Background: At this instant, automated cell separators are used for the purification of platelets from healthy donors but the donor’s safety should not be overlooked. Hence, the present study aimed to compare the effect of platelet apheresis (via mega unit) on hematological indices of healthy donors. Methodology: In this cross-sectional study, 100 samples of healthy plateletpheresis or thrombopheresis donors were included. The pre and post plateletphresis hematological parameters were assessed. The total platelet count (PLT), hemoglobin (Hb), white blood cells count (WBCs), red blood cells count (RBCs), mean cell volume (MCV), mean cell hemoglobin (MCH) or mean corpuscular hemoglobin concentration (MHCH) were estimated through complete blood count (CBC) analysis and platelet apheresis was done by Mega unit Haemonetics MCS+9000 Cell Separator and 994- CFE. Results: After platelet apheresis procedure, a significant decrement in PLT value (p-value <0.001). And with that a remarkable difference was also observed in the haemoglobin, MCH and MCHC.  Conclusion: In conclusion, the hematological parameters significant decree among donors after the procedure of platelet aphresis but the revealed characteristics of thrombocytopenia and anemia were not found.