Historically, there are inconsistencies in the calculation of whole-grain intake, particularly through use of highly variable whole-grain food definitions. The current study aimed to determine the impact of using a whole-grain food definition on whole-grain intake estimation in Australian and Swedish national cohorts and investigate impacts on apparent associations with CVD risk factors. This utilised the Australian National Nutrition and Physical Activity Survey 2011–2012, the Swedish Riksmaten adults 2010–2011 and relevant food composition databases. Whole-grain intakes and associations with CVD risk factors were determined based on consumption of foods complying with the Healthgrain definition (≥30 % whole grain (dry weight), more whole than refined grain and meeting accepted standards for ‘healthy foods’ based on local regulations) and compared with absolute whole-grain intake. Compliance of whole-grain containing foods with the Healthgrain definition was low in both Sweden (twenty-nine of 155 foods) and Australia (214 of 609 foods). Significant mean differences of up to 24·6 g/10 MJ per d of whole-grain intake were highlighted using Swedish data. Despite these large differences, application of a whole-grain food definition altered very few associations with CVD risk factors, specifically, changes with body weight and blood glucose associations in Australian adults where a whole-grain food definition was applied, and some anthropometric measures in Swedish data where a high percentage of whole-grain content was included. Use of whole-grain food definitions appears to have limited impact on measuring whole-grain health benefits but may have greater relevance in public health messaging.