title: Determination of ximelagatran, an oral direct thrombin inhibitor, its active metabolite melagatran, and the intermediate metabolites, in biological samples by liquid chromatography-mass spectrometry.
abstract: Analytical methods for the determination of ximelagatran, an oral direct thrombin inhibitor, its active metabolite melagatran, and intermediate metabolites, melagatran hydroxyamidine and melagatran ethyl ester, in biological samples by liquid chromatography (LC) positive electrospray ionization mass spectrometry (MS) using selected reaction monitoring are described. Isolation from human plasma was achieved by solid-phase extraction on octylsilica. Analytes and isotope-labelled internal standards were separated by LC utilising a C(18) analytical column and a mobile phase comprising acetonitrile-4 mmol/l ammonium acetate (35:65, v/v) containing 0.1% formic acid, at a flow-rate of 0.75 ml/min. Absolute recovery was approximately 80% for ximelagatran, approximately 60% for melagatran ethyl ester and >90% for melagatran and melagatran hydroxyamidine. Limit of quantification was 10 nmol/l, with a relative standard deviation <20% for each analyte and <5% above 100 nmol/l. Procedures for determination of these analytes in human urine and breast milk, plus whole blood from rat and mouse are also described.
authors.affiliation: DMPK and Bioanalytical Chemistry, AstraZeneca R&D Mölndal, S-431 83, Mölndal, Sweden. marita.s.larsson@astrazeneca.com
authors.forename: Marita
authors.initials: M
authors.lastname: Larsson
authors.forename: Martin
authors.initials: M
authors.lastname: Ahnoff
authors.forename: Anna
authors.initials: A
authors.lastname: Abrahamsson
authors.forename: Ulrika
authors.initials: U
authors.lastname: Logren
authors.forename: Christina
authors.initials: C
authors.lastname: Fakt
authors.forename: Irene
authors.initials: I
authors.lastname: Ohrman
authors.forename: Bengt Arne
authors.initials: BA
authors.lastname: Persson
mesh_terms: D000818:Animals; D000991:Antithrombins / Q000032:analysis* / Q000378:metabolism / Q000652:urine; D001384:Azetidines / Q000032:analysis* / Q000097:blood / Q000652:urine; D001596:Benzylamines; D002853:Chromatography, Liquid / Q000379:methods*; D005998:Glycine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D051379:Mice; D008895:Milk, Human / Q000737:chemistry; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D000991:Antithrombins; D001384:Azetidines; D001596:Benzylamines; C109573:melagatran; C426686:ximelagatran; D005998:Glycine