title: Quantitation and metabolism of mitoquinone, a mitochondria-targeted antioxidant, in rat by liquid chromatography/tandem mass spectrometry.
abstract: Mitoquinone (MitoQ10 mesylate) is a mitochondria-targeted antioxidant undergoing development for the treatment of neurodegenerative diseases. The aim of this study was to develop and validate an assay based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) to determine mitoquinone and to detect and identify the metabolites of MitoQ10 in rat plasma after an oral dose. After a simple protein precipitation step, plasma samples were analyzed by reversed-phase liquid chromatography using gradient elution with acetonitrile/water/formic acid. Electrospray ionization in the positive ion mode with multiple reaction monitoring (MRM) was used to analyze mitoquinone employing the deuterated compound (d3-MitoQ10 mesylate) as internal standard. The calibration curve for mitoquinone was linear over the concentration range 0.5-250 ng/mL with a correlation coefficient>0.995. The method was sensitive (limit of quantitation 0.5 ng/mL) and had acceptable accuracy (relative error<8.7%) and precision (intra- and inter-day coefficient of variation<12.4%). Recoveries of mitoquinone at concentrations of 1.5, 20 and 200 ng/mL were in the range 87-114%. The method was successfully applied to a pharmacokinetic study in rat after a single oral dose in which four metabolites of MitoQ10 were tentatively identified as hydroxylated MitoQ10, desmethyl MitoQ10 and the glucuronide and sulfate conjugates of the quinol form of MitoQ10.
authors.affiliation: School of Pharmacy, University of Otago, Dunedin, P.O. Box 913, New Zealand. yan.li@stonebow.otago.ac.nz
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Hu
authors.initials: H
authors.lastname: Zhang
authors.forename: J Paul
authors.initials: JP
authors.lastname: Fawcett
authors.forename: Ian G
authors.initials: IG
authors.lastname: Tucker
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics / Q000494:pharmacology; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D008297:Male; D008928:Mitochondria / Q000187:drug effects; D015394:Molecular Structure; D009943:Organophosphorus Compounds / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014451:Ubiquinone / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology / Q000031:analogs & derivatives
chemical_list: D000975:Antioxidants; D009943:Organophosphorus Compounds; D014451:Ubiquinone; C429014:mitoquinone