Tandem Laser-Induced Fluorescence and Mass Spectrometry Detection for High-Performance Liquid Chromatography Analysis of the in Vitro Metabolism of Doxorubicin

摘要

Structural characterization, identification, and quantificationnof xenobiotics and their metabolic products commonlynrequire the use of at least two different techniques.nThis has been the case in the analysis of metabolicnproducts of doxorubicin, a widely used fluorescent anthracyclinenfor the treatment of tumors and leukemia. Innthis work, we combine high-performance liquid chromatographyn(HPLC) with a tandem laser-induced fluorescencen(LIF) and mass spectrometry (MS) detection schemenfor the characterization of doxorubicin and its metabolitesnproduced in the postmitochondrial fraction prepared fromnFischer 344 rat liver. LIF detection allowed quantificationnof the metabolic compounds while MS detection aided innthe identification of the metabolites. Using this HPLC-LIFMSnmethodology, the disapperance of doxorubicin and thenappearance of 7-deoxydoxorubicinone and 7-deoxydoxorubicinolonenwere monitored over the course of 40 min.nThis application demonstrates the potential of the tandemnLIF-MS detection scheme in quantification and characterizationnof biotransformations of fluorescent xenobioticsnof biomedical and environmental relevance. Furthermore,nthis detection scheme would be particularly relevant innthe analysis of fluorescent analytes in complex samplesnand in validation of methods for the analysis of suchnsamples that typically rely only on LIF detectors.