Essential oil is a natural substance widely utilized in the cosmetics industry. However, there has been no study on the cosmetic bioactivity of essential oil from the peel of Citrus amblycarpa Hassk. Ochse (CAEO). This study aimed to investigate the potential of CAEO, obtained through NaCl hydrodistillation (HD) and microwave-assisted hydrodistillation (MAHD), as an anti-tyrosinase, anti-elastase, and antibacterial agent, along with analyzing its chemical constituents. GC-MS analysis identified β-pinene (28.3%) and limonene (27.1%) as the main compounds in CAEO (HD), while limonene (13.75%) and citronellal (7.49%) predominated in CAEO (MAHD). The CAEO obtained from HD with 25% NaCl as the solvent exhibited significantly higher (p < 0.05) anti-tyrosinase (IC50 = 235.94 ± 1.34 μg/mL) and anti-elastase (IC50 = 41.59 ± 0.47 μg/mL) activities compared to the CAEO from the MAHD method. This result was consistent with the in silico pharmacological study, which revealed that limonene, the major component of CAEO, showed good binding affinity to the active sites of elastase and tyrosinase. The CAEO obtained from both methods exhibited strong and selective antibacterial activity in a disc diffusion assay against two bacteria associated with skin acne: Propionibacterium acnes ATCC 11827 (CAEO HD = 24.08 ± 5.85 mm) and Staphylococcus aureus ATCC 25923 (CAEO MAHD = 20.5 ± 0.27 mm). These results pave the way for further studies on the bioassay-guided isolation of CAEO to enhance its effectiveness as a cosmetic agent. Moreover, this study provides the first report indicating the potential of CAEO for development as a cosmetic agent. © 2025 Har Krishan Bhalla & Sons.