Combination of c oxidase subunit I based deoxyribonucleic acid barcoding and HPLC techniques for the identification and quality evaluation of Pheretima aspergillum.

: This study aimed to identify Pheretima aspergillum (Guang-Pheretima) and its adulterants using the cytochrome c oxidase subunit I based deoxyribonucleic acid barcoding technology, and further to evaluate their quality using an optimized HPLC method. For deoxyribonucleic acid barcoding identification, the Kimura-2-Parameter model was used to analyze genetic distance, and phylogenetic neighbor-joining tree was constructed for specie identification of 20 labeled Guang-Pheretima samples. A HPLC method was developed for the simultaneous determination of seven nucleoside components for quality evaluation. Compared with the GenBank database, 10 samples were identified as real Guang-Pheretima (Pheretima aspergillum), and the others as the adulterants-Metaphire magna. The maximum intraspecific genetic distances of c oxidase subunit I sequence for P. aspergillum were smaller than the minimum interspecific genetic distances between Pheretima aspergillum and Metaphire magna. 10 P. aspergillum and 10 Metaphire magna samples were clearly clustered in the neighbor-joining tree. The contents of 7 nucleosides components in Pheretima aspergillum were significantly higher than that in its adulterant-Metaphire magna. The incidence of adulterants for Guang-Pheretima was high (up to 50%) with an alarming quality. This study provided a powerful idea for the quality evaluation of other highly-valuable plant- or animal-derived products for safety concerns to avoid misidentification. This article is protected by copyright. All rights reserved.