在线净化二维液相色谱快速检测食品中维生素A、D、E

Rapid detection of vitamin A,D and E in food by on-line purification two-dimensional high performance liquid chromatography

中文关键词:  维生素  在线净化  二维液相色谱

英文关键词:vitamin  on-line purification  two-dimensional liquid chromatography

中文摘要:

      目的 利用三泵两阀二维液相色谱系统, 建立食品中维生素A、D、E在线二维液相色谱快速检测方法。方法 采用中心切割方法搭建在线二维液相色谱分离系统, 以高聚合物为填料的具有耐强碱溶液能力的PLRP-S色谱柱作为净化样品皂化液的固相萃取(solid phase extraction, SPE)柱, 以C18为一维色谱柱分离维生素A和E, 以多环芳烃(polycyclic aromatic hydrocarbon, PAH)色谱柱为二维色谱柱分离维生素D2和D3。一维色谱确立维生素D的切割时间, 由C18小柱捕获维生素D, 建立在线二维液相色谱分析方法。利用样品和标品液对建立的方法进行方法学验证, 同时比较该方法与GB 5009.82-2016对实际样品的测试结果。结果 维生素A、D、E在试验标准曲线范围内线性系数r＞0.999, 定量限分别为30 μg/100g, 2 μg/100 g和120 μg/100 g, 与国标一致; 该方法3水平加标回收率为90.4%~101.7%, 可以满足目前食品中维生素A、D、E的检测需求。结论 本文建立的测试方法操作简单高效、自动化程度高、准确度高、重复性好, 适合于大批量样品的测试。

英文摘要:

      Objective To establish an on-line two-dimensional liquid chromatographic method for the determination of vitamins A, D and E in food by using a three-pump and two-valve two-dimensional liquid chromatographic system. Methods An on-line two-dimensional liquid chromatography separation system was set up by central cutting method. The PLRP-S column with high polymer as filler which had strong alkali resistance was used as the solid phase extraction (SPE) column for purifying saponification solution of samples. Vitamin A and E were separated by C18 as one-dimensional column, and vitamin D2 and D3 were separated by polycyclic aromatic hydrocarbon (PAH) as two-dimensional column. One-dimensional chromatography confirmed the cutting time of vitamin D, VD was captured by C18 column, and then established on-line two-dimensional liquid chromatography analysis method. Samples and standard solutions were used to conduct the validation of the established method, and the test results of actual samples were compared by using established method and GB 5009.82-2016. Results The linear coefficients of vitamin A, D and E were all more than 0.999 in the range of standard curves. The limits of quantitation were 30 μg/100 g, 2 μg/100 g and 120 μg/100 g, which were consistent with the national standard GB 5009.82-2016. The recoveries of vitamin A, D and E at 3 levels of this method were all between 90.4% and 101.7%, which could meet the detection requirements of vitamin A, D and E in food. Conclusion The method is simple, efficient, highly automated, accurate and reproducible, which is suitable for the determination of large quantities of samples.