Ab iosensor was created for the simultaneous monitoring of endogenous H 2 S n and H 2 Si nm ouse brains and exploring their roles in activation of the TRPA1 channel under two types of brain disease models:i schemia and Alzheimersd isease (AD). Based on DFT calculations and electrochemical measurements,t wo probes,3 ,4-bis((2-fluoro-5-nitrobenzoyl)oxy)-benzoic acid (M PS-1 )a nd N-(4-(2,5-dinitrophenoxy) phenyl)-5-(1, 2-dithiolan-3-yl)pentanamide (M HS-1 ), were synthesized for specific recognition of H 2 S n and H 2 S. Through co-assembly of the two probes at the mesoporous gold film with good anti-biofouling ability and electrocatalytic activity,this microsensor showed high selectivity for H 2 S n and H 2 Sa gainst potential biological interferences.T he biosensor can simultaneously determine the concentration of H 2 S n from 0.2 to 50 mm,a sw ell as that of H 2 Sf rom 0.2 to 40 mm.T he expression of TRPA1 protein positively correlated with levels of H 2 S n under both ischemiaa nd AD.