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Merck

G001056

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

application for SPE, application for HPLC

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About This Item

Methode(n)

HPLC: suitable
solid phase extraction (SPE): suitable

Testparameter

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

Eignung

application for HPLC
application for SPE

Anwendung(en)

forensics and toxicology

Hinweis zur Analyse

Barbiturates are commonly abused and among the most widely tested compounds in clinical, forensic, or therapeutic drug monitoring applications. Shown here is the baseline separation of a set of barbiturates on a Discovery C18 HPLC column after extraction from spikd serum with Discovery DSC-18Lt SPE. Highest grade HPLC solvents were used to supply low background interference and low particulate contamination for robust, trouble-free operation. Cerilliant and Sigma-Aldrich reference standards provided reliable identification and quantification. A Zymark® RapidTrace® SPE Workstation was used in this study.

Rechtliche Hinweise

Discovery is a registered trademark of Merck KGaA, Darmstadt, Germany
RapidTrace is a registered trademark of Zymark Corp.
Zymark is a registered trademark of Sotax Corporation

Analyt

Beschreibung

    Phenobarbital solution, 1 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
    Aprobarbital
    Butabarbital solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
    Mephobarbital, United States Pharmacopeia (USP) Reference Standard
    Pentobarbital solution, 1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®
    Secobarbital solution, 1 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®

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