HPLC Analysis of Tetracycline Antibiotics on Ascentis^® Phenyl

column

Ascentis Phenyl, 15 cm x 4.6 mm I.D., 5 μm particles (581616-U)

mobile phase

[A] 0.1% ammonium acetate; [B] acetonitrile with 0.1% ammonium acetate; (80:20, A:B)

flow rate

1.0 mL/min

column temp.

35 °C

detector

UV, 353 nm

injection

10 μL

sample

50 μg/mL each in 80:20, water:methanol