Untargeted metabolomics analysis

Introduction:

Untargeted metabolomics analysis is the study of small molecule metabolites in cell line, tissue, or biological fluids. With the advances in mass spectrometry technology, it is possible to identify and quantify a large number of metabolites. The ultimate goal of metabolomics is to analyze the complete set of metabolites – the metabolome, in a living system. Metabolites in a living system is typically complex, thus, strong extraction solvent (80-100% methanol) is applied to extract as many kinds of metabolites as possible and liquid chromatography separation is used to reduce complexity of the sample before mass spectrometric analysis. High-resolution mass spectrometer is used to acquire relative intensities of the measured metabolites. Dedicated data processing software, such as XCMS and Progenesis QI, are used to analyze the data. Untargeted metabolomics analysis can compare the relative abundances of metabolites in organisms under different physiological states and its applications cover diverse research areas like drug discovery and the study of human diseases.

Sample submission:

To ensure accuracy of experimental results, the samples submitted should meet the following requirements: at least 100~200 μL for body fluids (serum, plasma, cerebrospinal fluid, lymph, urine, saliva, etc.) , at least 5×106~1×107 for cells, 30~200 mg for animal tissue; 100 mg~1 g for plant tissue, and 1×107~1×108 for microbe.


Please provide detailed information about related literature on sample preparation and data analysis, so that BMSC can establish analysis methods accordingly.

Sample processing:

The samples such as blood, tissue, cell, and body fluid are extracted with an organic extraction solution (80%-100% methanol or acetonitrile). The metabolites are extracted by homogenization, ultrasonication, or shaking. After centrifuging, the supernatant can be directly analyzed (after dilution when needed) by mass spectrometry. The supernatant can also be concentrated or purified by solid phase extraction (SPE) before mass spectrometry analysis.

Result output:

The data are analyzed using multidimensional software, Compound Discoverer, TraceFinder, XCMS, Progenesis QI, and Xcalibur. Statistical analysis is used to assess the difference in metabolite abundances between different samples. The results will be sent by email. If you need the original raw data, please download from data storage server at HPC core facility.