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G005599

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

application for HPLC

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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

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.

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany

Analyte

Description

    Taurocholic acid sodium salt hydrate, ≥95% (HPLC)
    Glycoursodeoxycholic acid, ≥96.0% (TLC)
    Taurocholic acid (TCA)
    Glycocholic acid hydrate, synthetic, ≥97% (HPLC)
    Chenodeoxycholic acid
    D4-Taurodeoxycholic acid (D4-TDC)
    Sodium taurodeoxycholate hydrate, ≥95% (HPLC)
    Ursodeoxycholic acid, ≥99%
    Sodium glycochenodeoxycholate
    Glycodeoxycholic-2,2,4,4-d4 acid, ≥98 atom % D, ≥98% (CP)
    Glycodeoxycholic acid monohydrate, ≥97% (TLC)
    Cholic acid, from bovine and/or ovine, ≥98%
    Sodium taurolithocholate
    Glycolithocholic acid (GLC)
    Chenodeoxycholic acid (CDC)
    D4- Deoxycholic acid (D4-DC)
    Deoxycholic acid, ≥98% (HPLC)
    Lithocholic acid, ≥95%
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