Food Colour & Quality Laboratory. Department of Nutrition & Food Science. Universidad de Sevilla. Facultad de Farmacia, 41012, Sevilla, Spain.
Dept. Analytical Chemistry, Universidad de Sevilla. Facultad de Farmacia, 41012, Sevilla, Spain.
J Chromatogr A. 2019 Jan 4;1583:63-72. doi: 10.1016/j.chroma.2018.11.010. Epub 2018 Nov 3.
An analytical method was validated for the quantitative determination of isoprenoids compounds in faecal samples, based on liquid-liquid extraction from a small aliquot (0.3-0.5 g of sample) and subsequent analysis by Rapid Resolution Liquid Chromatography (RRLC) on a C30 column. An excellent linear response was observed over the range specified for all dietary isoprenoids, as confirmed by the correlation coefficient, which ranged from 0.9977 to 0.9999. LODs ranged from 0.002 μg to 0.036 μg for lutein and α-tocopherol, respectively. Depending on the compound, LOQs ranged from 0.001 μg (lutein) to 0.120 μg (α-tocopherol). For accuracy testing, spiking of faeces samples with trans-β-apo-8'-carotenal, α-tocopherol and chlorophyll a were performed (three concentration levels). Excellent recoveries were obtained in all levels (>90%). The intra-day RSD% ranged from 0.86 to 9.78%. The inter-day RSD% was not higher than 10%, except to α-tocopherol (11.34%). In order to assess the applicability of the method faecal samples from a baby fed with different purees formulated from various vegetables were analysed during a six month period. α-carotene, β-carotene, capsanthin, lycopene, lutein, phytoene, phytofluene, violaxanthin, zeaxanthin and ζ-carotene), and their isomers were identified and quantified using this method. Besides, 2 tocopherols and 9 chlorophylls and derivatives were identified and quantified in the faecal samples analysed. This method is suitable to determine dietary isoprenoids from complex matrices such as human faeces within 28 min.
建立了一种基于小体积（0.3-0.5g 粪便）液-液萃取，用 C30 柱快速分辨液相色谱法（RRLC）分析粪便中类异戊二烯化合物的定量方法。所有膳食类异戊二烯的线性范围都有很好的响应，相关系数在 0.9977-0.9999 之间。叶黄素和α-生育酚的检测限分别为 0.002μg 和 0.036μg。根据化合物的不同，LOQ 范围为 0.001μg（叶黄素）至 0.120μg（α-生育酚）。为了进行准确度测试，对粪便样品进行了 trans-β-apo-8'-胡萝卜素、α-生育酚和叶绿素 a 的添加实验（三个浓度水平）。所有水平的回收率均大于 90%。日内 RSD% 范围为 0.86-9.78%。除了 α-生育酚（11.34%）之外，日间 RSD% 均不高于 10%。为了评估该方法的适用性，对 6 个月期间用不同蔬菜制成的不同泥状食物喂养的婴儿的粪便样本进行了分析。使用该方法鉴定和定量了α-胡萝卜素、β-胡萝卜素、辣椒红素、番茄红素、叶黄素、phytoene、phytofluene、violaxanthin、玉米黄质和 ζ-胡萝卜素及其异构体。此外，还鉴定和定量了分析的粪便样品中的 2 种生育酚和 9 种叶绿素及其衍生物。该方法适用于在 28min 内从复杂基质（如人粪便）中测定膳食类异戊二烯。
