Trimethylamine-N-oxide (TMAO) has been shown to be associated with cardiovascular (CV) disease and cognitive impairment. The association between early stages of cognitive impairment and TMAO in a high CV risk population has not been previously investigated. This study aimed to investigate the association between the plasma TMAO level and cognitive function in a population with a high risk of CV disease. Participants at a high risk of CV were included. The cognition was evaluated using the Montreal Cognitive Assessment. A score lower than 25 out of 30 was used to indicate mild cognitive impairment (MCI). Blood samples of all participants (n = 233) were collected to measure the plasma levels of TMAO and other metabolic parameters, including fasting blood sugar and lipid profiles. Logistic regression was used to evaluate the association between MCI and high plasma TMAO levels, adjusted for confounding factors. Of 233 patients, the mean age of patients in this study was 64 years old (SD 8.4). The median TMAO level was 4.31 μM (IQR 3.95). The high TMAO level was an independent risk factor of MCI (aOR 2.36, 95% CI 1.02 to 5.47; p 0.046), when adjusted for age, gender, health care service scheme, smoking history, metabolic syndrome, and history of established CV events. The high TMAO level was associated with MCI, after adjustment for potential confounding factors. These findings demonstrate that plasma TMAO levels can serve for target prediction as an independent risk factor for MCI in this population.