Abstract
IntroductionPost-collection handling, storage and transportation can affect the quality of blood samples. Pre-analytical biases can easily be introduced and can jeopardize accurate profiling of the plasma metabolome. Consequently, a mouse study must be carefully planned in order to avoid any kind of bias that can be introduced, in order not to compromise the outcome of the study. The storage and shipment of the samples should be made in such a way that the freeze–thaw cycles are kept to a minimum. In order to keep the latent effects on the stability of the blood metabolome to a minimum it is essential to study the effect that the post-collection and pre-analytical error have on the metabolome.ObjectivesThe aim of this study was to investigate the effects of thawing on the metabolic profiles of different sample types. MethodsIn the present study, a metabolomics approach was utilized to obtain a thawing profile of plasma samples obtained on three different days of experiment. The plasma samples were collected from the tail on day 1 and 3, while retro-orbital sampling was used on day 5. The samples were analysed using gas chromatography time-of-flight mass spectrometry (GC TOF-MS).ResultsThe thawed plasma samples were found to be characterized by higher levels of amino acids, fatty acids, glycerol metabolites and purine and pyrimidine metabolites as a result of protein degradation, cell degradation and increased phospholipase activity. The consensus profile was thereafter compared to the previously published study comparing thawing profiles of tissue samples from gut, kidney, liver, muscle and pancreas.ConclusionsThe comparison between thawed organ samples and thawed plasma samples indicate that the organ samples are more sensitive to thawing, however thawing still affected all investigated sample types.
Highlights
Post-collection handling, storage and transportation can affect the quality of blood samples
The run order effects were analysed by creating an orthogonal projection to latent structures (OPLS) model using the run order as the response variable
The present study showed that cholesterol is fairly stable in plasma, but not in organ samples
Summary
Post-collection handling, storage and transportation can affect the quality of blood samples. In order to keep the latent effects on the stability of the blood metabolome to a minimum it is essential to study the effect that the post-collection and pre-analytical error have on the metabolome. Methods In the present study, a metabolomics approach was utilized to obtain a thawing profile of plasma samples obtained on three different days of experiment. Metabolomic studies involve several steps starting from the hypothesis or study design and resulting in a biological interpretation (Brown et al 2005; Krastanov 2010). The pipeline for a metabolomics approach often involves experimental planning, sampling, storage, pre-treatment, chemical analysis, data processing,. Steps that are often overlooked include the experimental planning, sampling and transportation of samples and the effects that these have on the final results and the biological interpretation
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
