Grün Christian H, van Dorsten Ferdi A, Jacobs Doris M, Le Belleguic Marie, van Velzen Ewoud J J, Bingham Max O, Janssen Hans-Gerd, van Duynhoven John P M
Unilever Food and Health Research Institute, Olivier van Noortlaan 120, 3133 AT, Vlaardingen, The Netherlands.
J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Aug 15;871(2):212-9. doi: 10.1016/j.jchromb.2008.04.039. Epub 2008 May 3.
Flavonoids, a subclass of polyphenols, are major constituents of many plant-based foods and beverages, including tea, wine and chocolate. Epidemiological studies have shown that a flavonoid-rich diet is associated with reduced risk of cardiovascular diseases. The majority of the flavonoids survive intact until they reach the colon where they are then extensively metabolized into smaller fragments. Here, we describe the development of GC-MS-based methods for the profiling of phenolic microbial fermentation products in urine, plasma, and fecal water. Furthermore, the methods are applicable for profiling products obtained from in vitro batch culture fermentation models. The methods incorporate enzymatic deconjugation, liquid-liquid extraction, derivatization, and subsequent analysis by GC-MS. At the level of individual compounds, the methods gave recoveries better than 80% with inter-day precision being better than 20%, depending on the matrix. Limits of detection were below 0.1 microg/ml for most phenolic acids. The newly developed methods were successfully applied to samples from human and in-vitro intervention trials, studying the metabolic impact of flavonoid intake. In conclusion, the methods presented are robust and generally applicable to diverse biological fluids. Its profiling character is useful to investigate on a large scale the gut microbiome-mediated bioavailability of flavonoids.
黄酮类化合物是多酚类的一个亚类，是许多植物性食品和饮料（包括茶、葡萄酒和巧克力）的主要成分。流行病学研究表明，富含黄酮类化合物的饮食与心血管疾病风险降低有关。大多数黄酮类化合物在到达结肠之前保持完整，然后在结肠中被广泛代谢成较小的片段。在此，我们描述了基于气相色谱-质谱联用（GC-MS）方法的尿液、血浆和粪便水中酚类微生物发酵产物的分析方法。此外，这些方法适用于分析从体外分批培养发酵模型获得的产物。这些方法包括酶解结合、液液萃取、衍生化以及随后的GC-MS分析。在单个化合物水平上，根据基质不同，这些方法的回收率高于80%，日间精密度优于20%。大多数酚酸的检测限低于0.1微克/毫升。新开发的方法已成功应用于来自人体和体外干预试验的样本，以研究黄酮类化合物摄入的代谢影响。总之，所提出的方法稳健且普遍适用于多种生物流体。其分析特性有助于大规模研究肠道微生物群介导的黄酮类化合物生物利用度。
