GC-MS-Based Metabolic Flux Analysis

GC-MS-Based Metabolic Flux Analysis

In addition to nuclear magnetic resonance (NMR)-based metabolic flux analysis (MFA), Creative Proteomics also offers mass spectrometry (MS)-based MFA, which is characterized by more efficiency, accuracy, and high sensitivity. Among them, the gas chromatography-mass spectrometry (GC-MS) technique is the preferred choice for MFA. Based on advanced instrument platforms and experienced experts, we are able to provide GC-MS/GC-MS/MS-based metabolic flux analysis to study fluxes in bacteria, fungi, mammalian cells, or intact tissues.

GC-MS—the most popular technique for labeling measurement in metabolic flux studies

The MS-based metabolic flux approach, which mainly utilizes ¹³C-labeling tracer substrates, has become a key technology in metabolic physiology and biotechnology. MS plays a central role in providing accurate ¹³C-labeling data for metabolites formed during the cultivation of the studied organisms on the tracer substrate. GC-MS is the most commonly used technology for metabolic flux measurements. GC-MS not only have high signal-to-noise ratios but also allow accurate and quick quantification of the ¹³C-labelling pattern of various amino acids from the cell protein. Because of the ease of access to proteins in almost all studied cells and the wealth of information available on amino acids, this type of analysis has become a routine method in many laboratories. To improve the flux resolution, parallel experiments can be performed with different tracers if necessary. The widespread use of GC-MS for metabolic flux analysis is mainly due to the following several advantageous characteristics.

Fig. 1 Strategy for 13C metabolic flux analysis. (Wittmann, Christoph., 2007)

Several advantageous characteristics of GC-MS

• high robustness, precision and sensitivity
• Great separation capacity for complex biological mixtures
• GC-MS provides information-rich labeling data to resolve important network structures of metabolic networks, such as fluxes through parallel, bidirectional or circular pathways
• Suitable for metabolic flux studies in many biological systems and culture conditions.

Service offering

Creative Proteomics provides GC-MS/GC-MS/MS-based metabolic flux analysis to help accelerate your research. Based on highly stable, reproducible and sensitive separation, characterization, identification, and quantitative analysis systems, we are committed to providing reliable, rapid, and cost-effective metabolic flue analysis services. Typically, our services include isotope-labeling culture, MS measurement, and data analysis.

• Isotope-labeling culture

-Experimental consulting and design

-¹³C-labeling experiment

-Cell harvest

-Sample pre-treatment

• GC-MS/GC-MS/MS measurement for labeling analysis
• Data analysis

-Metabolic modeling

-Metabolite Identification and ¹³C flux ratio analysis

-Metabolite absolute quantification

-Delivery of result reports and interpretation

Instrument platform

• Shimadzu GC-MS-QP2010 Ultra
• Sciex Triple TOF 6600
• Sciex Qtrap 5500

Application of our services

• Isotopic tracer experiments with GC-MS labeling analysis have been applied to various microorganisms, including bacteria, yeasts and fungi
• GC-MS-based metabolic flux analysis is utilized in the biomedical area to help understand the metabolic function of mammalian cell lines or organs or potential metabolism associated with disease

Why choose us?

• We accept a wide range of biological materials including, but not limited to, cells, tissues, organs, and biofluids
• A comprehensive platform with advanced instrumentation and data analysis software
• Provide a complete analysis report including method interpretation, experimental parameters, data and results analysis
• A team of experts with years of experience in metabolomics, bioinformatics, and statistics

MFA uses MS or NMR to monitor the fate of isotope tracers (such as ¹³C-glucose and ¹⁵N-glutamine), helping researchers to assess the contribution of specific metabolic pathways to the prevailing levels of specific metabolites. Creative Proteomics is a leading provider of metabolic flux analysis services. Our reliable and personalized service can accommodate any innovative research project. Please contact our experts to discuss your specific needs.

Reference

1. Wittmann, Christoph. "Fluxome analysis using GC-MS." Microbial cell factories 6.1 (2007): 1-17.