High-Performance Liquid Chromatography with Fluorescence Detection for Simultaneous Analysis of Phytosterols (Stigmasterol, β-Sitosterol, Campesterol, Ergosterol, and Fucosterol) and Cholesterol in Plant Foods
High-Performance Liquid Chromatography with Fluorescence Detection for Simultaneous Analysis of Phytosterols (Stigmasterol, β-Sitosterol, Campesterol, Ergosterol, and Fucosterol) and Cholesterol in Plant Foods
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DOI：
10.1007/s12161-017-0841-2
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发表时间：
2017-08-01
影响因子：
2.9
通讯作者：
Tanaka, Ryusuke
中科院分区：
文献类型：
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作者：
Ito, Meiko;Ishimaru, Mami;Tanaka, Ryusuke
建立了高效液相色谱-荧光检测同时分析植物甾醇（豆甾醇、β -谷甾醇、油菜甾醇、麦角甾醇、focusterol）和胆固醇的方法。为了使甾醇荧光化，它们被1-蒽基氰化物衍生到每个甾醇骨架的碳3羟基上。该方法采用C-30色谱柱，以丙酮/乙腈/己烷/水（71:20:4:5,v/v）为流动相的等压溶液为流动相，在1.0 mL min（-1）下进行荧光检测，激发波长为370 nm，发射波长为470 nm。分离出5种植物甾醇、胆固醇和1-六糖醇作为内标，具有足够的重现性和定量能力。该方法可对木耳、大豆、欧芹等陆生植物以及海苔、红藻、绿藻等海洋藻类的甾醇含量进行评价。通过对陆地植物的分析，木耳中含有大量麦角甾醇作为维生素D-2的前体。大豆中含有大量的豆甾醇、油菜甾醇和-谷甾醇。与木耳和大豆相比，欧芹含有少量的这些甾醇。在这些海藻中，Hiziki、Ogonori和Heraiwazuta分别含有大量的聚珠甾醇、胆固醇和-谷甾醇。海藻的组成与陆地植物不同。
A method based on high-performance liquid chromatography (HPLC) with fluorescence (FL) detection for the simultaneous analysis of phytosterols (stigmasterol, beta-sitosterol, campesterol, ergosterol, and fucosterol) and cholesterol was developed. To fluoresceinate the sterols, they were derivatized by 1-anthroyl cyanide to the hydroxyl group at carbon 3 of each sterol skeleton. This HPLC-FL method consists of a C-30 column, an isocratic solution using acetone/acetonitril/hexane/water (71:20:4:5, v/v) as the mobile phase at 1.0 mL min(-1) and fluorescence detection at an excitation of 370 nm and an emission of 470 nm. The separation of five phytosterols, cholesterol, and 1-hexacosanol as an internal standard was achieved with sufficient reproducibility and quantitative ability. Our method could evaluate the sterols of land plants such as wood ear fungus, soybean, and parsley, as well as marine algae such as Hiziki (Phaeophyta), Ogonori (Rhodophyta), and Heraiwazuta (Chlorophyta). As a result of the analysis of land plants, wood ear contained a large amount of ergosterol as a precursor of vitamin D-2. Soybean contained a large amount of stigmasterol, campesterol, and beta-sitosterol. Parsley contained small amounts of these sterols compared with wood ear and soybean. Among the marine algae, Hiziki, Ogonori, and Heraiwazuta contained large amounts of fucosterol, cholesterol, and beta-sitosterol, respectively. The compositions of marine algae differed from those of land plants.