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HPLC Analysis of Diazepam and Metabolites in Serum on Discovery® C18 after SPE using Discovery® DSC-8

HPLC Analysis of Diazepam and Metabolites in Serum on Discovery® C18 after SPE using Discovery® DSC-8 application for SPE, application for HPLC

Matériaux

Colonne analytique

Réf. du produit
Description
Tarif

Colonne HPLC Discovery® C18

5 μm particle size, L × I.D. 5 cm × 4.6 mm

CONDITIONS

sample preparation

SPE (Solid Phase Extraction)

sample/matrix

goat serum spiked with diazepam and metabolites at 0.5 μg/mL and diluted 1:1 in 25 mM ammonium formate, pH 7.1

SPE well plate

Discovery DSC-8 96-well SPE Plate, 100 mg/well (575627-U)

condition

1 mL methanol followed by 1 mL DI water.

sample addition

1 mL spiked goat serum

washing

1 mL 25% methanol in 25 mM ammonium formate, pH 2.75

elution

1 mL 60% methanol in 25 mM ammonium formate, pH 2.75

elution

Evaporate eluate with nitrogen purge (30 °C; ~15 min.). Reconstitute in 200 μL mobile phase.

column

Discovery C18, 5 cm x 4.6 mm I.D., 5 μm particles (504947)

mobile phase

[A] Methanol; [B]10 mM ammonium acetate, pH 4.5;(45:55, A:B)

flow rate

1.5 mL/min

column temp.

35 °C

detector

UV, 240 nm

injection

25 μL

Description

Informations légales

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