HPLC Analysis of Levothyroxine and Liothyronine on Ascentis^® Express ES-Cyano

column

Ascentis Express ES-Cyano, 10 cm x 2.1 mm I.D., 2.7 μm particles (53491-U)

mobile phase

[A] water with 0.05% phosphoric acid; [B] acetonitrile with 0.05% phophoric acid; (60:40, A:B)

flow rate

1.5 mL/min

pressure

3270 psi (225 bar)

column temp.

30 °C

detector

UV, 225 nm

injection

5 μL

sample

20 μg/mL in mobile phase