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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2013, Vol. 39 Issue (2): 233-236    DOI: 10.3785/j.issn.1008-9209.2012.11.619
论文     
植物甾醇高效液相色谱法正相和反相检测方法对比
阮慧娜， 刘松柏*
浙江大学生物系统工程与食品科学学院食品科学与营养系，杭州 310058
Comparison of normal and reverse phase column in phytosterol detection by high performance liquid chromatography (HPLC)
RUAN Huina, LIU Songbai*
Department of Food Science and Nutrition, School of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China
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摘要： 分别使用正相柱和反相柱，采用高效液相色谱紫外检测器(high performance liquid chromatographyultraviolet detector, HPLCUV)法测定植物甾醇，比较2种色谱柱法对植物甾醇检测的优缺点. 正相色谱条件为：以Hypersil SiO2 (4.6 mm×250 mm, 5 μm)为色谱柱，流动相为V(正己烷)∶V(异丙醇)=99∶1，柱温35 ℃，流速1.0 mL/min，检测波长205 nm；反相色谱条件为：以默克 RP18 (4.6 mm×250 mm, 5 μm)为色谱柱，流动相为纯甲醇，柱温35 ℃，流速1.0 mL/min，检测波长205 nm. 结果表明：混合甾醇在反相柱分离下出现3个峰，各种甾醇的分离度好；正相柱的甾醇只有1个峰，但样品在正相体系中的溶解性高于反相体系，而且正相体系甾醇出峰时间比反相体系快，可节约实验时间. 说明在不要求各种甾醇分离效果的前提下，正相柱更适合用于植物甾醇检测.
Abstract: Phytosterols are natural bioactive compounds, usually including stigmasterol, sitosterol, campesterol, and rapeseed sterol. They have attracted growing attention owing to their beneficial effects. High performance liquid chromatography (HPLC) is the most frequently used method to quantify phytosterols. Although several detectors such as ELSD (evaporative light scattering detector) and UV (ultraviolet detector) are available for HPLC analysis, but UV detector is believed to be much more effective and always preferred in laboratories. In the recent literature available, the reverse phase column was mostly applied to determine phytosterols in HPLCUV system. However, the performance of normal phase column in phytosterol analysis has not been well studied and compared. In this study, the performances of both reverse phase and normal phase columns in HPLCUV system for phytosterol analysis were compared, and their respective advantages were illustrated. Both normal phase and reverse phase columns in HPLCUV were applied to detect phytosterols, respectively. The normal phase chromatographic conditions were as follows: using hypersil SiO2（4.6 mm×250 mm, 5 μm) as column and V(hexane)∶V(isopropanol)=99∶1 as mobile phase, column temperature of 35 ℃, flow rate of 1.0 mL/min, and detection wavelength of 205 nm. Reverse phase chromatographic conditions were as follows: using Merck RP18 (4.6 mm×250 mm, 5 μm) as column and pure methanol as mobile phase, column temperature of 35 ℃, flow rate of 1.0 mL/min, and detection wavelength of 205 nm. As a result, high sensitivity was observed for phytosterol detection using UV detector in both systems. It was revealed that the normal phase system could not separate individual phytosterol compounds, while the reverse phase system exhibited good separation capacity. In terms of retention time, the normal phase system needed much less time as compared with reverse one. In addition, phytosterols were much more soluble in the eluent of the normal phase system. It is concluded that the normal phase column is more suitable than reverse one for quantification of total phytosterols despite of lower separation ability. Furthermore, an efficient method for quantification of phytosterols employing the normal phase column and the benzoate internal standard is developed.
出版日期: 2013-03-20
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阮慧娜
刘松柏*

引用本文:

阮慧娜， 刘松柏*. 植物甾醇高效液相色谱法正相和反相检测方法对比[J]. 浙江大学学报(农业与生命科学版), 2013, 39(2): 233-236.

RUAN Huina, LIU Songbai*. Comparison of normal and reverse phase column in phytosterol detection by high performance liquid chromatography (HPLC). Journal of Zhejiang University (Agriculture and Life Sciences), 2013, 39(2): 233-236.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2012.11.619    或    http://www.zjujournals.com/agr/CN/Y2013/V39/I2/233

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