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Background: Ethanol Extract of Mulberry Leaves (Morus alba L.) is known to have platelet anti-aggregation activity. Objective: This study aims to determine the anti-aggregation activity of platelets from the fraction of mulberry leaves. Materials and Methods: The study used human blood plasma taken from the Indonesian Red Cross (PMI) as a sample. The test was divided into five groups, namely the negative control group, the comparison group using acetosal 1 mg/ml, and 3 test groups using fractions of aquadest, N-hexane, and Ethyl acetate with a concentration of 0.849 mg/ml. This study was carried out in vitro with the parameters of the percentage of platelet aggregation formed. Results: The results of the statistical analysis of the data, there was an effect of treatment on the percentage of platelet aggregation (p <0.05). The ethyl acetate fraction group had a platelet aggregation percentage of 7.93% and did not differ significantly from the acetosal group by 8.14% (p> 0.05). Conclusion: Based on the results of these studies, the ethyl acetate fraction of mulberry leaves have the potential for platelet anti-aggregation, which is comparable to acetosal.