Choline is an essential dietary nutrient that plays a key role as one of the few sources of methyl donors in the diet. Additional roles include the neurotransmitter acetylcholine and lipids such as phosphatidylcholine and sphingomyelin. Studies in adults have shown that choline deficiency results in fatty liver. In developing animals, choline deficiency also leads to impaired neural development. Despite the importance of choline as a nutrient, there is limited data on dietary intake in relation to needs, or dietary choline and its connection to human development. This project aims to determine the choline intake and plasma status of choline among children 5 years and 9 months of age, and to explore the relationship plasma free choline and its metabolites and cognitive development. This study includes a cross-sectional study with a total of 200 children at 5 years and 9 months of age. Dietary intake was collected by FFQ, 3x24 hr recalls and 1x24 hr recall. Dietary choline intake was estimated using USDA database. Venous blood was collected and plasma free choline, betaine, dimethylglycine, homocysteine, methionine and cysteine were analyzed using LC-MS/MS, and plasma tB12 and folate were analyzed using microparticle enzyme immunoassay and ion capture assay, respectively. Cognitive development was evaluated using KABC-II, PPVT-4, and Beery-VIM. The median intakes of choline and betaine were 278 mg/d and 90.2 mg/d, respectively, when estimated using the FFQ. The mean±SD (IQR) for plasma free choline, betaine and dimethylglycine were 8.57±2.08 (7.20–9.60) µmol/L, 45.4±12.9 (37.0–53.1) µmol/L, and 3.26±0.95 (2.60–3.80) µmol/L, respectively. About 34% of the children consumed below the current DRI of 250 mg/d of choline. Significant positive association was found between dietary intake of choline and plasma free choline (r=0.198, P=0.014), but dietary intake of betaine was not related to plasma betaine (P=0.915). Plasma free choline was inversely associated with plasma homocysteine (P=0.013). No associations were found between plasma free choline, betaine or dimethylglycine and any of the cognitive test results (P>0.05). An inverse relationship between plasma free choline and homocysteine might indicate the presence of choline deficiency in the children in this study.