The Targeted Metabolomics and Proteomics Laboratory, University of Alabama at Birmingham, Birmingham, AL, USA.
AB Sciex, Concord, Ontario, Canada.
Arch Biochem Biophys. 2014 Oct 1;559:3-11. doi: 10.1016/j.abb.2014.06.014. Epub 2014 Jun 23.
This study was conducted to assess the value of a high resolution, high mass accuracy time-of-flight analyzer in combination with nanoliquid chromatography for the analysis of polyphenols and their metabolites. The goal was to create a method that utilizes small volumes of biological fluids and provides a significant improvement in sensitivity compared with existing methods. Accordingly, nanoLC-MS and nanoLC-pseudo-multiple reaction monitoring (MRM) methods were developed that had a lower limit of quantification of 0.5 nM for several polyphenols and were linear over 2-3 orders of magnitude (R(2)>0.999). Using urine samples, the ability to observe and quantify polyphenols in such a complex biological fluid depended on much narrower mass windows (0.050 amu or less) on a TOF analyzer than those used on a quadrupole analyzer (0.7 amu). Although a greater selectivity was possible with the low mass resolution of a triple quadrupole instrument using the MRM approach, for the daidzein metabolite O-DMA, a chromatographically resolvable second peak could only be substantially reduced by using a 0.01 amu mass window. The advantage of a TOF analyzer for product ion data is that the whole MSMS spectrum is collected at high mass accuracy and MRM experiments are conducted in silico after the analysis.
本研究旨在评估高分辨率、高质量精度飞行时间分析器与纳流液相色谱联用在多酚及其代谢物分析中的价值。目标是创建一种利用小体积生物流体的方法，并与现有方法相比显著提高灵敏度。因此，开发了纳流液相色谱-质谱和纳流伪多重反应监测（MRM）方法，几种多酚的定量下限为 0.5nM，线性范围为 2-3 个数量级（R²>0.999）。使用尿液样本，在如此复杂的生物流体中观察和定量多酚的能力取决于飞行时间分析器上的质量窗口比四极杆分析器上的窄得多（0.050amu 或更小）。尽管使用三重四极杆仪器的 MRM 方法可以实现更高的选择性，但对于大豆苷元代谢物 O-DMA，只有使用 0.01amu 的质量窗口才能大大减少可分辨的第二个色谱峰。飞行时间分析器在产物离子数据方面的优势在于，整个 MSMS 谱在高质量精度下进行采集，并且在分析后进行虚拟的 MRM 实验。
