Eriksen Jane N, Madsen Pia L, Dragsted Lars O, Arrigoni Eva
Agroscope, Competence Division Plants and Plant Products , Schloss 1, CH-8820 Wädenswil, Switzerland.
Department of Nutrition, Exercise and Sports, University of Copenhagen , Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark.
J Agric Food Chem. 2017 Feb 1;65(4):973-980. doi: 10.1021/acs.jafc.6b04925. Epub 2017 Jan 17.
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.
开发并验证了一种基于超高效液相色谱-二极管阵列检测（UHPLC-DAD）的改进方法，用于定量菠菜、血清、乳糜微粒和粪便中主要类胡萝卜素的含量。六种膳食类胡萝卜素和内标虾青素在12.5分钟内通过梯度洗脱实现分离。该方法为所有标准成分提供了大于1.1的分离度、覆盖目标范围的线性关系（R>0.99）、低于0.035 mg/L的定量限，以及日内和日间相对标准偏差分别小于2%和10%。在生物基质上测试了该方法的适用性。除番茄红素外，血清、乳糜微粒和粪便中类胡萝卜素定量的方法精密度（相对标准偏差%）在日内和日间分析中均低于10%。方法准确度一致，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%。此外，在两个机构对菠菜进行的实验室间验证研究表明，在四次评估中，叶黄素或β-胡萝卜素含量没有显著差异。
