Meconopsis integrifolia (Maxim.) Franch. is used extensively in traditional Tibetan medicine for its potent anti-inflammatory properties. In this study, six cyclooxygenase-2 (COX-2) inhibitors were purified from M. integrifolia using high-speed counter-current chromatography guided by ultrafiltration liquid chromatography (ultrafiltration-LC). First, ultrafiltration-LC was performed to profile the COX-2 inhibitors in M. integrifolia. The reflux extraction conditions were further optimized using response surface methodology, and the results showed that the targeted COX-2 inhibitors could be well enriched under the optimized extraction conditions. Then the six target COX-2 inhibitors were separated by high-speed countercurrent chromatography with a solvent system composed of ethyl acetate/n-butanol/water (4:1:4, v/v/v. Finally, the six COX-2 inhibitors, including 21.2 mg of 8-hydroxyluteolin 7-sophoroside, 29.6 mg of 8-hydroxyluteolin 7-[6′''-acetylallosyl-(1→2)-glucoside], 42.5 mg of Sinocrassoside D3, 54.1 mg of Hypolaetin 7-[6′′′-acetylallosyll-(l→2)-3′′-acetylglucoside, 30.6 mg of Hypolaetin 7-[6′′′-acetylallosyll-(l→2)-6′′-acetylglucoside and 17.8 mg of Hypolaetin were obtained from 500 mg of sample. Their structures were elucidated by ¹H-NMR spectroscopy. This study reveals that ultrafiltration-LC combined with high-speed counter-current chromatography is a robust and efficient strategy for target-guided isolation and purification of bioactive molecules. It also enhances the scientific understanding of the anti-inflammatory properties of M. integrifolia but also paves the way for its further medicinal applications.

中文翻译：

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从 Meconopsis integrifolia (Maxim.) Franch 中靶向分离和纯化 cyclooxygenase-2 抑制剂。高速逆流色谱结合超滤液相色谱

绿绒蒿 (Meconopsis integrifolia (Maxim.) Franch ) 由于其有效的抗炎特性，在传统藏药中被广泛使用。在本研究中，使用超滤液相色谱（超滤-LC）引导的高速逆流色谱从M. integrifolia中纯化了六种环氧合酶-2 (COX-2) 抑制剂。首先，进行超滤-LC 分析M. integrifolia中的 COX-2 抑制剂。利用响应面法进一步优化回流提取条件，结果表明，在优化的提取条件下，能够很好地富集靶向COX-2抑制剂。然后以乙酸乙酯/正丁醇/水(4:1:4, v/v/v)为溶剂体系，通过高速逆流色谱分离出六种目标COX-2抑制剂。抑制剂，包括 8-羟基木犀草素 7-槐糖苷 21.2 mg、8-羟基木犀草素 7-[6'''-乙酰别糖基-(1→2)-葡萄糖苷] 29.6 mg、Sinocrassoside D3 42.5 mg、Hypolaetin 7- 54.1 mg [6′′′-乙酰阿洛基-(1→2)-3′′-乙酰葡糖苷、30.6 mg 次乳酸素 7-[6′′′-乙酰阿洛基-(1→2)-6′′-乙酰葡糖苷和 17.8 mg 次乳酸素从 500 mg 样品中获得。通过¹ H-NMR 光谱阐明了它们的结构。这项研究表明，超滤-LC 与高速逆流色谱相结合是一种稳健而有效的策略，用于目标引导的生物活性分离和纯化它还增强了对M. integrifolia抗炎特性的科学认识，同时也为其进一步的医学应用铺平了道路。