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1. Precipitate Proteins by adding 100 µL plasma or serum to the HybridSPE-PPT plate followed by 300 µL 1% formic acid in acetonitrile. Add I.S. as necessary.
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2. Mix by vortexing/shaking HybridSPE-PPT plate or by aspirating/ dispensing with 0.5-1 mL pipette tip (e.g., TOMTEC Quadra liquid handler)
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| 3. |
Apply vacuum.
The packed-bed filter/frit assembly acts as a depth filter for the concurrent physical removal of precipitated proteins and chemical removal phospholipids. Small molecules (e.g., pharma compounds and metabolites) pass through unretained.
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4. Resulting filtrate/eluate is free of proteins and phospholipids and ready for immediate LC-MS-MS analysis; or it can be evaporated and reconstituted as necessary prior to analysis
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