Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University, Chuncheon 200-702, Republic of Korea.
Phytochem Anal. 2011 Sep-Oct;22(5):468-73. doi: 10.1002/pca.1306. Epub 2011 Apr 4.
Isoflavonoids from Belamcanda chinensis are known to have a number of physiological benefits including anti-inflammatory, anti-angiogenic and anti-mutagenic properties. However, there have been no reports on the effective isolation and purification of isoflavonoids from B. chinensis.
To develop an efficient method for the preparative isolation and purification of isoflavones from B. chinensis by high-speed counter-current chromatography (HSCCC).
A two-step HSCCC isolation method was developed using solvent system of n-hexane-ethyl acetate-2-propanol-methanol-water (5:6:2:3.5:6, v/v) and of ethyl acetate-methanol-water (10:2:9, v/v). FLASH purification system (45% methanol, isocratic) was also used for further purification. The purities and chemical structures of the isolated compounds were determined by high-performance liquid chromatography-photodiode array detection (HPLC-PDA), electrospray ionisation-mass spectrometry (ESI-MS), ¹H- and ¹³C-nuclear magnetic resonance spectrometry (NMR) and nuclear overhauser enhancement (NOE).
HSCCC was successfully used for the preparative separation and purification of seven isoflavones, including tectoridin (145.4 mg, 97.5%), iridin (77.9 mg, 94.0%), irilin D (42.0 mg, 92.0%), tectorigenin (294.1 mg, 98.6%), iristectorigenin A (86.8 mg, 93.4%), irigenin (141.8 mg, 95.8%) and irisflorentin (73.4 mg, 94.7%) from the rhizomes of B. chinensis.
Two isoflavone glycosides and five isoflavone derivatives were successfully isolated and purified from the crude methanol extract of dried rhizomes of the B. chinensis by HSCCC.
从射干中提取的异黄酮具有多种生理益处，包括抗炎、抗血管生成和抗突变特性。然而，目前还没有关于从射干中有效分离和纯化异黄酮的报道。
开发一种高效的高速逆流色谱（HSCCC）法从射干中分离和纯化异黄酮的方法。
采用正己烷-乙酸乙酯-2-丙醇-甲醇-水（5:6:2:3.5:6，v/v）和乙酸乙酯-甲醇-水（10:2:9，v/v）两步 HSCCC 分离法。还使用 FLASH 纯化系统（45%甲醇，等度洗脱）进行进一步纯化。通过高效液相色谱-光电二极管阵列检测（HPLC-PDA）、电喷雾电离质谱（ESI-MS）、¹H-和¹³C-核磁共振谱（NMR）和核Overhauser 增强（NOE）确定分离化合物的纯度和化学结构。
HSCCC 成功地用于七种异黄酮的制备分离和纯化，包括次野鸢尾黄素（145.4mg，97.5%）、鸢尾黄素（77.9mg，94.0%）、鸢尾苷元 D（42.0mg，92.0%）、次野鸢尾黄素（294.1mg，98.6%）、鸢尾苷元 A（86.8mg，93.4%）、染料木黄酮（141.8mg，95.8%）和鸢尾黄素（73.4mg，94.7%）。
通过 HSCCC 从射干干根茎的甲醇粗提物中成功分离和纯化了两种异黄酮糖苷和五种异黄酮衍生物。
