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.

Quantitative liquid chromatographic analysis of anthracyclines in biological fluids

“…The analytical aspects of the trace analysis of doxorubicin, epirubicin, daunorubicin and idarubicin in urine have primarily been described by Sottani and coworkers, who developed and validated two tandem mass spectrometric methods with minor differences [99,100]. Since larger starting volumes are used, solid phase extraction is preferred.…”

Selective determination of doxorubicin and doxorubicinol in rat plasma by HPLC with photosensitization reaction followed by chemiluminescence detection

“…To date, a number of publications have been reported of the determination of DXR and its metabolite DXR-ol including capillary electrophoresis (CE) with UV detection [4] or laser-induced fluorescence (LIF) detection [5][6][7], radioimmunoassay [8], high performance liquid chromatography (HPLC) with ultraviolet (UV) detection [9][10], fluorescence detection [11][12][13][14][15][16][17][18][19][20][21], chemiluminescence detection [22], electrochemical detection [23] and mass spectrometric detection [25][26][27][28][29]. However, many of these methods lack the enough sensitivity and selectivity which enable the determination of DXR and DXR-ol at small plasma concentration while the high cost and the need for technical experience limits the use of the others.…”

“…Some of them include time-consuming solid-phase extraction processes [23][24][25][26], or liquid-liquid extraction procedures [4,12,17,20,28]. The instability of DXR and DXR-ol in plasma also limits the utility of otherwise promising extraction strategies.…”

Antineoplastic drugs and their analysis: a state of the art review

“…320 lower than 0.01 ng mL −1 and 0.1 ng mL −1 were obtained, respectively, for the LC-MS analysis of anthracyclines in urine treated by SPE. 335 LC-LIF-MS was also developed for studying in vitro metabolism of doxorubicin: quantification was performed on the results obtained with LIF detection (LOD ∼ 1 µg mL −1 ) and metabolites identification was performed with MS detection. 339 LC-ELSD was also applied for the analysis of daunorubicin and its degradation products for a stability study of injectable solution of anthracycline (0.25 mg mL −1 was the lowest concentration of standard solutions tested), but the interest of ELSD for anthracyclines is not obvious since these molecules can be easily detected by UV, FD and MS. 340 As already mentioned, CE was another separation technique widely used for the determination of anthracyclines.…”