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This study represents the first part of a long-term research project aimed to assess the welfare status of two female Asian elephants hosted at Pistoia Zoo and to monitor their adaptation process to a new naturalistic enclosure through chemo-ethological analysis. Data were collected from October 2015 to May 2016 using a within-subject experimental design. Sampling days were randomly assigned over two periods P1 and P2, respectively before and after the new area was opened. Six 30-min observation sessions per day were collected using: (i) instantaneous scan sampling with 5-min interval; (ii) 15-min continuous focal animal sampling per individual. 2233 scan samples and 3780 minutes of focal observations were obtained for each elephant. Data were analysed using nonparametric and randomization tests, the Shannon Diversity Index and the Behavioural Competence Index. P1 activity budgets showed a lower percentage of inactivity and locomotion compared to previous studies performed in captive environments. Locomotion significantly increased in P2, suggesting that the access to the new area acted as a positive stimulus. Inter-individual differences were found in the frequencies of active behaviours with the youngest female showing a richer behavioural repertoire. Results also revealed a high level of behavioural competence even if the elephants displayed stereotyped behaviours. The analysis of faecal samples collected in P1 pointed out that mean cortisol concentration was lower in the youngest female but no significant difference was found between subjects. These findings will be compared with P2 cortisol concentrations to monitor the stress response after the opening of the new enclosure.