Introduction: Now many studies conducted on the drug substance from nature that can serve as an anticancer agent as a potential chemoprevention agent, such as Annona muricata Linn leaf escort chemotherapy, which was flaring. The cancer cell in humans was included the loss of p53 protein function due to mutations in the protein gene. Other causes are that the p53 proteins are not functioning due to an increase in protein misfolding event chaperones and degradation events ubiquitous as binding by viral protein. Method: Cytotoxicity assay performed on 24 well plate micro-cultures. HeLa cells are as 2 × 10⁴ cells in 100 mL in RPMI media. Created control is RPMI and solvent DMSO 0.25%. Cytotoxic Test performed by the method of calculation tryphan blue dye exclusion. Being fasted for 24 hours in the culture medium, then the cells are grown in micro-plate with media plus samples with a non-lethal concentration (LC50) of partition and fractionation Annona muricata Linn leaf. Sampling is performed at 24 hours. Each of these wells is calculated the number of living cells and made the curve of cell number and incubation time. Result: The results showed that HeLa cells are being LC50 partition of leaves Annona muricata Linn in ethyl acetate his cell death rate was higher (2000 μg/ml have 131.89%; 15.625 μg/ml have 11.37%) and in ethanol-distillate water his cell death rate was lower (2000 μg/ml have 35.80%; 15.625 μg/ml have 3.97%). Another results showed that HeLa cells are being LC50 fractionation of leaves Annona muricata Linn in chloroform his cell death rate was higher (2000 μg/ml have 91.86%; 15.625 μg/ml have 2.68%) and in ethyl acetate, his cell death rate was lower (2000 μg/ml have 23.79%; 15.625 <