Determination of the vitamin D analog EB 1089 (seocalcitol) in human and pig serum using liquid chromatography-tandem mass spectrometry.
A liquid chromatographic-tandem mass spectrometric assay in human and pig serum has been developed for quantitative analysis of EB 1089 (seocalcitol). EB 1089 is a novel vitamin D analog under development for the treatment of cancer. The analyte was extracted from serum after protein precipitation using an automated solid-phase extraction procedure involving both a reversed-phase and normal-phase procedure on a single C18 cartridge. The analytical chromatography was performed using a Symmetri C8 50x2.1 mm, 3.5 microm column. The mobile phase was a linear gradient from 75% to 99% methanol with a constant concentration of 2 mM ammonium acetate. EB 1089 and the internal standard [d6]-EB 1089 were detected by using MS-MS. The ion source was operated in the positive electrospray ionisation (ESI) mode. The assay is specific, sensitive, and has a capacity of more than 100 samples per day, with a limit of quantitation of 10 pg ml(-1) for a 1.0-ml sample aliquot. It is now used for routine analysis in connection with pharmacokinetic studies in humans and toxicokinetic studies in pigs.
Kissmeyer
A M
AM
Department of Pharmacokinetics and Metabolism, Leo Pharmaceutical Products, Ballerup, Denmark. anne-marie.kissmeyer@leo-pharma.com
Sonne
K
K
Binderup
E
E
D000818:Animals; D000970:Antineoplastic Agents / Q000032:analysis; D002117:Calcitriol / Q000031:analogs & derivatives* / Q000032:analysis; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D011786:Quality Control; D015203:Reproducibility of Results; D013552:Swine; D014807:Vitamin D / Q000031:analogs & derivatives / Q000032:analysis*
D000970:Antineoplastic Agents; D014807:Vitamin D; D002117:Calcitriol; C078903:seocalcitol