G001056

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

application for SPE, application for HPLC

Properties

• technique(s): HPLC: suitable; solid phase extraction (SPE): suitable
• test parameters: 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
• suitability: application for HPLC; application for SPE
• application(s): forensics and toxicology

Description

Analysis Note

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.

Legal Information

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

Analyte

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^®