title: Imitation of drug metabolism in human liver and cytotoxicity assay using a microfluidic device coupled to mass spectrometric detection.
abstract: In this work, we developed a microfluidic device for the imitation of drug metabolism in human liver and its cytotoxicity on cells. The integrated microfluidic device consists of three sections: (1) bioreactors containing poly(ethylene) glycol (PEG) hydrogel encapsulated human liver microsomes (HLMs); (2) cell culture chambers for cytotoxicity assay; and (3) integrated micro solid-phase extraction (SPE) columns to desalt and concentrate the products of enzymatic reaction. To verify the feasibility of the integrated microchip, we studied uridine 5'-diphosphate-glucuronosyltransferase (UGT) metabolism of acetaminophen (AP) and the cytotoxicity of products on HepG2 cells. The products of the reaction in one region of the device were injected into the cell culture chamber for cytotoxicity assay, while those in another region were directly detected online with an electrospray ionization quadrupole time-of-flight mass spectrometer (ESI-Q-TOF MS) after micro-SPE pre-treatment. Semiquantitative analysis achieved in the experiments could be related to the drug-induced HepG2 cell cytotoxicity. Total analysis time for one product was about 30 min and only less than 4 μg HLM protein was required for one reaction region. The results demonstrated that the established platform could be used to imitate drug metabolism occurring in the human liver, thereby replacing animal experiments in the near future. In addition, the integrated microchip will be a useful tool for drug metabolism studies and cytotoxicity assays, which are pivotal in drug development.
authors.affiliation: Beijing Key Laboratory of Microanalytical Method and Instrumentation, Department of Chemistry, Tsinghua University, Beijing, 100084, China.
authors.forename: Sifeng
authors.initials: S
authors.lastname: Mao
authors.forename: Dan
authors.initials: D
authors.lastname: Gao
authors.forename: Wu
authors.initials: W
authors.lastname: Liu
authors.forename: Huibin
authors.initials: H
authors.lastname: Wei
authors.forename: Jin-Ming
authors.initials: JM
authors.lastname: Lin
mesh_terms: D000082:Acetaminophen / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D019149:Bioreactors; D002470:Cell Survival / Q000187:drug effects; D004867:Equipment Design; D014453:Glucuronosyltransferase / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D008862:Microsomes, Liver / Q000378:metabolism*; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D052616:Solid Phase Extraction / Q000295:instrumentation; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D004364:Pharmaceutical Preparations; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D000082:Acetaminophen; C037386:acetaminophen glucuronide; D014453:Glucuronosyltransferase