Description
General description
HybridSPE-Phospholipid technology is a simple and generic sample prep platform designed for the gross level removal of endogenous protein and phospholipid interferences from biological plasma and serum prior to LC-MS or LC-MS/MS analysis. Biological plasma or serum is first subjected to protein precipitation via the addition and mixing of acidified acetonitrile. Precipitated proteins are then removed by centrifugation and the resulting supernatant is loaded on the HybridSPE-Phospholipid 96-well plate or cartridge which acts as a chemical filter that specifically targets the removal of endogenous sample phospholipids. The phospholipid retention mechanism is based on a highly selective Lewis acid-base interaction between the proprietary zirconia ions functionally bonded to the HybridSPE-Phospholipid stationary phase and the phosphate moiety consistent with all phospholipids. The resulting eluent is ready for immediate LC-MS or LC-MS-MS analysis.
The "In-well" and "In-cartridge" precipitation methods are available for the HybridSPE-Phospholipid 96-well version and HybridSPE-Phospholipid Ultra cartridge in which biological plasma/serum is first added to either the well or cartridge, followed by acidified acetonitrile (precipitation agent). After a brief mixing/vortexing step, vacuum is applied. Because the 96-well and Ultra cartridge versions contain a series of low porosity hydrophobic filters/frits, the packed-bed filter/frit assembly acts as a depth filter facilitating the concurrent removal of both phospholipids and precipitated proteins during the extraction process. Standard HybridSPE-Phospholipid cartridges require an "off-line" precipitation method.
Application
Less is more: a methodological assessment of extraction techniques for per- and polyfluoroalkyl substances (PFAS) analysis in mammalian tissues.: This study assesses various extraction methods, including HybridSPE®-Phospholipid, for analyzing PFAS in tissues, highlighting efficiency and reproducibility (Mertens et al., 2023).
Rapid analysis of 65 pharmaceuticals and 7 personal care products in plasma and whole-body tissue samples of fish using acidic extraction, zirconia-coated silica cleanup, and liquid chromatography-tandem mass spectrometry.: This paper describes a rapid analytical method incorporating HybridSPE®-Phospholipid for sample preparation, demonstrating its application in environmental toxicology (Tanoue et al., 2020).
Features and Benefits
For use in conjunction with off-line protein precipitation.
Merges the simplicity of protein precipitation and the selectivity of SPE via the targeted removal of phospholipids
Reduce ion-suppression through the complete removal of phospholipids and precipitated proteins
2-3 step generic procedure
Minimal to no method development
Available in 96-well and 1 mL cartridge dimensions
Legal Information
HybridSPE is a registered trademark of Merck KGaA, Darmstadt, Germany
