Little is known about the effect of serum amylase enzymatic activity on glucose metabolism. We investigated the association of serum amylase enzymatic activity with fasting plasma glucose, insulin resistance (IR), and the plasma glucose and insulin response to an oral starch test (OST) in Mexican children.Anthropometric data, glucose and insulin levels, and the serum enzymatic activity of total (AMYt), salivary (AMY1), and pancreatic (AMY2) amylase were analysed in 764 children (NA positive association between serum enzymatic activity of AMY2 and IR was observed in children with obesity (p = 0.018). Children with normal weight had lower plasma glucose and insulin response to OST than children with obesity (POur results suggest that AMY2 is positively associated with IR. A high level of AMYt is related to lower glucose and insulin responses to OST in Mexican children, regardless of their weight status.