Background Glycated hemoglobin (HbA1c) is often used to diagnose type 2 diabetes (T2D), but studies show that iron deficiency (ID) is associated with elevated HbA1c in the absence of hyperglycemia. It is unknown whether ID prevalence varies between sub-Saharan African populations living in different locations and whether ID influences HbA1c levels in these populations. Objectives We assessed the prevalence of ID among Ghanaian migrants in Europe and nonmigrant Ghanaians, and the influence of ID on HbA1c categories among Ghanaians without T2D. Methods We used the database from the cross-sectional RODAM (Research on Obesity and Diabetes among African Migrants) study. This contained data on 3377 Ghanaian men and women aged 25-70 y living in urban and rural Ghana and Ghanaian migrants living in Amsterdam, London, and Berlin. ID was defined as ferritin < 15 ng/mL or, if C-reactive protein was ≥5 mg/mL, as ferritin < 30 ng/mL according to the WHO. We used binary logistic regression to assess differences in ID between sites and its association with clinically defined HbA1c categories (<5.5%, ≥5.5% to <6.5%, ≥6.5%). Men and women were analyzed separately. Results The prevalence of ID was higher in migrant [28.4%; adjusted OR (aOR): 3.08; 95% CI: 2.04, 4.65)] and urban (23.2%; aOR: 2.37; 95% CI: 1.56, 3.59) women than in rural women (11.9%). Among women, ID was associated with higher odds of HbA1c ≥ 5.5% to <6.5% in the absence of hyperglycemia (aOR: 1.43; 95% CI: 1.08, 1.87). This association was not found in men. Conclusions Further research is needed to identify factors underlying the high prevalence of ID among urban and migrant Ghanaian women, and the association of ID with HbA1c ≥ 5.5% to <6.5% in women. In addition, our study reinforces the need to consider iron concentrations if interpreting HbA1c among African populations.