G003766
HPLC Analysis of Diazepam and Metabolites in Serum on Discovery® C18 after SPE using Discovery® DSC-8
application for SPE, application for HPLC
About This Item
technique(s)
HPLC: suitable
solid phase extraction (SPE): suitable
test parameters
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
suitability
application for HPLC
application for SPE
application(s)
clinical
forensics and toxicology
Legal Information
analytical column
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