São Paulo state is renowned for its extensive research on herpetofauna. However, there are still unexplored areas within the state, creating knowledge gaps in biodiversity. This study focused on the Itambé cave and waterfall in Altinópolis, São Paulo, which is an ecotone between the Cerrado and Atlantic Forest and popular among ecotourists. The research aimed to describe the herpetofauna composition in this area. Data collection occurred from April 2022 to January 2023, utilizing various methods. The study assessed the influence of climate variables and seasons on species richness and abundance. The findings identified eight herpetofauna species distributed across six genera. Among them, three lizard species and five anuran species were classified as generalists. Lizard richness peaked in winter, while anuran richness was highest in spring. Precipitation significantly influenced anuran abundance throughout the data collection period. Lizards were evenly distributed between the Cerrado and Atlantic Forest, while anurans were predominantly found in the Atlantic Forest. The research highlights the diverse herpetofauna present in the Itambé cave and waterfall region. However, all species encountered were generalists, suggesting potential human activity impacts on species with specific habitat requirements.