柠檬甾二烯醇、菜油甾醇、β-谷甾醇、Δ7-燕麦甾醇、Δ7-菜油甾醇和Δ7-谷甾醇。这些分离出的甾醇纯度为85%-98%。以胆甾烷醇作为真实的商业标准，计算了这些植物甾醇的相对响应因子（RRF）。这些RRF值用于通过气相色谱-质谱联用（GC-MS）对参考物质、油类和巧克力中的植物甾醇含量进行定量分析。
