Sound general and sports nutrition knowledge in athletes is essential for making appropriate dietary choices. Assessment of nutrition knowledge enables evaluation and tailoring of nutrition education. However, few well-validated tools are available to assess nutrition knowledge in athletes. The objective of the present study was to establish the validity of the Platform to Evaluate Athlete Knowledge Sports – Nutrition Questionnaire (PEAKS-NQ) for use in the United Kingdom and Irish (UK-I) athletes. To confirm content validity, twenty-three sports nutritionists (SNs) from elite, UK-I sports institutes provided feedback on the PEAKS-NQ via a modified Delphi method. After minor changes, the UK-I version of the PEAKS-NQ was administered to UK-I SN from the British Dietetic Association Sport and Exercise Nutrition Register, and elite athletes (EA) training at elite sports institutes in the UK and Ireland. Independent samples t-test and independent samples median tests were used to compare PEAKS-NQ total and subsection scores between EA and SN (to assess construct validity). Cronbach's alpha (good ≥ 0⋅7) was used to establish internal consistency. The SN achieved greater overall [SN (n 23) 92⋅3 (9⋅3) v. EA (n 154): 71⋅4 (10⋅0)%; P < 0⋅001] and individual section scores (P < 0⋅001) except Section B, Identification of Food Groups (P = 0⋅07). Largest knowledge differences between SN and EA were in Section D, Applied Sports Nutrition [SN: 88⋅5 (8⋅9) v. EA: 56⋅7 (14⋅5)%; P < 0⋅00]. Overall ES was large (2⋅1), with subsections ranging from 0⋅6 to 2⋅3. Cronbach's alpha was good (0⋅83). The PEAKS-NQ had good content and construct validity, supporting its use to assess nutrition knowledge of UK-I athletes.