Glyphosate, being the most commonly utilized herbicide globally, and a growing body of experimental research has linked its exposure to red blood cell impairment. However, the potential toxicity of glyphosate exposure on erythrocytes in the general population remains poorly understood. Therefore, we analyzed data from the 2013-2014 National Health and Nutrition Examination Survey (NHANES) of 1466 adults (≥ 18 years) to explore the potential relationship between glyphosate exposure and erythrocyte pro les. Our results indicated a signi cant negative association between urinary glyphosate levels and hemoglobin (Hb) and hematocrit (Hct) in multiple regression analysis, with ß coe cients of -0.157 (S.E. = 0.055, P = 0.012) and − 0.431 (S.E. = 0.195, P = 0.043), respectively. Additionally, the odds ratio showed a signi cant increase in individuals with anemia with a one-unit increase in ln-glyphosate levels (odds ratio = 1.523 (95% CI = 1.301-1.783), P < 0.001 in the nal model). The negative correlation between glyphosate and Hb was more pronounced in subjects aged over 60 years, non-Hispanic white ethnicity, lower income, and those with a body mass index (BMI) < 25 and ≥ 30. In conclusion, our results offer initial evidence of a plausible link between glyphosate exposure and anemia among the adult populace in the United States. However, further research is necessary to comprehend the underlying mechanisms and clinical implications of this link.