title: Trace determination of anthracyclines in urine: a new high-performance liquid chromatography/tandem mass spectrometry method for assessing exposure of hospital personnel.
abstract: Health-care workers handling antineoplastic agents may be exposed to extremely low doses of these drugs. Very sensitive and specific analytical methods are therefore needed for biological monitoring. The aim of this study was to develop and validate a method for trace level determination of doxorubicin, epirubicin, daunorubicin and idarubicin in human urine, using epi-daunorubicin as an internal standard. Solid-phase extraction (SPE) was used for sample preparation. Urine samples were loaded onto Bond Elut C18 cartridges. The analytes were eluted in methylene chloride/2-propanol (1:1, v/v) and then evaporated to dryness. The residue was reconstituted with the mobile phase prior to high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) analysis. Quantitation of each analyte was performed using the multiple reaction monitoring (MRM) method. The urine assay was linear over the range 0.1-2.0 microg/L, with a lower limit of quantification (LLOQ) of 0.10 microg/L for doxorubicin and epirubicin, and 0.03 microg/L for daunorubicin and idarubicin. The respective limits of detection (LODs) were 0.04 and 0.01 microg/L. The precision and accuracy of the assay were determined on three different days. The within-series precision was found to be always less than 13.9% for all the analytes. The overall precision expressed as relative standard deviation (RSD) was always less than 10.6%. The recovery of anthracyclines was assessed at two concentrations of the range tested (0.1 and 2.0 microg/L) and it ranged from 87.7% (daunorubicin) to 102.0% (doxorubicin) and from 79.1% (daunorubicin) to 90.7% (idarubicin) for the lower and the higher level, respectively, with a RSD always less than 9.1%. The uncertainty of the present assay was also evaluated and the combined uncertainty was always less than 20% over all the days of the validation study. This is the first method that makes use of LC/MS/MS for the biological monitoring of occupational exposure to anthracyclines.
authors.affiliation: Laboratory for Environmental and Toxicological Testing, S. Maugeri Foundation, IRCCS, via Ferrata 8, Pavia, Italy. csottani@fsm.it
authors.forename: Cristina
authors.initials: C
authors.lastname: Sottani
authors.forename: Giovanna
authors.initials: G
authors.lastname: Tranfo
authors.forename: Maurizio
authors.initials: M
authors.lastname: Bettinelli
authors.forename: Paolo
authors.initials: P
authors.lastname: Faranda
authors.forename: Mariangela
authors.initials: M
authors.lastname: Spagnoli
authors.forename: Claudio
authors.initials: C
authors.lastname: Minoia
mesh_terms: D018943:Anthracyclines / Q000652:urine*; D000970:Antineoplastic Agents / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D008832:Microchemistry / Q000379:methods*; D016273:Occupational Exposure / Q000032:analysis*; D010564:Personnel, Hospital; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D016482:Urinalysis / Q000379:methods*
chemical_list: D018943:Anthracyclines; D000970:Antineoplastic Agents