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In Ghana, prevalence of anaemia is higher than the worldwide average and contributes to deferral of blood donors. A cross‐sectional study was carried out as part of a pilot study aimed at improving haemoglobin levels and promoting repeat donations to retain donors who were deferred due to low haemoglobin. The copper sulphate test was used to determine low haemoglobin and anaemia assessed by the World Health Organization (WHO) gender‐specific criteria. Over sixteen months, 1213 donors were eligible, of which 826 (68%) were male and 78 (6.4%) were deferred for low haemoglobin. Among these 78 deferrals, 71 (91%) were female, 77 (99%) were first‐time donors and 77 (99%) were voluntary nonremunerated blood donors (VNRBDs). A total of 337 donors consented to provide a blood specimen out of which 325 donors met eligibility criteria and had complete FBC results. Of those, 189 ( N = 39 males; N = 150 females), or 58%, were classified as anaemic. Model‐based estimates which correct for selection bias in the enrolment process found that 61.6% of female donors (95% credible interval: [53.4%, 70.8%]) and 19.7% of male donors (95% credible interval: [11.5%, 33.8%]) were anaemic by WHO criteria. Among the 252 consenting donors with completed blood specimen analyses and haemoglobin levels meeting the threshold for blood donation, 118 (47%) were classified as anaemic according to WHO criteria. Population‐level estimates of anaemia using WHO criteria suggest anaemia is highly prevalent and the results generally matched donor deferral using the copper sulphate test among women blood donors. Trial Registration: ClinicalTrials.gov identifier: NCT04949165
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