Central Laboratory, Changchun Normal University, Changchun, China.
Guelph Research and Development Centre, Agriculture and Agri-Food Canada, Guelph, Ontario, Canada.
Phytochem Anal. 2021 Jul;32(4):640-653. doi: 10.1002/pca.3012. Epub 2020 Nov 25.
Radix Astragali, the dried root of Astragalus membranaceus (Fish.) Bge. (family Fabaceae), which is known as Huangqi in China, has been proven to be an immunostimulant, diuretic, antidiabetic, analgesic, and it has also been used as a health food supplement in some Asian populations and also serves as a lead herb in many traditional Chinese medicine formulations as well as in Chinese ethnic tonifying soups.
Screening and purification of bioactive compounds from natural products is challenging work due to their complexity. We present the first report on the use of pressurised liquid extraction and on-line two-dimensional counter current chromatography as an efficient medium for scaled-up extraction and separation of six bioactive compounds from Astragalus membranaceus.
We applied the established method with ultrafiltration-liquid chromatography to screen acetylcholinesterase inhibitors, which were then evaluated and confirmed for anti-Alzheimer activity using PC12 cell model.
Six major compounds, namely, calycosin-7-O-β-d-glucoside, pratensein-7-O-β-d-glucoside, formononetin-7-O-β-d-glucoside, calycosin, genistein, and formononetin, with acetylcholinesterase binding affinities were identified and isolated from the raw plant materials via two sets of n-hexane/ethyl acetate/0.2% acetic acid (first-stage counter current chromatography) and n-hexane/ethyl acetate/methanol/water (second-stage counter current chromatography) solvent systems: 1.87:1.0:1.33 and 5.62:1.0:2.42:5.25, v/v/v/v, which were optimised by a mathematical model.
Therefore, a useful platform for the large-scale production of bioactive and nutraceutical ingredients was developed herein. With the on-line system developed here, we present a feasible, selective, and effective strategy for rapid screening and identification of enzyme inhibitors from complex mixtures.
黄芪，是豆科植物膜荚黄芪或蒙古黄芪的干燥根，在中国被称为黄芪，已被证明具有免疫刺激、利尿、抗糖尿病、镇痛作用，也被一些亚洲人群用作保健品，并且还是许多中药配方以及中药滋补汤中的主要草药。
由于天然产物的复杂性，筛选和纯化生物活性化合物是一项具有挑战性的工作。我们首次报道了使用加压液体提取和在线二维逆流色谱作为从膜荚黄芪中大规模提取和分离六种生物活性化合物的有效介质。
我们应用超滤液-液相色谱法建立的方法筛选乙酰胆碱酯酶抑制剂，然后使用 PC12 细胞模型对其进行评估和确认抗阿尔茨海默病活性。
从原料中通过两组正己烷/乙酸乙酯/0.2%乙酸（第一阶段逆流色谱）和正己烷/乙酸乙酯/甲醇/水（第二阶段逆流色谱）溶剂系统：1.87:1.0:1.33 和 5.62:1.0:2.42:5.25，v/v/v/v，鉴定和分离出六种主要化合物，即毛蕊异黄酮-7-O-β-D-葡萄糖苷、紫堇灵-7-O-β-D-葡萄糖苷、芒柄花素-7-O-β-D-葡萄糖苷、毛蕊异黄酮、染料木黄酮和芒柄花素，它们与乙酰胆碱酯酶具有结合亲和力，通过数学模型进行了优化。
因此，本文开发了一种用于大规模生产生物活性和营养成分的有用平台。通过这里开发的在线系统，我们提出了一种可行、选择性和有效的策略，用于从复杂混合物中快速筛选和鉴定酶抑制剂。
