The study examined the association between circulating gut microbial metabolites and risk of coronary heart disease (CHD) in a multistage prospective metabolomic analysis of five cohorts. In a discovery phase with 896 cases and 896 controls, they identified 73 microbial metabolites associated with incident CHD (false discovery rate <0.10). Of the 61 metabolites validated in-silico in the ARIC (N=3,539, 663 cases) and MESA (N=3,860, 446 cases) cohorts, 24 were significantly associated (p<0.05) in the same direction. A targeted quantitative test in 864 cases and 864 controls confirmed five metabolites with a significant association: imidazole propionate, 3-hydroxy-2-ethylpropionate, 4-hydroxyphenylacetate, trans-4-hydroxyproline, and 3-hydroxybutyrate, with ORs per SD ranging from 1.18 to 1.27 after adjusting for age, sex, race, lifestyle, and BMI. The other four significant metabolites were trimethylamine-N-oxide, phenylacetyl-L-glutamine, 4-hydroxyhippuric acid and indole propionate. Associations were mostly consistent across subgroups by demographics and lifestyle, with some modification by race, age, obesity, and follow-up time (mean, 5.6–15.0 years). Major limitations include the observational design and limited coverage of metabolites in the assays.