Metabolomics & Biomarkers Group, Nestlé Research Center, Nestec Ltd., Vers-chez-les-Blanc, 1000 Lausanne, Switzerland.
J Chromatogr A. 2011 Jul 22;1218(29):4678-88. doi: 10.1016/j.chroma.2011.05.050. Epub 2011 May 26.
This study reports a liquid chromatography-mass spectrometry method for the detection of polyphenol-derived metabolites in human plasma without enzymatic treatment after coffee consumption. Separation of available standards was achieved by reversed-phase ultra performance liquid chromatography and detection was performed by high resolution mass spectrometry in negative electrospray ionization mode. This analytical method was then applied for the identification and relative quantification of circulating coffee metabolites. A total of 34 coffee metabolites (mainly reduced, sulfated and methylated forms of caffeic acid, coumaric acid, caffeoylquinic acid and caffeoylquinic acid lactone) were identified based on mass accuracy (<4 ppm for most metabolites), specific fragmentation pattern and co-chromatography (when standard available). Among them, 19 circulating coffee metabolites were identified for the first time in human plasma such as feruloylquinic acid lactone, sulfated and glucuronidated forms of feruloylquinic acid lactone and sulfated forms of coumaric acid. Phenolic acid derivatives such as dihydroferulic acid, dihydroferulic acid 4'-O-sulfate, caffeic acid 3'-O-sulfate, dimethoxycinnamic acid, dihydrocaffeic acid and coumaric acid O-sulfate appeared to be the main metabolites circulating in human plasma after coffee consumption. The described method is a sensitive and reliable approach for the identification of coffee metabolites in biological fluids. In future, this analytical method will give more confidence in compound identification to provide a more comprehensive assessment of coffee polyphenol bioavailability studies in humans.
本研究报告了一种无需在咖啡摄入后进行酶处理即可检测人血浆中多酚衍生代谢物的液相色谱-质谱法。通过反相超高效液相色谱实现了可用标准的分离，通过负离子电喷雾模式的高分辨质谱进行检测。然后，该分析方法用于鉴定和相对定量循环咖啡代谢物。基于质量精度（大多数代谢物 <4 ppm）、特定的碎片模式和共色谱（当有标准品时），共鉴定出 34 种咖啡代谢物（主要为咖啡酸、香豆酸、咖啡酰奎宁酸和咖啡酰奎宁酸内酯的还原、硫酸化和甲基化形式）。其中，19 种循环咖啡代谢物首次在人血浆中被鉴定出来，如阿魏酰奎宁酸内酯、阿魏酰奎宁酸内酯的硫酸化和葡萄糖醛酸化形式以及香豆酸的硫酸化形式。在咖啡摄入后，似乎主要的代谢物是对香豆酸、二氢阿魏酸、二氢阿魏酸 4'-O-硫酸盐、咖啡酸 3'-O-硫酸盐、二甲氧基肉桂酸、二氢咖啡酸和香豆酸 O-硫酸盐等酚酸衍生物。所描述的方法是一种灵敏可靠的方法，可用于鉴定生物液中的咖啡代谢物。在未来，这种分析方法将为化合物鉴定提供更多信心，从而更全面地评估人类咖啡多酚生物利用度研究。
