Identification of velvet antler and its mixed varieties by UPLC-QTOF-MS combined with principal component analysis

•The different velvet antler species were firstly compared using our chemical-assay.•UPLC-Q/TOF-MS, Progenesis QI and Makerlynx XS software were integrated to clarify the difference of chemical constituents from three velvet antler species.•The run time of each sample was 40 min by UPLC-Q-TOF/MS.•The appropriate multi-response monitoring(MRM) and the standard peptides of the marker peptides can be used integrately for the qualitative identification. Many species of velvet antler have been used as traditional medicine for thousands of years; however, as medicinal materials, velvet antler derived from different animals have different clinical effects. To distinguish the differences and homologies, ultra-performance liquid chromatography-quadruple-time of flight mass spectrometry (UPLC-QTOF-MS) coupled with principal component analysis (PCA) was developed and applied to identify these antler samples derived from Cervus nippon Temminck, Cervus elaphus Linnaeus and Rangifer tarandus Linnaeus, which were first tested and compared at the molecular level of protein. The UPLC-MS data of the trypsin digested samples were subjected to PCA, and the potential markers based on peptide were depicted to illustrate their differences. With the integrated strategy combining UPLC-QTOF-MS with PCA, the results from this study indicated that the proposed methods could be successfully applied to distinguish reindeer antler from sika deer antler and red deer antler, which were prescribed in the Chinese Pharmacopoeia (2015 edition).