Chan Wan, Poon Wing Tat, Chan Yan-Wo, Wan King-Yi, Cai Zongwei
Department of Chemistry, Hong Kong Baptist University, Kowloon, Hong Kong SAR, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Apr 1;877(10):848-52. doi: 10.1016/j.jchromb.2009.02.007. Epub 2009 Feb 11.
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.
本文介绍了一种用于测定肾毒性和致癌性马兜铃酸（AA）诱导的DNA加合物的灵敏的高效液相色谱荧光检测法（HPLC-FLD）。AAII的DNA加合物（dA-AAII）通过体外孵育合成，经制备型HPLC纯化，用荧光光谱和高分辨率质谱进行表征，并用作大鼠AA暴露的生物标志物。所建立的HPLC-FLD方法经过验证，并应用于大鼠肾组织中dA-AAII的测定。该方法的检测限为18.3飞摩尔，可检测单次口服AA后收集的大鼠肾组织样本中的dA-AAII。在分别以5mg/kg和30mg/kg剂量给予AA的大鼠肾DNA消化提取物中检测到dA-AAII，浓度分别为每10⁹正常dA中6.2±1.1和41.3±8.0个dA-AAII。
