HPLC Analysis of Piroxicam and 2-Aminopyridine in Urine on Discovery® HS F5 after SPE using Discovery® DSC-MCAX
CONDITIONS
sample preparation
SPE (Solid Phase Extraction)
sample/matrix
human urine spiked with piroxicam (4 μg/mL) and 2-aminopyridine (10 μg/mL)
SPE tube/cartridge
Discovery DSC-MCAX, 100 mg/3 mL (52783-U)
condition
1 mL methanol; 1 mL 10 mM potassium phosphate, pH 3
sample addition
1 mL
washing
1 mL 10 mM potassium phosphate, pH 3; 1 mL methanol
elution
1 mL 5% ammonium hydroxide in methanol
eluate post-treatment
evaporate to dryness with nitrogen at room temperature, reconstitute in 1 mL mobile phase
column
Discovery HS F5, 15 cm x 4.6 mm I.D., 5 μm particles (567516-U)
mobile phase
[A] 10 mM potassium phosphate, pH 6: [B] acetonitrile (85:15, A:B)
flow rate
2 mL/min
column temp.
25 °C
detector
UV, 220 nm
injection
10 μL
Description
Application
Sample Pre-Treatment:
Piroxicam and 2-aminopyridine (piroxicam′s polar metabolite) was spiked into human urine at the levels of 4 and 10 μg/mL, respectively. The urine sample was diluted 1:1 with 10 mM potassium phosphate, pH 3.
Legal Information
Discovery is a registered trademark of Merck KGaA, Darmstadt, Germany