Services - How we work
OWL strives to provide a top quality service for cutting-edge research, using strictly controlled analytical procedures and state-of-the-art facilities under expert supervision.
1. Initial Contact and experimental planning:
On receipt of a request, a qualified bioanalytical chemist from OWL will contact the client to discuss the details of the project. OWL metabolomics services include technical discussion and guidance from experts in the field, offering an optimal experimental design (sample collection, sample preparation procedures, analytical methodology and data processing methods) to provide a high quality, fast turnaround, cost efficient final product.
2. Metabolite extraction and LC/MS/MS analysis:
Sample preparation is a fundamental step in the metabolomics workflow, with a strong influence on metabolome coverage. Since thousands of endogenous compounds are typically present in biological samples, with large variations in concentration and physico-chemical properties, specific sample preparation methods are needed depending on the analysis requested. An optimal choice of method will be taken, depending on the sample type and portion of the metabolome considered by the prime interest (e.g lipids, aminoacids, ...etc).
UPLC-MS/MS based metabolomics offers selective, sensitive analyses with the potential to identify metabolites. The analytical platform at OWL includes several liquid-chromatography interfaced mass spectrometers ideally suited to metabolomics applications. Samples are typically analysed in positive and/or negative electrospray ionisation (ESI) modes, scanning from m/z 50-1000, on a reverse phase column packed with sub 2mm particles; the specific bonded phase (typically C8, C18 or HSS) depending on the application.
3. Data Analysis:
Data handling constitutes a challenging task and one of the biggest bottlenecks of the metabolomics workflow, being a time-consuming issue which usually requires detailed knowledge of bioinformatics, statistics and specialized software. However it is also the clue to accomplish valuable results.
Metabolomics data sets can only be managed by powerful informatics and statistical applications. OWL´s analytical platforms are supported by a state-of-the-art suite of software products complemented by visualization tools, which allow complex data sets to be transformed into informative charts and graphs. OWL applies chemometric tools such as principal component analysis (PCA), partial least squares discriminate analysis (PLS-DA), orthogonal least-squares to latent structures (OPLS), and univariate data analysis.
4. Report Writing:
The results package provided includes: a detailed report of the experimental methods, data analysis and results interpretations; an appendix containing scores plots and diagnostic parameters for the multivariate models calculated; full raw data set together with univariate statistical comparisons between the sample groups.
Additionally, OWL has developed OWL Stat App, an easy-to-use web application for metabolomics data analysis. It combines powerful univariate and multivariate data analysis with pathway mapping tools and visualization capacities to facilitate interpretation of the results. OWL Stat App is accessible independently of the operating system and without the need to install programs locally. This application combines the R-based analytical tools with metabolite identification and pathway mapping tools, overlaying the users data on the pathway mapping libraries of SMPDB (The Small Molecule Pathway Database) and pathway libraries originally developed in our laboratory.