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HPLC Analysis of Tetracycline Antibiotics on Ascentis® Express C18

HPLC Analysis of Tetracycline Antibiotics on Ascentis® Express C18


column Ascentis Express C18, 15 cm x 3.0 mm I.D., 2.7 µm particles (53816-U)
column temp. 30 C
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)
sample 100 ng/mL of each compound in acetontrile:water:formic acid (5:94.9:0.1)
injection 50 µl
detector UV, 355 nm


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.
Categories Analytical Chromatography, Antibiotics, Antimicrobials
Featured Industry Food and Beverages
Pharmaceutical (small molecule)
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for HPLC


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