Comprehensive metabolic profiling or “metabolomics” is a technology that can define the chemical phenotype of human subjects and animal and cell models. This unique chemical phenotyping allows for the discovery of biomarkers that predict disease incidence, severity, and progression, shedding light on the underlying mechanistic abnormalities of disease. The DMPI Metabolomics Laboratory is a leader in applying both targeted and non-targeted mass spectrometry (MS) based metabolic profiling in the quest to understand disease and its biological mechanisms. The lab is an integral component of the suite of technologies available to collaborators via the DMPI Storefront portal.
Figure1: Waters Xevo TQ-S Quadrupole MS
The DMPI Metabolomics Laboratory has a comprehensive suite of seven MS instruments (Waters, Agilent, and Thermo Fisher) that allow both targeted and non-targeted metabolomics assays to be conducted on a wide array of sample types including plasma, blood spots, tissue, urine, cerebral spinal fluid, and cell culture. For targeted analyses, the lab deploys four triple quadrupole (MS/MS) instruments (Figure 1) and two gas chromatography (GC)/MS instruments (Figure 2) to measure more than 200 specific metabolites across seven chemical classes: 1) 66 species of acylcarnitines; 2) 15 amino acids and urea cycle intermediates 3) 57 acyl CoAs species; 4) 20 organic acids; 5) 21 ceramides; 6) 30 sphingomyelins, and 7) 29 nucleotides and purine/pyrimidine pathway intermediates. Accurate quantitation of the metabolites in the targeted assays is achieved using isotope dilution or pseudo-isotope dilution techniques and quality control calibrators derived from biologic matrices and commercially available analytical standards.
Figure2: Thermo Scientific TRACE 1310 GC-MS
To complement the strong targeted platform, non-targeted metabolomic methods have been developed to greatly expand coverage of the metabolome across many biochemical classes of metabolites. The untargeted GC/MS-based platform involves chemical derivitization with electron-ionization mass spectrometry. In collaboration with Agilent Technologies, Inc., the laboratory has created one of the world’s largest retention-time-indexed spectral libraries of metabolites, which is used to identify metabolites that could reveal novel biochemical pathways and interactions. Another non-targeted approach under active development is based on high-performance liquid chromatography (HPLC)-MS. This method involves reverse-phase LC coupled to a quadrupole, time-of-flight mass spectrometer (Figure 3) which operates in both positive and negative ionization modes. This high resolution platform will provide a tremendous expansion in the coverage of the metabolome.
Figure 3: Agilent 6520 Quadrupole Time-of-Flight MS
A key feature of the DMPI metabolomics core laboratory is its long experience with internal and external collaborative projects, which provides a level of experience in data interpretation and translation not found in most other labs of its kind. The Storefront laboratory employs dedicated PhD-level personnel who are equipped to assist collaborators with interpretation of the various kinds of metabolomics data sets described above, and to help our colleagues to understand the data in a physiologic and pathophysiologic context. In addition, our Storefront scientists can draw upon the expertise of senior DMPI faculty when needed, thereby increasing the probability that a collaborative project with DMPI will have a productive outcome. Firmly supporting this statement, the DMPI Metabolomics core laboratory has so far co-authored >140 publications in leading scientific journals.
Director: Olga Ilkayeva, PhD
Office Phone: (919) 479-2370