Combination of Internal Transcribed Spacer 2 Barcoding and High‐Performance Liquid Chromatography Techniques for the Identification and Quality Evaluation of <i>Prismatomeris tetrandra</i>

ABSTRACT Prismatomeris tetrandra is a commonly used medicinal material for preventing and treating liver diseases. We use internal transcribed spacer 2 ( ITS2 ) barcode technology to provide molecular identification methods and sequences for P. tetrandra , optimize its high‐performance liquid chromatography (HPLC) fingerprint and multi‐component content determination method, and then evaluate the quality of different origins. In terms of ITS2 barcode identification, morphological and molecular identifications were performed on samples from 13 different origins, and a total of 28 ITS2 sequences were obtained, confirming that all the samples tested were authentic P. tetrandra . Since its sequences were not included in the GenBank database, we created 28 sequences and assigned a unique ID number to each sequence. A phylogenetic neighbor‐joining evolutionary tree was constructed, and its haplotypes were analyzed. The phylogenetic tree shows that all the P. tetrandra can be clearly clustered and identified, and are clearly distinguished from the other four medicinal materials of the same genus. In addition, a quality evaluation method for the simultaneous determination of three chemical components by HPLC was optimized and established. This method employed methanol as the extraction solvent, ultrasonic extraction for 40 minutes, a detection wavelength of 277 nm, and gradient elution using acetonitrile‐water as the mobile phase. The measured contents of scopolamine, 1‐rubiacin, and rubiacin were 0.009–0.359, 2.638–94.901, and 0.061–2.649 mg/g, respectively. It is speculated that the significant differences in content may be due to differences in growth environment and year. This study provides new insights for the accurate identification and quality evaluation of P. tetrandra .