Carreer William J, Flight Robert M, Moseley Hunter N B
Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY 40536, USA;
Metabolites. 2013 Sep 25;3(4):853-66. doi: 10.3390/metabo3040853.
New metabolomics applications of ultra-high resolution and accuracy mass spectrometry can provide thousands of detectable isotopologues, with the number of potentially detectable isotopologues increasing exponentially with the number of stable isotopes used in newer isotope tracing methods like stable isotope-resolved metabolomics (SIRM) experiments. This huge increase in usable data requires software capable of correcting the large number of isotopologue peaks resulting from SIRM experiments in a timely manner. We describe the design of a new algorithm and software system capable of handling these high volumes of data, while including quality control methods for maintaining data quality. We validate this new algorithm against a previous single isotope correction algorithm in a two-step cross-validation. Next, we demonstrate the algorithm and correct for the effects of natural abundance for both (13)C and (15)N isotopes on a set of raw isotopologue intensities of UDP-N-acetyl-D-glucosamine derived from a (13)C/(15)N-tracing experiment. Finally, we demonstrate the algorithm on a full omics-level dataset.
超高分辨率和高精度质谱的新代谢组学应用能够提供数千种可检测的同位素异构体，随着在诸如稳定同位素分辨代谢组学（SIRM）实验等更新的同位素示踪方法中使用的稳定同位素数量增加，潜在可检测的同位素异构体数量呈指数增长。可用数据的这种大幅增加需要能够及时校正SIRM实验产生的大量同位素异构体峰的软件。我们描述了一种能够处理这些大量数据的新算法和软件系统的设计，同时包括用于维持数据质量的质量控制方法。我们在两步交叉验证中针对先前的单同位素校正算法验证了这种新算法。接下来，我们展示该算法并校正来自（13）C/（15）N示踪实验的一组UDP-N-乙酰-D-葡萄糖胺原始同位素异构体强度上（13）C和（15）N同位素的自然丰度影响。最后，我们在完整的组学水平数据集上展示该算法。
