Optimization and application of high-throughput supported liquid extraction for simultaneous determination of carotenoids and fat-soluble vitamins in serum
Optimization and application of high-throughput supported liquid extraction for simultaneous determination of carotenoids and fat-soluble vitamins in serum
复制标题
高通量支持液体萃取同时测定血清中类胡萝卜素和脂溶性维生素的优化及应用
DOI：
10.1016/j.jchromb.2021.122672
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发表时间：
2021-05-11
影响因子：
3
通讯作者：
Yu, Yan
中科院分区：
文献类型：
--
作者：
Qi, Fei-Fei;Tao, Li-Mei;Yu, Yan
类胡萝卜素（CAR）和脂溶性维生素（FSV）的分析需求不断扩大，但目前使用的样品制备方法要么提取步骤复杂，要么样本量大，更不用说结果的可靠性。本研究旨在建立一种基于支持液萃取（SLE）的快速、高效、高通量的方法，用于高效液相色谱-二极管阵列检测器（HPLCDAD）分析前同时提取人血清中的FSV和CAR。采用基于Box-Behnken设计（BBD）的响应面法（RSM）优化SLE参数，包括血清-水-萃取溶剂比和洗脱液体积。在最佳条件下，该方法可接受检出限（LOD）为0.005 ~ 0.3 μ g/mL，回收率为89.6 ~ 110.9%，相对标准偏差（RSD）小于10.1%，所需血清样品较少（100 μ L），制样时间较少（2 min /份）。与液相萃取（LLE）相比，SLE萃取速度快，回收率高，重现性好，用于CAR和FSV分析的基质效应较低。该方法已成功应用于健康个体和年龄相关性黄斑变性（AMD）患者血清中CAR和FSV水平的量化，证明了该方法在流行病学和常规应用方面的可行性。
The demand for analysis of carotenoids (CAR) and fat-soluble vitamins (FSV) is continuously expanding, but currently used sample preparation methods either require complicated extraction procedure or large sample volume, let alone the reliability of the results. This study aimed to develop a fast, high-efficient, and highthroughput method based on supported liquid extraction (SLE) for the simultaneous extraction of FSV and CAR from human serum before using high-performance liquid chromatography-diode array detector (HPLCDAD) analysis. The optimization of SLE parameters was achieved through response surface methodology (RSM) based on the Box-Behnken design (BBD) and included serum-water-extraction solvent ratio and eluent volume. Under optimal conditions, the proposed method gives acceptable limits of detection (LOD) (0.005-0.3 mu g/mL), good recovery (89.6-110.9%) as well as relative standard deviation (RSD) of less than 10.1% by consuming lower serum sample (100 mu L) and less sample preparation time (2 min per sample). Compared with liquid-phase extraction (LLE), the SLE delivers rapid extraction with higher recovery, better reproducibility, and lower matrix effect for CAR and FSV analysis. The method has been successfully applied to quantify CAR and FSV levels in serum of healthy individuals and age-related macular degeneration (AMD) patients, demonstrating the feasibility of the proposed method for epidemiology and routine applications.