title: Development of LC-MS/MS method for the simultaneous analysis of the cardioprotective drug dexrazoxane and its metabolite ADR-925 in isolated cardiomyocytes and cell culture medium.
abstract: Dexrazoxane (DEX) is the only clinically used drug effective against anthracycline-induced cardiotoxicity and extravasation injury. However, the mechanism of its cardioprotective action still remains elusive. This paucity of comprehensive data is at least partially caused by the analytical difficulties associated with selective and sensitive simultaneous determination of the parent drug and its putative active metabolite ADR-925 in the relevant biological material. The aim of this study was to develop and validate the first LC-MS/MS method for simultaneous determination of DEX and ADR-925 in the isolated rat neonatal ventricular cardiomyocytes (NVCMs) and the cell culture medium. The analysis was performed on a Synergi Polar-RP column using the gradient profile of the mobile phase composed of 2mM ammonium formate and methanol. Electrospray ionization and ion trap mass analyzer were used as ionization and detection techniques, respectively. NVCMs were precipitated with methanol and the cell culture medium samples were diluted with the same solvent prior the LC-MS/MS analysis. The method was validated within the range of 4-80pmol/10(6) NVCMs and 7-70pmol/10(6) NVCMs for DEX and ADR-925, respectively, and at the concentrations of 8-100μM for both compounds in the culture cell medium. The practical applicability of this method was confirmed by the pilot analysis of NVCMs and the corresponding cell medium samples from relevant in vitro experiment. Hence, the LC-MS/MS method developed in this study represents a modern analytical tool suitable for investigation of DEX bioactivation inside the cardiomyocytes. In addition, the basic utility of the method for the analysis of DEX and ADR-925 in plasma samples was proved in a pilot experiment.
authors: Faculty of Pharmacy in Hradec Kralove, Charles University in Prague, Heyrovskeho 1203, 500 05 Hradec Kralove, Czech Republic. petra.kovarikova@faf.cuni.cz
authors: Petra
authors: P
authors: Kovarikova
authors: Ivana
authors: I
authors: Pasakova-Vrbatova
authors: Anna
authors: A
authors: Vavrova
authors: Jan
authors: J
authors: Stariat
authors: Jiri
authors: J
authors: Klimes
authors: Tomas
authors: T
authors: Simunek
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002317:Cardiovascular Agents / Q000493:pharmacokinetics; D002478:Cells, Cultured; D002853:Chromatography, Liquid / Q000379:methods*; D005029:Ethylenediamines / Q000493:pharmacokinetics*; D005998:Glycine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D032383:Myocytes, Cardiac / Q000378:metabolism; D010865:Pilot Projects; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D011929:Razoxane / Q000493:pharmacokinetics*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002317:Cardiovascular Agents; D005029:Ethylenediamines; D011929:Razoxane; C059866:ICRF 198; D005998:Glycine