Radix Gentianae Macrophyllae (RGM) is a well-known traditional Chinese medicine with anti-inflammatory, analgesia and gastric motility strengthening effects. The iridoids are believed to be the RGM bioactive ingredients, however, an effective strategy for its total iridoids extraction and purification is lacking. In addition, a reliable method for the bioactive iridoid constituents digesting needs to be developed. In the present work, followed by process optimization via response surface modeling and Box-Behnken analysis, the maximum yield of total RGM iridoids was predicted to be 7.75 % from rhizomes of Gentiana macrophylla Pall. via three cycles of extraction in 69.16 % ethanol at liquid-solid ratio 12.27 (mL/g) for 110.91 min. The crude iridoid extract was further purified using HPD-300 macroporous resin to give the enriched product with the iridoid content up to 73.27 %. The anti-inflammatory effects of refined iridoid extracts were quantitatively measured to validate the purification procedure. The refined extract was detected to significantly inhibit the expression of NO through expression inhibition of COX-2, iNOS in lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells with the IC₅₀ of 10.49 μg/mL, which was stronger than that of crude extract with the IC₅₀ of 51.11 μg/mL, indicating the iridoid compounds were enriched from crude extract through the purification process. Subsequently, an UPLC-QTOF-MS method was established for qualitative and quantitative analysis of the iridoid constituents in the crude and refined extracts. In addition, based on the UPLC-QTOF-MS data, 6 principle iridoids that give the most important support for the anti-inflammatory activities were digested by means of PCA and PLS analysis. The current study developed a new and effective strategy for optimal extraction, purification and identification of natural bioactive iridoids from Radix Gentianae Macrophyllae, which facilitates its further applications in pharmaceuticals and/or foods industry.

龙胆草（RGM）是一种著名的中药，具有抗炎，镇痛和促进胃动力的作用。人们认为该类虹膜是RGM生物活性成分，但是，缺乏一种有效的方法来对其总的类虹膜进行提取和纯化。此外，还需要开发一种可靠的生物活性类环烷类成分消化方法。在目前的工作中，通过响应面建模和Box-Behnken分析进行工艺优化，从龙胆的根茎中预测总RGM虹膜类化合物的最大收率为7.75％。颇尔 通过在69.16％的乙醇中以液-固比12.27（mL / g）萃取的三个循环进行110.91分钟。使用HPD-300大孔树脂进一步纯化粗制的鸢尾花提取物，得到的富含鸢尾酮含量的产品高达73.27％。定量测量了精制的鸢尾酮提取物的抗炎作用，以验证纯化程序。检测到精制提取物通过抑制脂多糖（LPS）处理的RAW264.7巨噬细胞中COX-2，iNOS的表达而显着抑制NO的表达，其IC ₅₀为10.49μg/ mL，比原油的IC ₅₀强。用IC ₅₀提取的51.11μg/ mL表明，通过纯化过程从粗提取物中富集了烯类化合物。随后，建立了UPLC-QTOF-MS方法，用于定性和定量分析粗制和精制提取物中的虹彩成分。此外，根据UPLC-QTOF-MS数据，通过PCA和PLS分析消化了6种对抗炎活性提供最重要支持的类虹彩。当前的研究开发了一种新的有效策略，用于从龙胆草中提取，纯化和鉴定天然生物活性虹彩化合物的最佳方法，从而促进了其在制药和/或食品行业中的进一步应用。