LC-HRMS-Based Metabolic Flux Analysis

LC-HRMS-Based Metabolic Flux Analysis

Metabolic flux analysis (MFA) quantitatively characterizes cellular fluxes to understand metabolic phenotypes and functional behavior in response to environmental and/or genetic perturbations. Over the past decade, the use of stable isotopes to determine in vitro and in vivo measurements of metabolic reactions in systems biology has become extremely important. To meet the growing demand for highly sensitive, reproducible and accurate results in metabolic flux analysis, we are proud to offer liquid chromatography-high resolution mass spectrometry (LC-HRMS)-based analytical services. This method combines the properties of liquid chromatography and high-resolution mass spectrometry to provide a wide range of information about the components of the detectable metabolites.

Overview of LC-HRMS in metabolic flux studies

When certain metabolites are incompatible with gas chromatography-mass spectrometry (GC-MS) analysis or require derivatization, in which case LC-MS is the method of choice. LC-MS, with its softer ionization and higher sensitivity, is usually applied to measure metabolic intermediates that are less stable and less abundant than proteinogenic amino acids in the cell. In comparison to NMR, LC-MS is the technique of choice for metabolomics and metabolic flux analysis due to its higher sensitivity and a broader range of detectable metabolites. Previous research studies have reported that better accuracy of mass isotope analysis can be achieved using high-resolution mass spectrometry (HRMS). The method developed in LC-HRMS allows the separation of free sugars and the quantification of the isotopic mass for the quasi-molecular ion [M-H]- of each free sugar. LC-HRMS easily analyzes the isotopomer distribution of free sugars labeled with carbon 13 without a derivatization procedure and is suitable for metabolic flux analysis.

Outstanding features of LC-HRMS

• Improves the accuracy of mass isotope analysis while saving sample preparation time
• Compared to GC-MS, LC-HRMS has good sensitivity and reproducibility to determine isotopic enrichment of free sugars

LC-HRMS-based metabolic flux analysis service

Creative Proteomics is a leading provider of metabolic flux analysis services. We are proud to offer LC-HRMS-based metabolic flux analysis services to our customers worldwide. This method does not require derivatization and provides better precision for mass isotopomer analysis than GC-MS.

¹³C-labeling experiments using modern LC-HRMS systems yield rich data sets, often covering hundreds or even thousands of endogenous molecules, whose isotopic status allows the identification of various active and inactive metabolic pathways.

In addition, we use software specifically designed to meet the challenges of LC-MS-based metabolic flux analysis. Target metabolites can be easily found in stable isotope labeling data and provide results for pathway visualization. High-quality images are obtained in a streamlined and fast data analysis workflow.

Workflow of our service

Sample requirement

• We accept a wide range of biological materials including, but not limited to, cells, tissues, organs, and biofluids
• Please contact us for specific sample requirements

Our metabolic flux analysis service is supported by the latest knowledge in biology, bioinformatics and advanced software. If you are interested in LC-HRMS-based metabolic flux analysis, please contact us and one of our professional experts will provide you with further knowledge or consulting services.

References

1. Acket, Sébastien, et al. "Data documenting the comparison between the theoretically expected values of free sugars mass isotopomer composition with standards using GC-MS and LC-HRMS for Metabolic Flux Analysis." Data in brief 12 (2017): 108-112.
2. Acket, Sébastien, et al. "13C labeling analysis of sugars by high resolution-mass spectrometry for Metabolic Flux Analysis." Analytical Biochemistry 527 (2017): 45-48.