Dendrobium nobile is an important medicinal food beneficial for human health, well known for polysaccharides and dendrobine. For fast, accurate, and comprehensive comparison of its quality, high performance liquid chromatography (HPLC) fingerprinting method was constructed. Firstly, spring frost stressed D. nobile herb was observed for assessment. Decreased leaf thickness, chlorophyll, and drying rate, and increased free-proline indicated heavy damages on growth. But, the content of polysaccharides increased significantly in during-frost (DF), and dropped significantly in after-frost (AF). The content of dendrobine accumulated significantly in AF. Then, low similarity among HPLC fingerprints of before-frost (BF), DF, and AF, and 75.82% of significantly variant peaks indicated the changing of much more components. Especially, some less-polar components increased significantly in DF, but not in AF. Moreover, the highest suppression rates (SRs) to A549 lung cancer cells were up to 33.08% in DF, but only 15.63% and 12.12% in BF and AF. After association analysis, eleven less-polar components were found to be significantly and positively correlated to SRs under relatively high concentration. The result shows that frost stress not only causes damages to plant growth, but also promotes the accumulation of some health-beneficial bioactive metabolites. HPLC based fingerprinting method shows good applicability on quality evaluation and bioactivity correlation analysis of complexed agricultural products.
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