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.
Wei Sheng Yan Jiu. 2023 Mar;52(2):272-279. doi: 10.19813/j.cnki.weishengyanjiu.2023.02.016.
To establish a method for the simultaneous determination of vitamins A, D and E in dishes by two-dimensional liquid chromatography.
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, and α-tocotrienols was achieved by using Alphasil pentafluorophenyl column(PFP, 150 mm×4.6 mm, 5 μm) 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-C_(18) column(150 mm×4.6 mm, 3.5 μm) as the two-dimensional column.
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 0.7 μg/100 g, and the limit of quantitation was 2.4 μg/100 g. The limits of detection and quantification of tocopherols ranged from 1.1 to 2.5 μg/100 g and 3.6 to 8.3 μg/100 g. The detection limit of ergocalciferol and cholecalciferol was 0.3 μg/100 g, and the limit of quantification was 1.0 μg/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%.
The 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.
建立一种用二维液相色谱法同时测定菜肴中维生素A、D和E的方法。
样品在抗氧化剂保护下皂化，用乙酸乙酯和正己烷（3∶2，V/V）混合液萃取，采用二维液相色谱法测定。以Alphasil五氟苯基柱（PFP，150 mm×4.6 mm，5 μm）为一维柱，以水和甲醇为流动相进行梯度洗脱，在荧光检测器下实现视黄醇、α-生育酚、β-生育酚、γ-生育酚、δ-生育酚和α-生育三烯酚的基线分离。以Alphasil VC-C₁₈柱（150 mm×4.6 mm，3.5 μm）为二维柱，在紫外检测器上实现麦角钙化醇和胆钙化醇与其他杂质的分离检测。
实现了视黄醇、麦角钙化醇、胆钙化醇和五种生育酚的基线分离检测。各化合物在设定范围内呈线性相关，相关系数均大于0.999。该方法的回收率在85.4%～106.4%之间。全反式视黄醇的检出限为0.7 μg/100 g，定量限为2.4 μg/100 g。生育酚的检出限和定量限分别为1.1～2.5 μg/100 g和3.6～8.3 μg/100 g。麦角钙化醇和胆钙化醇的检出限为0.3 μg/100 g，定量限为1.0 μg/100 g。最后，用该方法对干煎带鱼、红烧香菇、酱油蒸蛋、糖醋排骨等成品菜肴进行6次重复测定，相对标准偏差小于10%。
该方法操作简便、重复性好、准确度高，对操作人员和生态环境友好。可实现成品菜肴中维生素A、D和E的同时分型检测。
