|UPLC-MS for Metabolic Phenotyping: Advantages, Assays and Applications
|Advances in Clinical Analysis
|Dr Elizabeth Want
|Imperial College London
Abstract Information :
Ultra performance liquid chromatography coupled to mass spectrometry (UPLC-MS) has become increasingly prevalent in the field of metabolic phenotyping over the past decade. Applications to date have included clinical, toxicological, and epidemiological investigations, improving our understanding of drug metabolism and toxicity, as well as disease diagnosis and prognosis. UPLC has advantages over conventional HPLC, including increased speed and chromatographic resolution, resulting in deeper metabolome coverage with higher sample throughput. This makes it an ideal analytical tool for large scale epidemiological studies, as well as for in depth interrogation of individual biological sample types. In this talk, the field of metabolic phenotyping will be introduced, the typical analytical workflow will be discussed and factors which affect the metabolic profile will be examined. Applications of UPLC-MS in metabolic phenotyping will be reviewed, illustrating advances in biomarker discovery and our understanding of human health and disease.
Despite the 3-5 fold increase in sample throughput that UPLC-MS offers over HPLC-MS, researchers remain interested in further improving speed without compromising metabolite coverage, in order to accommodate large sample numbers and to reduce solvent usage. Improvements to sample throughput will be mentioned, such as rapid microbore metabolic profiling (RAMMP) technology and other miniaturisation approaches. As part of the metabolic phenotyping pipeline, targeted UPLC-MS assays are crucial for quantitation of key metabolites and biomarker validation. Therefore, targeted assays will be introduced, with examples of successful applications in disease diagnosis. Crucially, the importance of quality control measures in all of these assay types will be covered. Finally, the future of this exciting and ever growing area of research will be discussed.