What Are the Standard Procedures for Serum Sample Preparation in Metabolomics Studies?
The preparation of serum samples for metabolomics analysis typically involves the following steps:
1. Blood Collection
Venous blood is collected without the use of anticoagulants to allow for natural coagulation, which facilitates subsequent serum separation.
2. Clotting and Centrifugation
The collected blood is left undisturbed at room temperature for 30 to 60 minutes to ensure complete clotting. It is then centrifuged at approximately 2000–3000 × g for 10 minutes to separate the serum from the clot.
3. Serum Harvesting
The supernatant serum is carefully extracted, ensuring that the underlying clot is not disturbed during the collection process.
4. Protein Precipitation
To remove proteins, cold acetonitrile or methanol is added to the serum in a typical volume ratio of 1:3 or 1:4 (serum:precipitant). The mixture is briefly incubated at low temperature to enhance precipitation, followed by centrifugation to eliminate the precipitated proteins.
5. Aliquoting and Storage
The deproteinized serum is then aliquoted into sterile tubes and immediately stored at –80 °C until analysis, preserving sample integrity for downstream metabolomic profiling.
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