Introduction : Voluntary unpaid blood donation is a altruistic deed towards the sick by the healthy. The well–being and fitness of the blood donors are of major importance in current health care scenario. In most nations, strict guidelines have been established for the choice of blood donors that include criteria that serve to protect both the donors and recipients. Moreover, 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. Aim: This work is aimed at assessing the influence of frequency of blood donation on iron stores by assessing voluntary blood donors. Method: This study was conducted in Rama Medical College and research centre, Hapur. This is prospective and observational study and was performed between January 2018 and December 2019. On 115 subjects aged between 18 and 50 years. Donors were divided into four donor groups Blood samples from the study subjects were collected for hematological tests data were analysed using SPSS and simple calculation. Result: In our study haematological parameters in different donors groups was analyazed which showed, subjects in the group I and group II don’t have iron storage deficiency while 61.11% of the participant in group III was having iron storage deficiency and in the group IV 81.81% members has low serum ferritin level this is a clear indication that multiple donation effect iron store in the body. Conclusion: The present study demonstrates iron depletion in regular voluntary blood donors. Therefore, volunteers who repeatedly donate blood should be advised on eating balanced diet with special care of iron supplements and they should undergo ferritin screening.