[Objectives]  To accurately identify insects of the superfamily Acridoidea in the alpine grasslands of Gansu province. [Methods]  Sequences of the mitochondrial COⅠ gene with a length of 657 bp were obtained from 106 individuals of 20 species from the superfamily Acridoidea, and two homologous sequences of Oxya adentata were downloaded from GenBank as outgroups. A Maximum Likelihood (ML) phylogenetic tree was constructed using the Automatic Barcode Gap Discovery (ABGD) threshold method based on genetic distance classification, and the Molecular Defined Operational Taxonomic Units (jMOTU) threshold method based on sequence difference classification. [Results]  The phylogenetic tree placed Calliptamus abbreviatus, Calliptamus barbarus, Gomphocerus licenti, Compsorhipis davidiana and Epacromius coerulipes on two branches, and the other 15 species on separate branches. The results of jMOTU place Anaracris barabensis, Bryodema kozlovi, Bryodema nigroptera, Compsorhipis davidiana in one MOTU, and Filchnerella sunanensis and Filchnereua qilianshanensis in another MOTU which cannot be distinguished from each other. Identification of the other 14 species were consistent with the traditional morphological classification. ABGD divided the 108 samples into 28 groups, placing Angaracris barabensis, Bryodema kozlovi, Bryodema nigroptera, and Compsorhipis davidiana in one group. Identification of the other 16 species was consistent with morphological classification. [Conclusion]  DNA barcoding technology is an important auxiliary method for classifying and identifying locust species, improving the identification of species in the superfamily Acridoidea, especially closely-related species that cannot be reliably distinguished using traditional morphological methods.