Functions of Muramic Acid (MA)

MA is an important component of the peptidoglycan structure in the bacterial cell wall. It is a biomarker compound of prokaryotic microbial cells, constructed as 2-amino-3-O-(1-carboxyethyl)-2-deoxy-D-glucose. MA is an aminosugar with the general property of aminosugar. Under heating conditions, MA easily reacts with amino acids to produce a Maillard reaction and caramelization. MA activates many types of immune cells such as macrophages, T- and B-lymphocytes, as well as the production and activation of immune substances such as antibodies, complements, and cytokines. MA has a wide range of important effects on cellular and humoral immunity and produces specific effects with lectins with a high affinity.

Fig.1 Chemical structure of MA. (Liang, et al., 2009)

MA Analysis Service at CD BioGlyco

MA and N-acetylmuramic acid have been widely used as micro determinants and quantitative markers of bacterial and bacterial cell wall residues in complex biological matrices and as new targets for drug development such as antitumors. Therefore, the development of new techniques for the isolation and analysis of MA is necessary. At CD BioGlyco, we have developed a variety of different technological approaches applied to the comprehensive analysis of MA for the different demands of our clients.

• Liquid chromatography-tandem mass spectrometry (LC-MS/MS)

CD BioGlyco uses hydrophilic liquid-phase interaction chromatography (HILIC) with optimized solvent composition, buffer strength, pH, flow rate, and column temperature. The method does not require a derivatization step, rapidly separates MA and the precursor molecule N-acetylglucosamine, and the detection of analytes is performed by tandem mass spectrometry (MS/MS) and time-of-flight mass spectrometry (TOF-MS). The method is easily used for structural confirmation and quantification using electrospray ionization with MS. Especially in ¹³C tracers, increasing the use of these residues for stable carbon isotope analysis of microbial processes.

• Multivariate curve resolution (MCR) gas combined with chromatography-mass spectrometry (GC-MS)

The MCR method is combined with GC-MS technology to correct and reduce the background and noise in the GC-MS analyses. MA overlapping peaks are decomposed into their pure chromatographic and mass spectral analyses. We use the external products of chromatography and MS to reconstruct the bidirectional response of each component. Quantitatively determine it uses the total volume integral method.

Fig.2 Flowchart of MA analysis. (CD BioGlyco)

Publication

Paper Title: The use of multivariate curve resolution methods to improve the analysis of muramic acid as bacterial marker using gas chromatography-mass spectrometry: an alternative method to gas chromatography-tandem mass spectrometry.

Technology: GC-MS

Journal: Journal of Chromatography B

IF: 3.7

Published: 2014

Results: This paper focuses on the use of the MCR methods as complementary techniques for the GC-MS analysis of MA in complex mixtures. GC-MS/MS is used to analyze this acid when baseline mass channels, large noise, and or complexity of mixture are the main barriers to thorough analysis with the GC-MS.

Fig.3 Reconstructed muramic acid peak cluster by the outer product of the resolved elution profiles to the resolved mass spectra using the MCR alternating least squares method. (Moazeni-Pourasil, et al., 2014)

Applications

• To assess the relative roles of fungi and bacteria in the degradation of the stability and fate of soil organic carbon.
• The total amount of microorganisms is evaluated, such as detecting the content of detected bacteria, cyanobacteria, etc.
• Develop anti-tumor drugs and new aminoglycoside antibiotic drugs, immune promoters, etc.

Advantages

• We provide rapid separation testing services: ten-minute separation of MA.
• Our analytical methods are highly selective for the separation of analytes.
• MCR-GC-MS improves the sensitivity and detection limit of the analysis, reduces the signal-to-noise ratio, and corrects incorrect quantitative analysis compared to GC-MS.

CD BioGlyco has focused on research for many years in the field of Carbohydrate Metabolism Analysis. We provide fast and efficient Monosaccharide Analysis Services, Disaccharide Analysis Services, Oligosaccharide Analysis Services, and Polysaccharide Analysis Services. If you are interested in our service, please contact us further to introduce the specific service in detail.

References:

1. Liang, C.; et al. Reliability of muramic acid as a bacterial biomarker is influenced by methodological artifacts from streptomycin. Microbial Ecology. 2009, 57(3): 494-500.
2. Moazeni-Pourasil, R.S.; et al. The use of multivariate curve resolution methods to improve the analysis of muramic acid as bacterial marker using gas chromatography-mass spectrometry: an alternative method to gas chromatography-tandem mass spectrometry. Journal of Chromatography B. 2014, 949: 1-6.
3. Vandenboer, T.C.; et al. Quantitation of amino sugar stereoisomer and muramic acid biomarkers by hydrophilic interaction liquid chromatography-mass spectrometry. Journal of Chromatography A. 2020, 1618: 460843.