Objective:To isolate α-mangostin (AMG) from the peels of mangosteen (Garcinia mangostana L.),grown in Vietnam,and to investigate antibiofilm activity of this com-pound against three Staphylococcus aureus (S. aureus) strains,one of which was methicillin-resistant S. aureus (MRSA) and the other two strains were methicillin-sensitive S. aureus (MSSA). Methods:AMG in n-hexane fraction was isolated on a silica gel column and chemically analyzed by HPLC and NMR. The antibiofilm activity of this compound was investigated by using a 96-well plate model for the formation of biofilms. Biofilm biomass was quantified using crystal violet. The viability of cells was observed under confocal mi-croscopy using LIVE/DEAD BacLight stains. Biofilm composition was determined using specific chemical and enzyme tests for polysaccharide,protein and DNA. Membrane-damaging activity was assayed by measuring the hemolysis of human red blood cells in presence of AMG. Results:The results indicated that the isolated AMG,with a purity that exceeded 98％,had minimal inhibitory concentrations in the range of 4.6–9.2 μmol/L for the three strains tested. Interestingly,the MSSA strains were more sensitive to AMG than the MRSA strain. Minimal bactericidal concentrations were 2-fold higher than the minimal inhibitory concentration values for the three strains,indicating that AMG was a bactericidal com-pound. AMG also prevented biofilm formation effectively,albeit that again the MRSA strain was the most resistant. Interestingly,biofilms of the MRSA strain contained protein as a main component of the extracellular matrix,whereas this was polysaccharide in the MSSA strains. This might relate to the resistance of the MRSA 252 strain to AMG. Assays using human red blood cells indicated that AMG caused significant membrane damage with 50％ of cell lysis occurred at concentration of about 36 μmol/L.