A new approach for the sensitive determination of DNA adduct of aristolochic acid II by using high-performance liquid chromatography with fluorescence detection.
A sensitive high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) for the determination of DNA adducts induced by nephrotoxic and carcinogenic aristolochic acid (AA) is presented. The DNA adduct of AAII (dA-AAII) was synthesized by in vitro incubation, purified by preparative HPLC, characterized using fluorescence spectroscopy and high-resolution mass spectrometry, and was used as the biomarker for AA exposure in rats. The developed HPLC-FLD method was validated and applied for the determination of dA-AAII in rat kidney tissues. The method provided a detection limit of 18.3fmol, which allowed the detection of dA-AAII in the rat kidney tissue samples collected after a single oral dose of AA. dA-AAII was detected in the kidney DNA digestion extracts of the rats that were dosed with AA at 5mg/kg and 30mg/kg at concentrations of 6.2+/-1.1 and 41.3+/-8.0 dA-AAII per 10(9) normal dA, respectively.
Chan
Wan
W
Department of Chemistry, Hong Kong Baptist University, Kowloon, Hong Kong SAR, China.
Poon
Wing Tat
WT
Chan
Yan-Wo
YW
Wan
King-Yi
KY
Cai
Zongwei
Z
D000818:Animals; D034341:Aristolochic Acids / Q000008:administration & dosage / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018736:DNA Adducts / Q000032:analysis*; D007668:Kidney / Q000737:chemistry / Q000378:metabolism; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D034341:Aristolochic Acids; D018736:DNA Adducts; C042310:aristolochic acid II