The aim of this study was to evaluate the cytotoxicity of the prepared Gold Nanoparticles from Triphala Plant extract and of the indigenous mouthwash prepared from these Triphala based gold nanoparticles (AuNP’s) using Artemia salina test. Its objective was to assess whether the prepared mouthwash was safe and biocompatible to use as an alternative to commercial mouthwashes in orthodontic patients to reduce the plaque load around brackets and bands as an adjuvant to tooth brushing. 15 g of brine shrimps was hatched in salt solution after incubating them for 24 – h. Prepared AuNP’s at concentrations 0µL (microliter), 5µL, 10µL, 20µL, 30µL, 50µL were added using a micropipette in one of the microtiter plates and mouthwash prepared from these AuNP’s were added at concentrations 0µL, 10µL, 20µL, 30µL, 40µL, 50µL in another microtiter plate. 10 Hatched nauplii (brine shrimps) were added in each of these microtiter plates and incubated for 24 – h. All nauplii were alive at the end of 24 – h in the microtiter plate containing AuNP’s. At concentrations 0µL, 10µL, 20µL, 30µL nauplii were alive and at 40µL, 50µL concentrations no nauplii were alive at the end of 24 – h in the microtiter plate containing mouthwash. Triphala mediated AuNP’s showed no toxic effect on the brine shrimps. AuNP’s based mouthwash showed excellent compatibility at lower concentrations. Mild toxic effects were elicited at higher concentrations. Hence, AuNP’s could be used to prepare mouthwashes at 20- 30 µL concentrations.