植物甾醇的超高效液相色谱测定方法的研究  

DETERMINATION OF STEROLS BY ULTRA-HIGH PERFORMANCE LIQUID CHROMATOGRAPHY IN PLANT-ORIGIN FOODS

作　　者：尹丹阳[1] 赵莹 赵迪 张宇晨 胡佳薇[1] YIN Dan-yang;ZHAO Ying;ZHAO Di;ZHANG Yu-chen;HU Jia-wei(Center for Disease Control and Prevention of Shaanxi Province,Xi'an 710054,China)

机构地区：[1]陕西省疾病预防控制中心,西安710054

出　　处：《营养学报》2023年第3期301-305,共5页Acta Nutrimenta Sinica

基　　金：国家财政项目:国民营养素需要量评估、食物环境评价及应用(No.131031107000210003)。

摘　　要：目的 优化植物甾醇测定皂化以及萃取条件,建立反相超高效液相色谱法测定植物性食品中三种植物甾醇(β-谷甾醇、豆甾醇和菜油甾醇)的方法。并用此方法测定了几种陕西省特色食品中三种植物甾醇的含量,为准确衡量居民日常膳食中的植物甾醇摄入量提供了新的方法。方法 样品经过皂化和液液萃取后采用AgilentPoroshell120PheHex色谱柱(2.1 mm×100 mm,2.7μm)进行分离,以甲醇-水梯度洗脱。DAD检测器在190~400 nm范围内进行3D扫描,选择200nm作为三种植物甾醇的特征吸收波长进行检测。结果 三种植物甾醇在浓度为5.00~300μg/ml内呈良好线性关系,相关系数r>0.997;三种植物甾醇检出限为0.08 mg/100g,定量限为0.24 mg/100g;通过优化皂化条件和提取试剂,最终选择5 ml 50%KOH溶液用来皂化样品,选择乙酸乙酯:正己烷(1:1)作为植物甾醇的提取试剂。样品中三种植物甾醇高中低3个浓度水平的测量精密度为2.3%~4.7%、加标回收率为82.4%~102.0%。结论 本研究建立的反相超高效液相色谱方法检测植物性食品中三种植物甾醇灵敏准确,适用于植物性食品中三种植物甾醇的同时测定。Objective To optimize saponification and extraction conditions for the determination of sterols and establish a method for simultaneous analysis of three sterols(β-sitosterol,stigmasterol,campesterol)in plant-origin foods by reversed-phase ultra-performance liquid chromatography.Methods The samples were pretreated with saponification and liquid-liquid extraction,then separated on Agilent Poroshell 120 PheHex column(2.1mmx100mm,2.7μm)with gradient elution using methanol-water as mobile phase.The analytes were detect by DAD detector and 200 nm was selected as the characteristic absorption wavelength of three sterols.Results The concentration of three sterols showed good linearity in the range of 5.00-300μg/ml,respectively,with the correlation coefficients greater than 0.997.The limit of detection(LODs)was 0.08 mg/100g and limit of quantification(LOQs)0.24 mg/100g for three sterols.By optimizing the saponification and extraction conditions,5 ml of 50%KOH solution was finally selected to saponify the samples and ethyl acetate:n-hexane(1:1)selected as the extraction reagent.The recoveries at three levels in the matrix were 82.4%-102.0%,and the relative standard deviations(RSDs)were 2.3%-4.7%.Conclusion This reversed-phase ultra high-performance liquid chromatography method is sensitive,accurate and suitable for the analysis of three sterols in plant-origin foods.

关 键 词：超高效液相色谱法 Β-谷甾醇 豆甾醇 菜油甾醇 植物性食品 

分 类 号：R151.3[医药卫生—营养与食品卫生学]