Objectives Dietary nitrate (NO3) plays an important role in human's physiological processes. Nutrition professionals (NPs) have a crucial role in increasing the awareness of NO3 benefits across various health and academic settings. The aim of this study was to assess the level of knowledge towards the NO3 effects among NPs in Jeddah, Saudi Arabia based on their qualifications and years of experience. Methods A cross-sectional study was conducted among NPs graduated or working in Clinical Nutrition Departments of health centers and two universities. A validated online questionnaire with a total of 12 questions assessed the dietary nitrate's knowledge with regards to its health effects, dietary sources, recommendations, biomarkers of intake, and metabolism. A Nitrate Knowledge Index (NKI) score (out of 23 points) was used to evaluate the responses. Results A total of 89 female NPs out of 144 successfully completed the questionnaire. Most of them aged ≤30 years (73%), possessed undergraduate degree (BS) (70.8%), and had an experience of ≤3 years of experience (73%). Overall, poor knowledge score was observed among NPs in most questions as the median was 10 (7) out of 23. The majority (64%) perceived the NO3 as beneficial. However, the most did not know its benefits in lowering the blood pressure (68.5%) and were unsure about the nitrate effects on cognitive function (60.7%), kidney function (57.3%) and cancer risk (48.3%) as well as the acceptable daily intake of nitrate (67.4%). Almost half of the NPs had insufficient knowledge of NO3 sources as they incorrectly identified bacon and sausage as high NO3 food. They also were unsure about the mechanisms involved in the conversion of nitrate into nitrite in the mouth (48.3%). Knowledge of factors that affect food NO3 content were generally good. No significant differences, (P > 0.05), were observed in the median scores of NKI among the participants based on their level of education (BS: 8(6) vs. graduate degrees 10 (8.25)), nor based on years of experience (Experience ≤ 3 years: 10 (7) vs. >3 years: 8.5 (6)). Conclusions The authors recommend developing and implementing educational programs to enhance the knowledge and beliefs towards NO3 among NPs, who work in clinical and academic settings. Funding Sources King Saud bin Abdulaziz University for Health Sciences.