二维高效液相色谱法同时测定菜肴中维生素A、D和E  

Determination of vitamin A,D and E in dishes by two-dimensional liquid chromatography

作　　者：黄百芬[1] 张京顺[1] 王梦莉 陈苘[1] 徐维盛 张正雨 Huang Baifen;Zhang Jingshun;Wang Mengli;Chen Qing;Xu Weisheng;Zhang Zhengyu(Zhejiang Provincial Center for Disease Control and Prevention,Hangzhou 310051,China;National Institute for Nutrition and Health,Chinese Center for Disease Control and Prevention,Beijing 100050,China;Acchrom Technology Co.,Ltd,Dalian 116025,China)

机构地区：[1]浙江省疾病预防控制中心,杭州310051 [2]中国疾病预防控制中心营养与健康所,北京100050 [3]华谱科仪(北京)科技有限公司,大连116025

出　　处：《卫生研究》2023年第2期272-279,共8页Journal of Hygiene Research

基　　金：国家卫生健康委员会食品安全国家标准修订项目(No.spaq-2018-047);国家卫生健康委员会中国食物成分监测项目;中国营养学会-百胜餐饮健康基金(No.CNS-YUM2017A03)。

摘　　要：目的建立二维高效液相色谱法同时测定菜肴中维生素A、D、E的方法。方法试样在抗氧化剂保护下,经过皂化,乙酸乙酯-正己烷混合液(3∶2,V/V)提取,二维高效液相色谱仪测定。以华谱Alphasil五氟苯基柱(150 mm×4.6 mm,5μm)为一维色谱柱,用水和甲醇为流动相进行梯度洗脱,实现视黄醇、α-生育酚、β-生育酚、γ-生育酚、δ-生育酚、α-生育三烯酚的基线分离,荧光检测器检测;以华谱Alphasil VC-C18柱(4.6 mm×150 mm,3.5μm)为二维色谱柱,用含2%甲醇的乙腈溶液为流动相进行梯度洗脱,实现维生素D2、D3与其他杂质的分离,紫外检测器检测。结果本研究实现视黄醇、维生素D2和维生素D3、5种生育酚的基线分离检测,在预设的浓度范围内各目标化合物呈线性相关,相关系数>0.999;样品中三水平加标回收率为85.4%~106.4%,相对标准偏差<8%。对干炸带鱼、油焖香菇、酱油蒸蛋、红烧肉、糖醋排骨等常见成品菜肴进行6次重复测定,相对标准偏差均小于10%。视黄醇的检出限为0.7μg/100 g,定量限为2.4μg/100 g,5种生育酚的检出限为1.1~2.5μg/100 g,定量限为3.6~8.3μg/100 g;维生素D2、D3的检出限为0.3μg/100 g,定量限为1.0μg/100 g。应用本方法检测了41份成品菜肴中维生素A、D、E含量,发现在动物源性菜肴中可检出维生素A和D3,在所有菜肴中均可检出α-生育酚和γ-生育酚,在使用了植物油的菜肴中可检出5种生育酚。带鱼采用清蒸、红烧、干炸3种不同的烹饪方式,维生素A和D3的含量依次降低。结论本方法具有操作简单、重复性好、准确性高,对操作人员和生态环境友好等特点,可实现成品菜肴中维生素A、D、E的同时分型检测。OBJECTIVE To establish a method for the simultaneous determination of vitamins A,D and E in dishes by two-dimensional liquid chromatography.METHODS The samples were saponified and extracted with a mixture of ethyl acetate and n-hexane(3∶2,V/V)under the protection of antioxidants,and determined by two-dimensional liquid chromatography.The baseline separation of retinol,α-tocopherol,β-tocopherol,γ-tocopherol,δ-tocopherol,andα-tocotrienols was achieved by using Alphasil pentafluorophenyl column(PFP,150 mm×4.6 mm,5μm)as the one-dimensional column,and using water and methanol as mobile phases for gradient elution under the fluorescence detector.The separation detection of ergocalciferol and cholecalciferol from other impurities was achieved on the UV detector by Alphasil VC-C18column(150 mm×4.6 mm,3.5μm)as the two-dimensional column.RESULTS The baseline separation detection of retinol,ergocalciferol,cholecalciferol and five tocopherols was achieved.The compounds were linearly correlated within the set range,and the correlation coefficients were>0.999.The recovery rate of the method was between 85.4%and 106.4%.The detection limit of all-trans retinol was 0.7μg/100 g,and the limit of quantitation was 2.4μg/100 g.The limits of detection and quantification of tocopherols ranged from 1.1 to 2.5μg/100 g and 3.6 to 8.3μg/100 g.The detection limit of ergocalciferol and cholecalciferol was 0.3μg/100 g,and the limit of quantification was 1.0μg/100 g.In the end,finished dishes such as dry fried hairtail,braised mushroom,steamed egg with soy sauce,sweet and sour ribs were repeatedly measured by this method for six times,and the relative standard deviation was less than 10%.CONCLUSIONThe method has the characteristics of simple operation,good repeatability,high accuracy and friendly to operators and ecological environment.It can realize the simultaneous typing detection of vitamins A,D and E in finished dishes.

关 键 词：二维高效液相色谱法 菜肴 维生素A 维生素D 维生素E 

分 类 号：O657.71[理学—分析化学] R151.3[理学—化学]