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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

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About This Item

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

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

SPE tube or plate


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