Department of Chemistry, Purdue University, West Lafayette, Indiana 47907, USA.
Anal Chem. 2010 Mar 15;82(6):2303-9. doi: 10.1021/ac9024818.
An increased interest in metabolite profiling is driving the need for improved analytical techniques with greater performance for a variety of important applications. Despite their limited sensitivity, nuclear magnetic resonance (NMR) methods are attractive because of their simplicity, reproducibility, quantitative nature, and wide applicability. The use of chemoselective isotopic tags has the potential to advance the application of NMR for analyzing metabolites in complex biofluids by allowing detection of metabolites down to the low micromoalr level with high resolution and specificity. Here, we report a new (13)C-tagging method using (13)C-formic acid that delivers high sensitivity, good quantitation, and excellent resolution for (1)H-(13)C 2D NMR profiling of amino metabolites. High reproducibility (coefficient of variation (CV) = 2%) was observed for metabolites in urine with concentrations down to 10 microM. As amino compounds comprise an important class of metabolites and small molecules of biological roles, this new method therefore should be amenable to a variety of applications.
人们对代谢物分析的兴趣日益浓厚，这促使人们需要开发性能更好的分析技术，以满足各种重要应用的需求。尽管核磁共振（NMR）方法的灵敏度有限，但由于其简单、可重现、定量和广泛适用的特点，仍然具有吸引力。利用化学选择性同位素标记物有可能通过允许检测低至低毫摩尔水平的代谢物，实现高分辨率和特异性，从而推进 NMR 在分析复杂生物流体中的代谢物的应用。在这里，我们报告了一种使用(13)C-甲酸的新的(13)C 标记方法，该方法可用于(1)H-(13)C 2D NMR 对氨基酸代谢物进行高灵敏度、定量和高分辨率的分析。在浓度低至 10 微摩尔的尿液中，代谢物的重现性（变异系数 (CV) = 2%）非常好。由于氨基酸化合物是一类重要的代谢物和具有生物学作用的小分子，因此这种新方法应该适用于多种应用。
