Evaluation of Sample Preparation Methods for Inter-Laboratory Metabolomics Investigation of Streptomyces lividans TK24.
Howbeer MuhamadaliKenneth SimoensYun XuBart NicolaiKristel BernaertsRoyston GoodacrePublished in: Metabolites (2020)
In the past two decades, metabolomics has proved to be a valuable tool with many potential applications in different areas of science. However, there are still some challenges that need to be addressed, particularly for multicenter studies. These challenges are mainly attributed to various sources of fluctuation and unwanted variations that can be introduced at pre-analytical, analytical, and/or post-analytical steps of any metabolomics experiment. Thus, this study aimed at using Streptomyces lividans TK24 as the model organism in a cross-laboratory experiment in Manchester and Leuven to evaluate the reproducibility of a standard sample preparation method, and determine the optimal sample format (cell extract or quenched biomass) required to preserve the metabolic profile of the cells during cross-lab sample transportation and storage. Principal component analysis (PCA) scores plot of the gas chromatography-mass spectrometry (GC-MS) data from both laboratories displayed clear growth-dependent clustering patterns which was in agreement with the Procrustes analysis findings. In addition, the data generated in Manchester displayed tight clustering of cell pellets (quenched biomass) and metabolite extracts, confirming the stability of both sample formats during the transportation and storage period.
Keyphrases
- single cell
- mass spectrometry
- gas chromatography mass spectrometry
- liquid chromatography
- rna seq
- wastewater treatment
- oxidative stress
- cell therapy
- electronic health record
- public health
- blood brain barrier
- clinical trial
- molecularly imprinted
- mesenchymal stem cells
- cross sectional
- machine learning
- bone marrow
- solid phase extraction
- pi k akt
- endoplasmic reticulum stress