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G000594

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

application for SPE, application for HPLC

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About This Item

technique(s)

HPLC: suitable
solid phase extraction (SPE): suitable

test parameters

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
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: 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: 2 mL 5% methanol in water; dry tube 10 min with nitrogen stream
column: Discovery RP-AmideC16, 15 cm × 4.6 mm, 5 μm particles, preceded by a 2 cm RP-AmideC16 guard column and 0.5 μm frit filter (505013)
mobile phase: [A] Methanol; [B] 1% acetic acid (17:38, A:B)
flow rate: 1 mL/min
column temp.: 30 °C
detector: UV, 272 nm
injection: 20 μL reconstituted porcine serum extract

suitability

application for HPLC
application for SPE

application(s)

clinical
pharmaceutical (small molecule)

Legal Information

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

Analyte

Description

    Caffeine
    Theobromine
    Theophylline
    7-Methylxanthine, ≥98.0% (HPLC)
    1,7-Dimethylxanthine, ~98%, solid

Analytical column

SPE tube or plate


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