Case study

HeatStab

Development of an improved protocol for metabolomics analysis, based on heat inactivation of enzymes.

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Background

Based on their proprietary heat stabilization technology, Denator AB develops, markets and sells instruments and consumables to address needs in the life science and clinical research markets.

Challenges

In metabolomics, the quality of samples is critical. After sampling, some cellular pathways remain active or can be reactivated during sample preparation, leading to artefacts with no biological relevance.

Thus, it is fundamental to preserve sample stability and the common process to block residual enzymatic activities after sampling is quenching by sample snap freezing (SF) and/or by precooled solvents addition. However, such requirements are not easy to achieve in all Metabolomics labs and particularly in clinical environment.

Solution

An alternative method to Snap Freezing consists in permanently quenching enzymatic activities, based on fast conductive heating stabilization.

To evaluate this approach, we used the StabilizorTM system from Denator to preserve mice liver tissues, and compared its efficacy with the classical freezing procedure, for subsequent global metabolomics studies, based on NMR for polar metabolites and Mass Spectrometry for lipids.

Results

Untargeted lipidomics results based on non-discriminant & global discriminant analyses showed that heat stabilization and snap freezing give similar results in term of sample stabilization.

NMR analysis results showed globally the same tendency for polar metabolites, but the post-sampling preservation of very reactive metabolites, such ATP and derivatives, was superior in heat-stabilized samples.

Taken together, these results have shown that heat stabilization of tissues using the StabilizorTM T1 deliver results equivalent or superior to common snap freezing methods. Therefore, heat stabilization is a good alternative to the snap frozen approach to preserve samples stability in metabolomics.

Outlook

BIOASTER has developed an expertise around heat stabilization for sample preservation before metabolomics studies that can be applied in future project.

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