HPLC Analysis of Barbiturates in Serum on Discovery^® C18 after SPE using Discovery^® DSC-18Lt

技术

HPLC: suitable
solid phase extraction (SPE): suitable

试验参数

sample preparation: SPE (Solid Phase Extraction)
sample/matrix: 0.5 mL porcine serum spiked with 0.5 μg/mL or 1.0 μg/mL each analyte then diluted with 0.5 mL water
SPE tube/cartridge: Discovery DSC-18Lt, 500 mg/3 mL (52613-U)
condition: 2 mL methanol; 2 mL DI water
sample addition: 1 mL at 0.75 mL/min
washing: 2 mL 5% methanol, then vacuum or air dry for 5-10 min
elution: 1-2 mL methanol
eluate post-treatment: dry eluate with nitrogen purge (40 °C; 15-20 min), reconstitute in 20 μL mobile phase
column: Discovery C18, 15 cm × 4.6 mm, 5 μm preceded by a 2 cm C18 guard column and 0.5 μm frit filter (504955)
mobile phase: [A] methanol; [B] water (40:60, A:B)
flow rate: 1 mL/min
column temp.: 30 °C
detector: UV, 214 nm
injection: 30 μL, diluted porcine serum extract

适用性

application for HPLC
application for SPE

应用

forensics and toxicology