|
植物来源 |
|
|
生物活性 |
|
|
鉴定 |
熔点 |
300°C |
| 旋亮度 |
[α]20D-66.6° |
1HNMR
|
| 分析方法 |
|
| 仪器 |
硅胶F254 板及乙酸钠 |
| 流动相 |
乙酸乙酯: 己烷 (9: 1) (不饱和) 或100% 乙酸甲酯 (饱和) |
|
|
|
| 仪器 |
Agilent 1100 高校液相色谱仪 (Agilent Corporation, USA); 2000ES 蒸发光散射品质检测器 (ELSD) (Alltech Corporation) |
| 色谱柱 |
ODS-C 18柱 (4.6 mm × 250 mm, 5 µm) (Dalian Elite Analytical Instruments Co., Ltd.) |
| 流动相 |
甲醇-四氢呋喃-水 (V/V/V, 25: 5: 70) 恒组成溶剂洗脱系统, 1.0 mL/min |
|
|
|
| 仪器 |
Series 1100 仪 (Agilent Technologies, Palo Alto, CA) |
| 色谱柱 |
Gemini octadecylsilyl (C18) 色谱柱 (150 × 4.6 mm id, 5 µm; Phenomenex, Torrance, CA). 配备一个保护柱 (4 × 2.0 mm, Part No. KJO-4282, Phenomenex) |
| 流动相 |
A: 0.1% 甲酸水, B: 0.1% 甲酸乙腈, 0-20 min, 15-100% B, 0.5 mL/min |
|
|
|
| 仪器 |
ACQUITY UPLC™ 仪 (Waters Corp., Milford, MA), ACQUITY UPLC Shield RP18 色谱柱 (50 × 2.1 mm), 1.7 µm 球形多孔颗粒 |
| 色谱柱 |
ACQUITY 色谱柱配备 LC-18 保护柱 (Vanguard 2.1 × 5 mm, Waters Corp.) |
| 流动相 |
0.05% 甲酸水 (A), 0.05% 甲酸乙腈 (B), 0-7 min, 90-75% A, 0.3 mL/min |
|
|
|
| 仪器 |
ThermoFinnigan 气象色谱仪 (Rodano, Italy), 配备FID 检测器 (3 pg C/s, 线性10 6) 及两个注射进样器: 分流/不分流及冷柱头进样. 使用分流模式. |
| 色谱柱 |
19091J-413, 30 m x 0.25mm i.d., 固定相HP-5, 膜厚度0.25 µ m (J&W Scientific, Folsom, USA), Hamilton model no. 701注射器 (Bonaduz, Switzerland). 温度300°C, 分流比1: 10, 分流流率17 mL/min. 程序设计柱箱温度50°C (2 min) 至300°C (for 9.7 min)升温速率30°C min-1.FID基温度: 325°C |
| 流动相 |
载气氦气 5.0 纯级 (Linde Gaz, Poland). 过滤器 OT3-2, R&D 氧气干燥器, R&D 分离. 记录载气流率 1.7 mL/min (35 cm/s). 流入ñ合成气 (350 mL/min), 氢气 (35 mL/min) 和氮气 (补充气, 33 mL/min) 5.0纯级 (Linde Gaz, Poland) |
|
| 样品制备 |
|
方法一 |
|
|
乙酸乙酯 (5-10 mL) 提取的样品 (4.0 g) 上样硅胶柱 (100 g). 乙酸乙酯/己烷洗脱. 初始容积系统 EtOAc/hexanes (3.5: 6.5, 500 mL). EtOAc 分六个梯度增加 (4: 6, 500 mL; 4.5: 5.5, 500 mL; 5: 5, 500 mL; 5.5: 4.5, 500 mL; 6: 4, 400 mL; 6.5: 3.5, 100 mL). 收集: EtOAc/hexanes (45: 55): 银杏内酯 (0.4 g). (50: 50): 粗提 BB (1) 和 GA (2) 合并 GA/GB. (55: 45): GA/GB (1.1 g). (40: 60): GC/GJ (0.4 g) 包含 GA (2) 和 GB (3). Et2O 乙醚部内银杏内酯 (2 mL), Et2O (1 mL) 过滤冲洗得纯银杏内酯 (217 mg 1HnmR P 98%). |
|
|
银杏提取物 (25 g) 或银杏叶 (250 g) 煎煮 1 小时, 得 0.5 L (提取物) 或 2 L (银杏叶) 5% H2O2 溶液 (或其他氧化剂水溶液). EtOAc (250/125/75 mL) 提取三次. 水、80% NaCl水溶液 (稀释至 80%)、饱和盐溶液 (NaHCO3/ Na2S2O3, NaHCO3/ Na2S2O3, or Na2SO3) 冲洗有机层. 干燥后加入 50-60% 萜烯内酯. Dianon HP-20 30 或 Amberlite XAD-16 高分子树脂应用反相色谱以甲醇水溶液分离 (30-90%). 合并萜烯内酯馏分, 去除溶剂. 60% 甲醇溶液填充 WP C18 色谱柱两次洗脱去除银杏酸. |
|
|
己烷提取物 G 中分离标准品.上样硅胶色谱柱 (1% 甲醇氯仿), RP-18 硅胶柱 (0-100% 甲醇, 保留 100% 甲醇馏分), 半制备HPLC制备 (1 × 25 cm 5 µm YMC ODS-AM 色谱柱, H2O/MeOH/AcOH (100: 10: 1), 215 µm 检测波长) |
|
|
| 参考文献 |
|
[1]
|
Chang, T. K. H., et al. (2006). "Effect of Ginkgo biloba extract on procarcinogen-bioactivating human CYP1 enzymes: Identification of isorhamnetin, kaempferol, and quercetin as potent inhibitors of CYP1B1." Toxicology and Applied Pharmacology 213(1): 18-26. |
|
[2]
|
Bouaziz, N., et al. (2002). "Mitochondrial respiratory chain as a new target for anti-ischemic molecules." European Journal of Pharmacology 441(1-2): 35-45. |
|
[3]
|
Qin, H., Song, F., Han, H., Qu, H., Zhai, X., Qin, B., et al. (2011). "Bilobalide inhibits the expression of aquaporin 1, 4 and glial fibrillary acidic protein in rat brain tissue after permanent focal cerebral ischemia." Neural Regeneration Research, 2105-2111. |
|
[4]
|
Teris A. van Beek*, Gerrit P. Lelyveld, "Thin layer chromatography of bilobalide and ginkgolides A, B, C and J on sodium acetate impregnated silica gel." Phytochemical Analysis Volume 4, Issue 3, pages 109-114, May or June 1993 |
|
[5]
|
WANG Xin, L. J. (2012). "Study on Ginkgolides of Wild Ginkgo boliba L. Leaves from Three Gorges Reservoir Areas of Chongqing City." Medicinal Plant, p. 17-19. |
|
[6]
|
Leung, A.Y., & Foster, S. (1996) "Encyclopedia of Common Natural Ingredients Used in Food." Drugs, and Cosmetics, John Wiley & Sons, New York, NY |
|
[7]
|
Bharthi Avula, Yan-Hong Wang, and Troy J. Smille. (2009). "Column Liquid Chromatography/Electrospray Ionization-Time of Flight-Mass Spectrometry and Ultraperformance Column Liquid Chromatography Mass Spectrometry Methods for the Determination of Ginkgolides and Bilobalide in the Leaves of Ginkgo biloba and Dietary Supplements." Journal of AOAC International, pp. 645-652. |
|
[8]
|
Jan Krzek, Janusz S. Czekaj, Wlodzimierz Rzeszutoko and Radoslaw J. Ekiert. (2007)."Validation of Capillary Gas Chromatographic Method for Determination of Bilobalide and Ginkgolides A, B, C in Ginkgo Biloba Dry and Liquid Extracts." Polish Pharmaceutical Society, pp. 303-310 |
|
[9]
|
Stanislav Jaracz, S. M. (2004). "Isolation of ginkgolides A, B, C, J and bilobalide from G. biloba extracts." Phytochemistry, pp. 2897-2902. |
|
[10]
|
Dirk Lichtblau, J. M. (2002). "Efficient Extraction of Ginkgolides and Bilobalide from Ginkgo biloba Leaves." J. Nat. Prod., pp. 1501 - 1504. |
|
