Improved preparation and identification of aristolochic acid-DNA adductsby solid-phase extraction with liquid chromatography-tandemmass spectrometry
GAO Haiyan , GUO Fangqiu , FENG Feng , YIN Junfa , SONG Maoyong , WANG Hailin
DOI:
Received January 13, 2009,Revised April 14, 2009, Accepted , Available online
Volume 21,2009,Pages 1769-1776
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Aristolochic acid (AA) is a known nephrotoxin and potential carcinogen, which can form covalent DNA adducts after metabolic
activation in vivo and in vitro. A simple method for preparation and characterization of aristolochic acid-DNA adducts was developed.
Four AA-adducts were synthesized by a direct reaction of AAI/AAII with 2’-deoxynucleosides. The reaction mixture was first
cleaned-up and pre-concentrated using solid phase extraction (SPE), and further purified by a reversed-phase high performance liquid
chromatography (HPLC). By the application of developed SPE procedure, matrices and byproducts in reaction mixture could be greatly
reduced and adducts of high purity (more than 94% as indicated by HPLC) were obtained. The purified AA-DNA adducts were
identified and characterized with liquid-electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-Q-TOF-MS/MS)
and LC-Diode array detector-fluorescence (LC-DAD-FL) analysis. This work provides a robust tool for possible large-scale preparation
of AA-DNA adduct standards, which can promote the further studies on carcinogenic and mutagenic mechanism of aristolochic acids.
