HPLC Analysis of Amphetamines in Urine on Discovery® HS F5 after SPE using Discovery® DSC-MCAX and Standard C18
CONDITIONS
sample preparation
SPE (Solid Phase Extraction)
sample/matrix
human urine spiked amphetamine and methylamphetamine at with 2 mg/mL
SPE tube/cartridge
Discovery DSC-MCAX, 100 mg/3mL (52783-U)
condition
DSC-MCAX SPE tube: 1 mL methanol; 1 mL 50 mM ammonium acetate, pH 6.0; C-18 SPE tube: 1 mL methanol; 1 mL DI water
sample addition
1 mL
washing
DSC-MCAX SPE tube: 1 mL 50 mM ammonium acetate, pH 6.0; 1 mL 1M acetic acid; 1mL methanol; C-18 SPE tube: 1 mL DI water; 1 mL 20% methanol
elution
DSC-MCAX SPE tube: 1 mL 5% ammonium hydroxide in methanol; C-18 SPE tube: 1 mL methanol
eluate post-treatment
evaporate to dryness with nitrogen at room temperature, reconstitute in mobile phase
column
Discovery HS F5, 15 cm x 4.6 mm I.D., 5 μm particles (567516-U)
mobile phase
[A] 10 mM ammonium acetate, pH 4.5: [B] methanol; (35:65, A:B)
flow rate
2 mL/min
column temp.
40 °C
detector
UV, 210 nm
injection
10 μL
Popis
Application
Sample Pre-Treatment:
1 mL human urine was spiked with 2 mg/mL amphetamine and methylamphetamine. The spiked sample was diluted 1:1 with 1:1 with 50 mM ammonium acetate, pH 6.0.
Legal Information
Discovery is a registered trademark of Merck KGaA, Darmstadt, Germany