络石藤总黄酮的提取纯化工艺及抗炎镇痛药理作用的研究
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摘要
目的
研究络石藤中总黄酮的提取及纯化工艺;观察在最佳提取纯化工艺下得到的络石藤总黄酮对小鼠的抗炎镇痛作用。
方法
采用微波与超声联用辅助提取络石藤总黄酮,依次进行单因素实验和正交实验,应用紫外分光光度法测定络石藤总黄酮含量,并以其提取率为考察指标,运用统计学方法分析实验结果确定最佳提取工艺;采用大孔吸附树脂纯化络石藤总黄酮,通过静态-动态吸附解吸实验及纯化工艺的考察确定最优纯化树脂和最佳纯化工艺;通过Xyl致小鼠耳肿胀和CAR致大鼠足趾肿胀观察纯化后络石藤总黄酮抗炎作用;通过小鼠热板法和小鼠HAc扭体法观察其镇痛作用。
结果
应用微波与超声联用辅助提取法,按照最优提取工艺得到的络石藤总黄酮提取率高于传统提取方法,并能显著缩短提取时间;采用大孔吸附树脂纯化的络石藤总黄酮,其纯度可达到86.43%;纯化后的络石藤总黄酮在抗炎实验中具有显著的抑制小鼠耳肿胀和大鼠足跖肿胀的作用,具有明显的抗炎作用;在镇痛实验中,总黄酮能显著减少小鼠扭体次数,提高小鼠痛阈值,具有一定的镇痛作用。
结论
微波与超声波联用辅助提取络石藤总黄酮的方法是可行的。粗提物可经大孔吸附树脂有效的纯化,纯化后的络石藤总黄酮具有一定的抗炎镇痛作用。
Objective
To study the extraction and purification process of total flavonoids from Chinese Starjasmine Stem and anti-inflammatory and analgesic effect of total flavonoids in the best condition of extraction and purification process.
Methods
The single factor test and the orthogonal test were taken to study the best extraction conditions of total flavonoids from Chinese Starjasmine Stem by microwave-assisted method and ultrasonic wave cooperated method through measuring the extraction rate of the total flavonoids by UV spectrophotometry and using statistical Methods for analysis of experimental results. Macroporous adsorptive resins was used in puriting total flavonoids from Chinese Starjasmine Stem for confirming the optimal purification resins and the best purification process through the static-dynamic adsorption and desorption experiments and purpification experiment. The anti-inflammatory effect of total flavonoids from Chinese Starjasmine Stem was researched using ear edema induced by xylene and paw edema caused by carrageen; and analgesic effects of Chinese Starjasmine Stem were observed in pain models of mice induced by acetic acid or hot plate.
Results
The extraction rate of the total flavonoids from Chinese Starjasmine Stem using the best extraction conditions were higher than the rate of using traditional method by microwave-assisted method and ultrasonic wave cooperated method and could significantly shorten the extraction time. The purity of total flavonoids could reach 86.43% in the best condition of extraction and purification process. The total flavonoids has inhibitory effect on the swelling of mouse ear piece and rat paw edema. The total flavonoids from Chinese Starjasmine Stem have analgesic effects on the two pain models.
Conclusions
The method of microwave and ultrasonic extraction in the best condition was feasible. Absorbent resin could effictively increase the purity of total flavonoids. The puritied total flavonoids from Chinese Starjasmine Stem have anti-inflammatory and analgesic effects.
研究络石藤中总黄酮的提取及纯化工艺;观察在最佳提取纯化工艺下得到的络石藤总黄酮对小鼠的抗炎镇痛作用。
方法
采用微波与超声联用辅助提取络石藤总黄酮,依次进行单因素实验和正交实验,应用紫外分光光度法测定络石藤总黄酮含量,并以其提取率为考察指标,运用统计学方法分析实验结果确定最佳提取工艺;采用大孔吸附树脂纯化络石藤总黄酮,通过静态-动态吸附解吸实验及纯化工艺的考察确定最优纯化树脂和最佳纯化工艺;通过Xyl致小鼠耳肿胀和CAR致大鼠足趾肿胀观察纯化后络石藤总黄酮抗炎作用;通过小鼠热板法和小鼠HAc扭体法观察其镇痛作用。
结果
应用微波与超声联用辅助提取法,按照最优提取工艺得到的络石藤总黄酮提取率高于传统提取方法,并能显著缩短提取时间;采用大孔吸附树脂纯化的络石藤总黄酮,其纯度可达到86.43%;纯化后的络石藤总黄酮在抗炎实验中具有显著的抑制小鼠耳肿胀和大鼠足跖肿胀的作用,具有明显的抗炎作用;在镇痛实验中,总黄酮能显著减少小鼠扭体次数,提高小鼠痛阈值,具有一定的镇痛作用。
结论
微波与超声波联用辅助提取络石藤总黄酮的方法是可行的。粗提物可经大孔吸附树脂有效的纯化,纯化后的络石藤总黄酮具有一定的抗炎镇痛作用。
Objective
To study the extraction and purification process of total flavonoids from Chinese Starjasmine Stem and anti-inflammatory and analgesic effect of total flavonoids in the best condition of extraction and purification process.
Methods
The single factor test and the orthogonal test were taken to study the best extraction conditions of total flavonoids from Chinese Starjasmine Stem by microwave-assisted method and ultrasonic wave cooperated method through measuring the extraction rate of the total flavonoids by UV spectrophotometry and using statistical Methods for analysis of experimental results. Macroporous adsorptive resins was used in puriting total flavonoids from Chinese Starjasmine Stem for confirming the optimal purification resins and the best purification process through the static-dynamic adsorption and desorption experiments and purpification experiment. The anti-inflammatory effect of total flavonoids from Chinese Starjasmine Stem was researched using ear edema induced by xylene and paw edema caused by carrageen; and analgesic effects of Chinese Starjasmine Stem were observed in pain models of mice induced by acetic acid or hot plate.
Results
The extraction rate of the total flavonoids from Chinese Starjasmine Stem using the best extraction conditions were higher than the rate of using traditional method by microwave-assisted method and ultrasonic wave cooperated method and could significantly shorten the extraction time. The purity of total flavonoids could reach 86.43% in the best condition of extraction and purification process. The total flavonoids has inhibitory effect on the swelling of mouse ear piece and rat paw edema. The total flavonoids from Chinese Starjasmine Stem have analgesic effects on the two pain models.
Conclusions
The method of microwave and ultrasonic extraction in the best condition was feasible. Absorbent resin could effictively increase the purity of total flavonoids. The puritied total flavonoids from Chinese Starjasmine Stem have anti-inflammatory and analgesic effects.
引文
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5王杰松,张俊平,张珉,等.脂多糖诱导细胞产生一氧化氮及木黄酮对其的抑制作用.第二军医大学学报,2002,23(12):1361.
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2姚新生,吴立军,吴继洲·天然药物化学(第四版)·北京:人民卫生出版社,2004·101-105.
3高锦明·植物化学·北京:科技出版社,2003·57-58.
4 Tan XQ, Chen HS, Liu RH, et al. Lignans from Trachelospermum jasminoides. Planta Med, 2005, 71(1): 93.
5富乐,赵毅民,王金辉.络石藤黄酮类化学成分研究.解放军药学学报,2008,24(4):299-301.
6王杰松,张俊平,张珉,等.脂多糖诱导细胞产生一氧化氮及木黄酮对其的抑制作用.第二军医大学学报,2002,23(12):1361.
7 Lee MH, Lee JM, Jun SH. In-vitro and in-vivo anti-inflammatory action of the ethanol extract of Trachelospermi caulis. J Pharm Pharmacol, 2007, 59(1): 123.
8西部三省,韩英梅.络石藤中的木脂素类成分及其生物活性.中国民族民间医药杂志,2001, 48(5): 1.
9刘国凌,宁正祥,彭小红.凤尾草黄酮类化合物的水提工艺条件研究.食品科学,2008, 29(8):286-288.
10唐敏,刘耀,王渝,等.均匀试验优化金银花总黄酮提取工艺.中国药房,2008, 19(18):1375-1376.
11 Pan Yingming, He Chunhuan, Wang Hengshan, et al. Antioxidant activity of microwave-assisted extract of Buddleia officinalis and its major active component. Food Chemistry, 2010,121(2):497-502.
12赵兵.中药提取分离新技术.北方牧业,2007,6(3):8-9.
13刘增琪,景涛.中药提取分离技术的应用进展.天津药学,2003,4(15):64-67.
14胡军,周跃华.大孔吸附树脂在中药成分精制纯化中的应用.中成药,2002,24(2):127-130.
15任荣军.大孔树脂吸附技术在黄酮类成分分离纯化中的应用.中国药学,2008,17(16):71-73.
16闫磊,何再安,刘焱文.大孔吸附树脂在中药研究中的应用.时珍国医国药,2006,17(12):2585-2586.
17周群,傅海滢,白佳伟.紫外分光光度法测定络石藤中总黄酮含量.时珍国医国药,2007,18(2):352.
18陈双,史俊燕,钟洁.微波辅助提取小麦麸皮总黄酮技术研究.食品与药品, 2009,11(3):18-20.
19马陶陶,张群林,李俊,等.三氯化铝比色法测定中药总黄酮方法的探讨.时珍国药研究,2008,19(1):54-56.
20伍晓春.正交试验优选蔓荆子总黄酮微波辅助提取纯化工艺.中国药房,2009,20(24) :1867-1869.
21李华荣,孙鑫.木瓜中总黄酮提取方法研究进展.中国药业,2010,19(16):85.
22李青山,丁红·药学基本实验技术·北京:中国医药科技出版社,2006·279-284.
23王大林.正交试验在中成药制备工艺中研究应用的概况.中成药,1997,19(11):41.
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