College of Food Science and Engineering, Shandong Agricultural University, 61 Daizong Street, Taian, Shandong 271018, People's Republic of China.
Phytochem Anal. 2010 May-Jun;21(3):268-72. doi: 10.1002/pca.1196.
Flavonoids, the primary constituents of the petals of Nelumbo nucifera, are known to have antioxidant properties and antibacterial bioactivities. However, efficient methods for the preparative isolation and purification of flavonoids from this plant are not currently available.
To develop an efficient method for the preparative isolation and purification of flavonoids from the petals of N. nucifera by high-speed counter-current chromatography (HSCCC).
Following an initial clean-up step on a polyamide column, HSCCC was utilised to separate and purify flavonoids. Purities and identities of the isolated compounds were established by HPLC-PAD, ESI-MS, (1)H-NMR and (13)C-NMR.
The separation was performed using a two-phase solvent system composed of ethyl acetate-methanol-water-acetic acid (4 : 1 : 5 : 0.1, by volume), in which the upper phase was used as the stationary phase and the lower phase was used as the mobile phase at a flow-rate of 1.0 mL/min in the head-to-tail elution mode. Ultimately, 5.0 mg syringetin-3-O-beta-d-glucoside, 6.5 mg quercetin-3-O-beta-d-glucoside, 12.8 mg isorhamnetin-3-O-beta-d-glucoside and 32.5 mg kaempferol-3-O-beta-d-glucoside were obtained from 125 mg crude sample.
The combination of HSCCC with a polyamide column is an efficient method for the preparative separation and purification of flavonoids from the petals of N. nucifera.
荷叶中的黄酮类化合物是花瓣的主要成分，具有抗氧化和抗菌的生物活性。然而，目前还没有从这种植物中制备分离和纯化黄酮类化合物的有效方法。
开发一种高效的方法，通过高速逆流色谱（HSCCC）从荷花花瓣中制备分离和纯化黄酮类化合物。
在聚酰胺柱上进行初步清洗后，利用 HSCCC 分离和纯化黄酮类化合物。通过 HPLC-PAD、ESI-MS、（1）H-NMR 和（13）C-NMR 确定分离化合物的纯度和结构。
分离采用乙酸乙酯-甲醇-水-乙酸（4：1：5：0.1，体积比）两相溶剂体系，其中上相为固定相，下相为流动相，流速为 1.0 mL/min，采用首尾洗脱模式。最终，从 125 mg 粗样品中得到 5.0 mg 芹菜素-3-O-β-d-葡萄糖苷、6.5 mg 槲皮素-3-O-β-d-葡萄糖苷、12.8 mg 异鼠李素-3-O-β-d-葡萄糖苷和 32.5 mg 山奈酚-3-O-β-d-葡萄糖苷。
HSCCC 与聚酰胺柱相结合是从荷花花瓣中制备分离和纯化黄酮类化合物的有效方法。
