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

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

Materiali

Colonna analitica

N° Catalogo
Descrizione
Determinazione del prezzo

Colonna HPLC Discovery® C18

5 μm particle size, L × I.D. 15 cm × 4.6 mm

Colonnina o piastra per SPE

N° Catalogo
Descrizione
Determinazione del prezzo

Discovery® DSC-18Lt SPE Tube

bed wt. 500 mg, volume 3 mL, pk of 54

Standard

N° Catalogo
Descrizione
Determinazione del prezzo

Secobarbital solution

1 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®

Phenobarbital solution

1 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®

Butabarbital solution

1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®

Barbiturate Mix-5 solution

250 μg/mL each component in methanol, ampule of 1.0 mL, certified reference material, Cerilliant®

Pentobarbital solution

1.0 mg/mL in methanol, ampule of 1 mL, certified reference material, Cerilliant®

Mephobarbital

United States Pharmacopeia (USP) Reference Standard

CONDITIONS

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

Descrizione

Risultati analitici

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.

Note legali

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