LC/MS Analysis of Plasma Samples on Ascentis® Express C18 Showing Baseline Improvements after Phospholipid Removal using HybridSPE®-Phospholipid

LC/MS Analysis of Plasma Samples on Ascentis® Express C18 Showing Baseline Improvements after Phospholipid Removal using HybridSPE®-Phospholipid

Conditions

sample preparation SPE (Solid Phase Extraction)
sample/matrix rat plasma
SPE well plate HybridSPE-Precipitation 96-well Plate, 50 mg/well (575656-U)
sample addition 100 µL rat plasma followed by 300 µL 1% formic acid in acetonitrile
elution apply vacuum
column Ascentis Express C18, 5 cm x 2.1 mm I.D., 2.7 µm particles (53822-U)
column temp. 35 °C
mobile phase [A] 10 mM ammonium acetate; [B] 10 mM ammonium acetate in acetonitrile
gradient 0 min - 95% (A); 10 min - 50% (A); 18 min - 50% (A); 18.1 min - 95% (A); 22 min - 95% (A)
flow rate 0.5 mL/min
injection 5 µL
detector ABI 3200QT; ESI (+), MRM (184/103 m/z)

Description

Application HybridSPE-Precipitation:
Precipitate
proteins by first adding 100 μL rat plasma to the HybridSPE-PPT plate followed by 300 μL 1% formic acid in acetonitrile. Do not apply vacuum.
Mix by vortexing the HybridSPE-PPT plate briefly.
Apply vacuum using a 96-well vacuum manifold and collect the resulting eluate for analysis.

Standard Protein Precipitation:
Combine
100 μL rat plasma with 300 μL 1% formic acid in acetonitrile.
Mix by vortexing and centrifuge.
Isolate resulting supernatant for analysis.
Categories Analytical Chromatography, Clinical, SPE Applications, proteins, Pharmaceutical & Clinical applications, Proteins
Featured Industry Clinical
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC, HybridSPE is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for LC-MS, application for SPE

Materials

     
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