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

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

Conditions

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)
column temp. 40 °C
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
detector ESI(-), MRM Mode

Description

Analysis Note A fast robust LC-MS/MS method was developed on a C18 Ascentis Express Fused-CoreR Particle Column allowing the 15 bile acid species to be measured individually rather than the alternative measurement of a total concentration by colourimetric kinetic enzyme assays.
Categories Analytical Chromatography, Clinical, organic acids, Organic Acids
Featured Industry Clinical
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for HPLC

Materials

     
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