Development of Innovative and Special Analytics
The expertise of BGC in chemistry and analytics covers the whole analytical pipeline involving specific microsampling techniques, sample preparations, functional/structural analyses and bioinformatics. We develop tailored targeted and non-targeted analyses enabling the description of the chemodiversity of complex organic systems. The non-targeted deep metabotyping approach is the basis of the analysis of metabolites in various complex microbiomes of health- or disease-related projects.
The expertise of BGC in chemistry and analytics covers the whole analytical pipeline involving specific microsampling techniques, sample preparations, functional/structural analyses and bioinformatics. We develop tailored targeted and non-targeted analyses enabling the description of the chemodiversity of complex organic systems. The non-targeted deep metabotyping approach is the basis of the analysis of metabolites in various complex microbiomes of health- or disease-related projects.
Separation science is our expertise over the last decades. We initiated in the 1990s a series of developments around new chromatographic and electrophoretic monolithic materials. This included methods in capillary electrophoresis and its coupling to mass spectrometry for organic analysis.
Currently we are using liquid chromatography coupled to high resolution mass spectrometry (UHPLC-HRMS/MS) for targeted and non-targeted analysis. Used methods comprises the quantification of selected small molecules of interest (long and short chain fatty acids (SCFA), bile acids, D/L amino acids, lipids, nucleotides, peptides, etc.) and the development of non-targeted standardized methods for the analysis of polar, mid- to non-polar metabolites utilizing hydrophilic interaction liquid chromatography (HILIC) and reversed phase (RP) chromatographic separation. Furthermore, we are investigating new 2D-column couplings for increased metabolome coverage. Additional developments include new dopants for enhanced electrospray ionization, quantitative structure-selectivity relationship (QSSR) modeling of chromatographic behavior of metabolites in RP and HILIC separations, miniaturization designs of the technologies for minimal sample volumes and in-silico analysis of MS/MS fragmentation patterns.
Our developments are related to new micro ionization interfaces and methods based on electrospray ionizations as guided by our needs in exploring and describing the chemical diversity in various sample matrices (surface and solution analyses).
We are currently establishing methods bringing direct injection MRMS to high throughput metabolomics for thousands of samples, which involves routines and data processing strategies (Big Data). This aspect covers data conversation and interpretation of ultrahigh resolved mass information in terms of compositional space or structural parametrization. The understanding and visualization of the multidimensional dataspace in informative and interactive networks allows us to describe thousands of metabolites out of a minute range analysis time. The applications fields in health are model organisms (plants, C. elegans, mouse) and human cohorts (KORA, PLIS, Freising Infant Cohort, etc.).
Additionally, density function theorie (DFT) analysis and physicochemical modeling enables us gain information of molecular stability and interpret mass spectrometry fragmentation pattern to describe chemical structures of novel chemical compounds.
MRMS is integrated with lower resolution mass spectrometry in the frame of big data integration from our three operational metabolomics platforms (UHPLC-MS, MRMS and NMR).
NMR spectroscopy is a powerful tool for detecting, identifying and quantifying a wide range of high and low molecular weight compounds without any prior selection and with little sample preparation. Furthermore, it provides invaluable information about the detailed molecular and chemical structure of the measured compounds. NMR can be quantitative, if the experiment is carried out appropriately. We apply this technique to complex organic mixtures, such as human and animal biofluids or tissues.
1H NMR-based metabolomics has shown its usefulness in analyzing a variety of biofluids and tissue extracts. The absence of analytical suppression effects allows a quantitative and therefore inter-sample comparison of main metabolite quantities. Through the availability of high-field instruments and the use of cryogenic probes sensitivity increases and 2-dimensional experiments are possible in a feasible time. Especially 2D experiments (e.g. 1H-13C-HSQC, 1H-1H-TOCSY, J-resolved) are necessary to aid and confirm signal annotation to metabolites.
Our equipment and developments ensure the management of large biofluid sample sets, i.e. automated sample preparation and cooled autosampler for sample stability while 24-7 measurement. Our high resolution NMR spectrometer enables the elucidation of new molecules and constantly expands the number of metabolites detectable and identifiable in various biosamples.
