Determination of melagatran, a novel, direct thrombin inhibitor, in human plasma and urine by liquid chromatography-mass spectrometry.
Analytical methods for the determination of melagatran (H 319/68) in biological samples by liquid chromatography (LC)-positive electrospray ionization mass spectrometry using multiple reaction monitoring are described. Melagatran in plasma was isolated by solid-phase extraction on octylsilica, either in separate extraction tubes or in 96-well plates. Absolute recovery of melagatran from plasma was >92%. Melagatran and the internal standard, H 319/68 D2 13C2, were separated from other sample components by LC utilizing a C18 stationary phase and a mobile phase comprising 35% acetonitrile and 0.08% formic acid in 0.0013 mol/l ammonium acetate solution. After dilution, urine was injected directly onto the LC column and subjected to gradient LC. The relative standard deviation was 1-5% for concentrations above the limit of quantification, which was estimated for plasma at 10 or 25 nmol/l for sample volumes of 500 or 200 microl, respectively, and 100 nmol/l for urine.
Larsson
Marita
M
DMPK and Bioanalytical Chemistry, AstraZeneca R&D Mölndal, Sweden. marita_larsson17@hotmail.com
Logren
Ulrika
U
Ahnoff
Martin
M
Lindmark
Bo
B
Abrahamsson
Peter
P
Svennberg
Henrik
H
Persson
Bengt-Arne
BA
D001384:Azetidines; D001596:Benzylamines; D002853:Chromatography, Liquid / Q000379:methods; D005343:Fibrinolytic Agents / Q000032:analysis* / Q000097:blood / Q000652:urine; D005998:Glycine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D013058:Mass Spectrometry; D012015:Reference Standards; D012680:Sensitivity and Specificity
D001384:Azetidines; D001596:Benzylamines; D005343:Fibrinolytic Agents; C109573:melagatran; D005998:Glycine