Abstract: Bacillus is an important biocontrol microorganism which can produce a variety of antimicrobial substances. In order to mine the strain resources producing the antibacterial substance quinolines, a total of 56 Bacillus strains were used as the experimental materials and their fermentation products were detected by the liquid chromatography-quadrupole time of flight-mass spectrometry (LC-QTOF-MS), and the molecular characteristics were extracted from the mass spectrometric data and compared with the Metlin library. The results showed that, 6 strains of Bacillus producing quinacetol were screened and obtained, including Bacillus soli FJAT-14232, Bacillus subtilis FJAT-8784, Brevibacillus borstelensis FJAT-10006, Paenibacillus macerans FJAT-14204, Bacillus alcalophilus FJAT-10014, Aneurinibacillus migulanus FJAT-14205. The relative contents of quinacetol in the total metabolites of the six Bacillus strains were all higher than 1.17%, and the matching scores with Metlin library were all greater than 95.35. Among them, the relative content of quinacetol in Bacillus soli FJAT-14232 was the highest, accounting for 1.68% of the relative content of its total metabolite, and Bacillus alcalophilus FJAT-10014 had the highest score of 99.13 by library searching. This study could provide the strain resources and data support for the development and utilization of the quinoline bacteriostatic agent from microorganisms.