Department of Food Chemistry and Toxicology, Faculty of Chemistry, University of Vienna, 1090, Vienna, Austria; Doctoral School of Chemistry, University of Vienna, 1090, Vienna, Austria; Universität Wien, Fakultät für Chemie, Institut für Biophysikalische Chemie, 1090, Wien, Österreich.
Department of Food Chemistry and Toxicology, Faculty of Chemistry, University of Vienna, 1090, Vienna, Austria.
Anal Chim Acta. 2022 Jul 11;1216:339977. doi: 10.1016/j.aca.2022.339977. Epub 2022 May 23.
Establishing a method for human biomonitoring (HBM) of polyphenols enables the assessment of internal concentrations of these food bio-actives and the correlation with potential health effects such as antioxidant or anti-inflammatory properties. Thus, a targeted LC-MS/MS method for quantifying up to 90 analytes, representing the main polyphenol classes including flavanones, isoflavones, stilbenes, and phenolic acids, was developed for human urine, serum, and plasma. The method was established for low sample volumes and with a cost and time efficient sample preparation protocol for high-throughput, which is critical for its application in large cohort and exposome-wide association studies. On average, the sample preparation yielded extraction efficiencies of 98% for urine, 98% for serum, and 87% for plasma. Limits of detection were between 0.11 ng mL and 300 ng mL for urine, 0.12 ng mL and 190 ng mL for serum, and 0.12 ng mL and 340 ng mL for plasma, excluding one analyte. In-house validation revealed that 66, 49, and 64 analytes for urine, serum, and plasma, respectively, fulfilled all stringent requirements, that are usually utilized for tailored single analyte methods, at all evaluated concentration levels. After validation, this method was applied in a proof-of-principle study that detected 39 polyphenols in urine. Changes in the concentrations of the analytes after the ingestion of a high polyphenol smoothie was examined over 24 h. The study further confirmed that the majority of polyphenols detected were phenolic acids, and phase II conjugated metabolites were more abundant than their respective non-conjugated forms.
建立一种用于人体生物监测（HBM）的多酚方法，可以评估这些食物生物活性物质的内部浓度，并与潜在的健康影响（如抗氧化或抗炎特性）相关联。因此，开发了一种针对多酚类物质的靶向 LC-MS/MS 方法，可定量分析多达 90 种分析物，代表了主要的多酚类物质类别，包括黄烷酮类、异黄酮类、芪类和酚酸类，用于人尿液、血清和血浆。该方法适用于小体积样本，并且具有成本效益高、耗时短的样本制备方案，可实现高通量，这对于其在大型队列和暴露组全基因组关联研究中的应用至关重要。平均而言，尿液的样品制备提取效率为 98%，血清为 98%，血浆为 87%。尿液的检测限为 0.11ng/mL 至 300ng/mL，血清为 0.12ng/mL 至 190ng/mL，血浆为 0.12ng/mL 至 340ng/mL，排除一种分析物。内部验证显示，尿液、血清和血浆中分别有 66、49 和 64 种分析物满足所有严格要求，通常用于定制的单分析物方法，在所有评估的浓度水平下。验证后，该方法应用于一项原理验证研究，检测了尿液中的 39 种多酚。研究检测了 24 小时内高多酚冰沙摄入后分析物浓度的变化。该研究进一步证实，检测到的多酚大多数为酚酸，且Ⅱ相共轭代谢物比其相应的非共轭形式更为丰富。
