title: A strategy for identification of drug metabolites from dried blood spots using triple-quadrupole/linear ion trap hybrid mass spectrometry.
abstract: Discovery stage studies that address issues of absorption, distribution, metabolism and excretion (ADME) are vital for lead optimization resulting in new drug candidates. Often pharmacokinetics (PK) is assessed in these experiments without regard for the metabolism of the compound or the potential for metabolites to circulate in vivo. This work presents a strategy for drug level determination and detection of metabolites using dried blood spots for sample collection. Initially, metabolites are detected from microsomal incubations and characterized using tandem mass spectrometry. Data dependent enhanced MS and enhanced product ion (EMS-EPI) scanning with dynamic background subtraction was used on a hybrid quadruple linear ion trap mass spectrometer. On-the-fly background subtraction greatly improved the detection of metabolites. These data were used to build a multiple reaction monitoring (MRM) method for the parent and metabolites. MRM-EPI scanning was used to analyze the extracted dried blood spots from the PK study. Circulating metabolites were detected using MRM and their identities confirmed on the basis of fragment ion spectra collected simultaneously. The use of dried blood spots provides a means for re-analysis of PK samples for metabolite identification without the need for complex sample storage and preparation. Both parent compound and metabolite information can be collected in these studies, resulting in a savings of time and resources.
authors.affiliation: Merck Frosst Canada Inc., 16711 Trans Canada Hwy. Kirkland, Quebec, H9H 3L1, Canada.
authors.forename: Timo
authors.initials: T
authors.lastname: Mauriala
authors.forename: Nathalie
authors.initials: N
authors.lastname: Chauret
authors.forename: Renata
authors.initials: R
authors.lastname: Oballa
authors.forename: Deborah A
authors.initials: DA
authors.lastname: Nicoll-Griffith
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Bateman
mesh_terms: D000818:Animals; D001800:Blood Specimen Collection / Q000379:methods*; D056657:Cathepsin K; D002403:Cathepsins / Q000037:antagonists & inhibitors; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics; D008862:Microsomes, Liver / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D004791:Enzyme Inhibitors; D002403:Cathepsins; D056657:Cathepsin K; C534133:Ctsk protein, rat