title: Determination of dalcetrapib by liquid chromatography-tandem mass spectrometry.
abstract: The cholesteryl ester transfer protein modulator dalcetrapib is currently under development for the prevention of dyslipidemia and cardiovascular disease. Dalcetrapib, a thioester, is rapidly hydrolyzed in vivo to the corresponding thiophenol which in turn is further oxidized to the dimer and mixed disulfides (where the thiophenol binds to peptides, proteins and other endogenous thiols). These forms co-exist in an oxidation-reduction equilibrium via the thiol and cannot be stabilized without influencing the equilibrium, hence specific determination of individual components, i.e., in order to distinguish between the free thiol, the disulfide dimer and mixed disulfide adducts, was not pursued for routine analysis. The individual forms were quantified collectively as dalcetrapib-thiol (dal-thiol) after reduction under basic conditions with dithiothreitol to break disulfide bonds and derivatization with N-ethylmaleimide to stabilize the free thiol. The S-methyl and S-glucuronide metabolites were determined simultaneously with dal-thiol with no effect from the derivatization procedure. Column-switching liquid chromatography-tandem mass spectrometry provided a simple, fast and robust method for analysis of human and animal plasma and human urine samples. Addition of the surfactant Tween 80 to urine prevented adsorptive compound loss. The lower limits of quantitation (LLOQ) were 5 ng/mL for dal-thiol, and 5 ng/mL for the S-methyl and 50 ng/mL for the S-glucuronide metabolites. Using stable isotope-labeled internal standards, inter- and intra-assay precisions were each <15% (<20% at LLOQ) and accuracy was between 85 and 115%. Recovery was close to 100%, and no significant matrix effect was observed.
authors.affiliation: Non-Clinical Drug Safety, Pharma Research and Early Development, F. Hoffmann-La Roche Ltd., Basel, Switzerland.
authors.forename: Katja
authors.initials: K
authors.lastname: Heinig
authors.forename: Franz
authors.initials: F
authors.lastname: Bucheli
authors.forename: Olaf
authors.initials: O
authors.lastname: Kuhlmann
authors.forename: Manfred
authors.initials: M
authors.lastname: Zell
authors.forename: Axel
authors.initials: A
authors.lastname: Pähler
authors.forename: Elke
authors.initials: E
authors.lastname: Zwanziger
authors.forename: Günter
authors.initials: G
authors.lastname: Gross
authors.forename: Joseph
authors.initials: J
authors.lastname: Tardio
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Ishikawa
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamashita
mesh_terms: D000577:Amides; D000818:Animals; D000924:Anticholesteremic Agents / Q000493:pharmacokinetics*; D002853:Chromatography, Liquid / Q000379:methods*; D006224:Cricetinae; D004952:Esters; D020719:Glucuronides / Q000032:analysis; D006801:Humans; D057230:Limit of Detection; D008252:Macaca fascicularis; D008745:Methylation; D051379:Mice; D011136:Polysorbates / Q000737:chemistry; D011817:Rabbits; D051381:Rats; D015203:Reproducibility of Results; D013045:Species Specificity; D013438:Sulfhydryl Compounds / Q000378:metabolism / Q000493:pharmacokinetics*; D013501:Surface-Active Agents / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000577:Amides; D000924:Anticholesteremic Agents; D004952:Esters; D020719:Glucuronides; D011136:Polysorbates; D013438:Sulfhydryl Compounds; D013501:Surface-Active Agents; C411602:dalcetrapib