OBJECTIVE： To study the inhibition effect of spinosin on 7 subtypes （CYP2B6， CYP2C8， CYP2C9， CYP2D6， CYP1A1， CYP2C19 and CYP3A4） of cytochrome P450 （CYP450） enzymes from human liver microsomes in vitro. METHODS： Taking 200.00， 100.00， 50.00， 25.00， 12.50， 6.25， 3.13， 1.56， 0.78， 0.39 μmol/L spinosin and human liver microsomes for incubation， using daktarin， bupropion， amodiaquine hydrochloride， diclofenac sodium， mephenytoin， dextromethorphan hydrobromide and midazolam as the specific probe drugs for above-mentioned 7 subtypes of CYP450 enzymes. UPLC-Q-TOF-MS was conducted to detect generation amount of 7 probe drug metabolites， and the half inhibitory concentration （IC50） of spinosin on 7 subtypes of CYP450 enzymes from human liver microsomes was calculated. RESULTS： IC50 of spinosin on 7 subtypes of CYP450 enzymes from human liver microsomes were 1 714， 1 158， 226.1， 2 288， 80.59， 101.1， 1 119 μmol/L， respectively， which were higher than 50 μmol/L. CONCLUSIONS： Spinosin has no inhibition effect on above-mentioned 7 subtypes of CYP450 enzymes from human liver microsomes， with very low probability of inducing metabolic drug interactions.