Purpose Despite unequivocal evidence demonstrating high carbohydrate (CHO) availability improves endurance performance, athletes often report under-eating CHO during competition. Such findings may be related to a lack of knowledge though currently there are no practical or time-efficient tools to assess CHO knowledge in athletes. Accordingly, we aimed to validate a novel questionnaire to rapidly assess endurance athletes' knowledge of competition CHO guidelines. Methods The Carbohydrate for Endurance Athletes in Competition Questionnaire (CEAC-Q) was created by research-active practitioners, based on contemporary guidelines. The CEAC-Q comprised 25 questions divided into 5 subsections (assessing CHO metabolism, CHO loading, pre-event meal, during-competition CHO and recovery) each worth 20 points for a total possible score of 100. Results A between-group analysis of variance compared scores in three different population groups to assess construct validity: general population (GenP; n= 68), endurance athletes (EA; n= 145), and sports dietitians/nutritionists (SDN; n=60). Total scores were different (mean +/- SD) in all pairwise comparisons of GenP (17 +/- 20%), EA (46 +/- 19%) and SDN (76 +/- 10%, p <0 .001). Subsection scores were also significantly different between the groups, with mean subsection scores of 3.4 +/- 4.7% (GenP), 9.2 +/- 5.2% (EA) and 15.2 +/- 3.5% (SDN, p < 0.001). Test-retest reliability of the total CEAC-Q was determined in EA (r =0.742, p <0.001). Conclusion Taking similar to 10 min to complete, the CEAC-Q is a new psychometrically valid, practical and time-efficient tool for practitioners to assess athletes' knowledge of CHO for competition and guide subsequent nutrition intervention.