Chapter 2: Experimental design in metabolomics

by Allycia Y. Lee, Jacopo Troisi, Steven J.K. Symes


The pattern of abundances and molecular identifications of all small molecules within a biological system is referred to as the metabolome. Both targeted and untargeted approaches can be used to study differences in patterns between samples of interest. A wide variety of sample types and analytical platforms are employed in the field of metabolomics. Sample types include biofluids, tissues, and cellular extracts. Each has specific requirements regarding sample collection, storage, and metabolite extraction. The choice of analytical technique for metabolite identification and quantification directly influences sample preparation procedures. Common assays utilize either liquid or gas chromatography coupled to mass spectrometry. Multi-variable statistical analysis is often applied to the complex datasets generated in order to identify specific metabolites that differ between samples. Biological insight can be gained through study of the relevant biochemical pathways in which the differing metabolites are involved.

Allycia Y. Lee

Department of Chemistry and Physics, University of Tennessee at Chattanooga, Chattanooga, YN, United States

Allycia Lee graduated from the University of Tennessee at Chattanooga (UTC) in May 2021 with a Bachelor's of Science in Biochemistry. While at UTC, she conducted research on the development of a screening system for endometrial cancer using a metabolomics-based approach. She began medical school at the University of Tennessee Health Science Center College of Medicine in August 2021 as a member of their inaugural 3-Year MD program.


Jacopo Troisi

Department of Medicine, Surgery and Dentistry “Scuola Medica Salernitana” University; Theoreo srl – spin off company of the University of Salerno, Italy; Department of Chemistry and Biology “A. Zambelli”, University of Salerno, Italy 

Jacopo Troisi is visiting professor of Metabolomics in both Salerno University and University of Sannio in Italy. He is a Research Scientist and COO at the European Biomedical Research Institute of Salerno (EBRIS), co-founded by the Harvard University and the Salerno Medical School. He is also co-founder and CEO at Theoreo srl, a spin-off company of the University of Salerno. Prof. Troisi research is related to metabolomics. In particular, he studies the metabolomic fingerprint of several human diseases to develop diagnostic tools. He has received several international patents for diagnostic tests based on metabolomic profiling. Dr. Troisi has published widely in such peer reviewed journals as Science, Nature Microbiology, Nature Communications, Frontiers in Immunology, Nutrients, BMC Microbiology, Metabolomics, and Scientific Reports.


Steven J.K. Symes

Department of Chemistry and Physics, University of Tennessee at Chattanooga, Chattanooga, YN, United States; Department of Obstretics and Gynecology, University of Tennessee College of MEdicine, Chattanooga, TN, United States

Dr. Steven Symes is an analytical chemist specializing in chromatography and mass spectrometry.  He has taught at The University of Tennessee-Chattanooga since 1999 and has mentored numerous undergraduate and graduate students in his lab. Together with his students, they use LC-MS and GC-MS to investigate interesting questions within the fields of bacterial lipidomics and human metabolomics.