川渝产细毡毛忍冬的质量标准及指纹图谱研究
|
摘要
目的：细毡毛忍冬(Lonicera similis Hemsl)为忍冬属植物细毡毛忍冬的干燥花蕾或初开的花(习称长吊子银花)。具有抗菌消炎、清热解毒、凉散风热的功效,主要用于治疗痈肿疔疮、喉痹、丹毒、热毒血痢、风热感冒、温热发病等疾病。其主要有效成分为绿原酸、异绿原酸、总黄酮。细毡毛忍冬为川渝产金银花的主流品种。本课题将对川渝产细毡毛忍冬的生药学特征进行研究,对其进行薄层鉴别,并进行有效成分的含量测定,同时对其进行指纹图谱的研究,来提高和完善川渝产细毡毛忍冬的质量控制方法。方法：为了对川渝产细毡毛忍冬的质量标准进行全面控制,本课题采用原植物鉴定、性状鉴定、显微鉴定等方法对川渝产细毡毛忍冬进行生药学鉴别,采用薄层色谱法对绿原酸进行定性鉴别,采用紫外分光光度法测定了总黄酮和总绿原酸的含量,采用高效液相色谱法测定了绿原酸的含量,并采用高效液相色谱法对其指纹图谱进行了研究,为开发利用川渝产细毡毛忍冬提供科学依据。结果：原植物为多年生半常绿木质藤本,幼枝、叶柄和总花梗均被开展的淡黄褐色长糙毛、短柔毛和稀疏腺毛,双花单生于叶腋或少数集生枝端成总状花序。药材具有花蕾细棒槌状,表面为黄棕色、红棕色或绿色,非腺毛较稀疏,腺毛较多的特征。花粉显微特征具有两种腺毛细胞,厚壁非腺毛细胞,薄壁非腺毛单细胞,花粉粒：类球形,具3孔沟,还有棱角细尖的草酸钙簇晶。在薄层色谱中能检测到绿原酸；采用紫外分光光度法测定总黄酮和总绿原酸,测得川银花中总黄酮在0.01～0.08 mg/mL范围内具有良好的线性关系(R=0.9998),回收率为97.71%,RSD为1.48%。采用HPLC测定绿原酸,在0.8-8μg范围内具有良好的线性关系(r=0.9999),平均回收率为98.34%,RSD为0.91%。采用高效液相色谱法进行指纹图谱研究,测得药材有11个共有峰,共有峰的参数符合“中药注射剂指纹图谱技术要求”的有关规定。结论：薄层鉴别显示细毡毛忍冬中含有绿原酸成分；紫外分光光度法测定其总黄酮和总绿原酸含量,方法操作简便、快速、灵敏、具有良好的重复性和回收率,可作为细毡毛忍冬总黄酮和总绿原酸的含量测定方法。高效液相色谱法测定绿原酸的含量,方法操作简便、快速、具有良好的重复性和回收率,可作为川渝产细毡毛忍冬中绿原酸的含量测定方法。采用高效液相色谱法进行指纹图谱研究,方法准确,重复性好,建立的指纹图谱检测标准,可作为细毡毛忍冬质量评价与品种鉴别的重要依据之一。
objective:Lonicera similis Hemsl from sichuan and chongqing are dry bud or with The flowers open in early.It belongs Lonicera macranthoides Hand.-Mazz. And Lonicera hypoglauca Miq.and Lonicera confuse DC. Of Caprifoliaceae plants.The uses of which includes Antibacterial anti-inflammatory, heat detoxification, cold wind scattered the effect of heat.It is mainly used for the treatment of carbuncle swollen boils, Houbi, erysipelas, fever sera dysentery, Fengreganmao, incidence of diseases such as warm.Its main effective part is Chlorogenic acid, Iso-chlorogenic acid and total flavonoids and et. Lonicera similis Hemsl are the mainstream varieties of Flos Lonicerae from sichuan.This issue will study the Pharmacognostical features,and it will identificate Lonicera similis Hemsl by TLC, and determinate its effective components, while fingerprinting studies them to enhance and improve the quality of Lonicera similis Hemsl from sichuan and chongqing. Method:For controling the quality standards of Lonicera similis Hemsl from sichuan and chongqing fully, This study using the original plant identification, trait identification, microscopic identification and chemical identification (TLC) method to make a Pharmacognosy Identification.This paper qualitative identificates chlorogenic acid in Lonicera similis Hemsl from sichuan and chongqing using TLC,and determines the total chlorogenic acid and total flavonoids content by ultraviolet spectrophotometry. While using high-performance liquid chromatography (HPLC) to carry out the content of chlorogenic acid, and using HPLC to research its fingerprint. Those could provide a scientific basis for development and utilization of Lonicera similis Hemsl from sichuan and chongqing.Results:The original plants are semi-evergreen and perennial woody plant vines. Young'branches, petiole, and peduncle length were carried out in short brown coarse hair, short hair and sparse hairs soft. Double flowers grow solitary and axillary or terminal branches into a few clusters of racemes. Herbs have the features of small wooden club-shaped flower, yellow brown surface, red brown or green, non-glandular hairs and scattered glandular hairs. Pollen microscopic characteristics with the two kinds of glandular cells, thick-walled non-glandular cells, thin-walled single-celled non-glandular hairs, pollen grains are class spherical, with three-hole groove, there are sharp edges and corners small clusters of calcium oxalate crystal. Using thin-layer chromatography identificates of the legal nature of chlorogenic acid,chlorogenic acid can be detected In the thin-layer chromatography.Using ultraviolet spectrophotometry to detect total flavonoids and total chlorogenic acid, Total flavonoids can be measured in 0.01-0.08 mg/mL with good linearity within the scope relationship (R=0.9998), recovery was 97. 71%, and RSD was 1.48%. Using HPLC to determinate chlorogenic acid,while in the range of 0.8-8μg it has good linear relationship (r=0.9998), the average recovery was 98.34%, and RSD was 0.91%.Using HPLC to carry out fingerprint study.It have been measured a total of 11 peaks,and the parameters of a total of peak accord with "traditional Chinese medicine injection fingerprinting requirements" of the relevant provisions. Conclusion:The TLC show Lonicera similis Hemsl from sichuan and chongqing contains chlorogenic acid composition; Ultraviolet spectrophotometry detecting their total flavonoids and total chlorogenic acid content. The method is simple, rapid, sensitive and with good repeatability and recovery.It can be used as the content of total flavonoids and total chlorogenic acid determination of Lonicera similis Hemsl from sichuan and chongqing. Using HPLC to determinate chlorogenic acid, This method is simple, rapid,efficient, sensitive, accurate and with good repeatability and recovery, it can be used as a quantitative analysismethod for Lonicera similis Hemsl from sichuan and chongqing. Using HPLC carried out its fingerprint study,and the method is accurate and reproducible.The established standards of fingerprints can be used as fine macranthoin of quality evaluation and an important basis for identification of species.
objective:Lonicera similis Hemsl from sichuan and chongqing are dry bud or with The flowers open in early.It belongs Lonicera macranthoides Hand.-Mazz. And Lonicera hypoglauca Miq.and Lonicera confuse DC. Of Caprifoliaceae plants.The uses of which includes Antibacterial anti-inflammatory, heat detoxification, cold wind scattered the effect of heat.It is mainly used for the treatment of carbuncle swollen boils, Houbi, erysipelas, fever sera dysentery, Fengreganmao, incidence of diseases such as warm.Its main effective part is Chlorogenic acid, Iso-chlorogenic acid and total flavonoids and et. Lonicera similis Hemsl are the mainstream varieties of Flos Lonicerae from sichuan.This issue will study the Pharmacognostical features,and it will identificate Lonicera similis Hemsl by TLC, and determinate its effective components, while fingerprinting studies them to enhance and improve the quality of Lonicera similis Hemsl from sichuan and chongqing. Method:For controling the quality standards of Lonicera similis Hemsl from sichuan and chongqing fully, This study using the original plant identification, trait identification, microscopic identification and chemical identification (TLC) method to make a Pharmacognosy Identification.This paper qualitative identificates chlorogenic acid in Lonicera similis Hemsl from sichuan and chongqing using TLC,and determines the total chlorogenic acid and total flavonoids content by ultraviolet spectrophotometry. While using high-performance liquid chromatography (HPLC) to carry out the content of chlorogenic acid, and using HPLC to research its fingerprint. Those could provide a scientific basis for development and utilization of Lonicera similis Hemsl from sichuan and chongqing.Results:The original plants are semi-evergreen and perennial woody plant vines. Young'branches, petiole, and peduncle length were carried out in short brown coarse hair, short hair and sparse hairs soft. Double flowers grow solitary and axillary or terminal branches into a few clusters of racemes. Herbs have the features of small wooden club-shaped flower, yellow brown surface, red brown or green, non-glandular hairs and scattered glandular hairs. Pollen microscopic characteristics with the two kinds of glandular cells, thick-walled non-glandular cells, thin-walled single-celled non-glandular hairs, pollen grains are class spherical, with three-hole groove, there are sharp edges and corners small clusters of calcium oxalate crystal. Using thin-layer chromatography identificates of the legal nature of chlorogenic acid,chlorogenic acid can be detected In the thin-layer chromatography.Using ultraviolet spectrophotometry to detect total flavonoids and total chlorogenic acid, Total flavonoids can be measured in 0.01-0.08 mg/mL with good linearity within the scope relationship (R=0.9998), recovery was 97. 71%, and RSD was 1.48%. Using HPLC to determinate chlorogenic acid,while in the range of 0.8-8μg it has good linear relationship (r=0.9998), the average recovery was 98.34%, and RSD was 0.91%.Using HPLC to carry out fingerprint study.It have been measured a total of 11 peaks,and the parameters of a total of peak accord with "traditional Chinese medicine injection fingerprinting requirements" of the relevant provisions. Conclusion:The TLC show Lonicera similis Hemsl from sichuan and chongqing contains chlorogenic acid composition; Ultraviolet spectrophotometry detecting their total flavonoids and total chlorogenic acid content. The method is simple, rapid, sensitive and with good repeatability and recovery.It can be used as the content of total flavonoids and total chlorogenic acid determination of Lonicera similis Hemsl from sichuan and chongqing. Using HPLC to determinate chlorogenic acid, This method is simple, rapid,efficient, sensitive, accurate and with good repeatability and recovery, it can be used as a quantitative analysismethod for Lonicera similis Hemsl from sichuan and chongqing. Using HPLC carried out its fingerprint study,and the method is accurate and reproducible.The established standards of fingerprints can be used as fine macranthoin of quality evaluation and an important basis for identification of species.
引文
1.中华人民共和国药典(一部)[S].北京：化学工业出版社,2005.21.
2.苏维辉,王卫平.双波长法测定银菊冲剂中绿原酸含量[J].广东药学,1999,9 (1)： 17.
3.四川省卫生厅编.四川省中药材标准[S].1987年版,1987,146-150.
4.湖南省卫生厅编.湖南省中药材标准[S].1993年版,长沙：湖南科学技术出版社,1993,191-192.
5.贵州省卫生厅编.贵州省中药材质量标准[S].1988年版,贵阳：贵州人民出版社,1990,85-86.
6.舒胜辉.不同产地山银花有效成分绿原酸含量的比较研究[J].中医药导报,2006,12(2)：74-75.
7.曹冬红,张贺文,杨科,等.高效液相色谱法测定山银花中绿原酸的含量[J].中南药学,2007,5(1)：36-38.
8.何兵,王琳,田吉,等.青银注射液中总绿原酸的紫外含量测定[J].泸州医学院学报,2005,6(28)：497-499.
9.崔征.生药学[M].第1版.北京：中国医药科技出版社,1999,225-227.
10.金辉,吕文伟,崔新明,等.蒲黄总黄酮对急性心肌梗死犬血清中微量元素及心肌细胞超微结构的影响[J].中草药,2008,39(1)：97-99,122.
11.赵明,于影,邵慧杰,等.黄芪总黄酮对大鼠实验性心律失常的保护作用[J].中国心血管病研究,2007,5(12)：918-919.
12.张建华,陈志武,武征.杜鹃花总黄酮预处理对大鼠心肌缺血再灌注损伤的延迟相保护作用[J].中国中医基础医学杂志,2007,13(3)：192-194.
13.吕雄文,李俊,邹字宏,等.老鹰茶总黄酮降血糖作用的实验研究[J].中国中医药科技,2008,15(2)：119-121.
14.苏曼曼,于晓风,曲绍春,等.分蘖葱头总黄酮对实验性高脂血症大鼠血脂代谢的影响[J].中国药理学通报.2007,23(12)：1594-1597.
15.回瑞华,侯冬岩,郭华,等.抗病毒草药金银花的化学成分分析[J].鞍山师范学院学报,2002,5(6)：56-58.
16.何兵,冯文宇,田吉,等.双青咽喉片的薄层色谱鉴别[J].中国医院药学杂志,2008,28(2)：122-124.
17.陈玉娟,张宏桂,贺丽鹏,等.水曲柳HPLC指纹图谱研究[J].中国药房,2008,19(9)：673-675.
1.中华人民共和国药典(一部)[S].北京：化学工业出版社,2005.152
2.单爱莲,权菊香,钱丽奇,等对预防与治疗非典型肺炎的中药处方情况调查
分析[J].中国临床药理学杂志,2003,19 (30)：289-300.
3.张玲,彭广芳,钟方晓,等.山东金银花挥发油的化学成分分析[J].时珍国药研究,1996,7(2)：89-91.
4.张玲,彭广芳,林慧彬,等.山东金银花挥发油化学成分的研究[J].中国药学杂志,1995,30(11)：651.
5.张玲,彭广芳,于宗渊.山东金银花主流品种挥发油成分比较研究[J].中国药科大学学报,1994,25(3)：184.
6.吉力,潘炳兴,徐植灵.忍冬挥发油的C,C.MS分析[J].中国中药杂志,1990,15(11)：680.
7.高玉敏.金银花化学成分的研究[J].中草药,1995,26(11)：568.
8.刘祥兰,刘芳,张英,等.金银花中绿原酸提取工艺的比较和优化研究[J].中成药,2000,22(6)：402.
9.王天志,李永梅.金银花的研究进展[J].华西药学杂志,2000,15(4)292-298.
10.白瑞,王真真.指纹图谱在中药质量控制中的应用进展[J].安徽医药,2009,13(12)：1458-1462.
11.赵惠茹,王锐平,王燕.聚类分析法在金银花及其伪品鉴别分类中的应用[J].陕西中医,2005,26(1)：72-73.
12.王斌,王燕,孙文基.中药金银花GC法指纹图谱研究[J].陕西中医,2004,25(4).：364-365.
13.田吉,冯文字,邱华荣,等.金银花药材气相指纹图谱研究[J].泸州医学院学报,2002,25(1)：76.
14.王倩,王建新,于治国,等.金银花药材高效液相色谱指纹图谱研究[J].中成药,2005,7(7)：27.
15.刘艳娥,吴玫涵,吴玉田,等.金银花药材高效液相指纹图谱研究[J].解放军药学学报,2003,19(3)：161-164.
16.容蓉,闰斌,等.不同产地金银花药材的HPCE指纹图谱分析[J],2006,15(6)：66-68.
17.孔德鑫,黄庶识,黄荣韶,等.基于双指标分析法和聚类分析法的鸡骨草红外指纹图谱比较研究[J].光谱学与光谱分析.2010,30(1)：45-49.
18.宋海英,邱世翠,王志强.金银花的体外抑菌作用[J].时珍国医国药.2003,14(5)：269.
19.何晶.金银花的药理作用及临床应用[J].天津药学,2008,20,(5)：74-76.
20.郑虎占,黄泽宏,余靖.中药现代化研究与应用.第3卷.北京：学苑出版社,1998：2938-2956.
21.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,2008,20(1)：2.
22.马骏,陈凌,任远,等.抗菌消炎片主要药效学研究[J].中成药,2005,27(6)：687.
23.葛冰,卢向阳,易克,等.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.
24.王力川.金银花的化学成分及功效研究进展[J].安徽农业科学,2009,37(5)：2036-2037.
25.袁毅君,宋瑛.清热类中药的抗生育作用.天水师范学院学报,2001,21(5)：28.
26.高春梅,杜跃中,潘晓鹏,等.金银花药理作用的研究进展及应用[J].人参研究,2007,1(4)：16-18.
27.王晓东.金银花治疗卡他性结膜炎[J].辽宁中医杂志,2002,29(12)：29.
28.薛新民,许占军,戚建国,等.用中药金银花等制剂保存发炎活髓的疗效观察[J].宁夏医学杂志,2003,25(2)：112-113.
29.高春梅,杜跃中,潘晓鹏,等.金银花药理作用的研究进展及应用[J].人参研究,2007,4：16-18.
2.苏维辉,王卫平.双波长法测定银菊冲剂中绿原酸含量[J].广东药学,1999,9 (1)： 17.
3.四川省卫生厅编.四川省中药材标准[S].1987年版,1987,146-150.
4.湖南省卫生厅编.湖南省中药材标准[S].1993年版,长沙：湖南科学技术出版社,1993,191-192.
5.贵州省卫生厅编.贵州省中药材质量标准[S].1988年版,贵阳：贵州人民出版社,1990,85-86.
6.舒胜辉.不同产地山银花有效成分绿原酸含量的比较研究[J].中医药导报,2006,12(2)：74-75.
7.曹冬红,张贺文,杨科,等.高效液相色谱法测定山银花中绿原酸的含量[J].中南药学,2007,5(1)：36-38.
8.何兵,王琳,田吉,等.青银注射液中总绿原酸的紫外含量测定[J].泸州医学院学报,2005,6(28)：497-499.
9.崔征.生药学[M].第1版.北京：中国医药科技出版社,1999,225-227.
10.金辉,吕文伟,崔新明,等.蒲黄总黄酮对急性心肌梗死犬血清中微量元素及心肌细胞超微结构的影响[J].中草药,2008,39(1)：97-99,122.
11.赵明,于影,邵慧杰,等.黄芪总黄酮对大鼠实验性心律失常的保护作用[J].中国心血管病研究,2007,5(12)：918-919.
12.张建华,陈志武,武征.杜鹃花总黄酮预处理对大鼠心肌缺血再灌注损伤的延迟相保护作用[J].中国中医基础医学杂志,2007,13(3)：192-194.
13.吕雄文,李俊,邹字宏,等.老鹰茶总黄酮降血糖作用的实验研究[J].中国中医药科技,2008,15(2)：119-121.
14.苏曼曼,于晓风,曲绍春,等.分蘖葱头总黄酮对实验性高脂血症大鼠血脂代谢的影响[J].中国药理学通报.2007,23(12)：1594-1597.
15.回瑞华,侯冬岩,郭华,等.抗病毒草药金银花的化学成分分析[J].鞍山师范学院学报,2002,5(6)：56-58.
16.何兵,冯文宇,田吉,等.双青咽喉片的薄层色谱鉴别[J].中国医院药学杂志,2008,28(2)：122-124.
17.陈玉娟,张宏桂,贺丽鹏,等.水曲柳HPLC指纹图谱研究[J].中国药房,2008,19(9)：673-675.
1.中华人民共和国药典(一部)[S].北京：化学工业出版社,2005.152
2.单爱莲,权菊香,钱丽奇,等对预防与治疗非典型肺炎的中药处方情况调查
分析[J].中国临床药理学杂志,2003,19 (30)：289-300.
3.张玲,彭广芳,钟方晓,等.山东金银花挥发油的化学成分分析[J].时珍国药研究,1996,7(2)：89-91.
4.张玲,彭广芳,林慧彬,等.山东金银花挥发油化学成分的研究[J].中国药学杂志,1995,30(11)：651.
5.张玲,彭广芳,于宗渊.山东金银花主流品种挥发油成分比较研究[J].中国药科大学学报,1994,25(3)：184.
6.吉力,潘炳兴,徐植灵.忍冬挥发油的C,C.MS分析[J].中国中药杂志,1990,15(11)：680.
7.高玉敏.金银花化学成分的研究[J].中草药,1995,26(11)：568.
8.刘祥兰,刘芳,张英,等.金银花中绿原酸提取工艺的比较和优化研究[J].中成药,2000,22(6)：402.
9.王天志,李永梅.金银花的研究进展[J].华西药学杂志,2000,15(4)292-298.
10.白瑞,王真真.指纹图谱在中药质量控制中的应用进展[J].安徽医药,2009,13(12)：1458-1462.
11.赵惠茹,王锐平,王燕.聚类分析法在金银花及其伪品鉴别分类中的应用[J].陕西中医,2005,26(1)：72-73.
12.王斌,王燕,孙文基.中药金银花GC法指纹图谱研究[J].陕西中医,2004,25(4).：364-365.
13.田吉,冯文字,邱华荣,等.金银花药材气相指纹图谱研究[J].泸州医学院学报,2002,25(1)：76.
14.王倩,王建新,于治国,等.金银花药材高效液相色谱指纹图谱研究[J].中成药,2005,7(7)：27.
15.刘艳娥,吴玫涵,吴玉田,等.金银花药材高效液相指纹图谱研究[J].解放军药学学报,2003,19(3)：161-164.
16.容蓉,闰斌,等.不同产地金银花药材的HPCE指纹图谱分析[J],2006,15(6)：66-68.
17.孔德鑫,黄庶识,黄荣韶,等.基于双指标分析法和聚类分析法的鸡骨草红外指纹图谱比较研究[J].光谱学与光谱分析.2010,30(1)：45-49.
18.宋海英,邱世翠,王志强.金银花的体外抑菌作用[J].时珍国医国药.2003,14(5)：269.
19.何晶.金银花的药理作用及临床应用[J].天津药学,2008,20,(5)：74-76.
20.郑虎占,黄泽宏,余靖.中药现代化研究与应用.第3卷.北京：学苑出版社,1998：2938-2956.
21.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,2008,20(1)：2.
22.马骏,陈凌,任远,等.抗菌消炎片主要药效学研究[J].中成药,2005,27(6)：687.
23.葛冰,卢向阳,易克,等.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.
24.王力川.金银花的化学成分及功效研究进展[J].安徽农业科学,2009,37(5)：2036-2037.
25.袁毅君,宋瑛.清热类中药的抗生育作用.天水师范学院学报,2001,21(5)：28.
26.高春梅,杜跃中,潘晓鹏,等.金银花药理作用的研究进展及应用[J].人参研究,2007,1(4)：16-18.
27.王晓东.金银花治疗卡他性结膜炎[J].辽宁中医杂志,2002,29(12)：29.
28.薛新民,许占军,戚建国,等.用中药金银花等制剂保存发炎活髓的疗效观察[J].宁夏医学杂志,2003,25(2)：112-113.
29.高春梅,杜跃中,潘晓鹏,等.金银花药理作用的研究进展及应用[J].人参研究,2007,4：16-18.