HPLC Analysis of Tetracycline Antibiotics on Ascentis® Express C18
Materiales
Analytical column
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Guard column
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Ascentis® Express C18, 2.7 Micron Guard Cartridge
2.7 μm particle size, L × I.D. 5 mm × 3 mm, pkg of 3 eaStandard
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also commonly purchased with this product
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Screw Top Vial, Silane-Treated
volume 2 mL, amber glass vial, O.D. × H × I.D. 12 mm × 32 mm × 4.6 mmmobile phase component
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CONDITIONS
column
Ascentis Express C18, 15 cm x 3.0 mm I.D., 2.7 μm particles (53816-U)
mobile phase
[A] 10 mM oxalic acid in water; [B] 5 mM oxalic acid in methanol:acetonitrile (1:1)
gradient
15% B for 1 min; to 20% B in 0.1 min, held at 20% for 4 min; to 80% B in 5 min; to 15% B in 0.1 min, held at 15% B for 3 min
flow rate
0.5 mL/min
pressure
3597 psi (248 bar)
column temp.
30 °C
detector
UV, 355 nm
injection
50 μL
sample
100 ng/mL of each compound in acetontrile:water:formic acid (5:94.9:0.1)
Descripción
Analysis Note
Separation of tetracycline antibiotics from their epi-metabolites was achieved on an Ascentis® Express C18 HPLC column. Oxalic acid was added to the mobile phase to prevent the loss of these metal-binding compounds in the HPLC system.
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany