Abstract. Saudah, Zumaidar, Darusman, Firmawati, Rislim DI, Juliantari E. 2022. Molecular authentication of Etlingera spp. (Zingiberaceae) in Aceh Province, Indonesia using DNA barcoding. Biodiversitas 23: 3976-3983. Etlingera spp.or locally known as Etlingera spp.(Acehnese) already has strong basic evidence in the treatment of traditional tribes in Aceh such as cough medicine, fever and sprains, and used to food and spices. Etlingera spp. used in various places varies based on morphological characteristics. On the other hand, for the development of phytopharmaceutical, the use of Etlingera spp. as plant herb requires authentication about the names. This study aimed to authenticate Etlingera spp. from Aceh using DNA barcoding. Nuclear ITS and trnL-F DNA sequences were used to authenticate among species of Etlingera spp. In this study different species of Etlingera spp. can be differentiated effectively by comparing the DNA barcoding regions. ITS sequence data can identify two different species of Etlingera (E. elatior and E. hemisphaerica). The two identified Etlingera spp.species can be used as important information for further research related to their pharmacological studies. One marker and combined analysis of ITS and trnL-F data proved that the Etlingeraspp. is a monophyletic group and provided phylogenetic information in closely related species. The intraspecific and interspecific variation of ITS markers were more discriminatory regions, while trnL-F IGS were of lower divergences. Our findings indicated that DNA barcoding is efficient and powerful to authenticate herbal medicine Etlingera spp..