Institute of Nutrition and Functional Foods (INAF), Faculty of Agriculture and Food Sciences, Laval University, 2440 Boulevard Hochelaga, Québec, Québec G1V 0A6, Canada.
Nutrition, Health and Society Centre (NUTRISS), INAF, Laval University, 2440 boulevard Hochelaga, Québec, Québec G1V 0A6, Canada.
J Agric Food Chem. 2023 Nov 8;71(44):16787-16796. doi: 10.1021/acs.jafc.3c03888. Epub 2023 Oct 27.
Quantification of nutritional biomarkers is crucial to accurately assess the dietary intake of different classes of (poly)phenols in large epidemiological studies. High-throughput analysis is mandatory to apply this methodology in large cohorts. However, the current validated methods to quantify (poly)phenols metabolites in biological fluids use ultra performance liquid chromatography (UPLC), leading to analysis time of several minutes per sample. To significantly reduce the run time, we developed and validated a method to quantify in urine the flavan-3-ols biomarkers, phenyl-γ-valerolactones (PVLs), using laser diode thermal desorption (LDTD). This mass spectrometry source allows direct introduction of sample extracts, resulting in analysis time of less than 10 s per sample. Also, to encompass the problem associated with the cost and availability of sulfated and glucuronide analytical standards, urine samples were subjected to enzymatic hydrolysis. Creatinine was also quantified to normalize the results obtained from the urinary spot. Results obtained with LDTD-MS/MS were cross-validated by UPLC-MS/MS using 155 urine samples. Coefficient of correlation was above 0.975 for PVLs and creatinine. For all analytes, the accuracy was between 90% and 113% by LDTD-MS/MS. Altogether, sample preparation was fully automated to demonstrate the application potential of this method to large cohorts.
定量分析营养生物标志物对于在大型流行病学研究中准确评估不同类别的（多）酚类物质的膳食摄入量至关重要。高通量分析对于在大样本中应用这种方法是强制性的。然而，目前用于定量生物体液中（多）酚代谢物的验证方法使用超高效液相色谱（UPLC），导致每个样本的分析时间为几分钟。为了显著缩短运行时间，我们开发并验证了一种使用激光二极管热解吸（LDTD）定量尿液中黄烷-3-醇生物标志物苯丙氨酸-γ-缬草酸（PVL）的方法。这种质谱源允许直接引入样品提取物，从而使每个样本的分析时间少于 10 秒。此外，为了解决与硫酸化和葡萄糖醛酸分析标准品的成本和可用性相关的问题，尿液样本进行了酶水解。还定量了肌酐以标准化从尿液斑点中获得的结果。使用 155 个尿液样本通过 UPLC-MS/MS 对 LDTD-MS/MS 获得的结果进行了交叉验证。对于 PVL 和肌酐，相关系数均高于 0.975。通过 LDTD-MS/MS，所有分析物的准确度在 90%到 113%之间。总之，样品制备完全自动化，展示了该方法在大样本中的应用潜力。
