Integrated chemometric and spectrum-effect analysis discriminates Belamcanda chinensis and Iris tectorum in treating A549 lung cancer cells

Abstract

Belamcanda chinensis (, She Gan, SG) and Iris tectorum (, Chuan She Gan, CS), morphologically similar Iridaceae species with overlapping phytochemistry and respiratory applications, are frequently misused in traditional and modern practice. This study established a comprehensive differentiation strategy for SG and CS by integrating chemical characterization with pharmacological evaluation to identify bioactive markers. Twenty-nine herbal samples (11 CS, 18 SG) were analyzed for extraction yields, HPLC fingerprints, isoflavone quantification, multivariate chemometric analysis, in vitro cytotoxicity assessment, and spectrum-effect correlations. CS exhibited significantly higher extraction yields (44.8 % vs. 23.4 %) and total isoflavonoid content (168.7 vs. 42.6 mg/g) than SG, which correlated with stronger cytotoxicity against A549 cells. Chemometric analysis (PCA, PLS-DA and HCA) distinguished species-specific markers: tectoridin/ iristectorin A & B/ iristectorigenin B dominated CS, while irigenin characterized SG. In cytotoxicity assays using A549 cells at concentrations of 50-400 μg/mL, CS extracts showed significantly stronger dose-dependent cytotoxicity (93.6 % to 32.1 % viability reduction) compared to SG extracts (102.8 % to 68.3 %). PLSR-based spectrum-effect relationship analysis at 100/200 μg/mL linked eleven chromatographic peaks to bioactivity, with tectoridin, iristectorin A, and iristectorigenin B demonstrating strong correlations. Experimental validation confirmed these monomers as key cytotoxic agents. In summary, our integrated chemometric and spectrum-effect analysis showed stronger cytotoxicity of CS versus SG in A549 cells, with tectoridin, iristectorin A, and iristectorigenin B identified as potential quality markers. This study establishes a reliable chemometric-bioassay approach for bioactive marker discovery and herb-herb differentiation in traditional medicine. Further validation in diverse models and mechanistic studies is needed.

Keywords: Anticancer; Belamcanda chinensis; Iridaceae; Iris tectorum; Spectrum-effect relationship.