中药丁公藤、金银花活性成分的荧光分析法研究
|
摘要
研究了中药丁公藤(Caulis Erycibes)、金银花(Flos Lonicerae Japonicae)活性成分的荧光光谱,考察了实验条件对荧光光谱的影响,确立了活性成分荧光最强的环境条件,并以此为基础,建立了测定中药丁公藤中东莨菪内酯含量的溶液荧光法和薄层荧光扫描法,以及测定中药金银花中绿原酸含量的溶液荧光法。
论文工作主要分为五部分:
1、研究了中药丁公藤的活性成分东莨菪内酯和东莨菪甙的荧光光谱性质。实验发现溶液酸度对东莨菪内酯的荧光光谱有很大影响,而对东莨菪甙的荧光光谱影响不大。以硫酸奎宁为参比,测得东莨菪内酯的荧光量子产率为0.71,东莨菪甙的荧光量子产率为0.21。采用荧光分析法测定了东莨菪内酯的电离常数pKa = 7.32。
2、研究了中药丁公藤甲醇提取液的荧光光谱性质,利用东莨菪内酯和东莨菪甙在溶液酸度改变时荧光光谱的差异性,建立了测定中药丁公藤中东莨菪内酯含量的溶液荧光法和薄层荧光扫描法。
3、将东莨菪甙转化为东莨菪内酯,建立了测定中药丁公藤中总东莨菪内酯含量的溶液荧光法和薄层荧光扫描法。
4、绿原酸在溶液中的荧光很弱。探索了敏化绿原酸荧光的实验条件,发现硼砂和十六烷基三甲基溴化铵两种试剂共存时能够显著增强绿原酸荧光,确定了硼砂和十六烷基三甲基溴化铵的最佳浓度。研究了三元体系绿原酸-硼砂-十六烷基三甲基溴化铵的光谱性质,并以硫酸奎宁为参比,测得体系的荧光量子产率为0.010。
5、考察了金银花中的共存荧光组分,以硼砂-十六烷基三甲基溴化铵为增敏剂,建立了测定中药金银花中绿原酸含量的敏化荧光分析法。
Fluorescence spectra of active components in Caulis Erycibes (CE, Dinggongteng in Chinese) and Flos Lonicerae Japonicae (FLJ, Jinyinhua in Chinese) were studied in this thesis, and influences of experimental conditions on fluorescence spectra were also investigated to establish the best conditions. Solution fluorescent method and thin-layer fluorescence scanning method were established for the determination of scopoletin. The content of Chlorogenic Acid (CA) in FLJ was determined by solution fluorescent method.
Five parts were included in this thesis:
1. Fluorescence spectra of scopoletin and scopolin in CE were studied. It was found that solution acidity had great effect on the fluorescence spectra of scopoletin, and had no effect on scopolin. The fluorescence quantum yields of scopoletin and scopolin were measured to be 0.71 and 0.21 respectively, using quinine sulfate as a reference. The ionization constant of scopoletin was determined to be pKa = 7.32, by using a fluorimetric method.
2. Fluorescence spectra of the extract of CE were studied. Solution fluorescent method was established on the basis of difference between scopoletin and scopolin in the change of solution acidity. And thin-layer fluorescence scanning method was also established.
3. Fluorimetric method as well as thin-layer fluorescence scanning method was established to determine the total content of scopoletin in CE by transforming scopoletin into scopolin.
4. The fluorescence of CA was weak. So, method for obtaining sensitized fluorescence of CA was explored. It was found that the fluorescence of CA can be sensitized by coexisting with borax and CTAB. Fluorescence spectra of CA-borax -CTAB ternary system was then studied, and the fluorescence quantum yield of the ternary system was measured to be 0.010, using quinine sulfate as a reference.
5. The fluorescence components in FLJ were studied. Fluorimetric method for the determination of total content of CA was established using borax-CTAB as the sensitizing agent.
论文工作主要分为五部分:
1、研究了中药丁公藤的活性成分东莨菪内酯和东莨菪甙的荧光光谱性质。实验发现溶液酸度对东莨菪内酯的荧光光谱有很大影响,而对东莨菪甙的荧光光谱影响不大。以硫酸奎宁为参比,测得东莨菪内酯的荧光量子产率为0.71,东莨菪甙的荧光量子产率为0.21。采用荧光分析法测定了东莨菪内酯的电离常数pKa = 7.32。
2、研究了中药丁公藤甲醇提取液的荧光光谱性质,利用东莨菪内酯和东莨菪甙在溶液酸度改变时荧光光谱的差异性,建立了测定中药丁公藤中东莨菪内酯含量的溶液荧光法和薄层荧光扫描法。
3、将东莨菪甙转化为东莨菪内酯,建立了测定中药丁公藤中总东莨菪内酯含量的溶液荧光法和薄层荧光扫描法。
4、绿原酸在溶液中的荧光很弱。探索了敏化绿原酸荧光的实验条件,发现硼砂和十六烷基三甲基溴化铵两种试剂共存时能够显著增强绿原酸荧光,确定了硼砂和十六烷基三甲基溴化铵的最佳浓度。研究了三元体系绿原酸-硼砂-十六烷基三甲基溴化铵的光谱性质,并以硫酸奎宁为参比,测得体系的荧光量子产率为0.010。
5、考察了金银花中的共存荧光组分,以硼砂-十六烷基三甲基溴化铵为增敏剂,建立了测定中药金银花中绿原酸含量的敏化荧光分析法。
Fluorescence spectra of active components in Caulis Erycibes (CE, Dinggongteng in Chinese) and Flos Lonicerae Japonicae (FLJ, Jinyinhua in Chinese) were studied in this thesis, and influences of experimental conditions on fluorescence spectra were also investigated to establish the best conditions. Solution fluorescent method and thin-layer fluorescence scanning method were established for the determination of scopoletin. The content of Chlorogenic Acid (CA) in FLJ was determined by solution fluorescent method.
Five parts were included in this thesis:
1. Fluorescence spectra of scopoletin and scopolin in CE were studied. It was found that solution acidity had great effect on the fluorescence spectra of scopoletin, and had no effect on scopolin. The fluorescence quantum yields of scopoletin and scopolin were measured to be 0.71 and 0.21 respectively, using quinine sulfate as a reference. The ionization constant of scopoletin was determined to be pKa = 7.32, by using a fluorimetric method.
2. Fluorescence spectra of the extract of CE were studied. Solution fluorescent method was established on the basis of difference between scopoletin and scopolin in the change of solution acidity. And thin-layer fluorescence scanning method was also established.
3. Fluorimetric method as well as thin-layer fluorescence scanning method was established to determine the total content of scopoletin in CE by transforming scopoletin into scopolin.
4. The fluorescence of CA was weak. So, method for obtaining sensitized fluorescence of CA was explored. It was found that the fluorescence of CA can be sensitized by coexisting with borax and CTAB. Fluorescence spectra of CA-borax -CTAB ternary system was then studied, and the fluorescence quantum yield of the ternary system was measured to be 0.010, using quinine sulfate as a reference.
5. The fluorescence components in FLJ were studied. Fluorimetric method for the determination of total content of CA was established using borax-CTAB as the sensitizing agent.
引文
[1]国家药典委员会.中华人民共和国药典(2005年版一部)[M],北京:化学工业出版社,2005, 3.
[2]吴立宏,朱恩圆,张紫佳,王峥涛.广西产丁公藤原植物的调查及商品丁公藤主流品种的鉴定[J].中草药, 2005, 36(9):1398-1400.
[3]宋蔚,金蓉鸾,刘继华.光叶丁公藤化学成分的研究[J].中国中药杂志, 1997, 22(6):359-360.
[4]陆阳,姚天荣,陈泽乃.凹脉丁公藤化学成分的研究[J].药学学报, 1986, 21(11):829-835.
[5]王朴,姚天荣,陈泽乃.毛叶丁公藤化学成分的研究[J].植物学报, 1989, 31(8):616-619.
[6]郭福丽.丁公藤注射液治疗肾绞痛[J].四川中医1986, 4:32.
[7]周名璐,许少伟,程新敏.复方丁公藤胶囊的消炎镇痛作用[J].中成药, 1993, 15(7):29-30.
[8]杨柯,林启云,赵一,曾春晖.东莨菪素对家兔体温的影响[J].广西中医药, 2005, 28(1):62.
[9] María Gabriela Manuele, Graciela Ferraro, Maria Laura Barreiro Arcos, Paula López, Graciela Cremaschi, Claudia Anesini. Comparative immunomodulatory effect of scopoletin on tumoral and normal lymphocytes[J]. Life Sciences, 2006, 79: 2043-2048.
[10] Teresa Valle, JoséL. López, JoséM, Hernández, Purificación Corchete. Antifungal activity of scopoletin and its differential accumulation in Ulmus pumila and ulmus campestris cell suspension cultures infected with Ophiostoma ulmi spores[J]. Plant Science, 1997, 125: 97-101.
[11]刘明珠,李玲.用乙醇回流法从丁公藤中提取东莨菪素[J].第二军医大学学报, 1995, 16(5):483.
[12]秦泽平,黄振宇,汤浚溪.华山参两种提取法与酸度对东莨菪内酯含量的影响[J].基层中药杂志, 1995, 9(4):26-27.
[13] Tzu-Ching Tzeng, Young-Lung Lin, Ting-Ting Jong, Chieh-MingJ.Chang. Ethanol modified supercritical fluids extraction of scopoletin and artemisinin from Artemisia annua L[J]. Separation and Purification Technology, 2007, 56: 18-24.
[14] Wang Ketai, LiuHuitao, Chen Xingguo, Zhao Yunkun, Hu Zhide. Identification and determination of active components in Angelica dahurica Benth and its medicinal preparation by capillary electrophoresis[J]. Talanta, 2001, 54: 753-761.
[15]杨裕忠,罗干明. HPLC法测定丁公藤中东莨菪内酯的含量[J].中药新药与临床药理, 2004, 15(4):272-274.
[16]宋蔚,刘锁兰,李秀青,杨隽,王韵天. HPLC测定4种丁公藤类生药中东莨菪素及东莨菪苷的含量[J].中国中药杂志, 2004, 29(2):185-186.
[17]张志兰,项苏留,张俭,褚清俊.丁公藤注射液中东莨菪内酯含量的HPLC法测定[J].分析测试学报, 2002, 21(5):92-93 .
[18]曾洁英.东莨菪素的导数光谱测定[J].广州中医学院学报, 1989, 6(2):97-99.
[19]陆敏仪,黄西峰,马祥欢.薄层光密度法测定丁公藤注射液东莨菪素含量及其质量分析[J].中国中药杂志, 1993, 18(3):158-160.
[20]王勇,庆伟霞,李松武,李明静,刘秀华.薄层扫描法测定华山参中总东莨菪内酯的含量[J].中南药学, 2005, 3(5):274-276.
[21]李玲,刘明珠,徐卫明.薄层刮板-荧光分析法在中药复方制剂质量控制中的应用[J].中草药, 1996, 27(5):275-277.
[22]王宏军,吴国娟,孙建,蒋红.绿原酸的药效学研究[J].中兽医学杂志, 2005, 5:7-11.
[23]胡宗福,于文利,赵亚平.绿原酸清除活性氧和抗脂质过氧化的研究[J].食品科学, 2006, 27(2):128-130.
[24] Suresh K. Abraham, Lakshmi Sarma, P.C. Kesavan. Protective effects of chlorogenic acid, curcurmin andβ-carotene againstγ-radiation-induced in vivo chromosomal damage[J]. Mutation Research, 1993, 303: 109-112.
[25]盛卸晃,刘文谦,薛霞,张海艇,刘兆平.绿原酸体外抗单纯疱疹病毒作用[J].中国天然药物, 2008, 6(3): 232-234.
[26] L.C. Chiang, W. Chiang, M.Y. Chang, L.T. Ng, C.C. Lin. Antiviral activity of Plantago major extracts and related compounds in vitro[J]. Antiviral Research, 2002, 55: 53-62.
[27]胡克杰,孙考祥,薛霞,王璟璐,谢丽萍.绿原酸体外抗病毒作用研究[J].哈尔滨医科大学学报, 2001, 35(6):430-432.
[28]常翠青,陈吉棣,王香生.有机酸对人血管内皮细胞的保护作用[J].中华预防医学杂志, 2001, 35(2):79-82.
[29] Delcy V. Rodriguez de Sotillo, M. Hadley. Chlorogenic acid modifies plasma and liver concentrations of :cholesterol, triacylglycerol, and minerals in (fa/fa) Zucker rats[J]. Journal of Nutritional Biochemistry, 2002, 13:717-726.
[30]顾利红,朱品业.日光和温度对绿原酸供试液稳定性的影响[J].中成药, 1999, 21(11):568-569.
[31]王云峰,陆兔林,毛春芹,徐佳佳.通关藤浸膏中绿原酸的稳定性考察[J].中国药房, 2008, 19(12):916-918.
[32]林丽洋,贺英菊,罗巍伟,莫毅,陈艳,闫根全.增强绿原酸水溶液稳定性的方法研究[J].华西药学杂志, 2005, 20(3):225-227.
[33]陈芳平,李玉贤,孙德梅.β–环糊精对绿原酸稳定作用的研究[J].河南科学, 1999,17(2):166-167.
[34]聂凌鸿,岳淼,孙德梅.金银花中绿原酸提取工艺的优化[J].食品研究与开发, 2008, 29(1):51-55.
[35]周军,黄琼,李志光,谢磊,黄可龙.超声波法提取金银花中的绿原酸[J].化学与生物工程, 2008, 25(1):31-33.
[36]郭晓哥,赵俊廷.利用反胶束溶液萃取葵花籽中绿原酸的研究[J].中国油脂, 2008, 33(3):56-59.
[37]陈永胜,李志光,董建国.酶解法提取葵花粕绿原酸工艺研究[J].食品科学, 2008, 29(2):202-204.
[38]武雪芬,彭仁武,张村,张淑玲.β–环糊精在绿原酸分离中的应用研究[J].河南科学, 1999, 17(4):385-387.
[39]范全民,孙冬云,周慧娟. HPLC测定维C银翘片中绿原酸的含量[J].现代中药研究与实践, 2005, 19(6):385-387.
[40]黄平权,梁峰,黄燕萍.维C银翘颗粒中绿原酸的HPLC测定[J].中国医药工业杂志, 2004, 35(8):494-495.
[41]杨滨,屈巧玲,王谦鹏,易崇勤,肖永庆,王永炎.急支糖浆中原儿茶酸4种成分的含量测定[J].中国中药杂志, 2007., 32(9):858-860.
[42]金永新,要林青,杨晓冬,杨春兰. RP-HPLC法同时测定复方鱼腥草片中绿原酸、连翘苷、黄芩苷的含量[J].中国药师, 2004, 7(7):524-526.
[43]刘磊,何再安,于子川,刘超英,刘焱文.高效液相色谱法测定金银花提取物中绿原酸的含量[J].时珍国医国药, 2008, 19(10):524-526.
[44]唐启令,刘永俊,崔冬梅.薄层扫描法测定抗感颗粒中绿原酸的含量[J].中国医院药学杂, 2004, 24(2):2416-2417.
[45]张秀琨,宋喆，傅广振.薄层色谱-紫外分光光度法测定抗毒退热冲剂中绿原酸含量[J].中国医院药学杂志, 1996, 27(3):181-182.
[46]戈早川,余倩.胶束薄层色谱扫描法测定银黄片与银黄注射液中的黄芩苷与绿原酸[J].药物分析杂志, 1999, 19(5):348-350.
[47]彭友元,楚清脆,叶建农.毛细管电泳-电化学检测Vc银翘片中的有效成分[J].华东师范大学学报(自然科学版), 2003, (3):98-101.
[48]傅亮,楚清脆,黄宝康,郑汉臣,叶建农.毛细管电泳-电化学检测法测定蜘蛛香中多元酚类化合物[J].分析化学研究报告, 2005, 33(2):161-164.
[49]张兰,刘远环,何聿,,陈国南.毛细管电泳-电化学检测法用于中药石韦中绿原酸和槲皮素的同时测定[J].分析测试技术与仪器, 2005, 11(1):22-27.
[50]田秀慧,楚清脆,吴婷,江莲梅,叶建农.毛细管电泳-电化学检测法测定复方罗布麻片中的活性成分[J].化学世界, 2008, 144-147.
[51]袁华,孙炎彬,池汝安,吴元欣.络合显色法测定杜仲叶中的绿原酸[J].时珍国医国药, 2008, 19(9):2206-2208.
[52]周波林.二阶导数紫外光谱法测定双黄连口服液中总绿原酸的含量[J].安徽医药, 2007, 11(4):324-325.
[53]罗琼,金岚峰,张媛.近红外光谱法快速测定烟草中的绿原酸中的绿绿原酸、新绿原酸和芸香苷[J].烟草化学, 2008, (5):30-33.
[54]王柏强.差示分光光度法测定杜仲叶提取液中绿原酸的含量[J].川北医学院学报, 2008, 23(5):461-463.
[55]张广强,梁生旺,姜玉风.三波长分光光度法测定清热解毒注射剂中绿原酸的含量[J].中国中药杂志, 2008, 15(8):33-35.
[56]国家药典委员会.中华人民共和国药典(2005年版一部)[M],北京:化学工业出版社,2005,173.
[57]赵国玲,刘佳佳.金银花化学成分及药理研究进展[J].中药材, 2002, 25(10):762-763.
[58]毕岳峰,田野,裴姗姗,张聪,刘宏民.金银花化学成分分析[J].郑州大学学报(理学版), 2007, 39(2):184-186.
[59]黄喜茹,刘伟娜,曹冬.金银花的化学成分药理作用研究评析[J].中医药学刊, 2005, 23(3):418-419.
[60]吴树国,翟德翠.药用金银花的化学成分、药理作用研究及其进展[J].科技创新导报, 2008, 2.
[61]葛冰,卢向阳,易克,田云.金银花活性成分药理作用及其应用[J].中国野生植物资源, 2004, 23(5):13-16.
[62]王志洁,黄铁牛.金银花在体内外抗人Ⅰ型疱疹病毒的实验研究[J].中国中医基础医学杂志, 2003, 9(7):39-43.
[63]李佺，崔瑛.金银花药理作用比较[J].中药材, 1999, 22(1):37-39.
[64]刘慧纯,刘广南,陈历雄,丘卓翘等.金银花能降低小鼠血糖血脂水平[J].广州医药, 1998, 29(3):59-62.
[65]李斐,黎海芪.金银花水提物对卵清蛋白致敏小鼠的抗过敏作用研究[J].重庆医科大学学报, 2004, 29(3):288-291.
[66]关炳峰,谭军,周志娣.金银花提取物的抗氧化作用与其绿原酸含量的相关性研究[J].食品工业科技, 2007, 28(10):127-129.
[67]陈慧浓.金银花胶囊的功效观察及药理分析[J].浙江中医药大学学报, 2008, 32(2):249-250.
[68]段晓颖.金银花水提工艺中绿原酸变化的研究[J].中国野生植物资源中草药, 2007, 38(8):1189-1190.
[69] Ma Xihan, Wei Qin, Jing Qianping, Wang Yonghong. A Comparision on the Extraction of Chlorogenic Acid with Different Concentration of Ethanol from Honeysuckle Flower [J]. Journal of Northwest Forestry University, 2000, 15(4): 77-81.
[70]屈景年,莫运春,刘梦琴,李荣.金银花中绿原酸一步提取法及绿原酸抗菌活性[J].化学世界, 2005, 3:167-169.
[71]林云良,孙庆雷,祝贺,王晓,李福伟,耿岩玲,王晓东.超声波法提取金银花中绿原酸工艺参数优化研究[J].山东科学, 2008, 21(1):25-27.
[72]刘志平,韩伟,李想.金银花中绿原酸的微波辅助提取[J].华东理工大学学报(自然科学版), 2008, 34(4):543-546.
[73]王丽婷,赵堂,杨敏丽.金银花中黄酮类物质的提取工艺[J].华西药学杂志, 2006, 21(6):520-523.
[74]许永,向能军,桂永发,赵娟.金银花总黄酮的萃取及在卷烟中的应用研究[J].广东化工, 2008 , 35(186):109-111.
[75]刘磊,何再安,于子川,刘超英,刘焱文.高效液相色谱法测定金银花提取物中绿原酸的含量[J].时珍国医国药, 2008, 19(10):2416-2417.
[76]刘燕,易丽,梅艳.反相高效液相色谱法测定金银花及抗感颗粒中绿原酸的含量[J].中国现代应用药学杂志, 2006, 23(2):160-162.
[77]金蓉鸾,叶玉兰,单文碘.双波长薄层层析扫描法测定金银花中异绿原酸和绿原酸的含量[J].南京药学院学报, 1980, 1:15-18.
[78]颜流水,温振东,黄智敏,万益群,井晶.毛细管电泳法测定金银花中绿原酸、芦丁和槲皮素[J].药物分析杂志, 2007, 27(3):367-370.
[79]何报作.紫外光度法测定金银花中绿原酸含量的改进[J].广西中医药, 1992, 15(5):45-47.
[80]耿玉珍,刘葵,周杰.阻抑-催化褪色光度法测定痕量绿原酸[J].分析化学研究简报, 2002, 30(6):684-686.
[81]林黎明.差示导数光谱法测定金银花及银翘片中总绿原酸的含量[J].中国中药杂志, 1991, 16(5):282-284.
[82]李自林,郑莜梅,王琰.绿原酸的导数极谱法测定[J].化学研究与应用, 1998, 10(2):199-202.
[83]门孝英,张守雪,梁云爱.金银花中绿原酸的二阶导数差示脉冲极谱法定量研究[J].化学分析计量, 2001, 10(1):11-14.
[84] HE Caixia, CUI Hua, ZHAO Xiaoyu, ZHAO Huazhang, ZHAO Guiwen. Determination of Chlorogenic Acid by Flow Injection with Chemiluminescence Detection [J]. JOURNAL OF CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY, 1999, 29(5): 521-525.
[85]许金钩,王尊本.荧光分析法(第三版)[M],北京:科学出版社,2006.
[86]潘祖亭,关洪亮,原华平,李微,陈果.荧光光谱法在药物分析中的应用[J].吉首大学学报, 2005, 26(3):27-34.
[87]赵文武,中药丁香、天麻活性成分的光谱性质及荧光分析法研究[D],河北师范大学硕士学位论文, 2009.
[88]杨菲,硼、铝敏化荧光法测定中药成分丹皮酚、原儿茶酸和没食子酸[D],河北师范大学硕士学位论文, 2008.
[89]郑彦慧,黄连活性成分的光谱性质及中药中小檗碱的环糊精敏化荧光法测定[D],河北师范大学硕士学位论文, 2008.
[90]李恩来,潘秀芳.荧光分析法在中药鉴定中的应用[J].黑龙江中医药, 2002, (5):57-58.
[91] Pawe? Górnas, Gra?yna Neunert, Krzysztof Baczyński, Krzysztof Polewski. Beta-cyclodextrin complexes with Chlorogenic acid and caffeic acid from coffee brew:Spectroscopic,thermodynamic and molecular modelling study[J]. Food Chemistry, 2009, 114: 190-196.
[92]袁华,孙炎彬,池汝安,吴元欣.络和显色法测定杜仲叶中的绿原酸[J].时珍国医国药, 2008, 19(9):2206-2208.
[93]屈景年,刘义,莫运春,刘梦琴.绿原酸锌(Ⅱ)配合物的合成和抑菌活性研究[J].化学世界, 2006, 5:267-268.
[94]屈景年,李荣,莫运春,刘梦琴,曾荣英.绿原酸与La(Ⅱ)配合物的合成及抗菌活性研究[J].衡阳师范学院学报, 2004, 25 (6):49-50.
[95]查建蓬,林颖,杨星惠,侯海妮,魏铁军,陈兴丽.十六烷基三甲基溴化铵对脱水四环素和异四环素荧光增强作用研究[J].光谱学与光谱分析, 2002, 22 (3):430-432.
[96]田穗康.十六烷基三甲基溴化铵增敏光度法测定铬(Ⅵ)离子[J].延安大学学报(自然科学版), 2006, 25 (2):50-52.
[97]杨怀成.十六烷基三甲基溴化铵为增敏剂光度法测定抗坏血酸[J].福建分析测试, 2007, 16(3):54-57.
[98]杜娟.十六烷基三甲基溴化铵增敏二甲酚橙光度法测定化妆品中铅[J].日用化学工业, 2005, 35(1):55-57.
[99]许金钩,王尊本.荧光分析法(第三版)[M],北京:科学出版社,2006,59-60.
[100]许金钩,王尊本.荧光分析法(第三版)[M],北京:科学出版社,2006,28.
[101]金蓉鸾,叶玉兰,单文碘.双波长薄层层析扫描法测定金银花中异绿原酸和绿原酸的含量[J].南京药学院学报, 1980, 1:15-18.
[102]何报作.紫外光度法测定金银花中绿原酸含量的改进[J].实验研究, 1992, 15(5):45-47.
[103]钟方晓.高效液相及紫外分光光度法测定金银花中绿原酸和异绿原酸含量方法学比较[J].时珍国医国药, 2005, 16(3):212.
[2]吴立宏,朱恩圆,张紫佳,王峥涛.广西产丁公藤原植物的调查及商品丁公藤主流品种的鉴定[J].中草药, 2005, 36(9):1398-1400.
[3]宋蔚,金蓉鸾,刘继华.光叶丁公藤化学成分的研究[J].中国中药杂志, 1997, 22(6):359-360.
[4]陆阳,姚天荣,陈泽乃.凹脉丁公藤化学成分的研究[J].药学学报, 1986, 21(11):829-835.
[5]王朴,姚天荣,陈泽乃.毛叶丁公藤化学成分的研究[J].植物学报, 1989, 31(8):616-619.
[6]郭福丽.丁公藤注射液治疗肾绞痛[J].四川中医1986, 4:32.
[7]周名璐,许少伟,程新敏.复方丁公藤胶囊的消炎镇痛作用[J].中成药, 1993, 15(7):29-30.
[8]杨柯,林启云,赵一,曾春晖.东莨菪素对家兔体温的影响[J].广西中医药, 2005, 28(1):62.
[9] María Gabriela Manuele, Graciela Ferraro, Maria Laura Barreiro Arcos, Paula López, Graciela Cremaschi, Claudia Anesini. Comparative immunomodulatory effect of scopoletin on tumoral and normal lymphocytes[J]. Life Sciences, 2006, 79: 2043-2048.
[10] Teresa Valle, JoséL. López, JoséM, Hernández, Purificación Corchete. Antifungal activity of scopoletin and its differential accumulation in Ulmus pumila and ulmus campestris cell suspension cultures infected with Ophiostoma ulmi spores[J]. Plant Science, 1997, 125: 97-101.
[11]刘明珠,李玲.用乙醇回流法从丁公藤中提取东莨菪素[J].第二军医大学学报, 1995, 16(5):483.
[12]秦泽平,黄振宇,汤浚溪.华山参两种提取法与酸度对东莨菪内酯含量的影响[J].基层中药杂志, 1995, 9(4):26-27.
[13] Tzu-Ching Tzeng, Young-Lung Lin, Ting-Ting Jong, Chieh-MingJ.Chang. Ethanol modified supercritical fluids extraction of scopoletin and artemisinin from Artemisia annua L[J]. Separation and Purification Technology, 2007, 56: 18-24.
[14] Wang Ketai, LiuHuitao, Chen Xingguo, Zhao Yunkun, Hu Zhide. Identification and determination of active components in Angelica dahurica Benth and its medicinal preparation by capillary electrophoresis[J]. Talanta, 2001, 54: 753-761.
[15]杨裕忠,罗干明. HPLC法测定丁公藤中东莨菪内酯的含量[J].中药新药与临床药理, 2004, 15(4):272-274.
[16]宋蔚,刘锁兰,李秀青,杨隽,王韵天. HPLC测定4种丁公藤类生药中东莨菪素及东莨菪苷的含量[J].中国中药杂志, 2004, 29(2):185-186.
[17]张志兰,项苏留,张俭,褚清俊.丁公藤注射液中东莨菪内酯含量的HPLC法测定[J].分析测试学报, 2002, 21(5):92-93 .
[18]曾洁英.东莨菪素的导数光谱测定[J].广州中医学院学报, 1989, 6(2):97-99.
[19]陆敏仪,黄西峰,马祥欢.薄层光密度法测定丁公藤注射液东莨菪素含量及其质量分析[J].中国中药杂志, 1993, 18(3):158-160.
[20]王勇,庆伟霞,李松武,李明静,刘秀华.薄层扫描法测定华山参中总东莨菪内酯的含量[J].中南药学, 2005, 3(5):274-276.
[21]李玲,刘明珠,徐卫明.薄层刮板-荧光分析法在中药复方制剂质量控制中的应用[J].中草药, 1996, 27(5):275-277.
[22]王宏军,吴国娟,孙建,蒋红.绿原酸的药效学研究[J].中兽医学杂志, 2005, 5:7-11.
[23]胡宗福,于文利,赵亚平.绿原酸清除活性氧和抗脂质过氧化的研究[J].食品科学, 2006, 27(2):128-130.
[24] Suresh K. Abraham, Lakshmi Sarma, P.C. Kesavan. Protective effects of chlorogenic acid, curcurmin andβ-carotene againstγ-radiation-induced in vivo chromosomal damage[J]. Mutation Research, 1993, 303: 109-112.
[25]盛卸晃,刘文谦,薛霞,张海艇,刘兆平.绿原酸体外抗单纯疱疹病毒作用[J].中国天然药物, 2008, 6(3): 232-234.
[26] L.C. Chiang, W. Chiang, M.Y. Chang, L.T. Ng, C.C. Lin. Antiviral activity of Plantago major extracts and related compounds in vitro[J]. Antiviral Research, 2002, 55: 53-62.
[27]胡克杰,孙考祥,薛霞,王璟璐,谢丽萍.绿原酸体外抗病毒作用研究[J].哈尔滨医科大学学报, 2001, 35(6):430-432.
[28]常翠青,陈吉棣,王香生.有机酸对人血管内皮细胞的保护作用[J].中华预防医学杂志, 2001, 35(2):79-82.
[29] Delcy V. Rodriguez de Sotillo, M. Hadley. Chlorogenic acid modifies plasma and liver concentrations of :cholesterol, triacylglycerol, and minerals in (fa/fa) Zucker rats[J]. Journal of Nutritional Biochemistry, 2002, 13:717-726.
[30]顾利红,朱品业.日光和温度对绿原酸供试液稳定性的影响[J].中成药, 1999, 21(11):568-569.
[31]王云峰,陆兔林,毛春芹,徐佳佳.通关藤浸膏中绿原酸的稳定性考察[J].中国药房, 2008, 19(12):916-918.
[32]林丽洋,贺英菊,罗巍伟,莫毅,陈艳,闫根全.增强绿原酸水溶液稳定性的方法研究[J].华西药学杂志, 2005, 20(3):225-227.
[33]陈芳平,李玉贤,孙德梅.β–环糊精对绿原酸稳定作用的研究[J].河南科学, 1999,17(2):166-167.
[34]聂凌鸿,岳淼,孙德梅.金银花中绿原酸提取工艺的优化[J].食品研究与开发, 2008, 29(1):51-55.
[35]周军,黄琼,李志光,谢磊,黄可龙.超声波法提取金银花中的绿原酸[J].化学与生物工程, 2008, 25(1):31-33.
[36]郭晓哥,赵俊廷.利用反胶束溶液萃取葵花籽中绿原酸的研究[J].中国油脂, 2008, 33(3):56-59.
[37]陈永胜,李志光,董建国.酶解法提取葵花粕绿原酸工艺研究[J].食品科学, 2008, 29(2):202-204.
[38]武雪芬,彭仁武,张村,张淑玲.β–环糊精在绿原酸分离中的应用研究[J].河南科学, 1999, 17(4):385-387.
[39]范全民,孙冬云,周慧娟. HPLC测定维C银翘片中绿原酸的含量[J].现代中药研究与实践, 2005, 19(6):385-387.
[40]黄平权,梁峰,黄燕萍.维C银翘颗粒中绿原酸的HPLC测定[J].中国医药工业杂志, 2004, 35(8):494-495.
[41]杨滨,屈巧玲,王谦鹏,易崇勤,肖永庆,王永炎.急支糖浆中原儿茶酸4种成分的含量测定[J].中国中药杂志, 2007., 32(9):858-860.
[42]金永新,要林青,杨晓冬,杨春兰. RP-HPLC法同时测定复方鱼腥草片中绿原酸、连翘苷、黄芩苷的含量[J].中国药师, 2004, 7(7):524-526.
[43]刘磊,何再安,于子川,刘超英,刘焱文.高效液相色谱法测定金银花提取物中绿原酸的含量[J].时珍国医国药, 2008, 19(10):524-526.
[44]唐启令,刘永俊,崔冬梅.薄层扫描法测定抗感颗粒中绿原酸的含量[J].中国医院药学杂, 2004, 24(2):2416-2417.
[45]张秀琨,宋喆，傅广振.薄层色谱-紫外分光光度法测定抗毒退热冲剂中绿原酸含量[J].中国医院药学杂志, 1996, 27(3):181-182.
[46]戈早川,余倩.胶束薄层色谱扫描法测定银黄片与银黄注射液中的黄芩苷与绿原酸[J].药物分析杂志, 1999, 19(5):348-350.
[47]彭友元,楚清脆,叶建农.毛细管电泳-电化学检测Vc银翘片中的有效成分[J].华东师范大学学报(自然科学版), 2003, (3):98-101.
[48]傅亮,楚清脆,黄宝康,郑汉臣,叶建农.毛细管电泳-电化学检测法测定蜘蛛香中多元酚类化合物[J].分析化学研究报告, 2005, 33(2):161-164.
[49]张兰,刘远环,何聿,,陈国南.毛细管电泳-电化学检测法用于中药石韦中绿原酸和槲皮素的同时测定[J].分析测试技术与仪器, 2005, 11(1):22-27.
[50]田秀慧,楚清脆,吴婷,江莲梅,叶建农.毛细管电泳-电化学检测法测定复方罗布麻片中的活性成分[J].化学世界, 2008, 144-147.
[51]袁华,孙炎彬,池汝安,吴元欣.络合显色法测定杜仲叶中的绿原酸[J].时珍国医国药, 2008, 19(9):2206-2208.
[52]周波林.二阶导数紫外光谱法测定双黄连口服液中总绿原酸的含量[J].安徽医药, 2007, 11(4):324-325.
[53]罗琼,金岚峰,张媛.近红外光谱法快速测定烟草中的绿原酸中的绿绿原酸、新绿原酸和芸香苷[J].烟草化学, 2008, (5):30-33.
[54]王柏强.差示分光光度法测定杜仲叶提取液中绿原酸的含量[J].川北医学院学报, 2008, 23(5):461-463.
[55]张广强,梁生旺,姜玉风.三波长分光光度法测定清热解毒注射剂中绿原酸的含量[J].中国中药杂志, 2008, 15(8):33-35.
[56]国家药典委员会.中华人民共和国药典(2005年版一部)[M],北京:化学工业出版社,2005,173.
[57]赵国玲,刘佳佳.金银花化学成分及药理研究进展[J].中药材, 2002, 25(10):762-763.
[58]毕岳峰,田野,裴姗姗,张聪,刘宏民.金银花化学成分分析[J].郑州大学学报(理学版), 2007, 39(2):184-186.
[59]黄喜茹,刘伟娜,曹冬.金银花的化学成分药理作用研究评析[J].中医药学刊, 2005, 23(3):418-419.
[60]吴树国,翟德翠.药用金银花的化学成分、药理作用研究及其进展[J].科技创新导报, 2008, 2.
[61]葛冰,卢向阳,易克,田云.金银花活性成分药理作用及其应用[J].中国野生植物资源, 2004, 23(5):13-16.
[62]王志洁,黄铁牛.金银花在体内外抗人Ⅰ型疱疹病毒的实验研究[J].中国中医基础医学杂志, 2003, 9(7):39-43.
[63]李佺，崔瑛.金银花药理作用比较[J].中药材, 1999, 22(1):37-39.
[64]刘慧纯,刘广南,陈历雄,丘卓翘等.金银花能降低小鼠血糖血脂水平[J].广州医药, 1998, 29(3):59-62.
[65]李斐,黎海芪.金银花水提物对卵清蛋白致敏小鼠的抗过敏作用研究[J].重庆医科大学学报, 2004, 29(3):288-291.
[66]关炳峰,谭军,周志娣.金银花提取物的抗氧化作用与其绿原酸含量的相关性研究[J].食品工业科技, 2007, 28(10):127-129.
[67]陈慧浓.金银花胶囊的功效观察及药理分析[J].浙江中医药大学学报, 2008, 32(2):249-250.
[68]段晓颖.金银花水提工艺中绿原酸变化的研究[J].中国野生植物资源中草药, 2007, 38(8):1189-1190.
[69] Ma Xihan, Wei Qin, Jing Qianping, Wang Yonghong. A Comparision on the Extraction of Chlorogenic Acid with Different Concentration of Ethanol from Honeysuckle Flower [J]. Journal of Northwest Forestry University, 2000, 15(4): 77-81.
[70]屈景年,莫运春,刘梦琴,李荣.金银花中绿原酸一步提取法及绿原酸抗菌活性[J].化学世界, 2005, 3:167-169.
[71]林云良,孙庆雷,祝贺,王晓,李福伟,耿岩玲,王晓东.超声波法提取金银花中绿原酸工艺参数优化研究[J].山东科学, 2008, 21(1):25-27.
[72]刘志平,韩伟,李想.金银花中绿原酸的微波辅助提取[J].华东理工大学学报(自然科学版), 2008, 34(4):543-546.
[73]王丽婷,赵堂,杨敏丽.金银花中黄酮类物质的提取工艺[J].华西药学杂志, 2006, 21(6):520-523.
[74]许永,向能军,桂永发,赵娟.金银花总黄酮的萃取及在卷烟中的应用研究[J].广东化工, 2008 , 35(186):109-111.
[75]刘磊,何再安,于子川,刘超英,刘焱文.高效液相色谱法测定金银花提取物中绿原酸的含量[J].时珍国医国药, 2008, 19(10):2416-2417.
[76]刘燕,易丽,梅艳.反相高效液相色谱法测定金银花及抗感颗粒中绿原酸的含量[J].中国现代应用药学杂志, 2006, 23(2):160-162.
[77]金蓉鸾,叶玉兰,单文碘.双波长薄层层析扫描法测定金银花中异绿原酸和绿原酸的含量[J].南京药学院学报, 1980, 1:15-18.
[78]颜流水,温振东,黄智敏,万益群,井晶.毛细管电泳法测定金银花中绿原酸、芦丁和槲皮素[J].药物分析杂志, 2007, 27(3):367-370.
[79]何报作.紫外光度法测定金银花中绿原酸含量的改进[J].广西中医药, 1992, 15(5):45-47.
[80]耿玉珍,刘葵,周杰.阻抑-催化褪色光度法测定痕量绿原酸[J].分析化学研究简报, 2002, 30(6):684-686.
[81]林黎明.差示导数光谱法测定金银花及银翘片中总绿原酸的含量[J].中国中药杂志, 1991, 16(5):282-284.
[82]李自林,郑莜梅,王琰.绿原酸的导数极谱法测定[J].化学研究与应用, 1998, 10(2):199-202.
[83]门孝英,张守雪,梁云爱.金银花中绿原酸的二阶导数差示脉冲极谱法定量研究[J].化学分析计量, 2001, 10(1):11-14.
[84] HE Caixia, CUI Hua, ZHAO Xiaoyu, ZHAO Huazhang, ZHAO Guiwen. Determination of Chlorogenic Acid by Flow Injection with Chemiluminescence Detection [J]. JOURNAL OF CHINA UNIVERSITY OF SCIENCE AND TECHNOLOGY, 1999, 29(5): 521-525.
[85]许金钩,王尊本.荧光分析法(第三版)[M],北京:科学出版社,2006.
[86]潘祖亭,关洪亮,原华平,李微,陈果.荧光光谱法在药物分析中的应用[J].吉首大学学报, 2005, 26(3):27-34.
[87]赵文武,中药丁香、天麻活性成分的光谱性质及荧光分析法研究[D],河北师范大学硕士学位论文, 2009.
[88]杨菲,硼、铝敏化荧光法测定中药成分丹皮酚、原儿茶酸和没食子酸[D],河北师范大学硕士学位论文, 2008.
[89]郑彦慧,黄连活性成分的光谱性质及中药中小檗碱的环糊精敏化荧光法测定[D],河北师范大学硕士学位论文, 2008.
[90]李恩来,潘秀芳.荧光分析法在中药鉴定中的应用[J].黑龙江中医药, 2002, (5):57-58.
[91] Pawe? Górnas, Gra?yna Neunert, Krzysztof Baczyński, Krzysztof Polewski. Beta-cyclodextrin complexes with Chlorogenic acid and caffeic acid from coffee brew:Spectroscopic,thermodynamic and molecular modelling study[J]. Food Chemistry, 2009, 114: 190-196.
[92]袁华,孙炎彬,池汝安,吴元欣.络和显色法测定杜仲叶中的绿原酸[J].时珍国医国药, 2008, 19(9):2206-2208.
[93]屈景年,刘义,莫运春,刘梦琴.绿原酸锌(Ⅱ)配合物的合成和抑菌活性研究[J].化学世界, 2006, 5:267-268.
[94]屈景年,李荣,莫运春,刘梦琴,曾荣英.绿原酸与La(Ⅱ)配合物的合成及抗菌活性研究[J].衡阳师范学院学报, 2004, 25 (6):49-50.
[95]查建蓬,林颖,杨星惠,侯海妮,魏铁军,陈兴丽.十六烷基三甲基溴化铵对脱水四环素和异四环素荧光增强作用研究[J].光谱学与光谱分析, 2002, 22 (3):430-432.
[96]田穗康.十六烷基三甲基溴化铵增敏光度法测定铬(Ⅵ)离子[J].延安大学学报(自然科学版), 2006, 25 (2):50-52.
[97]杨怀成.十六烷基三甲基溴化铵为增敏剂光度法测定抗坏血酸[J].福建分析测试, 2007, 16(3):54-57.
[98]杜娟.十六烷基三甲基溴化铵增敏二甲酚橙光度法测定化妆品中铅[J].日用化学工业, 2005, 35(1):55-57.
[99]许金钩,王尊本.荧光分析法(第三版)[M],北京:科学出版社,2006,59-60.
[100]许金钩,王尊本.荧光分析法(第三版)[M],北京:科学出版社,2006,28.
[101]金蓉鸾,叶玉兰,单文碘.双波长薄层层析扫描法测定金银花中异绿原酸和绿原酸的含量[J].南京药学院学报, 1980, 1:15-18.
[102]何报作.紫外光度法测定金银花中绿原酸含量的改进[J].实验研究, 1992, 15(5):45-47.
[103]钟方晓.高效液相及紫外分光光度法测定金银花中绿原酸和异绿原酸含量方法学比较[J].时珍国医国药, 2005, 16(3):212.