title: Determination of 4-demethoxy-3'-deamino-3'-aziridinyl-4'-methylsulphonyldaunorubicin+ ++ and its 13-hydroxy metabolite by direct injection of human plasma into a column-switching liquid chromatography system with mass spectrometric detection.
abstract: A selective, sensitive and fully automated column-switching LC system using direct injection of human plasma followed by mass spectrometry (MS) detection was developed and validated to determine the concentrations of 4-demethoxy-3'-deamino-3'-aziridinyl-4'-methylsulphonyldaunorubicin++ + (PNU-159548) and its 13-hydroxy metabolite (PNU-169884). A 50-microl human plasma sample was directly introduced into a C4-alkyl-diol silica clean-up column separating analytes from proteins and polar endogenous compounds using water and methanol as the mobile phase. The fraction containing PNU-159548 and its metabolite was back-flushed and transferred to the analytical column. The compounds were separated using a Zorbax SB C8 column (150x4.6 mm, 5 microm) under gradient conditions with the mobile phase containing acetonitrile and 2 mM ammonium formate, pH 3.5. MS detection was by atmospheric pressure ionisation with multiple reaction monitoring in positive ion mode. Linearity was demonstrated over the calibration range of 0.051-10.291 ng/ml for PNU-159548 and 0.104-10.434 ng/ml for PNU-169884. The assay was validated with respect to accuracy, precision and analyte stability. On the basis of the validation data, the developed analytical method was found to be suitable for use in Phase I clinical studies.
authors.affiliation: Drug Metabolism Research, Pharmacia and Upjohn, Milan, Italy. massimo.breda@eu.pnu.com
authors.forename: M
authors.initials: M
authors.lastname: Breda
authors.forename: G
authors.initials: G
authors.lastname: Basileo
authors.forename: G
authors.initials: G
authors.lastname: Fonte
authors.forename: J
authors.initials: J
authors.lastname: Long
authors.forename: C A
authors.initials: CA
authors.lastname: James
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000097:blood*; D001331:Automation; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003630:Daunorubicin / Q000031:analogs & derivatives* / Q000097:blood; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: C402179:4-demethoxy-3'-deamino-3'-aziridinyl-4'-methylsulfonyldaunorubicin; D000903:Antibiotics, Antineoplastic; C402180:PNU 169884; D003630:Daunorubicin