Nutrition Literacy (NL) positively impacts diet quality and has the potential to promote health and prevent nutrition-related chronic diseases. Brazil is one of the countries with the highest rates of nutrition-related chronic diseases. Nevertheless, in Brazil, few studies have explored the NL levels of its population. To provide remote access to the Nutrition Literacy Assessment Instrument for Brazilians (NLit-Br) and assess Brazilian bank employees, we conducted a study to estimate the validity of the NLit-Br online and to investigate whether bank employees have an adequate NL level. In the first step, we randomly assigned 21 employees from three financial institution branches to two groups to complete NLit-Br paper and online versions. After an interval period, both groups completed the NLit-Br with an opposite delivery method (paper vs. online). We compared the validity of the digital and paper versions of the NLit-Br by the Intraclass Correlation Coefficient (ICC), and the reliability by Kuder–Richardson formula 20. Second, we evaluated 1174 bank employees using the NLit-Br online version. We found an excellent absolute agreement (ICC ≥ 0.75) between the paper and online versions. The questionnaire had good internal consistency (KR-20 = 0.64). The sample was characterized as mostly male (61.0%), married/cohabitant (73.8%), and white (69.8%), with high household income (85.2%), and graduated or postgraduate (97.4%). The mean age of the population was 42.1 (SD = 7.6) years. Subjects predominantly had possibly inadequate NL (62.3%). The online NLit-Br total score was significantly associated with gender, age, and household income (p< 0.05). Women and individuals with higher incomes had a higher degree of NL. Subjects over 50 years old had a lower degree of NL. There was no significant association between the NLit-Br score and the participants’ education. The NLit-Br online is a valid instrument to assess NL remotely. The population studied showed a high prevalence of inadequacy of the NL. Therefore, there is a need for targeted actions to improve the NL of bank employees.