Development of a plasma pseudotargeted metabolomics method based on ultra-high-performance liquid chromatography-mass spectrometry
| Autor: | Fujian Zheng, Xinjie Zhao, Zhongda Zeng, Qingqing Wang, Wangjie Lv, Guowang Xu, Lichao Wang |
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| Rok vydání: | 2019 |
| Předmět: |
0303 health sciences
Time Factors Computer science Selected reaction monitoring Repeatability High coverage Mass spectrometry General Biochemistry Genetics and Molecular Biology Mass Spectrometry 03 medical and health sciences Plasma 0302 clinical medicine Metabolomics Liquid chromatography–mass spectrometry Ultra high performance Biological system 030217 neurology & neurosurgery Chromatography High Pressure Liquid 030304 developmental biology Targeted metabolomics |
| Zdroj: | Nature protocols. 15(8) |
| ISSN: | 1750-2799 |
| Popis: | Untargeted methods are typically used in the detection and discovery of small organic compounds in metabolomics research, and ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) is one of the most commonly used platforms for untargeted metabolomics. Although they are non-biased and have high coverage, untargeted approaches suffer from unsatisfying repeatability and a requirement for complex data processing. Targeted metabolomics based on triple-quadrupole mass spectrometry (TQMS) could be a complementary tool because of its high sensitivity, high specificity and excellent quantification ability. However, it is usually applicable to known compounds: compounds whose identities are known and/or are expected to be present in the analyzed samples. Pseudotargeted metabolomics merges the advantages of untargeted and targeted metabolomics and can act as an alternative to the untargeted method. Here, we describe a detailed protocol of pseudotargeted metabolomics using UHPLC-TQMS. An in-depth, untargeted metabolomics experiment involving multiple UHPLC-HRMS runs with MS at different collision energies (both positive and negative) is performed using a mixture obtained using small amounts of the analyzed samples. XCMS, CAMERA and Multiple Reaction Monitoring (MRM)-Ion Pair Finder are used to find and annotate peaks and choose transitions that will be used to analyze the real samples. A set of internal standards is used to correct for variations in retention time. High coverage and high-performance quantitative analysis can be realized. The entire protocol takes ~5 d to complete and enables the simultaneously semiquantitative analysis of 800-1,300 metabolites. |
| Databáze: | OpenAIRE |
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