特异性相互作用毛细管电色谱法筛选中药活性成分
|
摘要
中药是中国千年传统文化的积淀,也是中华民族的瑰宝。但中药因其成分复杂,药效组分模糊而限制了其国际化、现代化的步伐。如何对目标药物进行筛选,找出药效明确的活性成分便成为中药现代化发展的关键。传统的药物筛选方法是以动物模型为基础,这种方法虽然能够得到较为可靠的目标活性物质,但是其周期很长,成本也非常高,还会受到环境的影响。受体学说的阐明,为药物筛选提供了新的思路,以受体为靶分子的药物筛选也就应运而生,这种方法可与多种分析手段结合,目的性更强,且能大大缩短筛选的周期,成为了当今药物筛选中较为重要的一种方法。
本研究采用血小板作为靶分子建立开管毛细管电色谱法,对中药丹参和三七中抗/促血小板活性成分进行筛选。为进一步评价血小板和活性成分之间相互作用力的强度,本研究首次推导出了适用于所建立方法的结合常数计算公式,从而能够定量评价复杂体系中活性成分与血小板的相互作用。本文的研究内容如下:
①建立了一种新的开管毛细管电色谱方法,利用物理吸附的方式将血小板固定于毛细管内壁,通过改变固定长度,研究样品在其中的迁移行为变化,并根据迁移淌度与血小板涂层长度之间的线性方程,得到目标组分与血小板相互作用的结合常数。通过对血小板浓度,洗脱剂,运行缓冲液等影响因素的考察和优化,得到了本方法的适用条件。最后将所建立的方法应用到了HSA与帕珠沙星相互作用的评价中,所得结合常数与文献值较为相符,从而证明了本方法能够用于药物-受体的相互作用强度评价。
②将此方法应用于阳性药物阿司匹林、阴性药物帕珠沙星与血小板的相互作用中,通过荧光光谱法和亲和萃取法的验证,证明了本方法所建立的结合常数计算公式,能够对特异性相互作用进行评价。
③将此方法应用于核苷混合物中抗/促血小板成分的筛选中,首先通过各核苷在不同血小板涂层长度下的峰型变化,定性筛选出了抗/促血小板的成分,然后再将这些成分单独与血小板进行实验,得到了其与血小板的结合常数,从而定量地评价了这些成分与血小板之间的相互作用力。
④将此方法应用于实际中药丹参和三七提取物中抗/促血小板活性成分的筛选,并计算出了各峰所对应的成分与血小板之间的结合常数,通过比较结合常数值的大小,得到了这些峰所对应的成分与血小板相互作用的强度,为进一步筛选提供了数据参考。
⑤对九种野生菌/食用菌中核苷成分进行了定性定量分析,得到了各野生菌/食用菌中核苷的种类及含量,通过对各野生菌中核苷成分的分析,初步预测了具有潜在抗/促血小板功能的野生菌/食用菌,为其药理作用的进一步探索奠定了基础。
Traditional Chinese Medicine (TCM) is a great contribution of Chinese people to the human being. But the internationalizing and modernizing of TCM was restricted because of its complex matrix and undefined active components. Therefore, screening of active components in TCM is benificial for its modernization. Although animal models are commonly used screening method, it requires a long cycle, high cost and is easily influenced by the surroundings. With the development of the receptor theory, some new approaches for drug screeing have been developed using receptors as target molecules. These methods can combine with some analytical techniques with definite goals. And what’s more it can shorten the screening period. So it becomes an important and intensively used method in modern drug screening.
In present study, platelet was used as receptor to establish an open tubular capillary electrochromatography (OT-CEC) method, and some TCM extracts including salvia and pseudo-ginseng were screened. Furthermore, a formula related to the binding constant was deduced, by which the interaction between receptor and ligand can be evaluated quantitatively. The main contents of present study were as follows:
①A new OT-CEC was established which was executed by physically attached of platelet onto the inner wall of the capillary. By varying the coating length of the platelet, the corresponding sample mobility can be obtained and the binding constant can be calculated by the deduced formula. By optimizing the platelet density, washing solution and running buffer, the working conditions was carefully selected. Finally this method was used to evaluate the interaction between HSA and pazufloxacin. The result is almost agreed with the records which proved the method can be used to evaluate the interaction between the target and receptor.
②The established method was used to evaluate the interaction between platelet and aspirin, pazufloxacin, respectively. The result was compared by fluorescence spectrum and affinity extraction which revealed that the established method can be applied to evaluate the specific interaction.
③Applied the established method to screen the anti/promote platelet compounds from the mixture of nucleosides. The compounds were picked out in preliminary screening depending on the change of peak shape. And then the binding constants for the selected compounds were calculated separately to evaluate the binding strength quantitatively.
④The developed method was applied for the screening of platelet active components in the TCM extracts, and the binding constants were calculated for each peak. The result provide useful information for the further screening studies.
⑤The contents of four nucleosides in nine mushroom samples were determined by CE. Combined with the screening result in④, the potential anti- or pro- platelet effects of the investigated mushrooms can be predicted, which may be beneficial for the further pharmacology research of those mushrooms.
本研究采用血小板作为靶分子建立开管毛细管电色谱法,对中药丹参和三七中抗/促血小板活性成分进行筛选。为进一步评价血小板和活性成分之间相互作用力的强度,本研究首次推导出了适用于所建立方法的结合常数计算公式,从而能够定量评价复杂体系中活性成分与血小板的相互作用。本文的研究内容如下:
①建立了一种新的开管毛细管电色谱方法,利用物理吸附的方式将血小板固定于毛细管内壁,通过改变固定长度,研究样品在其中的迁移行为变化,并根据迁移淌度与血小板涂层长度之间的线性方程,得到目标组分与血小板相互作用的结合常数。通过对血小板浓度,洗脱剂,运行缓冲液等影响因素的考察和优化,得到了本方法的适用条件。最后将所建立的方法应用到了HSA与帕珠沙星相互作用的评价中,所得结合常数与文献值较为相符,从而证明了本方法能够用于药物-受体的相互作用强度评价。
②将此方法应用于阳性药物阿司匹林、阴性药物帕珠沙星与血小板的相互作用中,通过荧光光谱法和亲和萃取法的验证,证明了本方法所建立的结合常数计算公式,能够对特异性相互作用进行评价。
③将此方法应用于核苷混合物中抗/促血小板成分的筛选中,首先通过各核苷在不同血小板涂层长度下的峰型变化,定性筛选出了抗/促血小板的成分,然后再将这些成分单独与血小板进行实验,得到了其与血小板的结合常数,从而定量地评价了这些成分与血小板之间的相互作用力。
④将此方法应用于实际中药丹参和三七提取物中抗/促血小板活性成分的筛选,并计算出了各峰所对应的成分与血小板之间的结合常数,通过比较结合常数值的大小,得到了这些峰所对应的成分与血小板相互作用的强度,为进一步筛选提供了数据参考。
⑤对九种野生菌/食用菌中核苷成分进行了定性定量分析,得到了各野生菌/食用菌中核苷的种类及含量,通过对各野生菌中核苷成分的分析,初步预测了具有潜在抗/促血小板功能的野生菌/食用菌,为其药理作用的进一步探索奠定了基础。
Traditional Chinese Medicine (TCM) is a great contribution of Chinese people to the human being. But the internationalizing and modernizing of TCM was restricted because of its complex matrix and undefined active components. Therefore, screening of active components in TCM is benificial for its modernization. Although animal models are commonly used screening method, it requires a long cycle, high cost and is easily influenced by the surroundings. With the development of the receptor theory, some new approaches for drug screeing have been developed using receptors as target molecules. These methods can combine with some analytical techniques with definite goals. And what’s more it can shorten the screening period. So it becomes an important and intensively used method in modern drug screening.
In present study, platelet was used as receptor to establish an open tubular capillary electrochromatography (OT-CEC) method, and some TCM extracts including salvia and pseudo-ginseng were screened. Furthermore, a formula related to the binding constant was deduced, by which the interaction between receptor and ligand can be evaluated quantitatively. The main contents of present study were as follows:
①A new OT-CEC was established which was executed by physically attached of platelet onto the inner wall of the capillary. By varying the coating length of the platelet, the corresponding sample mobility can be obtained and the binding constant can be calculated by the deduced formula. By optimizing the platelet density, washing solution and running buffer, the working conditions was carefully selected. Finally this method was used to evaluate the interaction between HSA and pazufloxacin. The result is almost agreed with the records which proved the method can be used to evaluate the interaction between the target and receptor.
②The established method was used to evaluate the interaction between platelet and aspirin, pazufloxacin, respectively. The result was compared by fluorescence spectrum and affinity extraction which revealed that the established method can be applied to evaluate the specific interaction.
③Applied the established method to screen the anti/promote platelet compounds from the mixture of nucleosides. The compounds were picked out in preliminary screening depending on the change of peak shape. And then the binding constants for the selected compounds were calculated separately to evaluate the binding strength quantitatively.
④The developed method was applied for the screening of platelet active components in the TCM extracts, and the binding constants were calculated for each peak. The result provide useful information for the further screening studies.
⑤The contents of four nucleosides in nine mushroom samples were determined by CE. Combined with the screening result in④, the potential anti- or pro- platelet effects of the investigated mushrooms can be predicted, which may be beneficial for the further pharmacology research of those mushrooms.
引文
[1]吕宝璋,卢建,安明榜.受体学[M].合肥:安徽科学技术出版社, 2000:6-7.
[2]翁维良,廖福龙,吴云鹏等.血液流变学研究方法及其应用[M].科学出版社, 1989:168-171.
[3] Michelson A D. Platelet[M]. 2nd ed. MA: Academic press, 2006: 117-134.
[4] Michelson A D. Platelet[M]. 2nd ed. MA: Academic press, 2006: 1194-1202.
[5]杜冠华.高通量筛选技术进展与新药开发策略[J].中国新药杂志, 2002, 11(1): 31-36.
[6] Maddry J. A., Chen X., Jonsson C. B., et al. Discovery of novel benzoquinazolinones and thiazoloimidazoles, inhibitors of influenza H5N1 and H1N1 viruses, from a cell-based high-throughput screen[J]. Journal of Biomolecular Screening, 2011, 16(1): 73-81.
[7] Shibata Y., Takahashi H., Ishii Y.. A convenient in vitro screening method for predicting invivo drug metabolic clearance using isolated hepatocytes suspended in serum [J]. Drug Metab Dispos, 2000, 28: 1518-1523.
[8] Ulrich R. G., Bacon J. A., Cramer C. T., et al. Cultured hepatocytes as investigational models for hepatic toxicity: practical applications in drug discovery and development [J]. Toxicology Letter, 1995, 82-83: 107–115.
[9] Wagstaff K. M., Rawlinson S. M., Hearps A. C., et al. An alphascreen(R)-based assay for high-throughput screening for specific inhibitors of nuclear import [J]. Journal of Biomolecular Screening, 2011, 16(2): 192-200.
[10]孟鹏,齐西珍,郑芳,等.α-葡萄糖苷酶抑制剂高通量筛选模型的建立及其应用[J].微生物学报, 2010, 50(8): 1080-1086.
[11] Crowther G. J., Napuli A. J., Gilligan J. H., et al. Identification of inhibitors for putative malaria drug targets among novel antimalarial compounds [J]. Molecular& Biochemical Parasitology, 2011, 175: 21-29.
[12] Kim J., Park H. C., Gedi V., et al. Yeast-hybrid based high-throughput assay for identification of anthrax lethal factor inhibitors [J]. Biochemical and Biophysical Research Communications, 2011, 404: 517-522.
[13]杜冠华.药物筛选新技术与中药现代化研究[J].世界科学技术-中医药现代化, 2000, 2(4):47-52.
[14] Dixit C. K., Vashist S. K., O’Neill F. T., et al. Development of a high sensitivity rapid sandvich ELISA procedure and its comparison with the conventional approach [J]. Analytical Chemistry, 2010, 82: 7049-7052.
[15] Mallik R., Yoo M. J., Chen S., et al. Studies of verapamil binding to human serum albumin by high-performance affinity chromatography [J]. Journal of Chromatography B, 2008, 876(1): 69-75.
[16]孔亮,邹汉法,汪海林,等.以人血清白蛋白为固定相的分子生物色谱分析几种中药活性成分的研究[J].高等学校化学学报, 2000, 21(1): 36-40.
[17] Wang H. L., Kong L., Zou H. F., et al. Screening and analysis of biologically active compound in Angelica sinensis by molecular biochromatography [J]. Chromatographia, 1999, 50(7/8):439-445.
[18] Wang H., Zou H., Ni J., et al. Comparative separation of biologically active components in Rhizoma chuanxiong by affinity chromatography withα-acid glycol protein and human serum al-bum in as stationary phases [J]. Chromatographia, 2000, 52(7/8): 459-464.
[19] Wang H., Zou H., Ni J., et al. Fractionation and analysis of Artemisia capillaries Thumb. By affinity chromatography with human serum album in as stationary phase [J]. Journal of Chromatogra A, 2000, 870: 501-510.
[20] Yang Q., Lundahl P.. Binding of lysozyme on the surface of entrapped phosphatidylserine phosphatidylcholine vesicles and an example of high-performance lipid vesicle surface chromatography [J]. Journal of Chromatogrphy A, 1990, 512: 377-386.
[21] Pidgeon C., Cai S. J., Bernal C.. Mobile phase effects on membrane protein elution during immobilized artificial membrane chromatography [J]. J Chromatogr A, 1996, 721: 213-230.
[22] Hanna M., Biasi V. D., Bond B., et al. Biomembrane lipids as components of chromatographic phase [J]. Chromatographia, 2000, 52(11/12): 710-720.
[23] Beigi F., Yang Q., Lundahl P.. Immobilized-liposome chromatographic analysis of drug partitioning into lipid bilayers [J]. Journal of Chromatography A, 1995, 704: 315-321.
[24]毛希琴,邹汉法,罗权舟,等.模拟生物膜色谱用于预测药物的小肠吸收[J].分析化学,2001, 10(29): 1135-1139.
[25]赵小娟,党高潮,杨广德,等.淫羊藿根与叶活性成分的分析和比较[J].分析化学, 2002, 30(2) : 195-197.
[26] Xiao J. H., Zhang H., Niu L. Y., et al. Efficient screening of a novel antimicrobial peptide from Jatropha curcas by cell membrane affinity chromatography [J]. Journal of Agriculture and Food Chemistry, 2011, 59:1145-1151.
[27] Bing X. Y., Jin H., Lang C. H., et al. Evaluation of drug-muscarinic receptor affinities using cell membrance chromatography and radioligand binding assay in guinea pig jejunum membrance [J]. Acta Pharmacologica Sinica, 2005, 26(1): 113-116.
[28]杨广德,贺浪冲,边晓莉,等.胰岛β细胞膜色谱模型建立及其生物亲和特性[J].科学通报, 2005, 50(16): 1709-1713.
[29]张宇洁,贺浪冲.用细胞膜色谱模型筛选长春七抑制HeLa细胞增殖的活性成分[J],中国药学杂志, 2005, 40(6): 463-465.
[30]岳宣峰,张延妮,张志琪,等.心肌细胞固定相色谱研究中药川芎提取液与受体的作用[J].中国中药杂志, 2005, 30(2): 129-133.
[31]朱荃.细胞膜固相色谱及其在中药效应-物质基础研究中的应用[J].南京中医药大学学报, 2006,26(1):8-13.
[32]樊宏伟,余黎,洪敏,等.血小板细胞膜固相色谱法的建立及其丹参效应物质的初步分析[J].中国药学杂志, 2004, 39(5): 375-378.
[33]董倩倩,李萍,宋越,等.靶细胞提取和高效液相色谱飞行时间质谱联用分析预测丹参中的活性成分[J].分析化学, 2007, 35(5): 648-652.
[34]董倩倩,李萍,闻晓东.丹参成分在Caco-2细胞中吸收的研究.中国天然药物, 2007, 5(1): 59-62.
[35] Nie H., Meng L. Z., Zhang H., et al. Analysis of anti-platelet aggregation components of Rhizoma Zingiberis using chicken thrombocyte extract and high performance liquid chromatography [J]. Chinese Medical Journal, 2008, 121(13): 1226-1229.
[36] Wang G., Huang Z. G., Cao H., et al. Screening of anti-platelet aggregation agents from Panaxn oto ginseng using human platelet extraction and HPLC–DAD–ESI-MS/MS [J]. Journal of separation science, 2008, 31: 1173-1180.
[37] Donga Z. B., Li S. P., Hong M.. Hypothesis of potential active components in Angelica sinensisby using biomembrane extraction and high performance liquid chromatography [J]. Journal of Pharmaceutical and Biomedical Analysis, 2005, 38: 664–669.
[38] Yua J., Zhao J., Zhua Q., et al. Macrophage biospecific extraction and high performance liquid chromatography for hypothesis of immunological active components in Cordyceps sinensis [J]. Journal of Pharmaceutical and Biomedical Analysis, 2007, 44: 439-443.
[39] Li L. S., Li P., Sheng L. H., et al. Live cell extraction and HPLC-MS analysis for predicting bioactive components of traditional Chinese medicine [J]. Journal of Pharmaceutical and Biomedical Analysis, 2006, 41: 576-581.
[40]樊宏伟,朱荃,洪敏,等.血小板细胞膜固相色谱法在脉络宁注射液效应物质分析中的应用[J].中国药学杂志, 2006, 41(1): 63-66.
[41] Heegaard N. H. H., Nillsson S., Guzman N. A.. Affinity capillary electrophoresis: important application areas and some recent development [J]. Journal of Chromatography B, 1998, 715: 29-54.
[42]王京兰,钱小红.亲和毛细管电泳技术及应用[J].色谱, 1999, 17(4): 342-345.
[43] Bielejewska A., Bylina A., Duszczyk K., et al. Evaluation of ligand-selector interaction from effective diffusion coefficient [J]. Analytical Chemistry, 2010, 82: 5463-5469.
[44]赵燕燕,杨更亮,范子琳,等.高效毛细管电泳前沿分析法研究酸性药物那格列奈与人血浆白蛋白的结合[J].分析化学, 2003, 31(7): 860-864.
[45] Vuignier K., Schappler J., Veuthey J. L., et al. Improvement of a capillary electrophoresis/frontal analysis(CE/FA) method for determination binding constants: discussion on relavent parameters [J]. Journal of Pharmaceutical and Biomedical Analysis, 2010, 53: 1288-1297.
[46]咸德玲,黄可龙,胡卫国,等.脂质体电动色谱法评价阿魏酸与生物膜的相互作用[J].分析化学, 2007, 35(10): 1521-1524.
[47] ?stergaard J., Moeller E. H.. Ghrelin-liposome interactions: characterization of liposomal formulations of an acylated 28-amino acid peptide using CE [J]. Electrophoresis, 2010, 31(2): 339-345.
[48] ?stergaard J., Jorgensen L., Thomsen A. E., et al. Drug-liposome distribution phenomena studied by capillary electrophoresis-frontal analysis [J]. Electrophoresis, 2008, 29(16): 3320-3324.
[49] Zhao H., Dai D. S., Li J. R., et al. Quantitative study of HIV-1 Tat peptide and TAR RNA interaction inhibited by poly(allylamine hydrochloride) [J]. Biochemical and biophysical Research Communications, 2003, 312: 351-354.
[50]凌笑梅,刘一,赖先银,张媛,等.采用毛细管电泳方法以凝血酶为靶点筛选天然药物提取化合物[J].高等学校化学学报, 2007, 28: 234-237.
[51]刘书朋,孙烨,郭薇,等.应用毛细管电泳表征噬菌体与丹酚酸B的相互作用[J].上海大学学报, 2008, 14(1): 96-99.
[52]邓玉林,刘艳清,马文,等.生物功能化色谱新概念及其方法学研究[J].生命科学仪器, 2004, 5(2): 9-11.
[53]厉红,屈锋,徐建栋,等.应用生物功能化色谱研究SSAO酶与底物和抑制剂之间的相互作用[J].北京理工大学学报, 2008, 28(12): 1117-1121.
[54] Luo G. A., Wen T., Wang Y. M., et al. Advance of immunoaffinity capillary electrophoresis [J]. Acta Pharmaceutica Sinica, 2002, 37(6): 479-484.
[55]米健秋,张新祥,常文保.毛细管电泳免疫分析的发展及动向[J].化学进展, 2003, 15(1): 31-40.
[56] Chen H. X., Zhang X. X.. Antibody development to testosterone and its application in capillary electrophoresis-based immunoassay [J]. Electrophoresis, 2008, 29: 3406-3413.
[57]屈峰,刘允,任肖敏,等.毛细管电泳在核算适配体研究及核算适配体筛选中的应用[J].化学进展, 2009, 21(7/8): 1576-1582.
[58] Mendonsa S. D., Bowser M. T.. In vitro selection of high-affinity DNA ligands for lgE using capillary electrophoresis [J]. Analytical Chemistry, 2004, 76: 5387-5392.
[59]徐华,李少华,郭磊,等.基于毛细管电泳的细胞SELEX方法的初步建立[J].军事医学科学院院刊, 2009, 33(5): 461-464.
[60] Esther T., Antonio M. E.. Molecular imprinting technology in capillary electrochromatography [J]. Journal of Separation Science, 2005, 28: 719-728.
[61] Lin J. M., Nakagama T., Uchiyama T., et al. Temperature effect on chiral recognition of some amino acids with molecularly imprinted polymer filled capillary electrochromatography [J]. Biomedical Chromatography, 1997, 11(5): 298-302.
[62] Schweitz L., Andersson L., Nilsson S.. Molded rigid polymer monoliths as separation media for capillary electrochromatography [J]. Analytical Chemistry, 1997, 69: 1179-1183.
[63] Schweitz L., Andersson L., Nilsson S.. Rapid electrochromatography enantiomer separations on short molecularly imprinted polymer monoliths [J]. Analytica Chemica Acta, 2001, 435: 43-47.
[64] Huang Y. C., Lin C., Liu C. Y.. Preparation and evaluation of molecularly imprinted polymers based on 9-ethyladenine for the recognition of nucleotide in capillary electrochromatography [J]. Electrophoresis, 2004, 25(4-5): 554-561.
[65] Milena Q., Ersilia D. L., Claudia S., et al. Molecularly imprinted polymer films grafted from porous or nonporous silica: Novel affinity stationary phases in capillary electrochromatography [J]. Electrophoresis, 2003, 24: 952-957.
[66] Yoshida M., Hatate Y., Uezu K., et al. Selectivity towards multiple predetermined targets in nanoparticle capillary electrochromatography [J]. Analytical Chemistry, 2003, 75: 6608-6613.
[67] Huang B.Y., Chen Y. C., Wang G. R., et al. Preparation and evaluation of a monolithic molecularly imprinted polymer for the chiral separation of neurotransmitters and their analogues by capillary electrochromatography [J]. Journal of Chromatography A, 2011, 1218: 849-855.
[68] Cacho C., Schweitz L., Turiel E., et al. Molecular imprinted capillary electrochromatography for selective determination of thiabendazole in citrus samples [J]. Journal of Chromatography A, 2008, 1179: 216-223.
[69] Phillips T. M., Benjamin F. D.. Analysis of recombinant cytokines in human body fluids by immunoaffinity capillary electrophoresis [J]. Electrophoresis, 1998, 19: 2991-2996.
[70]吕宝璋,卢建,安明榜.受体学[M].合肥:安徽科学技术出版社, 2000:10-11.
[71]张维冰.毛细管电色谱理论基础[M].北京:科学出版社, 2006:97-99.
[72]吕宝璋,卢建,安明榜.受体学[M].合肥:安徽科学技术出版社, 2000: 6-7.
[73] Michelson A D. Platelet [M]. 2nd ed. MA: Academic press, 2006: 118-133.
[74] Wang F., Huang W., Su L., et al. Spectrofluorimetric study of the binding of codeine to nucleic acids [J]. Journal of Moeclar Structure, 2009, 927(1-3): 1-6.
[75] Wu F. Y., Zhang L. N., Ji Z. J., et al. Spectroscopic investigation of the interaction between thiourea-zinc complex and serum albumin [J]. Journal of Luminescence, 2010, 130(7): 1280-1284.
[76] Wang H. L., Zou H. F., Ni J. Y., et al. Fractionation and analysis of Artemisia capillaries Thunb. by affinity chromatography with human serum albumin as stationary phases [J]. Journal of Chromatography A, 2000, 870(1-2): 501-510.
[77]孔亮,邹汉法,汪海林,倪坚毅,张玉奎.以人血清白蛋白为固定相的分子生物色谱分析几种中药活性成分的研究[J].高等学校化学学报, 2000, 21(1): 36-40.
[78] Mao X. Q., Kong L., Luo Q. Z., et al. Screening and analysis of permeable compounds in Radix Angelica Sinensis with immobilized liposome chromatography [J]. Journal of Chromatography B, 2002, 779(2): 331-339.
[79] Caldwell G. W., McDonnel P. A., Masucci J. A., et al. Using affinity capillary electrophoresis to study the interaction of the extracellular binding domain of erythropoietin receptor with peptides [J]. Journal of Biochemical and Biophysical Methods, 1999, 40(1-2): 17-25.
[80]夏之宁,李丽仙,杨靖,熊彩侨.药物与细胞膜相互作用的毛细管电泳方法研究[J].中国科学, B辑, 2009, 39(7): 634-639.
[81] ?stergaard J., Jorgensen L., Thomsen A. E., et al. Drug-liposome distribution phenomena studied by capillary electrophoresis-frontal analysis [J]. Electrophoresis, 2008, 29(16): 3320-3324.
[82] Pedersen J. T., ?stergaard J., Houen G., et al. Affinity capillary electrophoresis for identification and investigation of human Gc-globulin(vitamin D-binding protein) and its isoforms interaction with G-actin [J]. Electrophoresis, 2008, 29(8): 1723-1733.
[83] EI-Hady D., Kühne S., EI-Maali N., et al. Precision in affinity capillary electrophoresis for drug-protein binding studies [J]. Journal of Pharmceutical and Biochemical Anaysisl, 2010, 52(2): 232-241.
[84] Martma K., Fernaeus S. Z., Land T., et al. Novel capillary coatings in open tubular CEC as models for biological membranes [J]. Electrophoresis, 2010, 31(9): 1586-1589.
[85] Kuldvee R., Wiedmer S. K., ??rin K., et al. Human low-density lipoprotein-coated capillaries in electrochromatography [J]. Analytical Chemistry, 2005, 77(10): 3401-3405.
[86] Kuldvee R., D’ulivo L., Yohannes G., et al. Open tubular capillary electrochromatography:Technique for oxidation and interaction studies on human low-density lipoproteins [J]. Analytical Chemistry, 2006, 78(8): 2665-2671.
[87] Chen J., Fallarero A., M??tt?nen A., et al. Living cells of Staphylococcus aureus immobilized onto the capillary surface in electrochromatography: A tool for screening of biofilms [J]. Analytical Chemistry, 2008, 80(13): 5103-5109.
[88]熊彩侨,夏之宁,黄锐,等.区段-区段动力学毛细管电泳测定动力学参数的新方法及其应用[J].中国科学, B辑, 2008, 38(8): 705-709.
[89]张维冰.毛细管电色谱理论基础[M].北京:科学出版社, 2006. 94-97.
[90] Jin J., Zhang X.. Spectrophotometric stidies on the interaction between pazufloxacin mesilate and human serum albumin or lysozyme [J]. Journal of Luminescence, 2008, 128: 81-86.
[91] Mckee S. A., Sane D. C., Deliargyris E. N., et al. Aspirin resistance in cardiovascular disease: A review of prevalence, mechanisms, and clinical significance [J]. Thromb Haemost, 2002, 88(5): 711.
[92]杨信怡,游学甫.注射用帕珠沙星的药理研究和临床应用[J].国外医药:抗生素分册, 2003, 24(2): 80-87.
[93] Michelson A D. Platelet [M]. 2nd ed. MA: Academic press, 2007. 1099-1101.
[94] Bobert M. B.. The role of adenosine in the regulation of coronary blood flow [J]. Circulation Research, 1980, 47(6): 807-813.
[95]田谋利,孙海静,高欣,等.尿苷三磷酸对大鼠局灶性缺血再灌注损伤脑组织的保护作用[J].第二军医大学学报, 2010, 31(6): 608-611.
[96] Cheng T. O.. Cardiovascular effects of Danshen [J]. International Journal of Cardiology, 2007, 121: 9-22.
[97]刘陶世,黄耀洲.丹参成分、制剂和质量控制研究概况[J].南京中医药大学学报, 1998, 14(4): 255-256.
[98]吕志刚,徐列明.丹酚酸B的药理研究概况[J].医学研究杂志, 2008, 37(8): 114-116.
[99]李琦,叶蕴华,刑其毅.三七水溶性化学成分及其药理研究新进展[J].高等学校化学学报, 1996, 17(12): 1886-1892.
[100]可燕,蒋嘉烨,王视珍,等.三七根及花总皂苷抗肿瘤细胞诱导的血小板聚集研究[J].中药材, 2010, 33(1): 96-99.
[101]郭洁文,李丽明,邱光清,等.三七总皂苷对心梗后心室重建构大鼠ACE2与TNF-α表达的影响[J].中药材, 2010, 33(1): 89-92.
[102]黄赵刚.血小板生物亲和萃取结合HPLC法筛选三七抗血小板聚集活性皂苷成分[D].安徽:安徽医科大学, 2005.
[103]刘佳,陈晓辉,钱忠直,等. RP-HPLC法测定藤三七不同部位中腺苷的含量[J].沈阳药科大学学报, 2009, 26(9): 724-739.
[104]张冰,陈晓辉,毕开顺. HPLC法同时测定三七花中鸟苷和腺苷的含量[J].药物分析杂志, 2009, 29(12): 2113-2115.
[105]吴杰,刘旭.腺苷的临床应用[J].国外医学.心血管疾病分册, 2002, 29(3): 165-166.
[106] Gong Y. X., Li S. P., Li P., et al. Simultaneous determination of six main nucleosides and bases in natural and cultured Cordyceps by capillary electrophoresis [J]. Journal of Chromatography A. 2004, 1055: 215-221.
[107] Yu L., Zhao J., Li S. P., et al. Quality evaluation of Cordyceps through simultaneous determination of eleven nucleosides and bases by RP-HPLC [J]. Journal of Separation Science. 2006, 29, 953–958.
[108] Yuan J. P., Zhao S. Y., Wang J. H., et al. Distribution of nucleosides and nucleobases in edible fungi [J]. Journal of Food Chemistry, 2008, 56: 809-815.
[2]翁维良,廖福龙,吴云鹏等.血液流变学研究方法及其应用[M].科学出版社, 1989:168-171.
[3] Michelson A D. Platelet[M]. 2nd ed. MA: Academic press, 2006: 117-134.
[4] Michelson A D. Platelet[M]. 2nd ed. MA: Academic press, 2006: 1194-1202.
[5]杜冠华.高通量筛选技术进展与新药开发策略[J].中国新药杂志, 2002, 11(1): 31-36.
[6] Maddry J. A., Chen X., Jonsson C. B., et al. Discovery of novel benzoquinazolinones and thiazoloimidazoles, inhibitors of influenza H5N1 and H1N1 viruses, from a cell-based high-throughput screen[J]. Journal of Biomolecular Screening, 2011, 16(1): 73-81.
[7] Shibata Y., Takahashi H., Ishii Y.. A convenient in vitro screening method for predicting invivo drug metabolic clearance using isolated hepatocytes suspended in serum [J]. Drug Metab Dispos, 2000, 28: 1518-1523.
[8] Ulrich R. G., Bacon J. A., Cramer C. T., et al. Cultured hepatocytes as investigational models for hepatic toxicity: practical applications in drug discovery and development [J]. Toxicology Letter, 1995, 82-83: 107–115.
[9] Wagstaff K. M., Rawlinson S. M., Hearps A. C., et al. An alphascreen(R)-based assay for high-throughput screening for specific inhibitors of nuclear import [J]. Journal of Biomolecular Screening, 2011, 16(2): 192-200.
[10]孟鹏,齐西珍,郑芳,等.α-葡萄糖苷酶抑制剂高通量筛选模型的建立及其应用[J].微生物学报, 2010, 50(8): 1080-1086.
[11] Crowther G. J., Napuli A. J., Gilligan J. H., et al. Identification of inhibitors for putative malaria drug targets among novel antimalarial compounds [J]. Molecular& Biochemical Parasitology, 2011, 175: 21-29.
[12] Kim J., Park H. C., Gedi V., et al. Yeast-hybrid based high-throughput assay for identification of anthrax lethal factor inhibitors [J]. Biochemical and Biophysical Research Communications, 2011, 404: 517-522.
[13]杜冠华.药物筛选新技术与中药现代化研究[J].世界科学技术-中医药现代化, 2000, 2(4):47-52.
[14] Dixit C. K., Vashist S. K., O’Neill F. T., et al. Development of a high sensitivity rapid sandvich ELISA procedure and its comparison with the conventional approach [J]. Analytical Chemistry, 2010, 82: 7049-7052.
[15] Mallik R., Yoo M. J., Chen S., et al. Studies of verapamil binding to human serum albumin by high-performance affinity chromatography [J]. Journal of Chromatography B, 2008, 876(1): 69-75.
[16]孔亮,邹汉法,汪海林,等.以人血清白蛋白为固定相的分子生物色谱分析几种中药活性成分的研究[J].高等学校化学学报, 2000, 21(1): 36-40.
[17] Wang H. L., Kong L., Zou H. F., et al. Screening and analysis of biologically active compound in Angelica sinensis by molecular biochromatography [J]. Chromatographia, 1999, 50(7/8):439-445.
[18] Wang H., Zou H., Ni J., et al. Comparative separation of biologically active components in Rhizoma chuanxiong by affinity chromatography withα-acid glycol protein and human serum al-bum in as stationary phases [J]. Chromatographia, 2000, 52(7/8): 459-464.
[19] Wang H., Zou H., Ni J., et al. Fractionation and analysis of Artemisia capillaries Thumb. By affinity chromatography with human serum album in as stationary phase [J]. Journal of Chromatogra A, 2000, 870: 501-510.
[20] Yang Q., Lundahl P.. Binding of lysozyme on the surface of entrapped phosphatidylserine phosphatidylcholine vesicles and an example of high-performance lipid vesicle surface chromatography [J]. Journal of Chromatogrphy A, 1990, 512: 377-386.
[21] Pidgeon C., Cai S. J., Bernal C.. Mobile phase effects on membrane protein elution during immobilized artificial membrane chromatography [J]. J Chromatogr A, 1996, 721: 213-230.
[22] Hanna M., Biasi V. D., Bond B., et al. Biomembrane lipids as components of chromatographic phase [J]. Chromatographia, 2000, 52(11/12): 710-720.
[23] Beigi F., Yang Q., Lundahl P.. Immobilized-liposome chromatographic analysis of drug partitioning into lipid bilayers [J]. Journal of Chromatography A, 1995, 704: 315-321.
[24]毛希琴,邹汉法,罗权舟,等.模拟生物膜色谱用于预测药物的小肠吸收[J].分析化学,2001, 10(29): 1135-1139.
[25]赵小娟,党高潮,杨广德,等.淫羊藿根与叶活性成分的分析和比较[J].分析化学, 2002, 30(2) : 195-197.
[26] Xiao J. H., Zhang H., Niu L. Y., et al. Efficient screening of a novel antimicrobial peptide from Jatropha curcas by cell membrane affinity chromatography [J]. Journal of Agriculture and Food Chemistry, 2011, 59:1145-1151.
[27] Bing X. Y., Jin H., Lang C. H., et al. Evaluation of drug-muscarinic receptor affinities using cell membrance chromatography and radioligand binding assay in guinea pig jejunum membrance [J]. Acta Pharmacologica Sinica, 2005, 26(1): 113-116.
[28]杨广德,贺浪冲,边晓莉,等.胰岛β细胞膜色谱模型建立及其生物亲和特性[J].科学通报, 2005, 50(16): 1709-1713.
[29]张宇洁,贺浪冲.用细胞膜色谱模型筛选长春七抑制HeLa细胞增殖的活性成分[J],中国药学杂志, 2005, 40(6): 463-465.
[30]岳宣峰,张延妮,张志琪,等.心肌细胞固定相色谱研究中药川芎提取液与受体的作用[J].中国中药杂志, 2005, 30(2): 129-133.
[31]朱荃.细胞膜固相色谱及其在中药效应-物质基础研究中的应用[J].南京中医药大学学报, 2006,26(1):8-13.
[32]樊宏伟,余黎,洪敏,等.血小板细胞膜固相色谱法的建立及其丹参效应物质的初步分析[J].中国药学杂志, 2004, 39(5): 375-378.
[33]董倩倩,李萍,宋越,等.靶细胞提取和高效液相色谱飞行时间质谱联用分析预测丹参中的活性成分[J].分析化学, 2007, 35(5): 648-652.
[34]董倩倩,李萍,闻晓东.丹参成分在Caco-2细胞中吸收的研究.中国天然药物, 2007, 5(1): 59-62.
[35] Nie H., Meng L. Z., Zhang H., et al. Analysis of anti-platelet aggregation components of Rhizoma Zingiberis using chicken thrombocyte extract and high performance liquid chromatography [J]. Chinese Medical Journal, 2008, 121(13): 1226-1229.
[36] Wang G., Huang Z. G., Cao H., et al. Screening of anti-platelet aggregation agents from Panaxn oto ginseng using human platelet extraction and HPLC–DAD–ESI-MS/MS [J]. Journal of separation science, 2008, 31: 1173-1180.
[37] Donga Z. B., Li S. P., Hong M.. Hypothesis of potential active components in Angelica sinensisby using biomembrane extraction and high performance liquid chromatography [J]. Journal of Pharmaceutical and Biomedical Analysis, 2005, 38: 664–669.
[38] Yua J., Zhao J., Zhua Q., et al. Macrophage biospecific extraction and high performance liquid chromatography for hypothesis of immunological active components in Cordyceps sinensis [J]. Journal of Pharmaceutical and Biomedical Analysis, 2007, 44: 439-443.
[39] Li L. S., Li P., Sheng L. H., et al. Live cell extraction and HPLC-MS analysis for predicting bioactive components of traditional Chinese medicine [J]. Journal of Pharmaceutical and Biomedical Analysis, 2006, 41: 576-581.
[40]樊宏伟,朱荃,洪敏,等.血小板细胞膜固相色谱法在脉络宁注射液效应物质分析中的应用[J].中国药学杂志, 2006, 41(1): 63-66.
[41] Heegaard N. H. H., Nillsson S., Guzman N. A.. Affinity capillary electrophoresis: important application areas and some recent development [J]. Journal of Chromatography B, 1998, 715: 29-54.
[42]王京兰,钱小红.亲和毛细管电泳技术及应用[J].色谱, 1999, 17(4): 342-345.
[43] Bielejewska A., Bylina A., Duszczyk K., et al. Evaluation of ligand-selector interaction from effective diffusion coefficient [J]. Analytical Chemistry, 2010, 82: 5463-5469.
[44]赵燕燕,杨更亮,范子琳,等.高效毛细管电泳前沿分析法研究酸性药物那格列奈与人血浆白蛋白的结合[J].分析化学, 2003, 31(7): 860-864.
[45] Vuignier K., Schappler J., Veuthey J. L., et al. Improvement of a capillary electrophoresis/frontal analysis(CE/FA) method for determination binding constants: discussion on relavent parameters [J]. Journal of Pharmaceutical and Biomedical Analysis, 2010, 53: 1288-1297.
[46]咸德玲,黄可龙,胡卫国,等.脂质体电动色谱法评价阿魏酸与生物膜的相互作用[J].分析化学, 2007, 35(10): 1521-1524.
[47] ?stergaard J., Moeller E. H.. Ghrelin-liposome interactions: characterization of liposomal formulations of an acylated 28-amino acid peptide using CE [J]. Electrophoresis, 2010, 31(2): 339-345.
[48] ?stergaard J., Jorgensen L., Thomsen A. E., et al. Drug-liposome distribution phenomena studied by capillary electrophoresis-frontal analysis [J]. Electrophoresis, 2008, 29(16): 3320-3324.
[49] Zhao H., Dai D. S., Li J. R., et al. Quantitative study of HIV-1 Tat peptide and TAR RNA interaction inhibited by poly(allylamine hydrochloride) [J]. Biochemical and biophysical Research Communications, 2003, 312: 351-354.
[50]凌笑梅,刘一,赖先银,张媛,等.采用毛细管电泳方法以凝血酶为靶点筛选天然药物提取化合物[J].高等学校化学学报, 2007, 28: 234-237.
[51]刘书朋,孙烨,郭薇,等.应用毛细管电泳表征噬菌体与丹酚酸B的相互作用[J].上海大学学报, 2008, 14(1): 96-99.
[52]邓玉林,刘艳清,马文,等.生物功能化色谱新概念及其方法学研究[J].生命科学仪器, 2004, 5(2): 9-11.
[53]厉红,屈锋,徐建栋,等.应用生物功能化色谱研究SSAO酶与底物和抑制剂之间的相互作用[J].北京理工大学学报, 2008, 28(12): 1117-1121.
[54] Luo G. A., Wen T., Wang Y. M., et al. Advance of immunoaffinity capillary electrophoresis [J]. Acta Pharmaceutica Sinica, 2002, 37(6): 479-484.
[55]米健秋,张新祥,常文保.毛细管电泳免疫分析的发展及动向[J].化学进展, 2003, 15(1): 31-40.
[56] Chen H. X., Zhang X. X.. Antibody development to testosterone and its application in capillary electrophoresis-based immunoassay [J]. Electrophoresis, 2008, 29: 3406-3413.
[57]屈峰,刘允,任肖敏,等.毛细管电泳在核算适配体研究及核算适配体筛选中的应用[J].化学进展, 2009, 21(7/8): 1576-1582.
[58] Mendonsa S. D., Bowser M. T.. In vitro selection of high-affinity DNA ligands for lgE using capillary electrophoresis [J]. Analytical Chemistry, 2004, 76: 5387-5392.
[59]徐华,李少华,郭磊,等.基于毛细管电泳的细胞SELEX方法的初步建立[J].军事医学科学院院刊, 2009, 33(5): 461-464.
[60] Esther T., Antonio M. E.. Molecular imprinting technology in capillary electrochromatography [J]. Journal of Separation Science, 2005, 28: 719-728.
[61] Lin J. M., Nakagama T., Uchiyama T., et al. Temperature effect on chiral recognition of some amino acids with molecularly imprinted polymer filled capillary electrochromatography [J]. Biomedical Chromatography, 1997, 11(5): 298-302.
[62] Schweitz L., Andersson L., Nilsson S.. Molded rigid polymer monoliths as separation media for capillary electrochromatography [J]. Analytical Chemistry, 1997, 69: 1179-1183.
[63] Schweitz L., Andersson L., Nilsson S.. Rapid electrochromatography enantiomer separations on short molecularly imprinted polymer monoliths [J]. Analytica Chemica Acta, 2001, 435: 43-47.
[64] Huang Y. C., Lin C., Liu C. Y.. Preparation and evaluation of molecularly imprinted polymers based on 9-ethyladenine for the recognition of nucleotide in capillary electrochromatography [J]. Electrophoresis, 2004, 25(4-5): 554-561.
[65] Milena Q., Ersilia D. L., Claudia S., et al. Molecularly imprinted polymer films grafted from porous or nonporous silica: Novel affinity stationary phases in capillary electrochromatography [J]. Electrophoresis, 2003, 24: 952-957.
[66] Yoshida M., Hatate Y., Uezu K., et al. Selectivity towards multiple predetermined targets in nanoparticle capillary electrochromatography [J]. Analytical Chemistry, 2003, 75: 6608-6613.
[67] Huang B.Y., Chen Y. C., Wang G. R., et al. Preparation and evaluation of a monolithic molecularly imprinted polymer for the chiral separation of neurotransmitters and their analogues by capillary electrochromatography [J]. Journal of Chromatography A, 2011, 1218: 849-855.
[68] Cacho C., Schweitz L., Turiel E., et al. Molecular imprinted capillary electrochromatography for selective determination of thiabendazole in citrus samples [J]. Journal of Chromatography A, 2008, 1179: 216-223.
[69] Phillips T. M., Benjamin F. D.. Analysis of recombinant cytokines in human body fluids by immunoaffinity capillary electrophoresis [J]. Electrophoresis, 1998, 19: 2991-2996.
[70]吕宝璋,卢建,安明榜.受体学[M].合肥:安徽科学技术出版社, 2000:10-11.
[71]张维冰.毛细管电色谱理论基础[M].北京:科学出版社, 2006:97-99.
[72]吕宝璋,卢建,安明榜.受体学[M].合肥:安徽科学技术出版社, 2000: 6-7.
[73] Michelson A D. Platelet [M]. 2nd ed. MA: Academic press, 2006: 118-133.
[74] Wang F., Huang W., Su L., et al. Spectrofluorimetric study of the binding of codeine to nucleic acids [J]. Journal of Moeclar Structure, 2009, 927(1-3): 1-6.
[75] Wu F. Y., Zhang L. N., Ji Z. J., et al. Spectroscopic investigation of the interaction between thiourea-zinc complex and serum albumin [J]. Journal of Luminescence, 2010, 130(7): 1280-1284.
[76] Wang H. L., Zou H. F., Ni J. Y., et al. Fractionation and analysis of Artemisia capillaries Thunb. by affinity chromatography with human serum albumin as stationary phases [J]. Journal of Chromatography A, 2000, 870(1-2): 501-510.
[77]孔亮,邹汉法,汪海林,倪坚毅,张玉奎.以人血清白蛋白为固定相的分子生物色谱分析几种中药活性成分的研究[J].高等学校化学学报, 2000, 21(1): 36-40.
[78] Mao X. Q., Kong L., Luo Q. Z., et al. Screening and analysis of permeable compounds in Radix Angelica Sinensis with immobilized liposome chromatography [J]. Journal of Chromatography B, 2002, 779(2): 331-339.
[79] Caldwell G. W., McDonnel P. A., Masucci J. A., et al. Using affinity capillary electrophoresis to study the interaction of the extracellular binding domain of erythropoietin receptor with peptides [J]. Journal of Biochemical and Biophysical Methods, 1999, 40(1-2): 17-25.
[80]夏之宁,李丽仙,杨靖,熊彩侨.药物与细胞膜相互作用的毛细管电泳方法研究[J].中国科学, B辑, 2009, 39(7): 634-639.
[81] ?stergaard J., Jorgensen L., Thomsen A. E., et al. Drug-liposome distribution phenomena studied by capillary electrophoresis-frontal analysis [J]. Electrophoresis, 2008, 29(16): 3320-3324.
[82] Pedersen J. T., ?stergaard J., Houen G., et al. Affinity capillary electrophoresis for identification and investigation of human Gc-globulin(vitamin D-binding protein) and its isoforms interaction with G-actin [J]. Electrophoresis, 2008, 29(8): 1723-1733.
[83] EI-Hady D., Kühne S., EI-Maali N., et al. Precision in affinity capillary electrophoresis for drug-protein binding studies [J]. Journal of Pharmceutical and Biochemical Anaysisl, 2010, 52(2): 232-241.
[84] Martma K., Fernaeus S. Z., Land T., et al. Novel capillary coatings in open tubular CEC as models for biological membranes [J]. Electrophoresis, 2010, 31(9): 1586-1589.
[85] Kuldvee R., Wiedmer S. K., ??rin K., et al. Human low-density lipoprotein-coated capillaries in electrochromatography [J]. Analytical Chemistry, 2005, 77(10): 3401-3405.
[86] Kuldvee R., D’ulivo L., Yohannes G., et al. Open tubular capillary electrochromatography:Technique for oxidation and interaction studies on human low-density lipoproteins [J]. Analytical Chemistry, 2006, 78(8): 2665-2671.
[87] Chen J., Fallarero A., M??tt?nen A., et al. Living cells of Staphylococcus aureus immobilized onto the capillary surface in electrochromatography: A tool for screening of biofilms [J]. Analytical Chemistry, 2008, 80(13): 5103-5109.
[88]熊彩侨,夏之宁,黄锐,等.区段-区段动力学毛细管电泳测定动力学参数的新方法及其应用[J].中国科学, B辑, 2008, 38(8): 705-709.
[89]张维冰.毛细管电色谱理论基础[M].北京:科学出版社, 2006. 94-97.
[90] Jin J., Zhang X.. Spectrophotometric stidies on the interaction between pazufloxacin mesilate and human serum albumin or lysozyme [J]. Journal of Luminescence, 2008, 128: 81-86.
[91] Mckee S. A., Sane D. C., Deliargyris E. N., et al. Aspirin resistance in cardiovascular disease: A review of prevalence, mechanisms, and clinical significance [J]. Thromb Haemost, 2002, 88(5): 711.
[92]杨信怡,游学甫.注射用帕珠沙星的药理研究和临床应用[J].国外医药:抗生素分册, 2003, 24(2): 80-87.
[93] Michelson A D. Platelet [M]. 2nd ed. MA: Academic press, 2007. 1099-1101.
[94] Bobert M. B.. The role of adenosine in the regulation of coronary blood flow [J]. Circulation Research, 1980, 47(6): 807-813.
[95]田谋利,孙海静,高欣,等.尿苷三磷酸对大鼠局灶性缺血再灌注损伤脑组织的保护作用[J].第二军医大学学报, 2010, 31(6): 608-611.
[96] Cheng T. O.. Cardiovascular effects of Danshen [J]. International Journal of Cardiology, 2007, 121: 9-22.
[97]刘陶世,黄耀洲.丹参成分、制剂和质量控制研究概况[J].南京中医药大学学报, 1998, 14(4): 255-256.
[98]吕志刚,徐列明.丹酚酸B的药理研究概况[J].医学研究杂志, 2008, 37(8): 114-116.
[99]李琦,叶蕴华,刑其毅.三七水溶性化学成分及其药理研究新进展[J].高等学校化学学报, 1996, 17(12): 1886-1892.
[100]可燕,蒋嘉烨,王视珍,等.三七根及花总皂苷抗肿瘤细胞诱导的血小板聚集研究[J].中药材, 2010, 33(1): 96-99.
[101]郭洁文,李丽明,邱光清,等.三七总皂苷对心梗后心室重建构大鼠ACE2与TNF-α表达的影响[J].中药材, 2010, 33(1): 89-92.
[102]黄赵刚.血小板生物亲和萃取结合HPLC法筛选三七抗血小板聚集活性皂苷成分[D].安徽:安徽医科大学, 2005.
[103]刘佳,陈晓辉,钱忠直,等. RP-HPLC法测定藤三七不同部位中腺苷的含量[J].沈阳药科大学学报, 2009, 26(9): 724-739.
[104]张冰,陈晓辉,毕开顺. HPLC法同时测定三七花中鸟苷和腺苷的含量[J].药物分析杂志, 2009, 29(12): 2113-2115.
[105]吴杰,刘旭.腺苷的临床应用[J].国外医学.心血管疾病分册, 2002, 29(3): 165-166.
[106] Gong Y. X., Li S. P., Li P., et al. Simultaneous determination of six main nucleosides and bases in natural and cultured Cordyceps by capillary electrophoresis [J]. Journal of Chromatography A. 2004, 1055: 215-221.
[107] Yu L., Zhao J., Li S. P., et al. Quality evaluation of Cordyceps through simultaneous determination of eleven nucleosides and bases by RP-HPLC [J]. Journal of Separation Science. 2006, 29, 953–958.
[108] Yuan J. P., Zhao S. Y., Wang J. H., et al. Distribution of nucleosides and nucleobases in edible fungi [J]. Journal of Food Chemistry, 2008, 56: 809-815.