LC/MS Analysis of Bile Acids and Their Conjugates on Ascentis^® Express C18

sample/matrix

Plasma proteins were removed with addition of 900 μL of acetonitrile, containing deuterated internal standards, to 250 μL of human EDTA plasma. The mixture was vortexed, (centrifuged and the supermatant evaporated before being reconstituted in a 50:50 solution of methanol and water. 10 μL (corresponding to 8.57 μL of plasma) was injected into the HPLC.)

column

Ascentis Express C18, 15 cm x 4.6 mm I.D., 2.7 μm particles (53829-U)

mobile phase

[A]: Water (5 mM ammonium acetate and 0.012% formic acid); [B]: Methanol (5 mM ammonium acetate and 0.012% formic acid)

gradient

70 to 95% B in 10 minutes held 4 minutes

flow rate

0.6 mL/min

column temp.

40 °C

detector

ESI(-), MRM Mode