Optimized, Fast-Throughput UHPLC-DAD Based Method for Carotenoid Quantification in Spinach, Serum, Chylomicrons, and Feces.
Optimized, Fast-Throughput UHPLC-DAD Based Method for Carotenoid Quantification in Spinach, Serum, Chylomicrons, and Feces.
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DOI：
10.1021/acs.jafc.6b04925
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发表时间：
2017-01
影响因子：
6.1
通讯作者：
J. Eriksen;P. Madsen;L. Dragsted;E. Arrigoni
中科院分区：
文献类型：
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作者：
J. Eriksen;P. Madsen;L. Dragsted;E. Arrigoni
开发并验证了一种改进的基于 UHPLC-DAD 的方法，用于定量菠菜、血清、乳糜微粒和粪便中存在的主要类胡萝卜素。六种膳食类胡萝卜素和内标海胆酮通过梯度洗脱在 12.5 分钟内实现分离。对于所有标准组分，所提出的方法提供的分辨率 > 1.1、覆盖目标范围的线性 (R > 0.99)、LOQ < 0.035 mg/L，日内和日间 RSD 分别 < 2 和 10%。该方法的适用性在生物基质上进行了测试。对于日内和日间分析，血清、乳糜微粒和粪便中类胡萝卜素定量的方法精度 (RSD%) 低于 10%（番茄红素除外）。方法准确度与血清和粪便中除番茄红素外的所有类胡萝卜素的平均回收率分别为 78.8% 至 96.9% 和 57.2% 至 96.9% 一致。此外，两家机构对菠菜进行的实验室间验证研究显示，在四次评估后，叶黄素或 β-胡萝卜素含量没有显着差异。
An improved UHPLC-DAD-based method was developed and validated for quantification of major carotenoids present in spinach, serum, chylomicrons, and feces. Separation was achieved with gradient elution within 12.5 min for six dietary carotenoids and the internal standard, echinenone. The proposed method provides, for all standard components, resolution > 1.1, linearity covering the target range (R > 0.99), LOQ < 0.035 mg/L, and intraday and interday RSDs < 2 and 10%, respectively. Suitability of the method was tested on biological matrices. Method precision (RSD%) for carotenoid quantification in serum, chylomicrons, and feces was below 10% for intra- and interday analysis, except for lycopene. Method accuracy was consistent with mean recoveries ranging from 78.8 to 96.9% and from 57.2 to 96.9% for all carotenoids, except for lycopene, in serum and feces, respectively. Additionally, an interlaboratory validation study on spinach at two institutions showed no significant differences in lutein or β-carotene content, when evaluated on four occasions.