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Merck

HPLC Analysis of Verapamil and Metabolites in Rat Plasma on Ascentisยฎ Express C18 after Phospholipid Removal using HybridSPEยฎ-Phospholipid

HPLC Analysis of Verapamil and Metabolites in Rat Plasma on Ascentis® Express C18 after Phospholipid Removal using HybridSPE®-Phospholipid application for SPE, application for LC-MS

Materials

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HybridSPEยฎ-Phospholipid

96-well Plate, bed wt. 50 mg, volume 2 mL, pk of 1

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Ascentisยฎ Express C18, 2.7 ฮผm HPLC Column

2.7 μm particle size, L ร— I.D. 5 cm ร— 2.1 mm

CONDITIONS

SPE well plate

HybridSPE-Precipitation 96-well Plate, 50 mg/well (575656-U)

sample preparation

SPE (Solid Phase Extraction)

column

Ascentis Express C18, 5 cm x 2.1 mm I.D., 2.7 μm particles (53822-U)

sample/matrix

Rat plasma spiked with verapamil and metabolites at 10 ng/mL

mobile phase

[A] water; [B] acetonitrile, pH 2.55 adjust with formic acid (30:70, A:B)

flow rate

0.6 mL/min

column temp.

35 ยฐC

detector

TOF/MS

injection

1 μL

sample addition

100 μL spiked rat plasma followed by 300 μL 1% formic acid in acetonitrile

elution

apply vacuum

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