Bee venom (Apis mellifera L., BV) has been used as a cosmetic ingredient for antiaging, anti-inflammatory and antibacterial functions. The aim of this study was to access the skin sensitization of BV, a Buehler test was conducted fifty healthy male Hartley guinea pigs with three groups; Group G1 (BV-sensitization group, 20 animals), group G2 (the positive control-sensitization group, 20 animals), and group G3 (the ethyl alcohol-sensitization group, 10 animals). The exposure on the left flank for induction was repeated three times at intervals of one week. Two weeks after the last induction, the challenge was performed on the right flank. No treatment-related clinical signs or body weight changes were observed during the study period. The average skin reaction evaluated by erythema and edema on the challenge sites and sensitization rate in the BV-sensitization group at 30 hours were 0.0 and 0%, respectively, which are substantially low compared with in positive control group (average skin reaction: 0.55, sensitization rate: 40%) and identical with in vehicle control group, representing a weak sensitizing potential. The average skin reaction and sensitization rate observed at 54 hours were 0.0 and 0% in the BV-sensitization group, respectively, and 0.25 and 20% in the positive control group, respectively. It was concluded that BV classified to Grade I, induced no sensitization when tested in guinea pigs and may provide a developmental basis for a cosmetic ingredient or external application for topical uses.