Abstract: Cardiogenol C is an important medicine for treating cardiovascular diseases. To search for a biological source for the substance, a high throughput method base on LC-MS was carried out to screen the potential Bacillus-like strains. The molecular information on the target metabolites was collected using the MassHunter software. Accordingly, substances with their retention times and molecular weights that matched those on the Metlin database were identified. Bacillus amyloliquefaciens FJAT-17934 was, thus, isolated to show a high production level of cardiogenol C in its culture medium with a relative content of 1.02%. The matching score with the database for the cardiogenol C collected was 92.08%, and the retention time, 3.6517 min.