Chromatography and Mass Spectrometry

Mass spectrometry is an almost universally applicable technology for the qualitative and quantitative analysis of biomolecules especially proteins, metabolites and drugs. Incorporating chromatographic separation techniques and high sensitivities as well as experimental precision, it is suitable for both the directed quantification of individual analytes and the screening of biomarkers. 

The services offered by the Fraunhofer IZI Proteomics Unit include protein analysis by liquid chromatography-mass spectrometry (LC-MS)-coupled methods of various biological systems, including cell culture systems, biofluids (e.g. blood plasma), extracellular vesicles (from cell culture and biofluids) and tissues. The goal is to comprehensively quantify all proteins to identify biomarks and reveal molecular mode-of-actions. For this purpose, various label-based (e.g. tandem mass tags) as well as label-free methods are available. All methods can be performed in the central testing facility taking into account established SOPs. We will be happy to implement your individual requests in proteomics and advise you according to your individual needs.

In addition to protein analysis, there is the possibility of determining toxins, metabolites and active substances by means of mass spectrometry-based Multiple Reaction Monitoring (MRM) and DNA conjugates. Specific purification by means of preparative chromatography and directed identification/quantification is also offered by Fraunhofer IZI. We will be happy to provide you with an individual offer for this.

Recent references

• Biadglegne F, Schmidt JR, Engel KM, Lehmann J, Lehmann RT, Reinert A, König B, Schiller J, Kalkhof S, Sack U. Mycobacterium tuberculosis Affects Protein and Lipid Content of Circulating Exosomes in Infected Patients Depending on Tuberculosis Disease State. Biomedicines. 2022 Mar 27;10(4):783. doi: 10.3390/biomedicines10040783. PMID: 35453532
• Furesi G, de Jesus Domingues AM, Alexopoulou D, Dahl A, Hackl M, Schmidt JR, Kalkhof S, Kurth T, Taipaleenmäki H, Conrad S, Hofbauer C, Rauner M, Hofbauer LC. Exosomal miRNAs from Prostate Cancer Impair Osteoblast Function in Mice. Int J Mol Sci. 2022 Jan 24;23(3):1285. doi: 10.3390/ijms23031285. PMID: 35163219

The "Chemical analytics" laboratory focuses on the identification and quantification of biomolecules (proteins, peptides and small molecules) in complex biological matrices like biofluids and tissue homogenates. It operates different mass spectrometers (MALDI-TOF/TOF, ESI-/APCI triple quadrupole/linear ion trap) and liquid chromatography systems (LC-UV/Vis) that can be used either individually or in combination (LC-MS, LC-MS/MS or LC-MRM) depending on the question being examined. In addition, the laboratory has a comprehensive range of equipment (e. g. homogenisers, various SPE systems) for the processing of biological samples and is approved for biosafety level 2 (BSL-2). The laboratory is GLP (Good Laboratory Practice) certified and part of the GLP test facility of the Fraunhofer IZI.

The main activities of the "Chemical Analytics" laboratory include the development and validation of analytical methods for various types of biomolecules as well as analytical services for preclinical studies (GLP/non-GLP) in the fields of pharmacokinetics and toxicology.

Volatile organic compounds (VOCs) are metabolic byproducts produced by human cells, bacteria, fungi, yeasts, and other organisms. Since VOCs transition into the air phase at low temperatures, they naturally migrate from cells into their environment and ultimately into exhaled breath or the headspace of cell cultures and bioreactors.

Human exhaled breath typically contains several thousand VOCs, while cell culture exhaust gases contain several hundred. The analysis of these complex, characteristic patterns of VOCs, also known as volatilomics, offers a non-invasive and resource-efficient method for diagnosing diseases and monitoring health. It also enables the tracking of specific metabolic pathways in cell cultures and the monitoring of the progress of biotechnological manufacturing processes.

Gas chromatography coupled with ion mobility spectrometry (GC-IMS) provides fast, sensitive, and practical VOC analyses for research and quality assurance. The combination of gas chromatography and ion mobility produces high-resolution fingerprints in minutes – ideal for non-invasive exhaled breath studies (for disease detection and therapy monitoring), the characterization of cell culture headspace, and the rapid detection of bacterial signatures or contamination. The method is also impressive outside the field of biomedicine: In the food sector, we analyze aroma, freshness, and off-flavors; in the pharmaceutical and chemical industries, we monitor solvent residues, cleaning, and processes; and we also test emissions and material aging.

We support you from sampling to the results report: Appropriate sampling design (e.g., online measurements on site, Tedlar bags, headspace vials, pre-concentration), standardized measurement protocols, VOC identification, and sophisticated data analysis. Thanks to high selectivity and sensitivity down to the low ppb range, you get reliable results with minimal sample preparation and small sample quantities.

Whether it's a pilot study, method development, or routine screening: We advise, measure, and train tailored to your specific needs. Contact us – we will prepare an individual offer and a quick proof of concept for your samples.

Equipment

• GC-IMS (STEP Sensortechnik Pockau / Graupner Medical)
• GC-IMS (G.A.S. Dortmund)
• GC-FAIMS (Schumann Analytics), FAIMS (Owlstone)

Recent references

• Steppert I, Schönfelder J, Schultz C, Kuhlmeier D. Rapid in vitro differentiation of bacteria by ion mobility spectrometry. Appl Microbiol Biotechnol. 105 (2021), 10, Seite 4297-4307. doi: 10.1007/s00253-021-11315-w