甘草抗肝炎有效部位及其纳米粒的制备
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摘要
肝炎是危害人类健康的严重疾病之一,近年来中医药在治疗肝炎方面取得了长足的发展,从传统的中药中寻找有效部位治疗病毒性肝炎已成为研究的热点。豆科甘草属植物甘草是常用中药材,它的主要成分为甘草酸等三萜皂苷和甘草苷为主的甘草黄酮,近年的研究发现其具有解痉镇痛,抗病毒,抗炎症,抗癌等多种药理作用,而甘草治疗肝炎的应用在国内外均有报道。一般的抗肝炎药物靶向性差,而利用纳米粒作为药物载体可有效地促进药物进入肝细胞,从而增强药物的肝靶向性和疗效,因此可用于治疗病毒性肝炎。
本课题为甘草抗肝炎有效部位及其纳米粒的制备。首先为全面控制甘草药材的质量,为制剂提供稳定的药材来源,用高效液相色谱法建立了不同产地甘草药材的指纹图谱,然后提取纯化了甘草抗肝炎有效部位—甘草总皂苷,考察了甘草总皂苷的提取纯化工艺,接着以羧甲基壳聚糖为载体制备了甘草总皂苷纳米粒并对其进行了质量评价。主要研究内容和结论如下：
1.采用高效液相色谱法(HPLC)建立了不同产地甘草药材的指纹图谱。采用Hypersil ODS2C18 (4.6mm×250mm; 5μm)色谱柱,甲醇-0.1%磷酸水溶液为流动相梯度洗脱,流速为0.6 mL/min,检测波长254nm,采集时间为90min。结果得到了分离度较好的甘草药材HPLC指纹图谱,标定出了11个共有指纹峰,方法学考察结果符合指纹图谱技术要求。确定的指纹图谱研究方法稳定、可靠,可用于甘草药材的质量控制。
2.研究了提取纯化甘草总皂苷的工艺条件及参数。以正交试验考察了甘草总皂苷的提取工艺；以甘草总皂苷质量浓度为指标,考察了大孔树脂纯化工艺。结果确定甘草总皂苷的最佳提取工艺为：加水量8倍,煎煮时间2h,煎煮次数3次；采用HPD300型树脂的纯化工艺为上样液质量浓度为0.2g生药／mL,上样量为5倍柱体积,50%乙醇洗脱,用量为6倍柱体积。实验结果表明在所确定的工艺条件下,可较好地提取纯化甘草总皂苷。
3.制备了甘草总皂苷纳米粒并对其进行了质量评价。以正交试验设计法优化了甘草总皂苷纳米粒的制备条件,以透射电镜观察纳米粒形态,紫外分光光度仪测定纳米粒包封率和载药量,激光散射仪测定粒度分布,同时进行纳米粒的体外释放性能研究。结果显示纳米粒外观形态圆整,平均粒径为114nm,包封率为43.4%,载药量为16.4%。所确定的实验方法为甘草总皂苷纳米粒制剂的研究和.应用提供了参考。
Hepatitis is one of the serious disease which threaten to human health, and in recent years Traditional Chinese Medicine has made great progress on the treatment of Hepatitis. The present invention relates to a process for the treatment of viral hepatitis by searching for effective fraction from Traditional Chinese Medicine. Leguminosae Glycyrrhiza licorice is commonly used in traditional Chinese medicine, which is composed of Triterpenoid saponins such as Glycyrrhizin and FGU major in LQ. Recently all sources mention its use for antispasmodic analgesic, anti-virus, anti-inflammatory, anti-cancer and other pharmacological effects, in addition, the treatment of hepatitis using licorice have been reported at home and abroad. The general anti-hepatitis drug is lack of targeting, while making use of nanoparticles as drug carrier can effectively accelerate the drug into the liver cells, in further enhancing liver targeting of drugs and efficacy. Thus it can be used to the treatment to viral hepatitis.
This topic relates to a process for searching for an anti-hepatitis effective fraction from Licorice and the preparation of nanoparticles. First of all, we must make full control of the quality of licorice, provide a stable source of medicine for the preparation. Meanwhile we adopt high performance liquid chromatography to establish the fingerprint of Glycyrrhiza uralensis from different Fisch origin, and then extract from licorice and purify the effective parts of anti-hepatitis-total saponins from liquorice.. Furthermore, we study the process of the extraction and purification technology of total saponins from liquorice., use Carboxymethyl chitosan as drug carrier to prepare for nanoparticles,and make evaluation of its quality. The main contents and conclusions are as follows:
1. To establish the HPLC fingerprint of Glycyrrhiza from different habitats. Separation was performed on a Hypersil ODS2 C18 column(4.6mm×250mm; 5μm) with methanol-0.1% phosphate water as mobile phase using gradient elution at flow rate of 0.6mL/min. The detection wavelength was at 254 nm and the analysis time was 70 min. The HPLC fingerprints of Glycyrrhiza with good separation were set up. And there were 11 common peaks. The results of method validation was consistent with the technical standard of fingerprints. The method of fingerprints was stable and reliable,so as to apply to the quality control of Glycyrrhiza.
2. To investigate the technological parameters of the extraction and purification process of total saponins from liquorice. Studied the extraction of total saponins from liquorice with orthogonal test; The technical process for purification of total saponins with the macroporous resins was screened by yield of total saponins as index. The optimum technological conditions for extraction:the water consumption 8 times, the total time 2h, the decoction times 3 times; The optimum parameters with resin HPD300 were established as followings:5 BV sample extract whose concentration was 0.2g (crude drug)/mL was passed through the column,and then the column was eluted with 6 BV 50% ethanol. With definite technical process, the total saponins from liquorice could be well extracted and purified.
3. To prepare the nanoparticles of total saponins from liquorice and to evaluate its quality. The formulation of NP were optimized by the orthogonal test. The morphology was examined by the transmission electron microscope. The ultraviolet spectrophotometer was employed to determine the entrapment rate and drug-loading rate of NP. The diameter of NP were investigated by laser light scattering analysis instrument. And the release performance of NP in vitro was studied. The nanoparticles were round, smooth particles with average size of 114nm, entrapment rate 43.4% and drug-loading rate 16.4%. The result can provide a reference for studying the nanoparticles of total saponins from liquorice.
本课题为甘草抗肝炎有效部位及其纳米粒的制备。首先为全面控制甘草药材的质量,为制剂提供稳定的药材来源,用高效液相色谱法建立了不同产地甘草药材的指纹图谱,然后提取纯化了甘草抗肝炎有效部位—甘草总皂苷,考察了甘草总皂苷的提取纯化工艺,接着以羧甲基壳聚糖为载体制备了甘草总皂苷纳米粒并对其进行了质量评价。主要研究内容和结论如下：
1.采用高效液相色谱法(HPLC)建立了不同产地甘草药材的指纹图谱。采用Hypersil ODS2C18 (4.6mm×250mm; 5μm)色谱柱,甲醇-0.1%磷酸水溶液为流动相梯度洗脱,流速为0.6 mL/min,检测波长254nm,采集时间为90min。结果得到了分离度较好的甘草药材HPLC指纹图谱,标定出了11个共有指纹峰,方法学考察结果符合指纹图谱技术要求。确定的指纹图谱研究方法稳定、可靠,可用于甘草药材的质量控制。
2.研究了提取纯化甘草总皂苷的工艺条件及参数。以正交试验考察了甘草总皂苷的提取工艺；以甘草总皂苷质量浓度为指标,考察了大孔树脂纯化工艺。结果确定甘草总皂苷的最佳提取工艺为：加水量8倍,煎煮时间2h,煎煮次数3次；采用HPD300型树脂的纯化工艺为上样液质量浓度为0.2g生药／mL,上样量为5倍柱体积,50%乙醇洗脱,用量为6倍柱体积。实验结果表明在所确定的工艺条件下,可较好地提取纯化甘草总皂苷。
3.制备了甘草总皂苷纳米粒并对其进行了质量评价。以正交试验设计法优化了甘草总皂苷纳米粒的制备条件,以透射电镜观察纳米粒形态,紫外分光光度仪测定纳米粒包封率和载药量,激光散射仪测定粒度分布,同时进行纳米粒的体外释放性能研究。结果显示纳米粒外观形态圆整,平均粒径为114nm,包封率为43.4%,载药量为16.4%。所确定的实验方法为甘草总皂苷纳米粒制剂的研究和.应用提供了参考。
Hepatitis is one of the serious disease which threaten to human health, and in recent years Traditional Chinese Medicine has made great progress on the treatment of Hepatitis. The present invention relates to a process for the treatment of viral hepatitis by searching for effective fraction from Traditional Chinese Medicine. Leguminosae Glycyrrhiza licorice is commonly used in traditional Chinese medicine, which is composed of Triterpenoid saponins such as Glycyrrhizin and FGU major in LQ. Recently all sources mention its use for antispasmodic analgesic, anti-virus, anti-inflammatory, anti-cancer and other pharmacological effects, in addition, the treatment of hepatitis using licorice have been reported at home and abroad. The general anti-hepatitis drug is lack of targeting, while making use of nanoparticles as drug carrier can effectively accelerate the drug into the liver cells, in further enhancing liver targeting of drugs and efficacy. Thus it can be used to the treatment to viral hepatitis.
This topic relates to a process for searching for an anti-hepatitis effective fraction from Licorice and the preparation of nanoparticles. First of all, we must make full control of the quality of licorice, provide a stable source of medicine for the preparation. Meanwhile we adopt high performance liquid chromatography to establish the fingerprint of Glycyrrhiza uralensis from different Fisch origin, and then extract from licorice and purify the effective parts of anti-hepatitis-total saponins from liquorice.. Furthermore, we study the process of the extraction and purification technology of total saponins from liquorice., use Carboxymethyl chitosan as drug carrier to prepare for nanoparticles,and make evaluation of its quality. The main contents and conclusions are as follows:
1. To establish the HPLC fingerprint of Glycyrrhiza from different habitats. Separation was performed on a Hypersil ODS2 C18 column(4.6mm×250mm; 5μm) with methanol-0.1% phosphate water as mobile phase using gradient elution at flow rate of 0.6mL/min. The detection wavelength was at 254 nm and the analysis time was 70 min. The HPLC fingerprints of Glycyrrhiza with good separation were set up. And there were 11 common peaks. The results of method validation was consistent with the technical standard of fingerprints. The method of fingerprints was stable and reliable,so as to apply to the quality control of Glycyrrhiza.
2. To investigate the technological parameters of the extraction and purification process of total saponins from liquorice. Studied the extraction of total saponins from liquorice with orthogonal test; The technical process for purification of total saponins with the macroporous resins was screened by yield of total saponins as index. The optimum technological conditions for extraction:the water consumption 8 times, the total time 2h, the decoction times 3 times; The optimum parameters with resin HPD300 were established as followings:5 BV sample extract whose concentration was 0.2g (crude drug)/mL was passed through the column,and then the column was eluted with 6 BV 50% ethanol. With definite technical process, the total saponins from liquorice could be well extracted and purified.
3. To prepare the nanoparticles of total saponins from liquorice and to evaluate its quality. The formulation of NP were optimized by the orthogonal test. The morphology was examined by the transmission electron microscope. The ultraviolet spectrophotometer was employed to determine the entrapment rate and drug-loading rate of NP. The diameter of NP were investigated by laser light scattering analysis instrument. And the release performance of NP in vitro was studied. The nanoparticles were round, smooth particles with average size of 114nm, entrapment rate 43.4% and drug-loading rate 16.4%. The result can provide a reference for studying the nanoparticles of total saponins from liquorice.
引文
[1]王常松,刘清华.中医药治疗乙型肝炎必须面对的几个问题[J].光明中医,2010,25(3)：345-348.
[2]Susumu Ohwada. Severe acute cholestatic hepatitis of unknown etiology successfully treated with the Chinese herbal medicine [J]. World Journal of Gastroenterology,2009, 15(23):2927-2929.
[3]黄春燕.慢性乙型肝炎的中医药治疗概况[J].江西中医药,2009,21(2)：130-131.
[4]陈志杰.中药辨证治疗改善300例慢性乙型肝炎患者生活质量评估[J].福建中医药,2009,40(5)：11-12.
[5]Zhang QY, Ye M. Chemical analysis of the Chinese herbal medicine Gan-Cao (licorice)[J]. Journal of Chromatography A,2009,1954-1969.
[6]陶晡,刘晓清,屈振刚.甘草化学成分研究进展[J].河北农业科学,2009,13(3)：77-79.
[7]仇志坤,冯冰虹.甘草中主要有效成分的抗肿瘤和神经保护作用[J].中国神经肿瘤杂志,2009,7(1)：70-76.
[8]国家药典委员会.中国药典(一部)[S].北京：中国化学工业出版社,2005：59-60.
[9]陈红.甘草药理作用概述[J].海峡药学,2005,17(4)：37-41.
[10]王敏.甘草研究综述[J].齐鲁药事,2005,24(10)：614-616.
[11]Hosseinzadeh Hossein. Review of pharmacological effects of Glycyrrhiza sp and its bioactive compounds [J]. Phytotherapy Research,2008,22(6):709-724.
[12]梁冰,杨爱馥,黄凤兰,胡宝忠.甘草属化学成分及药理作用研究进展[J].东北农业大学学报,2006,37(1)：115-119.
[13]Cristina Fiore. Antiviral effects of Glycyrrhiza species [J]. Phytotherapy Research,2008, 22(2):141-148.
[14]崔英杰.肝靶向制剂的研究进展[J].山西医药杂志,2006,35(11)：988-992.
[15]许红霞,高晓黎.肝靶向给药系统的研究进展[J].西北药学杂志,2006,21(3)：140-141.
[16]Chen QH,Lu WG,Ge QH, al. Study on targeting drug delivery system--the characteristics of methotrexate microsphere and experimental treatment of hepatic tumor in rats by arterial embolization[J]. Yao Xue Xue Bao,1991,26(4):293-8.
[17]Murakami Y, Tabata Y, Ikada Y. Tumor accumulation of poly (ethylene glycol) with different molecular weights after intravenous injections[J]. Drug Deliv,1997,4:23.
[18]袁东锋,易以木.肝靶向纳米粒的研究进展[J].医药导报,2003,22(2)：113-114.
[19]李强,李超.中药指纹图谱研究综述[J].齐鲁药事,2010,29(3)158-161.
[20]国家药品监督管理局.中药注射剂指纹图谱研究的技术要求(暂行)[J].中成药,2000,22(10)：671-675.
[21]赵德玉,毕桂宏.中药指纹图谱的不同方法特点及应用[J].中华中医药学刊,2007, 25(3)：601-603.
[22]张丁华,王艳丰.中药指纹图谱技术及应用研究进展[J].中兽医医药杂志,2010,29(1)：26-28.
[23]高鹏,代龙.薄层色谱法在中药指纹图谱研究中的应用[J].山东医药工业,2002,21(6)：23-25.
[24]杨先国,陈四保,等.玉竹中黄酮类成分的薄层色谱指纹图谱研究[J].中国中药杂志,2005,30(2)：104-106.
[25]郑琦方悦.气相色谱技术在中药指纹图谱中的应用[J].浙江中西医结合杂志,2008,18(11)：713-714.
[26]张丹,王霆,等.广枣四香滴丸中挥发性成分的GC指纹图谱研究[J].广东药学院学报,2009,25(3)：253-255.
[27]许一平,罗启恩.高效液相色谱技术在中药指纹图谱研究中的应用[J].中国科技信息,2007,12：179-180.
[28]TANRui, LIU Bo-ping. Fingerprint Analysis of PolygonumavculareL. Using High-Performance Liquid Chromatography[J]. Journal of Southwest Jiaotong University, 2009,17(2):139-142.
[29]顾铭,欧阳藩,等.高速逆流色谱法分离纯化丹参并尝试制订中药指纹图谱[J].生物工程学报,2003,19(6)：740-744.
[30]张军,檀华蓉,等.采用高效毛细管电泳法建立不同桑品种桑叶黄酮类化合物指纹图谱的试验[J].蚕业科学,2008,34(1)：119-123.
[31]赵大伟,孙丽梅,等.中药指纹图谱技术在中药研究中的应用[J].黑龙江科技信息,2009,30：251-251.
[32]Guo-Hua Zheng, Gui-Hong Wang.Development of a fingerprint of Melilotus suaveolens Ledeb by high-performance liquid chromatography [J].Journal of Chinese Pharmaceutical Sci ences,2009,01:20-29.
[33]周欣,陈华国,等.蓬莪术药材的指纹图谱研究[J].时珍国医国药,2008,19(6)：1359-1360.
[34]XU Ya-ping, YAO Tong-wei. Study and Application of Fingerprints of Ginkgo biloba Leaves Preparations[J]. Journal of Chinese Pharmaceutical Sciences,2005,14(1):43-50.
[35]李滔,王维剑,等.乌檀注射剂中间体指纹图谱研究[J].中国中药杂志,2005,30(15)：1156-1158.
[36]王冬梅,刘朝燊,等.指纹图谱技术对贯叶连翘提取物制备工艺过程的评价[J].中国中药杂志,2006,31(10)：800-804.
[37]吕春平,关铭.中药指纹图谱的应用[J].天津药学,2004,16(2)：58-62.
[38]孙波,孙涛.高效液相色谱法在中药指纹图谱中的应用现状及分析[J].时珍国医国药,2006,17(1)：109-110.
[39]孙磊,乔善义.中药指纹图谱应用研究进展[J].国际药学研究杂志,2009,36(3)：194-197.
[40]刘博,张智博,等.新技术在现代中药提取中的研究进展[J].黑龙江科技信息,2009, 6：167.
[41]董立丽.中药提取分离新技术的进展[J].海峡药学,2009,21(12)：107-109.
[42]廖国庆.中药提取技术进展分析[J].医学信息,2009,1(9)：159-160.
[43]胡道德,顾磊,等.中药提取及优化的研究进展[J].医药导报,2009,28(1)：80-83.
[44]葛云初,黎阳.超临界流体萃取技术及其在中药提取中的应用[J].现代药物与临床,2009,24(5)：279-281.
[45]陈栋,周永传.酶法在中药提取中的应用和进展[J].中国中药杂志,2007,32(2)：99-101.
[46]韩章远,韩永龙,等.酶解技术在中药提取中的应用进展[J].广州化工,2009,37(6)：12-17.
[47]师奇松,刘太奇.中药有效成分分离技术新进展[J].科技创新导报,2009,15：9.
[48]刘光喜.中药有效成分的分离纯化技术研究概况[J].中医药导报,2008,14(7)：118-120.
[49]李敏杰,邓启刚,等.中药有效成分分离纯化工艺概述[J].齐齐哈尔大学学报,2006,22(2)：7-10.
[50]王凯,岳宣峰.中药有效成分分离纯化技术应用进展[J].榆林学院学报,2005,15(3)：21-23.
[51]李平华,王兴文.大孔吸附树脂在中药有效成分分离纯化中的研究进展[J].云南中医学院学报,2003,26(3)：43-48.
[52]刘鄂湖,鄢丹,等.大孔吸附树脂技术在中药中应用现存问题分析与探讨[J].中草药,2007,38(5)：1003-1005
[53]富玉.天然产物中有效成分分离及测试方法概述[J].中国测试技术,2007,33(2)：22-25.
[54]孙君辉,孟秀芬.大孔树脂的应用及其提取纯化中药有效成分的影响因素[J].山东商业职业技术学院学报,2008,8(6)：94-96.
[55]李崇明,熊富良.新药研究中大孔树脂型号与规格的选择应用[J].中成药,2008,30(8)：1208-1210.
[56]刘彬果,郭文勇,等.大孔树脂吸附技术在中药制剂中的应用[J].解放军药学学报,2003,19(6)：452-455.
[57]王一敏,任晓蕾.大孔树脂的应用研究[J].中医药信息,2008,25(4)：26-28.
[58]董珂.大孔树脂技术在中药研究中的应用概况[J].药学实践杂志,2006,24(1)：13-16.
[59]Li Huayue, Jae-Hwa Lee. Effective Purification of Ginsenosides from Cultured Wild Ginseng Roots, Red Ginseng, and White Ginseng with Macroporous Resins[J]. Journal of microbiology and biotechnology,2008,18(11):1789-1791.
[60]Yu Kong, Ming-Ming Yan. Preparative enrichment and separation of astragalosides from Radix Astragali extracts using macroporous resins[J]. Journal of Separation Science 2010 33(15):2278-2286.
[61]ZHANG Ying, LI Shufen. Macroporous Resin Adsorption for Purification of Flavonoids in Houttuynia cordata Thunb[J]. Chin. J. Chem. Eng,2007,15(6):872-876.
[62]Jun Wang, Fu An Wu. Isolation of flavonoids from mulberry (Morus alba L.) leaves with macroporous resins[J]. African Journal of Biotechnology,2008,7(13):2147-2155.
[63]Xuegong Qin, Yuanjin Yuan. Separation of Alkaloids from Sophora alopecuroides L. by Adsorption Using Macroporous Resins[J]. Chem. Eng. Japan,2007,40:93-97.
[64]潘道东,林璐.DA201-C大孔吸附树脂静态吸附ACE抑制肽的研究[J].食品科学,2009,30(5)：20-23.
[65]Zhang L, Granick S. How to stabilize phospholipid liposomes-using nanoparticles[J]. Nano Lett,2006,6:694-8.
[66]Bangham A, Standish M. Diffusion of univalent ions across the lamellae of swollen phospholipids[J]. J. Mol Biol,1965,13:238-52.
[67]Bangham AD. Liposome Letters[M]. Academic Press London,1983.
[68]连佳芳,张三奇.固体脂质纳米粒研究进展[J].第四军医大学学报,2005,26(17)：1621-1623.
[69]Zur Muhlen A, Schwarz C. Solid lipid nanoparticles for controlled drug delivery-drug release and release mechanism[J]. J Pharm Biopharm,1998,45:149-55.
[70]Gupta A, Cooper E. Update in antifungal therapy of dermatophytosis[J]. Mycopathologia, 2008,166:353-67.
[71]尚青,郑和堂.聚合物纳米粒子载药体系的研究进展[J].河北工业科技,2005,22(1)：38-43.
[72]Cheng J, Teply BA, Sherifi I, Sung J, Luther G, Gu F X, et al. Formulation of functionalized PLGA-PEG nanoparticles for in vivo targeted drug delivery[J]. Biomaterials,2007, 28:869-76.
[73]Gu F X, Zhang L, Teply BA, Mann N, Wang A, Langer R, et al. Precise engineering of targeted nanoparticles by using self-assembled biointegrated block copolymers[J]. Proc. Natl. Acad. Sci,2008,105:2586-91.
[74]马国,邓盛齐.纳米技术在药学中的研究应用进展[J].国外医药抗生素分册,2004,25(5)：233-237.
[75]Chen X, Young TJ, Sarkari M, et al. Preparation of cyclosporine A nanoparticles by evaporative precipitation into aqueous solution[J]. Int J Pharm,2002,242(1-2):3-14.
[76]贺玲,李健和.肝靶向给药系统的研究进展[J].中国当代医药,2010,17(9)：15-18.
[77]田洁,逄秀娟.顺铂固体脂质纳米粒的制备及其在大鼠体内的分布[J].中国药剂学杂志,2008,6(4)：141-149.
[78]余克富,逄秀娟,等.布洛芬固体脂质纳米粒的制备及性质考察[J].中国药剂学杂志,2009,7(3)：91-99.
[79]陈莹,平其能.口服载药纳米粒的研究进展[J].药学进展,2004,28(10)：451-455.
[80]唐仕炜,袁晓佳,等.胸腺五肽三甲基壳聚糖口服纳米粒的研究[J].四川大学学报,2007,38(5)：885-888.
[81]李春,冯神飞,等.纳米粒的制备方法在药物制剂中的应用概况[J].天津药学,2009 21(6)：59-60.
[82]Mende S, Stenger F, Peukert W, Schwedes J. Mechanical production and stabilization of submicron particles in stirred media mills[J]. Powder Technology,2003,132:64-73.
[83]Merisko-Liversidge E, Liversidge GG, Cooper ER. Nanosizing:a formulation approach for poorly-water-soluble compound[J]. European Journal of Pharmaceutical Sciences,2003,18: 113-120.
[84]刘志挺.溶剂蒸发法在微球制备中的应用及研究进展[J].广东药学院学报,2007,23(5)：596-600.
[85]Jung Yoon Jang, Byung Soo Kwon, Hea Eun Lee.Preparation of Biodegradable PLGA Nanospheres Employing a Fast Solvent Evaporation Method[J]. J.Ind.Eng.Chem,2007, 13(6):1043-1046.
[86]Allemann E, Gurny R, Doelker E. Drug-loaded nanoparticles-preparation methods and drug targeting issues[J]. Eur J Pharm Biopharm,1993,39(5):173.
[87]王琪,丛龙波,等.纳米粒载药系统研究进展[C].2007年中华中医药学会中成药学术研讨会论文集：91-97.
[88]任慧,丁一刚.超临界流体技术在超微粒子制备中的应用[J].化工进展,2001,20(9)：30-32.
[89]银建中,张宪阵.超临界流体沉积技术在纳米复合材料制备中的应用[J].化学进展,2009,21(4)：606-614.
[90]李秀凤,殷辉安,等.纳米中药研究进展[J].中医药通报,2004,3(5)：53-56.
[91]刘平,吉爱国.壳聚糖载药纳米粒的制备方法及应用展望[C].第二届中国资源生物技术与糖工程学术研讨会论文集：157-161.
[92]Muzzarelli R. Biochemical significance of exogenous chitins and chitosans in animals and patients[J]. Carbohyd Polym,1993,20:7-16.
[93]Tang ZX, Qian JQ, Shi LE. Preparation of chitosan nanoparticles as carrier for immobilized enryme[J]. Appl Biochem Biotechnol,2007,136(1):77-96.
[94]Roller S, Covill N. The antimicrobial properties of chitosan in may onnaise and may onnaise based shrimp salads [J]. Food Prot,2000,63 (2):202-209.
[95]Sugamori T, Iwase H. Local hemostatic effects of microcrystalline partially deacetylated chitin hydrochloride [J]. Biomed Mater Res,2000,49 (2):225-232.
[96]周少华,陈勇,等.壳聚糖的纳米化及其生物学效应[J].中国组织工程研究与临床康复,2007,11(26)：5190-5193.
[97]L Illum. Chitosan and its use as a pharmaceutical excipient [J]. Pharm Res,1998,15(9): 1326-1331.
[98]Grenha A, Seijo B. Microencapsulated chitosan nanoparticles for lung protein delivery [J]. Eur J Pharm Sci,2005,25 (4):427-437.
[99]李玉慧,彭婷婷,等.羧甲基壳聚糖磁性纳米粒子的合成及应用[J].应用化学,2010, 27(1)：87-91.
[100]李志峰,陈钟,等.载肝细胞生长因子聚乳酸-O-羧甲基壳聚糖纳米粒子制备、表征及其对培养大鼠肝细胞活力的影响[J].中国组织工程研究与临床康复,2008,12(32)：6251-6256.
[101]杨祥良,徐辉碧,谢长生,等.基于纳米技术的中药基础问题研究[J].华中理工大学报,2000,28(12)：104-105.
[102]咎旺.纳米中药制剂研究进展[J].成都医学院学报,2008,3(3)：232-234.
[103]马应龙,李淑红.纳米中药研究进展[J].安徽农学通报,2007,13(9)：192-194.
[104]刘丽芳.纳米中药研究与中药现代化[J].中国现代医学杂志,2009,19(8)：1186-1190.
[105]丁志平,乔延江.纳米技术与纳米中药[J].北京中医药大学学报,2003,26(4)：43-46.
[106]寇正涌,任凤梅,等.纳米中药-中药现代化的新途径[J].长春师范学院学报(自然科学版,2007,26(2)：96-98.
[107]唐致远,凌国维.纳米中药研究进展[J].中草药,2007,38(4)：627-629.
[108]段天璇,于密密,刘春生,等HPLC法同时测定甘草指纹图谱暨甘草苷、甘草酸含量[J].中成药,2006,28(2)：161-165.
[109]樊天宇,王跃飞,过科家,等.内蒙古产甘草药材HPLC指纹图谱研究[J].中药新药与临床药理,2007,18(1)：44-46.
[110]兰霞,王洪新.比色法测定甘草中总皂苷的含量[J].时珍国医国药,2007,18(4)：886-887.
[111]林爱华,平其能.壳聚糖载药纳米粒研究进展[J].中国药业,2006,15(21)：25-27.
[2]Susumu Ohwada. Severe acute cholestatic hepatitis of unknown etiology successfully treated with the Chinese herbal medicine [J]. World Journal of Gastroenterology,2009, 15(23):2927-2929.
[3]黄春燕.慢性乙型肝炎的中医药治疗概况[J].江西中医药,2009,21(2)：130-131.
[4]陈志杰.中药辨证治疗改善300例慢性乙型肝炎患者生活质量评估[J].福建中医药,2009,40(5)：11-12.
[5]Zhang QY, Ye M. Chemical analysis of the Chinese herbal medicine Gan-Cao (licorice)[J]. Journal of Chromatography A,2009,1954-1969.
[6]陶晡,刘晓清,屈振刚.甘草化学成分研究进展[J].河北农业科学,2009,13(3)：77-79.
[7]仇志坤,冯冰虹.甘草中主要有效成分的抗肿瘤和神经保护作用[J].中国神经肿瘤杂志,2009,7(1)：70-76.
[8]国家药典委员会.中国药典(一部)[S].北京：中国化学工业出版社,2005：59-60.
[9]陈红.甘草药理作用概述[J].海峡药学,2005,17(4)：37-41.
[10]王敏.甘草研究综述[J].齐鲁药事,2005,24(10)：614-616.
[11]Hosseinzadeh Hossein. Review of pharmacological effects of Glycyrrhiza sp and its bioactive compounds [J]. Phytotherapy Research,2008,22(6):709-724.
[12]梁冰,杨爱馥,黄凤兰,胡宝忠.甘草属化学成分及药理作用研究进展[J].东北农业大学学报,2006,37(1)：115-119.
[13]Cristina Fiore. Antiviral effects of Glycyrrhiza species [J]. Phytotherapy Research,2008, 22(2):141-148.
[14]崔英杰.肝靶向制剂的研究进展[J].山西医药杂志,2006,35(11)：988-992.
[15]许红霞,高晓黎.肝靶向给药系统的研究进展[J].西北药学杂志,2006,21(3)：140-141.
[16]Chen QH,Lu WG,Ge QH, al. Study on targeting drug delivery system--the characteristics of methotrexate microsphere and experimental treatment of hepatic tumor in rats by arterial embolization[J]. Yao Xue Xue Bao,1991,26(4):293-8.
[17]Murakami Y, Tabata Y, Ikada Y. Tumor accumulation of poly (ethylene glycol) with different molecular weights after intravenous injections[J]. Drug Deliv,1997,4:23.
[18]袁东锋,易以木.肝靶向纳米粒的研究进展[J].医药导报,2003,22(2)：113-114.
[19]李强,李超.中药指纹图谱研究综述[J].齐鲁药事,2010,29(3)158-161.
[20]国家药品监督管理局.中药注射剂指纹图谱研究的技术要求(暂行)[J].中成药,2000,22(10)：671-675.
[21]赵德玉,毕桂宏.中药指纹图谱的不同方法特点及应用[J].中华中医药学刊,2007, 25(3)：601-603.
[22]张丁华,王艳丰.中药指纹图谱技术及应用研究进展[J].中兽医医药杂志,2010,29(1)：26-28.
[23]高鹏,代龙.薄层色谱法在中药指纹图谱研究中的应用[J].山东医药工业,2002,21(6)：23-25.
[24]杨先国,陈四保,等.玉竹中黄酮类成分的薄层色谱指纹图谱研究[J].中国中药杂志,2005,30(2)：104-106.
[25]郑琦方悦.气相色谱技术在中药指纹图谱中的应用[J].浙江中西医结合杂志,2008,18(11)：713-714.
[26]张丹,王霆,等.广枣四香滴丸中挥发性成分的GC指纹图谱研究[J].广东药学院学报,2009,25(3)：253-255.
[27]许一平,罗启恩.高效液相色谱技术在中药指纹图谱研究中的应用[J].中国科技信息,2007,12：179-180.
[28]TANRui, LIU Bo-ping. Fingerprint Analysis of PolygonumavculareL. Using High-Performance Liquid Chromatography[J]. Journal of Southwest Jiaotong University, 2009,17(2):139-142.
[29]顾铭,欧阳藩,等.高速逆流色谱法分离纯化丹参并尝试制订中药指纹图谱[J].生物工程学报,2003,19(6)：740-744.
[30]张军,檀华蓉,等.采用高效毛细管电泳法建立不同桑品种桑叶黄酮类化合物指纹图谱的试验[J].蚕业科学,2008,34(1)：119-123.
[31]赵大伟,孙丽梅,等.中药指纹图谱技术在中药研究中的应用[J].黑龙江科技信息,2009,30：251-251.
[32]Guo-Hua Zheng, Gui-Hong Wang.Development of a fingerprint of Melilotus suaveolens Ledeb by high-performance liquid chromatography [J].Journal of Chinese Pharmaceutical Sci ences,2009,01:20-29.
[33]周欣,陈华国,等.蓬莪术药材的指纹图谱研究[J].时珍国医国药,2008,19(6)：1359-1360.
[34]XU Ya-ping, YAO Tong-wei. Study and Application of Fingerprints of Ginkgo biloba Leaves Preparations[J]. Journal of Chinese Pharmaceutical Sciences,2005,14(1):43-50.
[35]李滔,王维剑,等.乌檀注射剂中间体指纹图谱研究[J].中国中药杂志,2005,30(15)：1156-1158.
[36]王冬梅,刘朝燊,等.指纹图谱技术对贯叶连翘提取物制备工艺过程的评价[J].中国中药杂志,2006,31(10)：800-804.
[37]吕春平,关铭.中药指纹图谱的应用[J].天津药学,2004,16(2)：58-62.
[38]孙波,孙涛.高效液相色谱法在中药指纹图谱中的应用现状及分析[J].时珍国医国药,2006,17(1)：109-110.
[39]孙磊,乔善义.中药指纹图谱应用研究进展[J].国际药学研究杂志,2009,36(3)：194-197.
[40]刘博,张智博,等.新技术在现代中药提取中的研究进展[J].黑龙江科技信息,2009, 6：167.
[41]董立丽.中药提取分离新技术的进展[J].海峡药学,2009,21(12)：107-109.
[42]廖国庆.中药提取技术进展分析[J].医学信息,2009,1(9)：159-160.
[43]胡道德,顾磊,等.中药提取及优化的研究进展[J].医药导报,2009,28(1)：80-83.
[44]葛云初,黎阳.超临界流体萃取技术及其在中药提取中的应用[J].现代药物与临床,2009,24(5)：279-281.
[45]陈栋,周永传.酶法在中药提取中的应用和进展[J].中国中药杂志,2007,32(2)：99-101.
[46]韩章远,韩永龙,等.酶解技术在中药提取中的应用进展[J].广州化工,2009,37(6)：12-17.
[47]师奇松,刘太奇.中药有效成分分离技术新进展[J].科技创新导报,2009,15：9.
[48]刘光喜.中药有效成分的分离纯化技术研究概况[J].中医药导报,2008,14(7)：118-120.
[49]李敏杰,邓启刚,等.中药有效成分分离纯化工艺概述[J].齐齐哈尔大学学报,2006,22(2)：7-10.
[50]王凯,岳宣峰.中药有效成分分离纯化技术应用进展[J].榆林学院学报,2005,15(3)：21-23.
[51]李平华,王兴文.大孔吸附树脂在中药有效成分分离纯化中的研究进展[J].云南中医学院学报,2003,26(3)：43-48.
[52]刘鄂湖,鄢丹,等.大孔吸附树脂技术在中药中应用现存问题分析与探讨[J].中草药,2007,38(5)：1003-1005
[53]富玉.天然产物中有效成分分离及测试方法概述[J].中国测试技术,2007,33(2)：22-25.
[54]孙君辉,孟秀芬.大孔树脂的应用及其提取纯化中药有效成分的影响因素[J].山东商业职业技术学院学报,2008,8(6)：94-96.
[55]李崇明,熊富良.新药研究中大孔树脂型号与规格的选择应用[J].中成药,2008,30(8)：1208-1210.
[56]刘彬果,郭文勇,等.大孔树脂吸附技术在中药制剂中的应用[J].解放军药学学报,2003,19(6)：452-455.
[57]王一敏,任晓蕾.大孔树脂的应用研究[J].中医药信息,2008,25(4)：26-28.
[58]董珂.大孔树脂技术在中药研究中的应用概况[J].药学实践杂志,2006,24(1)：13-16.
[59]Li Huayue, Jae-Hwa Lee. Effective Purification of Ginsenosides from Cultured Wild Ginseng Roots, Red Ginseng, and White Ginseng with Macroporous Resins[J]. Journal of microbiology and biotechnology,2008,18(11):1789-1791.
[60]Yu Kong, Ming-Ming Yan. Preparative enrichment and separation of astragalosides from Radix Astragali extracts using macroporous resins[J]. Journal of Separation Science 2010 33(15):2278-2286.
[61]ZHANG Ying, LI Shufen. Macroporous Resin Adsorption for Purification of Flavonoids in Houttuynia cordata Thunb[J]. Chin. J. Chem. Eng,2007,15(6):872-876.
[62]Jun Wang, Fu An Wu. Isolation of flavonoids from mulberry (Morus alba L.) leaves with macroporous resins[J]. African Journal of Biotechnology,2008,7(13):2147-2155.
[63]Xuegong Qin, Yuanjin Yuan. Separation of Alkaloids from Sophora alopecuroides L. by Adsorption Using Macroporous Resins[J]. Chem. Eng. Japan,2007,40:93-97.
[64]潘道东,林璐.DA201-C大孔吸附树脂静态吸附ACE抑制肽的研究[J].食品科学,2009,30(5)：20-23.
[65]Zhang L, Granick S. How to stabilize phospholipid liposomes-using nanoparticles[J]. Nano Lett,2006,6:694-8.
[66]Bangham A, Standish M. Diffusion of univalent ions across the lamellae of swollen phospholipids[J]. J. Mol Biol,1965,13:238-52.
[67]Bangham AD. Liposome Letters[M]. Academic Press London,1983.
[68]连佳芳,张三奇.固体脂质纳米粒研究进展[J].第四军医大学学报,2005,26(17)：1621-1623.
[69]Zur Muhlen A, Schwarz C. Solid lipid nanoparticles for controlled drug delivery-drug release and release mechanism[J]. J Pharm Biopharm,1998,45:149-55.
[70]Gupta A, Cooper E. Update in antifungal therapy of dermatophytosis[J]. Mycopathologia, 2008,166:353-67.
[71]尚青,郑和堂.聚合物纳米粒子载药体系的研究进展[J].河北工业科技,2005,22(1)：38-43.
[72]Cheng J, Teply BA, Sherifi I, Sung J, Luther G, Gu F X, et al. Formulation of functionalized PLGA-PEG nanoparticles for in vivo targeted drug delivery[J]. Biomaterials,2007, 28:869-76.
[73]Gu F X, Zhang L, Teply BA, Mann N, Wang A, Langer R, et al. Precise engineering of targeted nanoparticles by using self-assembled biointegrated block copolymers[J]. Proc. Natl. Acad. Sci,2008,105:2586-91.
[74]马国,邓盛齐.纳米技术在药学中的研究应用进展[J].国外医药抗生素分册,2004,25(5)：233-237.
[75]Chen X, Young TJ, Sarkari M, et al. Preparation of cyclosporine A nanoparticles by evaporative precipitation into aqueous solution[J]. Int J Pharm,2002,242(1-2):3-14.
[76]贺玲,李健和.肝靶向给药系统的研究进展[J].中国当代医药,2010,17(9)：15-18.
[77]田洁,逄秀娟.顺铂固体脂质纳米粒的制备及其在大鼠体内的分布[J].中国药剂学杂志,2008,6(4)：141-149.
[78]余克富,逄秀娟,等.布洛芬固体脂质纳米粒的制备及性质考察[J].中国药剂学杂志,2009,7(3)：91-99.
[79]陈莹,平其能.口服载药纳米粒的研究进展[J].药学进展,2004,28(10)：451-455.
[80]唐仕炜,袁晓佳,等.胸腺五肽三甲基壳聚糖口服纳米粒的研究[J].四川大学学报,2007,38(5)：885-888.
[81]李春,冯神飞,等.纳米粒的制备方法在药物制剂中的应用概况[J].天津药学,2009 21(6)：59-60.
[82]Mende S, Stenger F, Peukert W, Schwedes J. Mechanical production and stabilization of submicron particles in stirred media mills[J]. Powder Technology,2003,132:64-73.
[83]Merisko-Liversidge E, Liversidge GG, Cooper ER. Nanosizing:a formulation approach for poorly-water-soluble compound[J]. European Journal of Pharmaceutical Sciences,2003,18: 113-120.
[84]刘志挺.溶剂蒸发法在微球制备中的应用及研究进展[J].广东药学院学报,2007,23(5)：596-600.
[85]Jung Yoon Jang, Byung Soo Kwon, Hea Eun Lee.Preparation of Biodegradable PLGA Nanospheres Employing a Fast Solvent Evaporation Method[J]. J.Ind.Eng.Chem,2007, 13(6):1043-1046.
[86]Allemann E, Gurny R, Doelker E. Drug-loaded nanoparticles-preparation methods and drug targeting issues[J]. Eur J Pharm Biopharm,1993,39(5):173.
[87]王琪,丛龙波,等.纳米粒载药系统研究进展[C].2007年中华中医药学会中成药学术研讨会论文集：91-97.
[88]任慧,丁一刚.超临界流体技术在超微粒子制备中的应用[J].化工进展,2001,20(9)：30-32.
[89]银建中,张宪阵.超临界流体沉积技术在纳米复合材料制备中的应用[J].化学进展,2009,21(4)：606-614.
[90]李秀凤,殷辉安,等.纳米中药研究进展[J].中医药通报,2004,3(5)：53-56.
[91]刘平,吉爱国.壳聚糖载药纳米粒的制备方法及应用展望[C].第二届中国资源生物技术与糖工程学术研讨会论文集：157-161.
[92]Muzzarelli R. Biochemical significance of exogenous chitins and chitosans in animals and patients[J]. Carbohyd Polym,1993,20:7-16.
[93]Tang ZX, Qian JQ, Shi LE. Preparation of chitosan nanoparticles as carrier for immobilized enryme[J]. Appl Biochem Biotechnol,2007,136(1):77-96.
[94]Roller S, Covill N. The antimicrobial properties of chitosan in may onnaise and may onnaise based shrimp salads [J]. Food Prot,2000,63 (2):202-209.
[95]Sugamori T, Iwase H. Local hemostatic effects of microcrystalline partially deacetylated chitin hydrochloride [J]. Biomed Mater Res,2000,49 (2):225-232.
[96]周少华,陈勇,等.壳聚糖的纳米化及其生物学效应[J].中国组织工程研究与临床康复,2007,11(26)：5190-5193.
[97]L Illum. Chitosan and its use as a pharmaceutical excipient [J]. Pharm Res,1998,15(9): 1326-1331.
[98]Grenha A, Seijo B. Microencapsulated chitosan nanoparticles for lung protein delivery [J]. Eur J Pharm Sci,2005,25 (4):427-437.
[99]李玉慧,彭婷婷,等.羧甲基壳聚糖磁性纳米粒子的合成及应用[J].应用化学,2010, 27(1)：87-91.
[100]李志峰,陈钟,等.载肝细胞生长因子聚乳酸-O-羧甲基壳聚糖纳米粒子制备、表征及其对培养大鼠肝细胞活力的影响[J].中国组织工程研究与临床康复,2008,12(32)：6251-6256.
[101]杨祥良,徐辉碧,谢长生,等.基于纳米技术的中药基础问题研究[J].华中理工大学报,2000,28(12)：104-105.
[102]咎旺.纳米中药制剂研究进展[J].成都医学院学报,2008,3(3)：232-234.
[103]马应龙,李淑红.纳米中药研究进展[J].安徽农学通报,2007,13(9)：192-194.
[104]刘丽芳.纳米中药研究与中药现代化[J].中国现代医学杂志,2009,19(8)：1186-1190.
[105]丁志平,乔延江.纳米技术与纳米中药[J].北京中医药大学学报,2003,26(4)：43-46.
[106]寇正涌,任凤梅,等.纳米中药-中药现代化的新途径[J].长春师范学院学报(自然科学版,2007,26(2)：96-98.
[107]唐致远,凌国维.纳米中药研究进展[J].中草药,2007,38(4)：627-629.
[108]段天璇,于密密,刘春生,等HPLC法同时测定甘草指纹图谱暨甘草苷、甘草酸含量[J].中成药,2006,28(2)：161-165.
[109]樊天宇,王跃飞,过科家,等.内蒙古产甘草药材HPLC指纹图谱研究[J].中药新药与临床药理,2007,18(1)：44-46.
[110]兰霞,王洪新.比色法测定甘草中总皂苷的含量[J].时珍国医国药,2007,18(4)：886-887.
[111]林爱华,平其能.壳聚糖载药纳米粒研究进展[J].中国药业,2006,15(21)：25-27.
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