Determination of aristolochic acid I and its metabolites in cell culture with a hyphenated high-performance liquid chromatographic technique for cell toxicology.
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.
Yuan
Jinbin
J
State Key Laboratory of Chemo/Biosensing & Chemometrics, Chemistry & Chemical Engineering College, Hunan University, Changsha 410082, China.
Luo
Xizhen
X
Guo
Manli
M
Wu
Jiangang
J
Yang
Wuliang
W
Yu
Riyue
R
Yao
Shouzhuo
S
D034341:Aristolochic Acids / Q000032:analysis* / Q000378:metabolism*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D008099:Liver / Q000166:cytology; D014116:Toxicology / Q000295:instrumentation / Q000379:methods
D034341:Aristolochic Acids; C000228:aristolochic acid I