Ultra Performance Liquid Chromatography-Triple-Time of Flight Mass Spectrometer (UPLC-Triple-TOF/MS) was used for non-target metabolomics analysis of metabolites in Rhodococcus opacus PD630 in the presence of 1-propanol. Then a multivariate statistical analysis as well as an identification of differential metabolites was conducted for it.The results showed that there were 54 differential metabolites between the experimental group and the control group at 48 h, and 61 differential metabolites at 72 h. The decrease of short peptides and the increase in the degree of unsaturation of phospholipids were major metabolite changes. Meanwhile, a significant increase in propionyl CoA was observed(P＜0.001). The fatty acid composition was also determined by gas chromatography. The content of odd carbon fatty acids increased from 32.91% to 79.66%, with an increase of 2.4 times. This indicated that 1-propanol also had a great influence on the synthesis pathway of fatty acids.