LC/MS/MS Analysis of Tetracycline Antibiotics on Kromasil® Eternity™ C18

LC/MS/MS Analysis of Tetracycline Antibiotics on Kromasil® Eternity™ C18


column Kromasil Eternity C18, 10 cm x 2.1 mm I.D., 2.5 µm particles (K08670895)
column temp. 30 °C
mobile phase [A] 0.5% acetonitrile and 0.1% formic acid in water; [B] 0.1% formic acid in methanol:acetonitrile (1:1)
gradient Held at 10% B for 0.1 min; to 20% B in 1.9 min; held at 20% B for 3.5 min; to 80% B in 0.1 min; held at 80% B for 1.0 min; to 10% B in 0.1 min; held at 10% B for 3.3 min
flow rate 0.2 mL/min
pressure 3070 psi (212 bar)
sample each compound 50 ng/mL in acetonitrile:water (11:89)
injection 10 µL
detector MS/MS


Analysis Note The analysis of tetracycline antibiotics can be problematic via liquid chromatography-mass spectrometry due to their chelating properties resulting in compounds’ irreversible retention. This application uses the organosilane-coated silica stationary phase that prevents unwanted binding of compounds and provides good peak shape with an LC/MS-compatible mobile phase. The method also delivers good resolution of compounds from their isobaric epi-metabolites that cannot be resolved via mass spectrometry alone.
Categories Analytical Chromatography, Antibiotics, Antimicrobials
Featured Industry Food and Beverages
Pharmaceutical (small molecule)
Legal Information Kromasil is a registered trademark of EKA Chemicals AB, Eternity is a trademark of EKA Chemicals AB
suitability application for LC-MS


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