Background: Essential oils (EO) are considered as safe and sustainable alternatives of synthetically produced industrial raw materials. While EO are renewable resources their production is traced to land use, therefore employing nonrenewable resources. This fact is often neglected during market up-take, which is established on EO bioactivity efficacy. Methods: Present study is aiming this knowledge gap through an innovative algorithm that employs spatial yield, bioactivity performance and fundamental experimentation details to calculate the land footprint. The proposed methodology is tested upon a concise pool of 54 EO, of which 9 originate from 8 culinary herbs, 27 from 3 juniper taxa, and 18 from 6 Citrus sp. crops. All 54 EO were subjected to repellent evaluation and 44 of them also to larvicidal, encompassing in the protocol both choice and no-choice bioassays. Results: Based on these bioprospecting data the proposed protocol effectively calculated the land footprint for all EO and bioassays. The repellent land footprint indicated as more sustainable the EO from savory, oregano, tarhan, thyme, Greek sage, and juniper berries for which each application corresponds to 3.97, 4.74, 7.33, 7.66, 8.01 and 8.32 m{}^2 respectively. The larvicidal assessment suggested as more sustainable the EOs from savory, oregano, fennel, thyme, tarhan, and rue with land footprints of 1.56, 1.79, 2.16, 2.89, 3.70 and 4.30 m{}^2 respectively. Conclusions: The proposed protocol managed to calculate the land footprint for each EO and bioactivity and indicated the more sustainable EO per use based on widely available bioprospecting data.