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.
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