Abstract Quinoa (Chenopodium quinoa Willd.) is traditionally grown in the mountain regions of the Andes where frost is common. However, the physiological mechanisms responsible for the frost resistance observed in quinoa are largely unknown. For this reason, a study on cultivars of quinoa originating from the Andean highlands and from the inter-Andean valleys was performed. Frost tolerance was determined by measuring the average lethal temperature of 50% of the leaf tissues (LT50) by ion leakage, and supercooling activity was assessed by thermal analysis using thermocouples. Quinoa demonstrated supercooling capacity (a mechanism that prevents immediate damage by freezing temperatures) of 5 °C. Ice nucleation temperature was always lower than the LT50. This indicates that the main survival mechanism of quinoa to frost is avoidance of ice formation by moderate supercooling. The study revealed that quinoa has a high soluble sugar content, which may cause a lowering of the freezing point and therefore contributing to lower the LT50. It is suggested that the content of proline and soluble sugars, such as sucrose, may serve as indicators of frost tolerance in quinoa breeding material.