Hand hygiene is an important factor that needs to be observed in controlling the spread of diseases transmitted through hand-to-hand contact. Synthesis of silver nanoparticles from tembelekan (Lantana camara) and paku laut (Acrostichum aureum) using the green synthesis method has good antibacterial activity against Staphylococcus aureus and Escherichia coli bacteria. Therefore, a preparation formulation was made, namely hand sanitizer, which is still rarely used. Formulations that have successfully entered the evaluation stage include organoleptic tests, homogeneity tests, spreadability tests, adhesion tests, viscosity tests, pH tests, accelerated stability tests, and irritation tests. Antibacterial activity was evaluated against bacteria Staphylococcus aureus and Escherichia coli. The hand sanitizer is formulated to contain 5% tembelekan AgNPs (F1); paku laut AgNPs 5% (F2); and a combination of 2.5% paku laut AgNPs and 2.5% tembelekan AgNPs. The resulting hand sanitizer has good organoleptic characteristics, except for the color of the preparation, which changed during the accelerated stability test. Test results for pH, adhesion, spreadability, viscosity, and homogeneity of hand sanitizer meet the requirements of a good test. Irritation tests on ten volunteers showed no irritation reaction. Antibacterial tests show that hand sanitizer has bacterial antibacterial activity with an average ± standard deviation of the inhibition zone Staphylococcus aureus is 6.605±0.459(F1); 6.665±0.615(F2); 6.380±0.282(F3) dan Escherichia coli namely 6.575 ± 0.219 (F1); 6.860 ± 0.155 (F2); 6.810 ± 0.056 (F3). Making hand sanitizer AgNPs-based ingredients from plants can be used as hand sanitizer, but stabilizers are required to prevent color changes during storage.