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

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)
column temp. 35 °C
mobile phase [A] Methanol; [B]10 mM ammonium acetate, pH 4.5;(45:55, A:B)
flow rate 1.5 mL/min
injection 25 µL
detector UV, 240 nm

Description

Categories Analytical Chromatography, Pharmaceutical Analysis, SPE Applications, Forensics and Toxicology, Clinical, antiepileptic/antiseizure, Anticonvulsant/antiepiletic/antiseizure, Pharmaceutical & Clinical applications, anxiolytics/sedatives, Antiepileptic/antiseizure
Featured Industry Clinical
Forensics and Toxicology
Legal Information Discovery is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for SPE, application for HPLC

Materials

     
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