Biomolecular Mass Spectrometry Unit, Department of Parasitology, Leiden University Medical Center, NL-2300 RC Leiden, The Netherlands.
J Proteome Res. 2010 Sep 3;9(9):4869-74. doi: 10.1021/pr100634d.
Metabolic profiling of biological samples is increasingly used to obtain more insight into the pathophysiology of diseases. For translational studies, biological samples from animal models are explored; however, the volume of these samples can be a limiting factor for metabolic profiling studies. For instance, only a few microliters of urine is often available from small animals like mice. Hence, there is a need for a tailor-made analytical method for metabolic profiling of volume-limited samples. In the present study, the feasibility of capillary electrophoresis time-of-flight mass spectrometry (CE-ToF-MS) for metabolic profiling of urine from mice is evaluated. Special attention is paid to the analytical workflow; that is, such aspects as sample preparation, analysis, and data treatment are discussed from the metabolomics viewpoint. We show that metabolites belonging to several chemical families can be analyzed in mouse urine with the CE-ToF-MS method using minimal sample pretreatment and an in-capillary preconcentration procedure. This exemplifies the advantages of CE-ToF-MS for metabolic profiling of volume-limited samples as loss of material is minimized. The feasibility of the CE-ToF-MS-based workflow for metabolic profiling is illustrated by the analysis of urine samples from wild-type as well as from TTD mutant mice, which are a model for the accelerated aging, with osteoporosis being one of the main hallmarks.
生物样本的代谢组学分析越来越多地用于深入了解疾病的病理生理学。对于转化研究，探索了来自动物模型的生物样本；然而，这些样本的体积可能是代谢组学研究的一个限制因素。例如，从小鼠等小动物中通常只能获得几微升的尿液。因此，需要针对体积有限的样本进行代谢组学分析的定制化分析方法。在本研究中，评估了毛细管电泳飞行时间质谱（CE-ToF-MS）用于分析小鼠尿液代谢组学的可行性。特别关注分析工作流程；也就是说，从代谢组学的角度讨论了样品制备、分析和数据处理等方面。我们表明，使用 CE-ToF-MS 方法，通过最小的样品预处理和毛细管内浓缩程序，可分析尿液中属于几个化学家族的代谢物。这说明了 CE-ToF-MS 用于体积有限的样本代谢组学分析的优势，因为最大限度地减少了物质的损失。通过分析野生型和 TTD 突变型小鼠的尿液样本，证明了基于 CE-ToF-MS 的工作流程进行代谢组学分析的可行性，TTD 突变型小鼠是加速衰老的模型，骨质疏松症是其主要特征之一。
