Electronic nose (E-nose) combined with gas chromatography–mass spectrometry (GC-MS) was used to analyze the volatile components of silver wormwood from different habitats, and the antibacterial activity of essential oils was also studied, to provide a scientific basis for quality control of silver wormwood and rational utilization of their essential oils. In this study, the total content of essential oils in silver wormwood was determined by steam distillation; the volatile components were conducted in an overall analysis by E-nose combined with chemometrics; the volatile components were analyzed and identified by GC-MS; and two G-negative bacteria and one Gram-positive bacteria were used as test bacteria to determine the antibacterial activity of the essential oils from silver wormwood. The results showed that principal component analysis (PCA) and linear discriminant analysis (LDA) of E-nose could distinguish the essential oils of silver wormwood from different habitats, and the odor difference of essential oils was obvious. A total of 87 volatile components were identified by GC-MS, and there were significant differences in components and contents in silver wormwood from different habitats; PCA and hierarchical cluster analysis (HCA) could effectively distinguish silver wormwood from different habitats. The essential oils from silver wormwood from different habitats all had a certain inhibitory effect on Bacillus subtilis, Staphylococcus aureus, and Escherichia coli. Therefore, the combination of E-nose and GC-MS could quickly distinguish silver wormwood from different habitats and provide a reference for quality control, drug selection, and comprehensive utilization of silver wormwood.