Comparison of blood plasma sample preparation methods for combined LC–MS lipidomics and metabolomics

• Folch, Matyash, and Ostro plate preparations were compared for lipid coverage.
• Matyash method provides recovery and reproducibility across lipid classes.
• The Waters Ostro plate is beneficial for targeted lipidomics by class.
• Matyash provides recovery of polar compounds as use in biphasic extraction.

The goal of this research was to find the most comprehensive lipid extraction of blood plasma, while also providing adequate aqueous preparation for metabolite analysis. Comparisons have been made previously of the Folch, Bligh–Dyer, and Matyash lipid extractions; furthermore, this paper provides an additional comparison of a phospholipid removal plate for analysis. This plate was used for lipid extraction rather than its intended use in lipid removal for polar analysis, and it proves to be robust for targeted lipid analysis. Folch and Matyash provided reproducible recovery over a range of lipid classes, however the Matyash aqueous layer compared well to a typical methanol preparation for polar metabolite analysis. Thus, the Matyash method is the best choice for an untargeted biphasic extraction for metabolomics and lipidomics in blood plasma.