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

technique(s)

HPLC: suitable

test parameters

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

suitability

application for HPLC

application(s)

clinical