SPE (Solid Phase Extraction)
acidified horse serum spiked with indapamide at 10 mg/mL
Discovery DSC-18, 50 mg/1 mL (52601-U)
1 mL methanol; 1 mL 10 mM potassium phosphate, pH 3
1 mL
10 mM potassium phosphate, pH 3
1 mL 60% methanol in DI water.
Discovery C18, 15 cm x 4.6 mm I.D., 5 μm particles (504955)
[A] 10 mM KH2PO4, pH 3 (adjusted with H3PO4): [B] acetonitrile; (60:40, A:B)
1 mL/min
35 °C
UV, 234 nm
10 μL
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.
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