HPLC Analysis of Bronchodilators Theophylline and Other Caffeine Metabolites in Serum on Discovery® RP-AmideC16 after SPE using Discovery® DSC-18

HPLC Analysis of Bronchodilators Theophylline and Other Caffeine Metabolites in Serum on Discovery® RP-AmideC16 after SPE using Discovery® DSC-18

Conditions

sample preparation 1 mL porcine serum spiked with 0.1 µg/mL, 0.50 µg/mL, or 1.0 µg/mL each analyte
sample preparation 2 mL 5% methanol in water; dry tube 10 min with nitrogen stream
sample preparation 1 mL methanol; evaporate to dryness with nitrogen stream at room temperature; reconstitute in 200 µL mobile phase containing 0.2 µg/mL or 7-methylxanthine (IS)
sample preparation SPE (Solid Phase Extraction)
SPE tube/cartridge Discovery® DSC-18, 500 mg/3 mL (52603-U)
condition 2 mL methanol, then 2 mL water
column Discovery RP-AmideC16, 15 cm x 4.6 mm, 5 µm particles, preceded by a 2 cm RP-AmideC16 guard column and 0.5 µm frit filter (505013)
column temp. 30 °C
mobile phase [A] Methanol; [B] 1% acetic acid (17:38, A:B)
flow rate 1 mL/min
injection 20 µLreconstituted porcine serum extract
detector UV, 272 nm

Description

Categories Analytical Chromatography, SPE Applications, Bronchodilators, Pharmaceutical & Clinical applications
Featured Industry Clinical
Pharmaceutical (small molecule)
Legal Information Discovery is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for SPE, application for HPLC

Materials

     
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