G004243

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

application for SPE, application for LC-MS

Properties

• technique(s): LC/MS: suitable; solid phase extraction (SPE): suitable
• test parameters: 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); 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; column temp.: 35 °C; detector: ABI 3200QT; ESI (+), MRM (184/103 m/z); injection: 5 μL
• suitability: application for LC-MS; application for SPE
• application(s): clinical

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.

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany

HybridSPE is a registered trademark of Merck KGaA, Darmstadt, Germany