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Background Belamcanda chinensis (L.) DC. (BC) belongs to the family of Iridaceae and is widely cultivated and used in many Chinese patent medicine and Chinese medicinal formulae. However, due to the high similarities in appearance such as color and shape to Iris tectorum Maxim (ITM), another plant from the same family, BC is often confused or even misused with ITM. Methods Therefore, in order to distinguish the chemical constituents, qualities and biological activities of BC and ITM, multiple technologies including plant metabolomics, digital reference standard (DRS) analyzer and biological activities assay were employed to provide a sufficient basis for their practical applications. Results In plant metabolomics, the PCA and OPLS-DA score plot indicated the obvious differences in chemical profiling between BC and ITM and 6 compounds were successfully identified to contribute to the differences. In DRS study, the fingerprints of 10 and 8 compounds in BC and ITM were developed based on DRS analyzer, respectively, involving relative retention time (RRT) method and linear calibration using two reference substances (LCTRS) technique. The DRS analyzer also accurately identified 10 and 8 compounds from BC and ITM, respectively, by using only two reference standards. In biological activities assay, BC had a better anticancer effect than ITM due to the high abundance of irigenin, while ITM showed stronger hepatoprotective activity than BC because of the high abundance of tectoridin. Conclusions Therefore, due to the significant differences of B. chinensis and I. dichotoma in chemical composition and biological activities, the current studies strongly proved that these two medicinal plants could not be mixed in industrial production and clinical medication.

A new amide (1), two new phenylpropanoid derivatives (2, 3), along with three new natural products, including three nitrogen chirality compounds, N‐(3‐methoxy‐1,3‐dioxopropyl)‐D‐phenylalanine methyl ester (4), N‐(3‐methoxy‐1,3‐dioxopropyl)‐L‐phenylalanine methyl ester (5), and N‐acetyl‐L‐phenylalanine methyl ester (6), as well as dimethyl (2R,3R)‐2‐hydroxy‐3‐(((E)‐3‐(4‐hydroxyphenyl)acryloyl)oxy)succinate (7) and dimethyl (S,E)‐2‐((3‐(4‐hydroxy‐3‐methoxyphenyl)acryloyl)oxy)succinate (8) were isolated from Delphinium kamaonense Hunth. Their structures were elucidated by extensive analysis of 1D and 2D NMR, and HR‐ESI‐MS experiments, and the absolute configurations were determined by comparative analysis of specific optical rotation. Compound 1 exhibited a moderate cytotoxicity effect against Hep‐3B cancer cell lines (IC50 41.39±0.13 μM) and an excellent antioxidant activity (IC50 0.527±0.06 μM in ABTS assay, and 1.235±0.09 μM in DPPH assay, respectively), which was superior to vitamin C in ABTS (IC50 1.670±0.07 μM) and DPPH (IC50 19.10±0.40 μM) methods.

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