Background and Objectives: Regular and repeat blood donations may cause reduced body iron stores (BIS), which may lead to a shrinking donor pool. Furthermore, hemoglobin (Hb) itself is quite an inadequate measure to detect iron deficiency. Hence, our primary aim was to understand the effect of repeated blood donations on BIS in regular blood donors. Study Design and Setting: We conducted a prospective study on voluntary whole-blood donors (September 2015 through August 2017). Donor demographics were noted. Donors were further divided into four groups, depending on their number of donations. Their samples were tested for Hb, red cell indices, and serum ferritin to check for reduced BIS. The data obtained were analyzed using SPSS version 20 (IBM, USA). Results: A total of 374 regular voluntary blood donors were included in the study. The mean serum ferritin levels in males and females were 75.4 and 36.2µg/L, respectively. There was a 45% decrement in the mean serum ferritin values in Group I (101.57 µg/L) in contrast to Group III (56.69 µg/L) (p < 0.0032). In the study, 9.8, 11.2, and 4.8% of the donors were in Stage 1, Stage 2, and Stage 3 of reduced BIS, respectively. Of the donors, 11.2% were in Stage 2 of reduced BIS. Further donations in such cases can compromise donor safety. Conclusion: The present study demonstrates iron depletion in regular voluntary blood donors. In 11.2% of the donors, iron deficiency erythropoiesis was noted. A ferritin screening after the first donation followed up at the tenth donation might help detect iron-deficient individuals. Iron supplementation for all regular blood donors and female donors in particular will help prevent the shrinking donor pool due to iron deficiency.