The invention discloses a preparation method of a 1, 8-naphthalimide hydrogen sulfide fluorescent molecular probe and application of the 1, 8-naphthalimide hydrogen sulfide fluorescent molecular probe in hydrogen sulfide recognition and detection. The fluorescent molecular probe with double recognition sites is obtained by using N-(p-benzyl)-4-hydroxy-1, 8-naphthalimide as a fluorescent matrix and 2, 4-dinitrophenyl ether as a recognition group, hydrogen sulfide performs thiolysis on the 2, 4-dinitrophenyl ether, so that hydroxyl is exposed out of a fluorophore, and the fluorescence intensity is enhanced (by more than 30 times). The fluorescent molecular probe has the advantages of stable photochemical properties, high fluorescence quantum yield, low price and the like, specifically recognizes hydrogen sulfide, is not interfered by ions such as Na<+>, Ca<2+>, NO2<->, H2O2, ClO<->, HSO4<->, CN2H4S, HSO3<->, S2O5<2-> and S2O8<2->, has a linear detection range increased to 0-40 [mu] mmol/L, can be used for quantitative detection of high-concentration hydrogen sulfide in river water, rainwater, drinking water, drilling fluid, marsh liquid and the like, and has potential practical application value.