Yuan Jinbin, Luo Xizhen, Guo Manli, Wu Jiangang, Yang Wuliang, Yu Riyue, Yao Shouzhuo
State Key Laboratory of Chemo/Biosensing & Chemometrics, Chemistry & Chemical Engineering College, Hunan University, Changsha 410082, China.
Talanta. 2009 May 15;78(3):1141-7. doi: 10.1016/j.talanta.2009.01.037. Epub 2009 Jan 24.
Aristolochic acid I (AA I), a major component of the carcinogenic plant extract aristolochic acid (AA), is known to be nephrotoxic, carcinogenic and mutagenic. A simple, rapid and sensitive high-performance liquid chromatography-diode array detection-fluorescence detection (HPLC-DAD-FLD) method was developed and validated for the analysis of AA I and its metabolites in cell culture medium for the first time. The samples were prepared with ethyl acetate liquid-liquid extraction (LLE). Good separation was obtained on an ODS C(18) analytical column with 0.2% HAc/methanol gradient solution. Linearities of about three orders of magnitude were gained with correlation coefficients exceeding 0.9990. The method appears to be a suitable tool for the cellular toxicokinetic study with acceptable precisions and recoveries. Cytotoxicity of AA I on human liver cells (L-02) was investigated with morphological observation and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide) assay, cytotoxicity increased in AA I concentration-dependent manner. AA I and its metabolites were monitored with the proposed chromatographic analysis, and some preliminary toxicokinetics were investigated.
马兜铃酸I（AA I）是致癌植物提取物马兜铃酸（AA）的主要成分，已知具有肾毒性、致癌性和致突变性。首次开发并验证了一种简单、快速且灵敏的高效液相色谱 - 二极管阵列检测 - 荧光检测（HPLC - DAD - FLD）方法，用于分析细胞培养基中的AA I及其代谢产物。样品采用乙酸乙酯液 - 液萃取（LLE）进行制备。在ODS C(18)分析柱上，使用0.2% HAc/甲醇梯度溶液获得了良好的分离效果。线性范围约为三个数量级，相关系数超过0.9990。该方法似乎是进行细胞毒代动力学研究的合适工具，具有可接受的精密度和回收率。通过形态学观察和MTT（3 - (4,5 - 二甲基噻唑 - 2 - 基) - 2,5 - 二苯基四氮唑溴盐）法研究了AA I对人肝细胞（L - 02）的细胞毒性，细胞毒性呈AA I浓度依赖性增加。采用所提出的色谱分析法监测AA I及其代谢产物，并研究了一些初步的毒代动力学。
