title: Development and validation of a LC-MS assay for the quantification of ikh12 a novel anti-tumor candidate in rat plasma and tissues and its application in a pharmacokinetic study.
abstract: IKH12 is a novel histone deacetylase 6 selective inhibitor. A rapid and sensitive liquid chromatography tandem mass spectrometry method was developed and validated for the quantification of IKH12 in rat plasma and tissue with kendine 91 as internal standard (IS). The samples were prepared by liquid-liquid extraction with tert-butyl methyl ether. The chromatographic separation was accomplished by using a Zorbax Extend C18 4.6 × 150 mm, 5 µm column, with a mobile phase consisting of methanol and 0.1% formic acid (75:25 v/v). Multiple reaction monitoring, using electrospray ionization in positive ion mode, was employed to quantitatively detect IKH12 and IS. The monitored transitions were set at m/z 418 → 252 and 444 → 169 for IKH12 and kendine 91, respectively. The calibration curve was linear over the concentration range 2-1000 ng mL(-1) . The intra- and inter-assay precision and accuracy of the quality controls and the limit of quantification were satisfactory in all cases (according to European Medicines Agency guidelines). Stability studies showed that plasma samples were stable in the chromatography rack for 24 h and at -80°C for 2 months and also after three freeze-thaw cycles. This method was successfully applied to a pharmacokinetic study of IKH12 in rat.
authors.affiliation: Ikerchem S.L, Paseo Mikeletegi 69, San Sebastián, Spain.
authors.forename: Dorleta
authors.initials: D
authors.lastname: Otaegui
authors.affiliation: Ikerchem S.L, Paseo Mikeletegi 69, San Sebastián, Spain.
authors.forename: Carme
authors.initials: C
authors.lastname: Masdeu
authors.affiliation: Ikerchem S.L, Paseo Mikeletegi 69, San Sebastián, Spain.
authors.forename: Eneko
authors.initials: E
authors.lastname: Aldaba
authors.affiliation: Ikerchem S.L, Paseo Mikeletegi 69, San Sebastián, Spain.
authors.forename: Yosu
authors.initials: Y
authors.lastname: Vara
authors.affiliation: Ikerchem S.L, Paseo Mikeletegi 69, San Sebastián, Spain.|Organic Chemistry Department, University of the Basque Country UPV/EHU, San Sebastián, Spain.
authors.forename: Aizpea
authors.initials: A
authors.lastname: Zubia
authors.affiliation: Ikerchem S.L, Paseo Mikeletegi 69, San Sebastián, Spain.|Applied Chemistry Department - Inorganic Chemistry, Chemistry Faculty, University of the Basque Country UPV/EHU, San Sebastián, Spain.
authors.forename: Eider
authors.initials: E
authors.lastname: San Sebastian
authors.affiliation: Ikerchem S.L, Paseo Mikeletegi 69, San Sebastián, Spain.
authors.forename: Maria
authors.initials: M
authors.lastname: Alcalá
authors.affiliation: Ikerchem S.L, Paseo Mikeletegi 69, San Sebastián, Spain.
authors.forename: Sergio
authors.initials: S
authors.lastname: Villafruela
authors.affiliation: Organic Chemistry Department, University of the Basque Country UPV/EHU, San Sebastián, Spain.
authors.forename: Fernando P
authors.initials: FP
authors.lastname: Cossío
authors.affiliation: Pharmacy and Pharmaceutical Technology Department, University of the Basque Country UPV/EHU, Vitoria, Spain.
authors.forename: Alicia
authors.initials: A
authors.lastname: Rodriguez-Gascón
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056572:Histone Deacetylase Inhibitors / Q000097:blood*; D006877:Hydroxamic Acids / Q000097:blood*; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction / Q000379:methods; D008297:Male; D008738:Methyl Ethers / Q000737:chemistry; D011392:Proline / Q000031:analogs & derivatives* / Q000097:blood; D017208:Rats, Wistar; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000970:Antineoplastic Agents; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; C000604166:IKH 12; D008738:Methyl Ethers; C043243:methyl tert-butyl ether; D011392:Proline
keywords: IKH12; analytical validation; histone deacetylase inhibitor; liquid chromatography; mass spectrometry; pharmacokinetics; selective inhibitor