This study aimed to analyze the effect of tidal time and/or depth of mating media on hormonal conditions related to puberty molting on mating behavior and molting latency of mud crabs S. serrata. A two-factorial completely randomized design was used to assess the effects of tidal conditions, with treatments including no tide (P1), 12-hour high tide followed by 1-hour low tide (P2), and 24-hour low tide followed by 1-hour high tide (P3). The water depth consisted of 3 treatment levels, namely a depth of 30cm (K1), a depth of 60cm (K2), and a depth of 90cm (K3). The treatment was a combination of factors from all levels with three replications. Observation parameters included ecdysteroid titer, molting latency period, and absolute carapace width. A two-way ANOVA analysis was used to analyze the data and continued with Duncan's test. The results showed that the highest ecdysteroid concentration value, 1,573.12± 20.85ngmL‒1 , was observed at P3K3. The shortest molting latency period was 14.17± 1.04 days in P3K3. Moreover, the total carapace width was not significantly different. It was concluded that the mating medium with a high tide of 24 hours, a low tide of 1.0 hour with a rearing water depth of 90cm affected the hormonal conditions and molting latency of the mud crabs S. serrata.