Background and Objectives: Gangliosides (GAs) are important components of neural tissue and cell membrane. This study aims to investigate the association between toddlers’ neurodevelopment, dietary GA intake, and serum GA concentration. Methods and Study Design: A cross-sectional study was conducted in Beijing and Xuchang, Henan Province in China. 110 eligible healthy toddlers aged 24–48 months were recruited. Food frequency ques- tionnaire (FFQ) and 24-h dietary recall were used to collect dietary information. Blood serum samples obtained from participants were used to perform GA composition analysis with high-performance liquid chromatography- mass spectrometry (HPLC–MS). The neurodevelopment level was assessed with the Gesell Developmental Scale (GDS). Results: Dietary ganglioside GD3, total GA, and seafood intake were identified to be associated with the gross motor developmental quotient (DQ). An inverse association was revealed between the fine motor DQ and fruit intake. No correlation was detected between serum GA concentration and DQ. Conclusions: Dietary GA in- take but not serum GA concentration is associated with neurodevelopment. Further prospective studies are need- ed to probe the relationships between the recommended dietary GA intake and toddlers.