title: Determination of emodin in L-02 cells and cell culture media with liquid chromatography-mass spectrometry: application to a cellular toxicokinetic study.
abstract: The emodin-involved hepatotoxicity has been gaining increasing attention. The purpose of the present study was to evaluate the cytotoxicity of emodin on cultured human liver cells (L-02) and predict the possible relation between its cytotoxicity and cellular toxicokinetics. Cell viability and cell damage were assessed by Cell Counting Kit-8 (CCK-8) assay and phase-contrast microscopy, respectively. Cytotoxicity tests demonstrated a concentration- and time-dependent toxic effect of emodin on L-02 cells. Furthermore, emodin at concentration of 30μM led to a significant apoptosis in a time-dependent manner supported by the morphological changes of drug-treated cells. In addition, to elucidate the toxicokinetic characteristics of emodin, a highly sensitive and selective liquid chromatography-mass spectrometry (LC-MS) method was employed and validated for detecting the dynamic alteration of emodin in cells and cell culture media. The proposed method appeared to be suitable for the analysis of emodin with desirable linearity (r(2)>0.99), and satisfying precision being less than 8.7%. The range of recoveries of this method was 90.2-101.9%. The preliminary cellular toxicokinetic study revealed a time-dependent intracellular accumulation of emodin, which was consistent with its in vitro toxic effects. These findings confirmed the cytotoxicity of emodin against L-02 cells and displayed the cytotoxic manner of emodin in terms of its cellular uptake and accumulation in L-02 cells.
authors.affiliation: State Key Laboratory of Natural Medicines (China Pharmaceutical University), No. 24 Tong jia Lane, Nanjing 210009, China.
authors.forename: Cui-Li
authors.initials: CL
authors.lastname: Li
authors.forename: Jiang
authors.initials: J
authors.lastname: Ma
authors.forename: Li
authors.initials: L
authors.lastname: Zheng
authors.forename: Hui-Jun
authors.initials: HJ
authors.lastname: Li
authors.forename: Ping
authors.initials: P
authors.lastname: Li
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002853:Chromatography, Liquid / Q000379:methods*; D003470:Culture Media / Q000737:chemistry*; D004642:Emodin / Q000032:analysis* / Q000493:pharmacokinetics / Q000633:toxicity*; D006801:Humans; D008099:Liver / Q000187:drug effects; D013058:Mass Spectrometry / Q000379:methods*; D008858:Microscopy, Phase-Contrast / Q000379:methods; D010599:Pharmacokinetics
chemical_list: D003470:Culture Media; D004642:Emodin