油脂类药用辅料橄榄油中甾醇谱的研究

目的:分析油脂类辅料橄榄油中甾醇组成及质量分数。方法:首先从橄榄油中提取不皂化物,运用高效液相色谱法从不皂化物中分离并收集甾醇的馏分。液相色谱条件:采用Kromasil 60-5 SIL硅胶柱(250 mm×4.6 mm,5μm),以异丙醇-正己烷(1:99,v/v)为流动相等度洗脱,流速1.0 mL·min^-1,检测波长210 nm,进样量20 μL。然后对馏分进行衍生化,采用气相色谱法检测各甾醇的组成及百分含量;气相色谱条件:采用DB-5毛细管柱(30 m×0.25 mm×0.25μm),进样口温度:290℃;进样量:2~3 μL;分流比为25:1;载气:氦气;流速1.8 mL·min^-1;程序升温:起始柱温260℃,保持50 min,再以5℃·min^-1的速率升温至290℃,保持5 min。结果:橄榄油中含有多种甾醇,如胆甾醇、菜子甾醇;菜油甾醇、豆甾醇、△7-菜油甾醇、△5,23-豆甾二烯醇、赤桐甾醇、β-谷甾醇、谷甾烷醇、△5-燕麦甾醇、△5,24-豆甾二烯醇、△7-豆甾醇等,其中β-谷甾醇的含量比较高,占总甾醇量的80%以上,其次是菜油甾醇,△5,24-豆甾二烯醇等。结论:本法可以对油脂类辅料橄榄油中的甾醇谱进行分析,为油脂类药用辅料植物油中甾醇谱的研究提供了参考。

Objective: To analyze composition and percentages of sterols in pharmaceutical lipid excipient such as olive oil. Methods: Unsaponifiable matters were extracted from olive oil, and unsaponified sterol fractions were separated and collected by HPLC. HPLC conditions were as follows:the analysis was achieved using a Kromasil 60-5 SIL (250 mm×4.6 mm,5 μm) with isocratic elution of isopropyl alcohol-hexane (1:99,V/V), the flow rate was 1.0 mL·min^-1, the detection wavelength was 210 nm, and the injection size was 20 μL. Then the sterol fractions were derived before analysis by gas chromatography to obtain the composition and content of sterol. GC conditions were as follows: an DB-5 capillary column (30 m×0.25 mm×0.25 μm) was adopted, the inlet temperature was controlled at 290℃, the injection volume was 2-3 μL, the split ratio was 25:1, the carrier gas was nitrogen at a flow rate of 1.8 mL·min^-1, the inlet temperature was controlled at 260℃, the oven temperature was initially held at 260℃ for 50 min and was then programmed to 290℃ at the rate of 5℃·min^-1 and held for 5 min. Results: A variety of sterols were detected in olive oil, such as cholesterol, brassicasterol, campesterol, stigmasterol, △7-campesterol, △5,23-stigmastadienol, clerosterol, β-sitosterol, sitostanol, △5-avenasterol, △5,24-stigmastadienol, △7-stigmastenol and so on. The content of β-sitosterol was more than 80%, followed by campesterol, △5,24-stigmastadienol and other compenents. Conclusion: The proposed method is applicable for the determination of sterols in olive oil, and can be used for the research of sterols in vegetable oils.