Identification of Baiying (Herba Solani Lyrati) commodity and its toxic substitute Xungufeng (Herba Aristolochiae Mollissimae) using DNA barcoding and chemical profiling techniques

► Baiying (Solanum lyratum) was adulterated by Xungufeng (Aristolochia mollissima). ► Molecular identification by DNA barcodes (ITS, matK, rbcL, trnH-psbA and trnL-trnF). ► Chemical identification by TLC and HPLC using aristolochic acids as markers. ► Biological cytotoxicity assay on HEK-293 and Vero kidney cell lines. ► An integrated molecular, chemical and biological approach for better quality control. Baiying derived from Solanum lyratum Hance is a commonly consumed natural product for ethnomedical treatment of cancer. One of the substitutes present in the market is a carcinogenic aristolochic acids-containing herb Xungufeng derived from Aristolochia mollissima Thunb. The purpose of this study is to establish DNA barcodes, thin layer chromatography (TLC), high performance liquid chromatography (HPLC) and cytotoxicity assay to differentiate Baiying from Xungufeng. A total of 30 DNA sequences from five DNA barcodes (ITS, matK, rbcL, trnH-psbA and trnL-trnF) were generated to differentiate S. lyratum from A. mollissima and authenticate ten samples of Baiying and Xungufeng commodities. Using aristolochic acids as standard markers, TLC and HPLC analyses also successfully authenticated these commodities. In vitro cytotoxicity assay using HEK-293 and Vero cells demonstrated that Xungufeng was significantly more toxic than Baiying. This is the first study applying an integrated molecular, chemical and biological approach to differentiate traditional Chinese medicine from Aristolochia adulterant.