Central Laboratory, Changchun Normal University, Changchun, China.
Department of Traditional Chinese Medicine, The Fifth Medical Center, Chinese Pla General Hospital, Beijing, China.
Phytochem Anal. 2020 Nov;31(6):937-947. doi: 10.1002/pca.2966. Epub 2020 Jul 12.
Radix Saposhnikoviae is an excellent source of chromones, the consumption of which has been shown to positively affect human health, such as anti-inflammatory and antioxidant activities in vivo, and it also has been used to treat headache, inflammation, and cancer as a herbal medicine or functional soup.
Isolation and screening of lipoxidase (LOX) inhibitors from Radix Saposhnikoviae. As a result, the metabolism of the active ingredients in cytochrome P450 (CYP450) in vitro was identified.
Screen and identify the LOX inhibitors from Radix Saposhnikoviae by ultrafiltration-liquid chromatography-mass spectrometry (UF-LC-MS). Then take the isolation and purification of the inhibitors by high-speed counter-current chromatography (HSCCC) and semi-preparative high-performance liquid chromatography (HPLC). Furthermore, the active ingredients were used to study the metabolism in vitro in CYP450.
Subsequently, the specific binding ligands, prim-O-glucosylcimifugin, cimifugin, and 5-O-methylvisamminol (with purities of 95.28, 92.33%, and 97.89, respectively) were separated by HSCCC. Semi-preparative HPLC was applied to separate the four major compounds in Radix Saposhnikoviae and identify them as prim-O-glucosylcimifugin, 5-O-methylvisamminol, sec-O-glucosylhamaudol, and hamaudol, which were obtained with relatively high purity. Furthermore, the three chromone possible metabolism pathways were obtained.
The results demonstrated that UF-LC-MS combined with HSCCC and semi-preparative HPLC is not only a powerful tool for screening and isolating LOX inhibitors in complex samples, but also a useful platform for discovering bioactive compounds for anti-inflammatory activity. This study provides information regarding the active ingredients of chromones in Radix Saposhnikoviae and a framework for anti-inflammatory activity treatment.
防风是一种极好的色原酮来源，其消费已被证明对人体健康有积极影响，如体内的抗炎和抗氧化活性，它也被用作草药或功能性汤来治疗头痛、炎症和癌症。
从防风中分离和筛选脂肪氧合酶（LOX）抑制剂。结果，鉴定了细胞色素 P450（CYP450）体外活性成分的代谢。
通过超滤-液相色谱-质谱联用（UF-LC-MS）筛选和鉴定防风中的 LOX 抑制剂。然后采用高速逆流色谱（HSCCC）和半制备高效液相色谱（HPLC）对抑制剂进行分离和纯化。此外，还使用活性成分研究 CYP450 体外代谢。
随后，通过 HSCCC 分离出具有特定结合配体的原-O-葡萄糖基升麻素、升麻素和 5-O-甲基维斯阿米醇（纯度分别为 95.28%、92.33%和 97.89%）。半制备 HPLC 用于分离防风中的四种主要化合物，并鉴定为原-O-葡萄糖基升麻素、5-O-甲基维斯阿米醇、sec-O-葡萄糖基哈马醇和哈马醇，它们的纯度相对较高。此外，还获得了三种色原酮可能的代谢途径。
结果表明，UF-LC-MS 结合 HSCCC 和半制备 HPLC 不仅是筛选和分离复杂样品中 LOX 抑制剂的有力工具，也是发现具有抗炎活性的生物活性化合物的有用平台。本研究提供了防风中色原酮活性成分的信息，为抗炎活性治疗提供了框架。
