银杏组织、细胞悬浮培养及银杏黄酮和萜内酯的提取、纯化和检测方法研究
|
摘要
银杏(Ginkgo biloba L．)又称白果树，是地球上生长着的最古老的高等植物之一。银杏叶中主要含有黄酮类和萜内酯类两大类有效成分，它们具有扩张血管、增加血流量、改善血液循环动力障碍、降低脑血管阻力、抑制血栓形成及清除人体自由基等多种作用。尤其银杏内酯B是目前发现的最有效的特异的血小板活化因子(PAF)拮抗剂，已成为世界多国医药专家的研究热点。
目前人们主要从银杏叶中提取银杏黄酮和萜内酯，但由于银杏黄酮和银杏内酯的含量很低，尤其银杏叶中最主要的有效成分银杏内酯B主要存在于根中，在银杏叶中含量极低(＜0.1％)，且是在生长状态的叶中含量最高，采叶影响树的生长；又因银杏叶中主要的有效成分受多种因素的影响又有很大差别，银杏叶革质化、蜡质化程度高，使得国内银杏叶提取物的质量很难达到国际标准。同时，银杏的大面积栽植占据了大量珍贵的耕地面积。为此，本研究利用植物细胞工程技术建立了银杏幼叶愈伤组织无性系和细胞悬浮培养系，并对银杏细胞悬浮培养系进行了多代筛选、驯化，对其培养条件的生理、生化调控进行了大量研究，建立起了生长旺盛且具有高黄酮和内酯含量的优良的银杏细胞悬浮培养系。
实验中发现基因型、外植体类型、树龄和季节等诸多因素对银杏愈伤组织培养物中的黄酮和萜内酯的含量都有较大影响。应选择树体自身黄酮和内酯含量高的树种，在4月中下旬银杏叶刚萌发时取3～5年生的幼树的嫩叶为外植体进行愈伤组织的诱导。通过对培养基种类、多种不同浓度的激素组合、紫外线照射、培养过程的温度和光照等条件的研究，采用分光光度法和HPLC法相结合的方法筛选出了银杏黄酮和萜内酯含量都较高的银杏幼叶愈伤组织无性系。
由于在固体培养基上进行培养的愈伤组织生长速率较慢，不适于培养物中目标次生代谢产物的工业化生产，实验中又进一步以固体培养基上的愈伤组织无性系为材料建立了银杏细胞悬浮培养系。对银杏细胞悬浮培养系的生理、生化调控研究结果表明：以B5培养基为基本培养基，附加NAA0.5～1.0mg／L、BA0.2～0.5mg／L、30g／L蔗糖和15g／L葡萄糖，培养液pH值为5.8～6.2，培养基中Cu~(2+)提高十倍，Fe~(2+)浓度增加0.5倍，氨态氮加倍，硝态氮减半，以30～40FWg／L的接种量接种于装有50ml培养液的150ml三角瓶中，置于25±1℃、3000Lux连续光照条件下进行悬浮培养，
大连理工大学博士学位论文
在培养的第15天加人0．ito．2叭的苯丙氨酸，在培养的第20天收获，最有利于银
杏细胞悬浮培养系中黄酮和格内酯的生产。细胞悬浮培养物中黄酮含量可达3．35％，
前内酯含量可达0．0758％。
本实验还对银杏叶及银杏组织细胞培养物中有效成分的提取、纯化和检测方法
进行了研究，发现将银杏叶粉碎成 40～60目，在料液比为 1：15的情况下以 70％
乙醇为提取溶剂，先湖预处理205，然后用超声波强牌取stnin，重复操作2次，
银杏叶中有效成分提取率可达98．5％。GBE粗提液经液一液法去杂后，将提取
液中黄酮浓度稀释为1．ul．Zmtw左右，再以10倍柱体积的提取液控制吸附流速
为 4倍树脂柱体规用 HPD 00大孔吸附树脂吸附，吸附完全后依次用去高于水、
20％乙测涤，再用流速为2倍树脂柱体肌的90％乙醇解吸的方法获得的GBE
精提物经分光光度法和 HPLC法检测黄酮着为 26．l％，蔬内酯为 6．2％，达到国际标
准。
对银杏黄酮提取液采用微波强化水解、HP＊C检测：对银杏内酯采用10％甲醇
回流提取、单柱层析纯化、y－pLC分析和二极管阵列检测器检测，以上方法减少
了样品的预纯化时间，且操作简便，易于掌握，可在短时间内检测很多样品。这将为
以银杏叶提取物为主要原料的厂家对其产品中有效成分含量的测定提供极大的方
便。
Ginkgo biloba L. is also called white fruitage tree, and it is one of the higher plants, which have the longest life. The leaves from Ginkgo biloba L. mainly included some effective constituents, such as flavonoids and ginkgolides, which can aggrandize the blood vessel, increase the outflow of blood, improve the kinetic of blood-circulating, decrease the resisting force of cerebrovascular, suppress thrombisis and clean free radical of human body. Ginkgolide B especially is the most effective agonist of plaque activation factor (PAF) and now become a hot research issue in the medicine field of the world.
Now, Flavonoids and ginkgolides were mainly extracted from the leaves of Ginkgo biloba., but the leaves often contain high content of corious and cereous, and the effective constituent is very different under the various conditions. In addition, the constituent of ginkgolides is very low in the leaves, and thus the quality of extraction is difficult to reach the international standards. The large area plantation of Ginkgo biloba occupied the rare cropland. In order to solve the problem above, in this study, the callus and cell suspension culture clone were constructed using plant cell engineering technology. After several generation selection and domestication, the cell suspension clone, which can vigorous grow and contained higher content of flavonoides and ginkgolides, were constructed.
The results showed the contents of flavone and ginkgolides in the callus culture substances were affected by the factors, such as genotype, expalnt category, season and so on. Thus, the trees with higher content of flavone and ginkgolides should be selected at first time.
After a ten days in April, the germination leaves, which grow on 3-5 years old tree, was selected as explant inducting callus. After research on the kinds of culture media .various content of hermone, ultraviolet irridation, temperature and intensity of light in the culture, the callus clone with higher content of flavonoids and ginkgolides of Ginkgo biloba spiraster screened using photometer and HPLC combined.
As the growth rate of callus is slow in the solid medium in the process of culture, callus is not fit for industrialization of secondary metabolite. The cell suspension culture line was established using the callus in the solid medium. The results of physiology and biochemical modulation on cell suspension culture line show that the primary medium was B5, pH value of culture was 5.8-6.2, and then NAA 0.5-1 .Omg/L. 6-BA 0.2~0.5mg/L, sucrose 30g/L and 15g/L glucose were added. In the experiment, the
content of Cu + was increased ten times; the content of Fe + was increased half of one time; the content of ammoniacal nitrogen was increased one time and the content of nitrate nitrogen was decreased half of one time. The inoculation weight was 30-40 FW/L, and inoculate the 150ml flask which contained 50ml culture liquid. At 25? 癈, SOOOlux continuous light treatment, conducted suspension cell culture, at Hie 15th day, add 0.1~0.2g/L phenylalanine, after 20 days, the product will be harvest, which is the optimium condition of producing flavone and ginkgolides in the suspension cell culture solution. In the production, the content of flavonoides can reach 3.35%, the content of ginkgolides can reach 0.0758%.
In this experiment, active constituents in the leaves, tissue and cell culture substration of Ginkgo biloba were extracted, purified and detected. The leaves of Ginkgo biloba were pulverized in 40-60 orders, and were extracted in 70% ethanol solution at the rate of material and solution 1:15, pretreated 20s by microwave, and extracted 5min by ultrasonic, and repeated 2 times above. The extraction rate of active constituent can reach 98.5%. After the extraction by liquid-liquid method diluted the concentration of flavone to 1.0~1.2mg/ml and using 10-times of volume colume at the flow velocity of 4 times of colume volume per hour absorbed by HPD 100, and then washed by distilled water and 20% ethanol, and using 3-times of volume colume at the f
目前人们主要从银杏叶中提取银杏黄酮和萜内酯，但由于银杏黄酮和银杏内酯的含量很低，尤其银杏叶中最主要的有效成分银杏内酯B主要存在于根中，在银杏叶中含量极低(＜0.1％)，且是在生长状态的叶中含量最高，采叶影响树的生长；又因银杏叶中主要的有效成分受多种因素的影响又有很大差别，银杏叶革质化、蜡质化程度高，使得国内银杏叶提取物的质量很难达到国际标准。同时，银杏的大面积栽植占据了大量珍贵的耕地面积。为此，本研究利用植物细胞工程技术建立了银杏幼叶愈伤组织无性系和细胞悬浮培养系，并对银杏细胞悬浮培养系进行了多代筛选、驯化，对其培养条件的生理、生化调控进行了大量研究，建立起了生长旺盛且具有高黄酮和内酯含量的优良的银杏细胞悬浮培养系。
实验中发现基因型、外植体类型、树龄和季节等诸多因素对银杏愈伤组织培养物中的黄酮和萜内酯的含量都有较大影响。应选择树体自身黄酮和内酯含量高的树种，在4月中下旬银杏叶刚萌发时取3～5年生的幼树的嫩叶为外植体进行愈伤组织的诱导。通过对培养基种类、多种不同浓度的激素组合、紫外线照射、培养过程的温度和光照等条件的研究，采用分光光度法和HPLC法相结合的方法筛选出了银杏黄酮和萜内酯含量都较高的银杏幼叶愈伤组织无性系。
由于在固体培养基上进行培养的愈伤组织生长速率较慢，不适于培养物中目标次生代谢产物的工业化生产，实验中又进一步以固体培养基上的愈伤组织无性系为材料建立了银杏细胞悬浮培养系。对银杏细胞悬浮培养系的生理、生化调控研究结果表明：以B5培养基为基本培养基，附加NAA0.5～1.0mg／L、BA0.2～0.5mg／L、30g／L蔗糖和15g／L葡萄糖，培养液pH值为5.8～6.2，培养基中Cu~(2+)提高十倍，Fe~(2+)浓度增加0.5倍，氨态氮加倍，硝态氮减半，以30～40FWg／L的接种量接种于装有50ml培养液的150ml三角瓶中，置于25±1℃、3000Lux连续光照条件下进行悬浮培养，
大连理工大学博士学位论文
在培养的第15天加人0．ito．2叭的苯丙氨酸，在培养的第20天收获，最有利于银
杏细胞悬浮培养系中黄酮和格内酯的生产。细胞悬浮培养物中黄酮含量可达3．35％，
前内酯含量可达0．0758％。
本实验还对银杏叶及银杏组织细胞培养物中有效成分的提取、纯化和检测方法
进行了研究，发现将银杏叶粉碎成 40～60目，在料液比为 1：15的情况下以 70％
乙醇为提取溶剂，先湖预处理205，然后用超声波强牌取stnin，重复操作2次，
银杏叶中有效成分提取率可达98．5％。GBE粗提液经液一液法去杂后，将提取
液中黄酮浓度稀释为1．ul．Zmtw左右，再以10倍柱体积的提取液控制吸附流速
为 4倍树脂柱体规用 HPD 00大孔吸附树脂吸附，吸附完全后依次用去高于水、
20％乙测涤，再用流速为2倍树脂柱体肌的90％乙醇解吸的方法获得的GBE
精提物经分光光度法和 HPLC法检测黄酮着为 26．l％，蔬内酯为 6．2％，达到国际标
准。
对银杏黄酮提取液采用微波强化水解、HP＊C检测：对银杏内酯采用10％甲醇
回流提取、单柱层析纯化、y－pLC分析和二极管阵列检测器检测，以上方法减少
了样品的预纯化时间，且操作简便，易于掌握，可在短时间内检测很多样品。这将为
以银杏叶提取物为主要原料的厂家对其产品中有效成分含量的测定提供极大的方
便。
Ginkgo biloba L. is also called white fruitage tree, and it is one of the higher plants, which have the longest life. The leaves from Ginkgo biloba L. mainly included some effective constituents, such as flavonoids and ginkgolides, which can aggrandize the blood vessel, increase the outflow of blood, improve the kinetic of blood-circulating, decrease the resisting force of cerebrovascular, suppress thrombisis and clean free radical of human body. Ginkgolide B especially is the most effective agonist of plaque activation factor (PAF) and now become a hot research issue in the medicine field of the world.
Now, Flavonoids and ginkgolides were mainly extracted from the leaves of Ginkgo biloba., but the leaves often contain high content of corious and cereous, and the effective constituent is very different under the various conditions. In addition, the constituent of ginkgolides is very low in the leaves, and thus the quality of extraction is difficult to reach the international standards. The large area plantation of Ginkgo biloba occupied the rare cropland. In order to solve the problem above, in this study, the callus and cell suspension culture clone were constructed using plant cell engineering technology. After several generation selection and domestication, the cell suspension clone, which can vigorous grow and contained higher content of flavonoides and ginkgolides, were constructed.
The results showed the contents of flavone and ginkgolides in the callus culture substances were affected by the factors, such as genotype, expalnt category, season and so on. Thus, the trees with higher content of flavone and ginkgolides should be selected at first time.
After a ten days in April, the germination leaves, which grow on 3-5 years old tree, was selected as explant inducting callus. After research on the kinds of culture media .various content of hermone, ultraviolet irridation, temperature and intensity of light in the culture, the callus clone with higher content of flavonoids and ginkgolides of Ginkgo biloba spiraster screened using photometer and HPLC combined.
As the growth rate of callus is slow in the solid medium in the process of culture, callus is not fit for industrialization of secondary metabolite. The cell suspension culture line was established using the callus in the solid medium. The results of physiology and biochemical modulation on cell suspension culture line show that the primary medium was B5, pH value of culture was 5.8-6.2, and then NAA 0.5-1 .Omg/L. 6-BA 0.2~0.5mg/L, sucrose 30g/L and 15g/L glucose were added. In the experiment, the
content of Cu + was increased ten times; the content of Fe + was increased half of one time; the content of ammoniacal nitrogen was increased one time and the content of nitrate nitrogen was decreased half of one time. The inoculation weight was 30-40 FW/L, and inoculate the 150ml flask which contained 50ml culture liquid. At 25? 癈, SOOOlux continuous light treatment, conducted suspension cell culture, at Hie 15th day, add 0.1~0.2g/L phenylalanine, after 20 days, the product will be harvest, which is the optimium condition of producing flavone and ginkgolides in the suspension cell culture solution. In the production, the content of flavonoides can reach 3.35%, the content of ginkgolides can reach 0.0758%.
In this experiment, active constituents in the leaves, tissue and cell culture substration of Ginkgo biloba were extracted, purified and detected. The leaves of Ginkgo biloba were pulverized in 40-60 orders, and were extracted in 70% ethanol solution at the rate of material and solution 1:15, pretreated 20s by microwave, and extracted 5min by ultrasonic, and repeated 2 times above. The extraction rate of active constituent can reach 98.5%. After the extraction by liquid-liquid method diluted the concentration of flavone to 1.0~1.2mg/ml and using 10-times of volume colume at the flow velocity of 4 times of colume volume per hour absorbed by HPD 100, and then washed by distilled water and 20% ethanol, and using 3-times of volume colume at the f
引文
[1] 王成章，沈兆邦，陈祥，银杏叶中聚戊烯醇的化学研究，林产化学与工业，1992，12(4)：279～283．
[2] 丁青龙，王云征，顾卫忠等，银杏叶及其制剂的质量研究，中国现代应用药学杂志，1998，15(5)，11～13．
[3] 于荣敏，赵鸿莲，郑玉果等，银杏细胞悬浮培养及银杏内酯生产的研究，生物工程学报，1999，15(1)：52～58．
[4] 方晓江，沈琴，戴瑞鸿，银杏叶提取物对心血管的作用，中成药，1998，20(10)43～44
[5] 王伏雄，陈祖铿，李宪章，银杏幼胚离体培养的研究Ⅰ，蜂皇浆对幼胚生长的影响，植物学报，1963，11(3)：217～224．
[6] 王伏雄，陈祖铿，李宪章，银杏幼胚离体培养的研究Ⅱ，椰乳汁对幼胚生长的影响，植物学报，1963，13(3)：224～239．
[7] 王兴鹏，袁耀宇，徐家袼，中国药理学通报，1995，11(3)：199～203．
[8] 王成章，陈祥，谭卫红等，银杏叶中黄酮类化合物及其分析方法，林产化学与工业，1998，18(1)：83～88．
[9] 邓启焕，高勇，第二类超临界流体萃取银杏叶有效成分的研究，中草药，1999，30(6)：419～422．
[10] 史清文，刘素云，张文素等，银杏叶的研究开发概况，天然产物研究与开发，1995，7(1)：73～76，
[11] 田卉南，王颉，厉尽忠等，高效液相色普法蒸发激光散射检测器测定银杏叶提取物中内酯的含量，药物分析杂志，1997，17(4)：282～285．
[12] 刑建民，查丽杭，李佐虎，等，植物细胞大规模培养生物反应器研究概况，生物工程进展，1997，5：49～53．
[13] 刘佳佳，郭勇，郑穗平等，高产黄酮苷银杏悬浮培养细胞系选育和继代培养稳定性的研究，生物工程学报，2001，17(1)94～97．
[14] 刘春朝，王玉春，赵兵等，生物诱导子调节植物组织次生代谢的研究，植物学通报，1999，16(2)：131～137．
[15] 刘重芳，吴志荣，方青汉，银杏叶总黄酮提取工艺探讨，中成药，1992，14(7)：7～8．
[16] 植物学(上册)，华东师范大学，上海师范大学，南京师范学院编，1991．
[17] 庄向平，虞杏英，杨更生等，银杏叶中黄酮含量的测定和提取方法，中草药，1992，23(3)：122～124．
[18] 齐藤和季，等，第十一回植物组织培养学大会讲演要旨集，罔山．1989．
[19] 吴伯平，1996年美国最风行的10种中草药，中国医药情报，1996，2(4)255～257．
[20] 李兆龙，前景诱人的银杏叶制剂，植物杂志，1996，4：7～8．
[21] 李兆龙，从银杏叶中提取黄酮类化合物的方法，中成药，1992，14(6)：5～8．
[22] 李兆龙，胡季强，卢耀明编著，银杏叶的开发利用，上海科技文献出版社，1996．
[23] 李俊，李健，苏小建，银杏叶提取物杂质去除方法研究，化学世界，1998，5：243～247．
[24] 李嵘，金美芳，微波法提取银杏黄酮甙的新工艺，食品科学，2000，21(2)：39～41．
[25] 李新岗，韦玮，陈薇，富含银杏内酯的银杏叶干浸膏的制备，中国医药工业杂志，1998，29(1)：8～9．
[26] 肖崇厚主编，中药化学，上海科学技术出版社，1998．
[27] 苍宁，银杏叶制品的研究开发及市场，中国医药情报，1995，1(4)217～222．
[28] 黄酮类化合物，Harborne著，戴凯伦译，科学出版社，北京，1983．
[29] 陈冲，罗思齐，银杏叶提取物的生产工艺条件研究，中草药，1997，28(7)402～404．
[30] 陈维军，华中农业大学硕士学位论文，1996．
[31] 陈维军，谢笔钧，胡慰望，银杏内酯的分光光度计法测定研究，中草药，1998，29(增刊)：45～48．
[32] 杨尔宁，谢笔钧，胡慰望，HPLC法分析银杏黄酮水解条件的研究，中草药(增刊)，1998，29：45～50
[33] 周立刚，郑光植，王世林等，滇紫草培养细胞中色素合成的代谢调节，生物工程学报，1992，8(4)：353～356．
[34] 林启寿编著，中草药成分化学，科学出版社，1977，p85．
[35] 苑可武，孟宪惠，徐文豪，银杏叶中黄酮含量的季节性变化，中草药，1997，28(4)：211～213．
[36] 郁青，沈兆邦，陈祥等，银杏内酯含量的变化规律研究，林产化学与工业．1998，18(4)：1～6．
[37] 姚胃溪，超临界萃取和HPLC测定银杏叶中黄酮，第四届全国中药和天然药物学术会议．北
京：1994-04．
[38] 姚渭溪，杨春，田宇等，银杏内酯的快速测定方法研究，药物分析杂志，1999，19(1)38～41．
[39] 胡敏，甘路，姜发堂等，银杏叶中黄酮类化合物最佳提取工艺研究，食品工业科技，1997，5：49～51．
[40] 倪德祥，邓志龙，植物激素对基因表达的调控，植物生理学通讯，1992，28(6)：461～466．
[41] 钱庭宝主编，吸附树脂及其应用，化学工业出版社，1990．
[42] 梅田诚一，日本公开特许公报，1992：平4-182434
[43] 黄桂宽，李毅，谢荣仿等，银杏多糖的分离纯化和鉴定，中国生化药物杂志，1996，17(4)：157～160．
[44] 彭芳，黄酮类化合物的生物活性，国外医药-植物药分册，1998，13(5)207～210．
[45] 游松，姚新生，陈英杰，银杏叶化学成分和药理研究，沈阳药学院学报，1988，5(2)：142～147。
[46] 詹祥灿，植物组织培养中的光自养生长，植物生理学通讯，1981，2：12～16．
[47] 廖亮，银杏叶总黄酮提取方法研究，食品科学，1994，8：33～35．
[48] 翟中和主编，细胞生物学，高等教育出版社，1996，P317．
[49] 潘瑞炽，植物激素的作用机理，植物杂志，1982，5：1～5．
[50] 潘瑞炽，董禺得编，植物生理学(下册)．1987，p63．
[51] 潘瑞炽，董愚得，植物生理学(上册)．1990：64．
[52] 戴均贵，朱蔚华，吴蕴祺等，不同培养基、激素、碳源和氮源对银杏愈伤组织生长及银杏内酯形成的影响．中草药(增刊)，1998，(29)：63～67．
[53] Abte S, et al., Plant Science Letters. 1979, 16:129～134.
[54] Agnoli A, et al, Paper Presented at the International Symposium Held in New York, 22nd September 1984.
[55] Antoni G, Lech C. The effect of platelet activating factor antagonist (BN52021) on cerulein-induced acute pancreatitis with reference to oxygen radicals. Int J Pancreatol, 1991, 8(1):1.
[56] Ball E. Growth of the embryo of Ginkgo biloba under experimental codition Ⅰ.Origin of the first root of the seedling in vitro. Amer. Jour. Bot. 1956,43:488～495.
[57] Ball E. Growth of the embryo of Ginkgo biloba under experimental codition Ⅱ. Effects
of a longitudinal split in the tip of the hypocotyl. Amer Jour Bot. 1956, 43:802～810.
[58] Bergmann L. Planta. 1967, 74: 243～249.
[59] Braquet P. The effects of PAF-acether on the cardiovascular system and their inhibition by a now highly specific PAF-accther receptor antagonist BN52021 Phermacol Res Communi, 1986, 18(8) :717.
[60] Braquet P. In Ginkgolides: Chemistry, Biology, Pharmacology and Clinical Perspectives. Vol Ⅱ. Barcelona: J.R. Prous Science, 1990:613
[61] Briancon- scheid Francoise. et al. Journal of Chromatography, 1982, 245:261-268.
[62] Brunello. at al, Pharmacol Res Commun. 1985, 17(11):1063.
[63] Capite L. Action of light and temperature on growth of plant tissue culture in vitro[J]. Amer J Bot, 1955(45) :869～873.
[64] Carrier D.J, et al. Nutritional and hormonal requirements of Ginkgo biloba embreyo-derived callus and suspention cell culture. Plant Cell Reports, 1990, 8(11): 635～638.
[65] Carrier D. J, et al. Detection of ginkgolide A in Ginkgo biloba cell culture. Plant Cell Reports, 1991, 10(5) 256～259.
[66] Chijnacki T, et al. Chemica Scripta, 1987, 27:209. 142～147
[67] Corcoran P, Wang Y, Katz N, et al., Platelet activating factor antugonist enhances lung preservation. J Sarg Res, 1992,52(6):615.
[68] Cordts J . et al. Effects of carbohydrates and nitrogen of the development of canthocyanins of a red leaf peach in vitro. Plant cell, tissue organ culture. 1987,9(2)103.
[69] Desquand S, lefort J, et al, Interforence of BN52021 with the bronchopulmonery effects of PAF-acether in the guinea pig. Eur J Pharmacol, 1986,127(1):83.
[70] Dolores Hernanz Vila, Fco Jose Heredia Mira ,Rafael Beltran Lucena,et al. Optimation of an extraction method of aroma compounds in white wine using ultrasound. Talana, 1999,50,143.
[71] Donenburg H and Knorr D, Enzyme Microbial. 1995,17:674.
[72] Droy - Lefaix M. Free radical scavenger properties of EGB 761 on functional disorders induced by exper mandal - diabetic retinopathy. Oxid Stress Aging. 1995:277-286.
[73] Edelmann J and Hanson A D , Planta. 1971, 98:150～156.
[74] Etienne Annie, et al, J Pharmacol . 1982, 13(2)291.
[75] Fitzl G, Welt, K, Schaffaremietz L, et al, EXP Tosicol Patthol, 1996, 48(1) 33.
[76] Foegh M, Prolongation of catdiac allograft survival with BN52021. Transplantation, 1986,42(1):86.
[77] Fowler M, In:The Proceeding of the 3ND Asia-Pasific Conference on Biotechnoloqy. Singapore. 1994.
[78] Fowler M, et al.,Plant tissue culture .1982,5:225.
[79] Fowler M. In:Plant Biotechnology .Editid by S. Melltelland H. Smith. Cambrige Univ. Press. 1983.
[80] Fumimaro T ,et al.,EP Patent 0166436.
[81] Gebner B, Voelp A. Study of the long term action of a ginkgo biloba extract on vigilance and mental performance as determined by means of quantitative pharmaco EEG and psychometric measurements. Arznei-Forsch/Drug Res. 1985,35(9):1459.
[82] Gellerman Joanne L, et al. Phytochemistry, 1976,15:1959. 17(4): 157～160.
[83] Guinot P. Effects of the PAF antagonists, BN52063, BN52021 in various clinical indications. J Lipid Mediators Cell Signalling, 1994, 10(1～2):141.
[84] Haslar A, Sticher O, et al. High-performance liquid chromatographic determination of five widespread aglycones. J Chromatogr. 1990,508:236.
[85] Haslar A, Sticher O, et al. Identification and determination of the falconoid From Ginkgo biloba by HPLC. J Chromatogr. 1992,605:41.
[86] Holden M A ,et al.,Manipulating secondary metabolism in culture. Robins J and Rhodes M, ed . Cambridge Univ. Press, Cambridge, 1988, 57-65.
[87] Huh Hoon, et al. Ontogeneic aspects of ginkgolide production in Ginkgo biloba Plant Med. 1993,59:232～239.
[88] Ibata Koichi, et al, Biochem J ,1983,213:305.
[89] Jacob L. Review Thermal and Non-thermal Interaction of Microwave Radiation with Materials. Journal of Materials Science, 1995, (30):5321～5327
[90] Jeon M. et al. Culture of Ginkgo biloba effect of nutrition and hormonal factors on thegroeth ofcultured cells derived from Ginkgo biloba. Archives of Pharmacal Research (Seoul), 1993, 16(3) :244～250.
[91] Kaul K and Sabharwal P S, Plant Physiol. 1971, 47, 691～695.
[92] Kimersley A , et al. W. Alton Jones Cell Science Centre Annual Report. PP lake placide. W. Alton Jones Cell Scince Centre , 1981.
[93] Kircher Henvy W, et al., Phytochemistry, 1970, 9:1879.
[94] Koltai M, Hosford D, Guinot P, et al. PAF: A review of its effects, antagonists and possible future clinical implications (part Ⅱ). Ear J Pharmacol, 1989, 164(2) :293.
[95] Kose K, Dogan P, J Int Med Res, 1995,23(1): 9.
[96] Krieglstein J Nanoprotective effective of Ginkgo Liloda Constituents, Eur J Pharmacol Sci, 1995, 3(1)391.
[97] Larsen R, et al, Therapie, 1978, 33(5): 651～655.
[98] Laurain D, et al., Phytochemistry, 1997, 46(1): 127～132.
[99] Laurain D, et al., Plant cell, Tissueand organ culture. 1996, 4:19.
[100] Levinel, Mol Pharmacol, 1988, 34(6) 793.
[101] Li J, T. (李继桐), The development of the enbryo of Ginkgo biloba, Net. Tsing Hua Univ, Sci. Rep. 1934, 132:29～35.
[102] Li J, T. and Shen, T(沈同), The effect of "Pantothenic acid" on the growth of the radical of Ginkgo embryos in artificial media, Net. Tsing Hua Univ, Sci. Rep. 1934, 132: 53～60.
[103] Lobstein A, et al. Seasonal Variations of the flavoniod Content from Ginkgo biloba. Leaves. Planta Med, 1991, 57:430～433..
[104] Lohn M , Lin F, Tseng C, et al, J Formos Med Assoc, 1994;93(7) :592.
[105] Makino, et al. Jpn Kokai Tokkyo Koho, 1991, JP 03, 275:629.
[106] Maruyama M, et al. The ginkgolides isolation and characterization of the various
grpups . Tetrahedron Letters, 1967, 4:299 .
[107]Mavituna F. Recent Advances in Biotecnology, Kluwger Academinc Publishers, Neitherland. 1992.
[108]Meruyama M, et al, Am Heart J, 1990, 120 (3) :510.
[109]Morimoto T, Agricultural and Biological Chemistry. 1988, 52(7)1835.
[110]Muller K, Borger H. Arbeiten aus der Biologischen Bundesanstalt fur Land-und For stwirtschaft (Berlin) . 1940, 23:189.
[111]Nakanish K. Structure of bilobelide, a rare tert-butyl containing sesquiterpenoid Related to the C20-ginkgolides. J Am Chem Soc, 1971, 93(14): 3544
[112]Ogihari Y and Tsunewakik. Bot Mag Tokyo. 1979, 92:163～171.
[113]Pal Nirupam, Korfiatis George P, Patel Vijay. Sonochemical extraction and biological treatmet of pentachlorophenol contaminated wood, Jounal of Hazardous Materials. 1997, 53: 165.
[114]Pamplim E J and Champan J M.J Exp Bot. 1975, 26:212～220.
[115]Pennkck J F , et al. Nature (London), 1960, 186:470.
[116]Pietta P, et al. Analysis of terpenes from Ginkgo biloba L. Extracts by Reversed hase High-Performance Liquid Chromatogrophy. Chiomatographia, 1990, 29(5)251～253.
[117]Puhan Z S, Martin M, Progress in Industrial Microbiology. 1971,9:13.
[118]Punkt K, Schaffranietl, et, al. Acta Histchem, 1995:97(1):67.
[119]Ranansky J, et al. Biologia Bratislava, 1996, 51(4) :471～477.
[120]Rong Y. et al. Free Radie Biomed. 1996, 20(1): 121.
[121]S. Furukawa, Scientific Papers of The Institute of Physical and Chemical Research , 1932, 19: 27-31.
[122]Salisova, Toma S, Mason T J. Comparison of conventional and ultrasonically assisted Extractions of pharmaceutically active compounds from Salvia officinalis . Ultrasonics Sonochemistry. 1997, 4:131
[123]Schemidt H, et al. Tetrahedron Letters. 1971, 44:4151. 353～356.
[124]Schennen A, et al. Plant Medical . 1986, 235,
[125] Seibert M, Kadade P. Enviromental factor :a light in:staba E jed. Plant tissue culture as a source of biochemicals[M]. Florida: CRC Press, 1980.
[126] Shen J G, Biochen Mol Bilint, 1995:35(1):125.
[127] Siron Al, et al, Am J Bhysiol, 1989, 257(1, Pt2): H25.
[128] Stetler D and Lastische V M, Science. 1965, 149:1387～1388.
[129] Sticher O. Quality of ginkgo preparation Plant Med. 1993, 59(1): 2～11.
[130] Sunderland N, Ann. Bot. 1966, 30:253～268.
[131] Szoeke E, et al., Fiziol Rast. 1977, 24:832～840.
[132] Tabata M and Fujita Y, In "Biotecnology in plant Science", New York: Academic Press, Inc., 1985, p207.
[133] Takeda J. light-induced synthesis of anthocyanin in carrot cells in suspension J. EXP. Bot. 1988, 39(205):1065～1077.
[134] Tisobe T C, et al. Plant Physiology. 1973, 51(4): 805～816.
[135] Tosaki A, et al., Coron Artery Dis, 1994,5(5)443
[136] Tosaki A, Pali T, etal, Diabetologia, 1996; 39(11): 1225.
[137] Touvay C, Etienne A, Braquetp, Inhibition of antigen-induced lung anaphy laxis in the quinea pig by BN52021 a new specific PAF isolated from Ginkgo biloha Agents Actions, 1985, 17(3):371.
[138] Tulecke W. A haploid tissue cultre from the female gametophyte of Ginkgo biloha L . Nature. 1964, 204(4): 94～95.
[139] Tulecke W. R., A tissue derived from the pollen of Ginkgo, Science. 1953, 117:599～600.
[140] Tulecke W. R., Arginine-requiring strains of tissue obttained from Ginkgo biloba, Plant physiol. 1960, 35:19～24.
[141] Tulecke W. R., The pollen of Ginkgo biloba in vitro culture and tissue formation, Amer, J, Bot. 1957, 44:602～608.
[142] Van Beek T A et al. Determination of ginkgolides and bilobalide in ginkgo biloba leaves and phytopharma ceuticals. J. Chromatogr. 1991, 543: 375.
[143] Vasil I and Hildebrandt A . Planta. 1966, 68:69～82.
[144]Villamediana L. Effects of the PAF antagcnist BN52021 on the hemodyrnamics of rats with experimentovl cirrhosis of the liver Life Sci, 1986, 39(3):201.
[145]Villbauer E B, etal, J org Chom, 1987, 52(7): 1186.
[146]Vinatoru, Maricela Toma, Otilia Radu, et al. The use of ultrasound for the extraction of bioactive principles from plant materials." . Ultrasonics Sonochemistry. 1997,4:135.
[147]Wanger H, et al. Dtsch Apoth Ztg. 1989, 129:2421.
[148]Weinges K et al. Liebigs Ann Chem., 1969, 724:261
[149]Weinges K, et al, Arzneimitted - Forschung, 1968, 18(5): 537～544.
[150]Yamakawa T et al. Bio. Chem. 1983, 47:2185～2191.
[151]Yeoman M, et al. Diffirentiation in vitro 4th Symposium British Society Cell Biology. Cambrige Uni. Press. Cambrige. 1981, 65～81.
[152]Zenk M H, et al. Plant tissue culture and Its Biotecknological Application, Edited by Zenk, M. H, Springer-Verlag, Berlin, Heideberry. New York. 1977.
[2] 丁青龙，王云征，顾卫忠等，银杏叶及其制剂的质量研究，中国现代应用药学杂志，1998，15(5)，11～13．
[3] 于荣敏，赵鸿莲，郑玉果等，银杏细胞悬浮培养及银杏内酯生产的研究，生物工程学报，1999，15(1)：52～58．
[4] 方晓江，沈琴，戴瑞鸿，银杏叶提取物对心血管的作用，中成药，1998，20(10)43～44
[5] 王伏雄，陈祖铿，李宪章，银杏幼胚离体培养的研究Ⅰ，蜂皇浆对幼胚生长的影响，植物学报，1963，11(3)：217～224．
[6] 王伏雄，陈祖铿，李宪章，银杏幼胚离体培养的研究Ⅱ，椰乳汁对幼胚生长的影响，植物学报，1963，13(3)：224～239．
[7] 王兴鹏，袁耀宇，徐家袼，中国药理学通报，1995，11(3)：199～203．
[8] 王成章，陈祥，谭卫红等，银杏叶中黄酮类化合物及其分析方法，林产化学与工业，1998，18(1)：83～88．
[9] 邓启焕，高勇，第二类超临界流体萃取银杏叶有效成分的研究，中草药，1999，30(6)：419～422．
[10] 史清文，刘素云，张文素等，银杏叶的研究开发概况，天然产物研究与开发，1995，7(1)：73～76，
[11] 田卉南，王颉，厉尽忠等，高效液相色普法蒸发激光散射检测器测定银杏叶提取物中内酯的含量，药物分析杂志，1997，17(4)：282～285．
[12] 刑建民，查丽杭，李佐虎，等，植物细胞大规模培养生物反应器研究概况，生物工程进展，1997，5：49～53．
[13] 刘佳佳，郭勇，郑穗平等，高产黄酮苷银杏悬浮培养细胞系选育和继代培养稳定性的研究，生物工程学报，2001，17(1)94～97．
[14] 刘春朝，王玉春，赵兵等，生物诱导子调节植物组织次生代谢的研究，植物学通报，1999，16(2)：131～137．
[15] 刘重芳，吴志荣，方青汉，银杏叶总黄酮提取工艺探讨，中成药，1992，14(7)：7～8．
[16] 植物学(上册)，华东师范大学，上海师范大学，南京师范学院编，1991．
[17] 庄向平，虞杏英，杨更生等，银杏叶中黄酮含量的测定和提取方法，中草药，1992，23(3)：122～124．
[18] 齐藤和季，等，第十一回植物组织培养学大会讲演要旨集，罔山．1989．
[19] 吴伯平，1996年美国最风行的10种中草药，中国医药情报，1996，2(4)255～257．
[20] 李兆龙，前景诱人的银杏叶制剂，植物杂志，1996，4：7～8．
[21] 李兆龙，从银杏叶中提取黄酮类化合物的方法，中成药，1992，14(6)：5～8．
[22] 李兆龙，胡季强，卢耀明编著，银杏叶的开发利用，上海科技文献出版社，1996．
[23] 李俊，李健，苏小建，银杏叶提取物杂质去除方法研究，化学世界，1998，5：243～247．
[24] 李嵘，金美芳，微波法提取银杏黄酮甙的新工艺，食品科学，2000，21(2)：39～41．
[25] 李新岗，韦玮，陈薇，富含银杏内酯的银杏叶干浸膏的制备，中国医药工业杂志，1998，29(1)：8～9．
[26] 肖崇厚主编，中药化学，上海科学技术出版社，1998．
[27] 苍宁，银杏叶制品的研究开发及市场，中国医药情报，1995，1(4)217～222．
[28] 黄酮类化合物，Harborne著，戴凯伦译，科学出版社，北京，1983．
[29] 陈冲，罗思齐，银杏叶提取物的生产工艺条件研究，中草药，1997，28(7)402～404．
[30] 陈维军，华中农业大学硕士学位论文，1996．
[31] 陈维军，谢笔钧，胡慰望，银杏内酯的分光光度计法测定研究，中草药，1998，29(增刊)：45～48．
[32] 杨尔宁，谢笔钧，胡慰望，HPLC法分析银杏黄酮水解条件的研究，中草药(增刊)，1998，29：45～50
[33] 周立刚，郑光植，王世林等，滇紫草培养细胞中色素合成的代谢调节，生物工程学报，1992，8(4)：353～356．
[34] 林启寿编著，中草药成分化学，科学出版社，1977，p85．
[35] 苑可武，孟宪惠，徐文豪，银杏叶中黄酮含量的季节性变化，中草药，1997，28(4)：211～213．
[36] 郁青，沈兆邦，陈祥等，银杏内酯含量的变化规律研究，林产化学与工业．1998，18(4)：1～6．
[37] 姚胃溪，超临界萃取和HPLC测定银杏叶中黄酮，第四届全国中药和天然药物学术会议．北
京：1994-04．
[38] 姚渭溪，杨春，田宇等，银杏内酯的快速测定方法研究，药物分析杂志，1999，19(1)38～41．
[39] 胡敏，甘路，姜发堂等，银杏叶中黄酮类化合物最佳提取工艺研究，食品工业科技，1997，5：49～51．
[40] 倪德祥，邓志龙，植物激素对基因表达的调控，植物生理学通讯，1992，28(6)：461～466．
[41] 钱庭宝主编，吸附树脂及其应用，化学工业出版社，1990．
[42] 梅田诚一，日本公开特许公报，1992：平4-182434
[43] 黄桂宽，李毅，谢荣仿等，银杏多糖的分离纯化和鉴定，中国生化药物杂志，1996，17(4)：157～160．
[44] 彭芳，黄酮类化合物的生物活性，国外医药-植物药分册，1998，13(5)207～210．
[45] 游松，姚新生，陈英杰，银杏叶化学成分和药理研究，沈阳药学院学报，1988，5(2)：142～147。
[46] 詹祥灿，植物组织培养中的光自养生长，植物生理学通讯，1981，2：12～16．
[47] 廖亮，银杏叶总黄酮提取方法研究，食品科学，1994，8：33～35．
[48] 翟中和主编，细胞生物学，高等教育出版社，1996，P317．
[49] 潘瑞炽，植物激素的作用机理，植物杂志，1982，5：1～5．
[50] 潘瑞炽，董禺得编，植物生理学(下册)．1987，p63．
[51] 潘瑞炽，董愚得，植物生理学(上册)．1990：64．
[52] 戴均贵，朱蔚华，吴蕴祺等，不同培养基、激素、碳源和氮源对银杏愈伤组织生长及银杏内酯形成的影响．中草药(增刊)，1998，(29)：63～67．
[53] Abte S, et al., Plant Science Letters. 1979, 16:129～134.
[54] Agnoli A, et al, Paper Presented at the International Symposium Held in New York, 22nd September 1984.
[55] Antoni G, Lech C. The effect of platelet activating factor antagonist (BN52021) on cerulein-induced acute pancreatitis with reference to oxygen radicals. Int J Pancreatol, 1991, 8(1):1.
[56] Ball E. Growth of the embryo of Ginkgo biloba under experimental codition Ⅰ.Origin of the first root of the seedling in vitro. Amer. Jour. Bot. 1956,43:488～495.
[57] Ball E. Growth of the embryo of Ginkgo biloba under experimental codition Ⅱ. Effects
of a longitudinal split in the tip of the hypocotyl. Amer Jour Bot. 1956, 43:802～810.
[58] Bergmann L. Planta. 1967, 74: 243～249.
[59] Braquet P. The effects of PAF-acether on the cardiovascular system and their inhibition by a now highly specific PAF-accther receptor antagonist BN52021 Phermacol Res Communi, 1986, 18(8) :717.
[60] Braquet P. In Ginkgolides: Chemistry, Biology, Pharmacology and Clinical Perspectives. Vol Ⅱ. Barcelona: J.R. Prous Science, 1990:613
[61] Briancon- scheid Francoise. et al. Journal of Chromatography, 1982, 245:261-268.
[62] Brunello. at al, Pharmacol Res Commun. 1985, 17(11):1063.
[63] Capite L. Action of light and temperature on growth of plant tissue culture in vitro[J]. Amer J Bot, 1955(45) :869～873.
[64] Carrier D.J, et al. Nutritional and hormonal requirements of Ginkgo biloba embreyo-derived callus and suspention cell culture. Plant Cell Reports, 1990, 8(11): 635～638.
[65] Carrier D. J, et al. Detection of ginkgolide A in Ginkgo biloba cell culture. Plant Cell Reports, 1991, 10(5) 256～259.
[66] Chijnacki T, et al. Chemica Scripta, 1987, 27:209. 142～147
[67] Corcoran P, Wang Y, Katz N, et al., Platelet activating factor antugonist enhances lung preservation. J Sarg Res, 1992,52(6):615.
[68] Cordts J . et al. Effects of carbohydrates and nitrogen of the development of canthocyanins of a red leaf peach in vitro. Plant cell, tissue organ culture. 1987,9(2)103.
[69] Desquand S, lefort J, et al, Interforence of BN52021 with the bronchopulmonery effects of PAF-acether in the guinea pig. Eur J Pharmacol, 1986,127(1):83.
[70] Dolores Hernanz Vila, Fco Jose Heredia Mira ,Rafael Beltran Lucena,et al. Optimation of an extraction method of aroma compounds in white wine using ultrasound. Talana, 1999,50,143.
[71] Donenburg H and Knorr D, Enzyme Microbial. 1995,17:674.
[72] Droy - Lefaix M. Free radical scavenger properties of EGB 761 on functional disorders induced by exper mandal - diabetic retinopathy. Oxid Stress Aging. 1995:277-286.
[73] Edelmann J and Hanson A D , Planta. 1971, 98:150～156.
[74] Etienne Annie, et al, J Pharmacol . 1982, 13(2)291.
[75] Fitzl G, Welt, K, Schaffaremietz L, et al, EXP Tosicol Patthol, 1996, 48(1) 33.
[76] Foegh M, Prolongation of catdiac allograft survival with BN52021. Transplantation, 1986,42(1):86.
[77] Fowler M, In:The Proceeding of the 3ND Asia-Pasific Conference on Biotechnoloqy. Singapore. 1994.
[78] Fowler M, et al.,Plant tissue culture .1982,5:225.
[79] Fowler M. In:Plant Biotechnology .Editid by S. Melltelland H. Smith. Cambrige Univ. Press. 1983.
[80] Fumimaro T ,et al.,EP Patent 0166436.
[81] Gebner B, Voelp A. Study of the long term action of a ginkgo biloba extract on vigilance and mental performance as determined by means of quantitative pharmaco EEG and psychometric measurements. Arznei-Forsch/Drug Res. 1985,35(9):1459.
[82] Gellerman Joanne L, et al. Phytochemistry, 1976,15:1959. 17(4): 157～160.
[83] Guinot P. Effects of the PAF antagonists, BN52063, BN52021 in various clinical indications. J Lipid Mediators Cell Signalling, 1994, 10(1～2):141.
[84] Haslar A, Sticher O, et al. High-performance liquid chromatographic determination of five widespread aglycones. J Chromatogr. 1990,508:236.
[85] Haslar A, Sticher O, et al. Identification and determination of the falconoid From Ginkgo biloba by HPLC. J Chromatogr. 1992,605:41.
[86] Holden M A ,et al.,Manipulating secondary metabolism in culture. Robins J and Rhodes M, ed . Cambridge Univ. Press, Cambridge, 1988, 57-65.
[87] Huh Hoon, et al. Ontogeneic aspects of ginkgolide production in Ginkgo biloba Plant Med. 1993,59:232～239.
[88] Ibata Koichi, et al, Biochem J ,1983,213:305.
[89] Jacob L. Review Thermal and Non-thermal Interaction of Microwave Radiation with Materials. Journal of Materials Science, 1995, (30):5321～5327
[90] Jeon M. et al. Culture of Ginkgo biloba effect of nutrition and hormonal factors on thegroeth ofcultured cells derived from Ginkgo biloba. Archives of Pharmacal Research (Seoul), 1993, 16(3) :244～250.
[91] Kaul K and Sabharwal P S, Plant Physiol. 1971, 47, 691～695.
[92] Kimersley A , et al. W. Alton Jones Cell Science Centre Annual Report. PP lake placide. W. Alton Jones Cell Scince Centre , 1981.
[93] Kircher Henvy W, et al., Phytochemistry, 1970, 9:1879.
[94] Koltai M, Hosford D, Guinot P, et al. PAF: A review of its effects, antagonists and possible future clinical implications (part Ⅱ). Ear J Pharmacol, 1989, 164(2) :293.
[95] Kose K, Dogan P, J Int Med Res, 1995,23(1): 9.
[96] Krieglstein J Nanoprotective effective of Ginkgo Liloda Constituents, Eur J Pharmacol Sci, 1995, 3(1)391.
[97] Larsen R, et al, Therapie, 1978, 33(5): 651～655.
[98] Laurain D, et al., Phytochemistry, 1997, 46(1): 127～132.
[99] Laurain D, et al., Plant cell, Tissueand organ culture. 1996, 4:19.
[100] Levinel, Mol Pharmacol, 1988, 34(6) 793.
[101] Li J, T. (李继桐), The development of the enbryo of Ginkgo biloba, Net. Tsing Hua Univ, Sci. Rep. 1934, 132:29～35.
[102] Li J, T. and Shen, T(沈同), The effect of "Pantothenic acid" on the growth of the radical of Ginkgo embryos in artificial media, Net. Tsing Hua Univ, Sci. Rep. 1934, 132: 53～60.
[103] Lobstein A, et al. Seasonal Variations of the flavoniod Content from Ginkgo biloba. Leaves. Planta Med, 1991, 57:430～433..
[104] Lohn M , Lin F, Tseng C, et al, J Formos Med Assoc, 1994;93(7) :592.
[105] Makino, et al. Jpn Kokai Tokkyo Koho, 1991, JP 03, 275:629.
[106] Maruyama M, et al. The ginkgolides isolation and characterization of the various
grpups . Tetrahedron Letters, 1967, 4:299 .
[107]Mavituna F. Recent Advances in Biotecnology, Kluwger Academinc Publishers, Neitherland. 1992.
[108]Meruyama M, et al, Am Heart J, 1990, 120 (3) :510.
[109]Morimoto T, Agricultural and Biological Chemistry. 1988, 52(7)1835.
[110]Muller K, Borger H. Arbeiten aus der Biologischen Bundesanstalt fur Land-und For stwirtschaft (Berlin) . 1940, 23:189.
[111]Nakanish K. Structure of bilobelide, a rare tert-butyl containing sesquiterpenoid Related to the C20-ginkgolides. J Am Chem Soc, 1971, 93(14): 3544
[112]Ogihari Y and Tsunewakik. Bot Mag Tokyo. 1979, 92:163～171.
[113]Pal Nirupam, Korfiatis George P, Patel Vijay. Sonochemical extraction and biological treatmet of pentachlorophenol contaminated wood, Jounal of Hazardous Materials. 1997, 53: 165.
[114]Pamplim E J and Champan J M.J Exp Bot. 1975, 26:212～220.
[115]Pennkck J F , et al. Nature (London), 1960, 186:470.
[116]Pietta P, et al. Analysis of terpenes from Ginkgo biloba L. Extracts by Reversed hase High-Performance Liquid Chromatogrophy. Chiomatographia, 1990, 29(5)251～253.
[117]Puhan Z S, Martin M, Progress in Industrial Microbiology. 1971,9:13.
[118]Punkt K, Schaffranietl, et, al. Acta Histchem, 1995:97(1):67.
[119]Ranansky J, et al. Biologia Bratislava, 1996, 51(4) :471～477.
[120]Rong Y. et al. Free Radie Biomed. 1996, 20(1): 121.
[121]S. Furukawa, Scientific Papers of The Institute of Physical and Chemical Research , 1932, 19: 27-31.
[122]Salisova, Toma S, Mason T J. Comparison of conventional and ultrasonically assisted Extractions of pharmaceutically active compounds from Salvia officinalis . Ultrasonics Sonochemistry. 1997, 4:131
[123]Schemidt H, et al. Tetrahedron Letters. 1971, 44:4151. 353～356.
[124]Schennen A, et al. Plant Medical . 1986, 235,
[125] Seibert M, Kadade P. Enviromental factor :a light in:staba E jed. Plant tissue culture as a source of biochemicals[M]. Florida: CRC Press, 1980.
[126] Shen J G, Biochen Mol Bilint, 1995:35(1):125.
[127] Siron Al, et al, Am J Bhysiol, 1989, 257(1, Pt2): H25.
[128] Stetler D and Lastische V M, Science. 1965, 149:1387～1388.
[129] Sticher O. Quality of ginkgo preparation Plant Med. 1993, 59(1): 2～11.
[130] Sunderland N, Ann. Bot. 1966, 30:253～268.
[131] Szoeke E, et al., Fiziol Rast. 1977, 24:832～840.
[132] Tabata M and Fujita Y, In "Biotecnology in plant Science", New York: Academic Press, Inc., 1985, p207.
[133] Takeda J. light-induced synthesis of anthocyanin in carrot cells in suspension J. EXP. Bot. 1988, 39(205):1065～1077.
[134] Tisobe T C, et al. Plant Physiology. 1973, 51(4): 805～816.
[135] Tosaki A, et al., Coron Artery Dis, 1994,5(5)443
[136] Tosaki A, Pali T, etal, Diabetologia, 1996; 39(11): 1225.
[137] Touvay C, Etienne A, Braquetp, Inhibition of antigen-induced lung anaphy laxis in the quinea pig by BN52021 a new specific PAF isolated from Ginkgo biloha Agents Actions, 1985, 17(3):371.
[138] Tulecke W. A haploid tissue cultre from the female gametophyte of Ginkgo biloha L . Nature. 1964, 204(4): 94～95.
[139] Tulecke W. R., A tissue derived from the pollen of Ginkgo, Science. 1953, 117:599～600.
[140] Tulecke W. R., Arginine-requiring strains of tissue obttained from Ginkgo biloba, Plant physiol. 1960, 35:19～24.
[141] Tulecke W. R., The pollen of Ginkgo biloba in vitro culture and tissue formation, Amer, J, Bot. 1957, 44:602～608.
[142] Van Beek T A et al. Determination of ginkgolides and bilobalide in ginkgo biloba leaves and phytopharma ceuticals. J. Chromatogr. 1991, 543: 375.
[143] Vasil I and Hildebrandt A . Planta. 1966, 68:69～82.
[144]Villamediana L. Effects of the PAF antagcnist BN52021 on the hemodyrnamics of rats with experimentovl cirrhosis of the liver Life Sci, 1986, 39(3):201.
[145]Villbauer E B, etal, J org Chom, 1987, 52(7): 1186.
[146]Vinatoru, Maricela Toma, Otilia Radu, et al. The use of ultrasound for the extraction of bioactive principles from plant materials." . Ultrasonics Sonochemistry. 1997,4:135.
[147]Wanger H, et al. Dtsch Apoth Ztg. 1989, 129:2421.
[148]Weinges K et al. Liebigs Ann Chem., 1969, 724:261
[149]Weinges K, et al, Arzneimitted - Forschung, 1968, 18(5): 537～544.
[150]Yamakawa T et al. Bio. Chem. 1983, 47:2185～2191.
[151]Yeoman M, et al. Diffirentiation in vitro 4th Symposium British Society Cell Biology. Cambrige Uni. Press. Cambrige. 1981, 65～81.
[152]Zenk M H, et al. Plant tissue culture and Its Biotecknological Application, Edited by Zenk, M. H, Springer-Verlag, Berlin, Heideberry. New York. 1977.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |