Objective To apply microfluidic technology to the human hepatoma （HepG2） cell model construction and potential in vitro safety assessment of aurantio-obtusin.Methods A simulated human microenvironment was constructed with rat tail collagen type Ⅰ （1.3 mg/mL） + gelatin （7.5%）， and acetaminophen （APAP） was used as a positive control. The proliferation activity， live/dead cell staining， and functional biochemical indicators of HepG2 cells were assessed for cell safety using different concentrations of aurantio-obtusin in vitro.Results The stable culture and application of HepG2 cells were achieved with the platform； after 72 h of culture， HepG2 cells proliferated in clusters. After 48 h of continuous treatment with aurantio-obtusin and an increase in the concentration of 0， 50， 100， and 200 μmol/L， the cell viability was 100.0%， 95.3%， 90.3%， and 81.6%， respectively. Compared with the blank control group， the cell inhibitory effect was especially significant at 200 μmol/L （P<0.05）； the number of red-marked dead cells gradually increased with the increase in concentration， and the green-marked number of cells gradually decreased. The content of blood urea nitrogen （BUN） decreased significantly with the increase in concentration. Compared with the blank control group， the difference was significant in the 50-200 μmol/L group （P<0.05）， and the content in the 200 μmol/L group was close to that in the APAP group. No significant difference was observed in the activities of alanine transaminase， aspartate transaminase， and lactate dehydrogenase at different concentrations.Conclusion The microfluidic platform HepG2 cell model has good applicability provides a new approach for combining microfluidic technology and the safety assessment of medicine and food homology substances， and has complementary feasibility in toxicological research methods in vitro.