LC/MS Analysis of Conjugated Bile Acids on Ascentis^® Express RP-Amide

column

Ascentis Express RP-Amide, 5 cm x 4.6 mm I.D., 2.7 μm particles (53922-U)

mobile phase

[A] 10 mM ammonium acetate, pH 3 with acetic acid; [B] acetonitrile

gradient

25 to 40% B in 10 min

flow rate

1.0 mL/min (postcolumn split to the MS)

column temp.

35 °C

detector

MS, ESI(-), SIR

injection

5 μL

sample

10 mg/L in mobile phase A