Validation of a dual LC-HRMS platform for clinical metabolic diagnosis in serum, bridging quantitative analysis and untargeted metabolomics.
Validation of a dual LC-HRMS platform for clinical metabolic diagnosis in serum, bridging quantitative analysis and untargeted metabolomics.
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DOI：
10.1007/s11306-013-0582-1
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发表时间：
2014-04-04
期刊：
影响因子：
3.6
通讯作者：
Barshop, Bruce A.
中科院分区：
文献类型：
--
作者：
Gertsman, Ilya;Gangoiti, Jon A.;Barshop, Bruce A.
关键词：
基于质谱的代谢组学是研究和诊断领域快速发展的领域。一般来说，用于临床和其他绝对定量实验的方法和仪器类型不同于用于生物标志物发现和非靶向分析的仪器，因为前者需要最佳的灵敏度和动态范围，而后者需要高分辨率和高质量准确度。我们使用具有两种不同类型五氟苯基 (PFP) 固定相的 Q-TOF 质谱仪，采用正电离和负电离，以开发和验证使用 LC-HRMS 的混合定量和发现平台。该双 PFP LC-MS 平台可对血清中 50 多种临床相关代谢物进行定量（使用 MS 和 MS/MS 采集），同时收集高分辨率和高质量准确度全扫描，以监测所有其他共洗脱非靶向分析物。我们证明，许多代谢物（包括氨基酸、有机酸、酰基肉碱和嘌呤/嘧啶）定量的线性、准确性和精密度结果在各自的生理范围内达到或超过正常的生物分析标准。该色谱法可在反相条件下解析高极性和疏水性分析物，从而能够分析多种化学物质，这是非靶向代谢组学实验所必需的。尽管之前的 LC-HRMS 方法已经证明了对各种药物和小分子化合物的定量能力，但本研究提供了针对代谢疾病量身定制的 HRMS 定量/定性平台；并涵盖多种不同的代谢物，包括通常通过单独仪器组合进行定量的化合物。
Mass spectrometry-based metabolomics is a rapidly growing field in both research and diagnosis. Generally, the methodologies and types of instruments used for clinical and other absolute quantification experiments are different from those used for biomarkers discovery and untargeted analysis, as the former requires optimal sensitivity and dynamic range, while the latter requires high resolution and high mass accuracy. We used a Q-TOF mass spectrometer with two different types of pentafluorophenyl (PFP) stationary phases, employing both positive and negative ionization, to develop and validate a hybrid quantification and discovery platform using LC-HRMS. This dual-PFP LC-MS platform quantifies over 50 clinically relevant metabolites in serum (using both MS and MS/MS acquisitions) while simultaneously collecting high resolution and high mass accuracy full scans to monitor all other co-eluting non-targeted analytes. We demonstrate that the linearity, accuracy, and precision results for the quantification of a number of metabolites, including amino acids, organic acids, acylcarnitines and purines/pyrimidines, meets or exceeds normal bioanalytical standards over their respective physiological ranges. The chromatography resolved highly polar as well as hydrophobic analytes under reverse-phase conditions, enabling analysis of a wide range of chemicals, necessary for untargeted metabolomics experiments. Though previous LC-HRMS methods have demonstrated quantification capabilities for various drug and small molecule compounds, the present study provides an HRMS quant/qual platform tailored to metabolic disease; and covers a multitude of different metabolites including compounds normally quantified by a combination of separate instrumentation.
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影响因子：
4.2
作者：
Dejong, Cornelis H. C.;van de Poll, Marcel C. G.;Damink, Steven W. M. Olde
通讯作者：
Damink, Steven W. M. Olde
影响因子：
--
作者：
Cooper, Helen M;Paterson, Yvonne
通讯作者：
Paterson, Yvonne
影响因子：
2
作者：
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通讯作者：
Rochat, Bertrand
DOI：
10.1007/978-1-61737-985-7_3
发表时间：
2011-01-01
期刊：
METABOLIC PROFILING: METHODS AND PROTOCOLS
影响因子：
--
作者：
Millington, David S.;Stevens, Robert D.
通讯作者：
Stevens, Robert D.
影响因子：
3.8
作者：
Forni, Sabrina;Fu, Xiaowei;Sweetman, Lawrence
通讯作者：
Sweetman, Lawrence