HPLC Analysis of Furosemide (Lasix) in Horse Serum on Discovery® C18 after SPE using Discovery® DSC-18
CONDITIONS
sample preparation
SPE (Solid Phase Extraction)
sample/matrix
acidified horse serum spiked with indapamide at 10 mg/mL
SPE tube/cartridge
Discovery DSC-18, 50 mg/1 mL (52601-U)
condition
1 mL methanol; 1 mL 10 mM potassium phosphate, pH 3
sample addition
1 mL
washing
10 mM potassium phosphate, pH 3
elution
1 mL 60% methanol in DI water.
column
Discovery C18, 15 cm x 4.6 mm I.D., 5 μm particles (504955)
mobile phase
[A] 10 mM KH2PO4, pH 3 (adjusted with H3PO4): [B] acetonitrile; (60:40, A:B)
flow rate
1 mL/min
column temp.
35 °C
detector
UV, 234 nm
injection
10 μL
Description
Application
Sample Pre-treatment:
Neat horse serum was acidified with 10 mL 6 M HCl/mL serum. Internal standard (I.S.) blank was prepared by spiking acidified horse serum with indapamide at 10 mg/mL. Sample A = 10 mg/mL furosemide in I.S. blank. Sample B = 5 mg/mL furosemide in I.S. blank. Sample C = 0.5 mg/mL furosemide in I.S. blank. Sample D = 0.1 mg/mL furosemide in I.S. blank. Sample E = 0.05 mg/mL furosemide in I.S. blank.
Legal Information
Discovery is a registered trademark of Merck KGaA, Darmstadt, Germany