Staphylococcus aureus and Escherichia coli are a major cause of various humans and animal infections. Washing hands is a simple matter to eliminate and minimize germs on the hands with antiseptic hand sanitizer. Pecut kuda (Stachytarpheta jamaicensis (L) Vahl) contains a flavonoid compound known as antibacterial activity. Flavonoids are generally soluble in polar solvents. The type of solvent used for extraction influences which compounds are isolated. The purpose of the study was to asses the solvent effect of hand sanitizer formulation from pecut kuda extract. METHOD. Extract of pecut kuda was made by used three combination solvent of water and ethanol. There were 0:1; 1;1; and 1:2 for water and ethanol. Every formula was tested antibacterial activity against E.coli and Staph aureus. The highest antibacterial activity was made hand sanitizer gel by used optimization from HPMC K100M and PEG used Design-Expert software. The antiseptic test was carried out on the best formula. RESULT. The result showed the best antibacterial activity was 0:1 composition of water and ethanol. The best formula of hand sanitizer was made of 11,7% HPMC K 100M and 88,3% PEG. That formula had antiseptic activity as 69,19%.