title: HPLC-MS/MS determination of a peptide conjugate prodrug of doxorubicin, and its active metabolites, leucine-doxorubicin and doxorubicin, in dog and rat plasma.
abstract: A HPLC-MS/MS Electrospray (ESI) method was developed and validated to quantify a peptide conjugate prodrug of doxorubicin (Dox-Con) and its active metabolites leucine-doxorubicin (Leu-Dox) and doxorubicin (Dox) in dog and rat plasma. The analytes were extracted from plasma by solid-phase extraction on a Bond Elut C8 cartridge and eluted with chloroform-methanol (2:1). Eluates were evaporated and reconstituted in acetonitrile-5 microM sodium trifluoroacetate in 0.1% aqueous formic acid (20:80) and injected onto a Waters Oasis HLB column. Analytes were eluted from the column with a solvent gradient into the mass analyzer. The ions were quantified in the selected reaction-monitoring mode (SRM), using positive ions, on a triple quadrupole mass spectrometer. The lower limits of quantification for Dox-Con, Leu-Dox, and Dox in plasma, were approximately 5, 1 (dog)/6 (rat), and 0.5 ng/ml, respectively. Intra- and inter-assay accuracy (% of nominal concentration) and precision (%CV) for all analytes were within 15 and 16%, respectively.
authors.affiliation: Laboratoires Merck-Sharp and Dohme-Chibret Route de Marsat--Riom, 63963 Clermont-Ferrand Cedex 9, France. claude_mazuel@merck.com
authors.forename: Claude
authors.initials: C
authors.lastname: Mazuel
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Grove
authors.forename: Geneviève
authors.initials: G
authors.lastname: Gerin
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Keenan
mesh_terms: D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004285:Dogs; D004317:Doxorubicin / Q000097:blood* / Q000378:metabolism; D004355:Drug Stability; D005260:Female; D005615:Freezing; D007930:Leucine / Q000097:blood*; D008297:Male; D010455:Peptides / Q000097:blood* / Q000378:metabolism; D011355:Prodrugs / Q000032:analysis / Q000378:metabolism*; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D010455:Peptides; D011355:Prodrugs; D004317:Doxorubicin; D007930:Leucine