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Capsaicin is a bioactive compound with poor solubility in water. Transfersomes can increase the solubility and thus penetration of capsaisin. Transfersomes has an advantage of being an ultra-deformable vesicles and is made from thin layer hydration method. The aims of this research are to know the effect of hydrophilicity surfactants on characterization and in vitro penetration ability of capsaisin transfersomes in gels using non-ionic surfactants with different HLB, which are span 80, a mixture of span 80 and tween 80, and tween 80. Best characterization result of transfersome is the formula using tween 80. Total cumulative amount of capsaisin penetrated from the transfersome gel using tween 80 is 1663.89 ± 1.58 mg/cm², percentage is 57.96 ± 0.05 % and the flux is 166.38 ± 0.15 mg/cm².hour^-1; from the transfersome gel using a mixture of span 80 and tween 80 is 1539.8 ± 21.23 mg/cm², percentage is 54.47 ± 0.75% and the flux is 153.98 ± 2.12 mg/cm².hour^-1; while from the transfersome gel using span 80 is 1395.10 ± 7.23 mg/cm², percentage is 50.80 ± 0.26 % and the flux is 139.51 ± 0.72 mg/cm².hour^-1.