绵马贯众的质量控制方法研究
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摘要
本论文分为三个部分
第一部分文献综述
本论文通过系统查阅2012年以前国内外的相关文献,对间苯三酚类成分的植物来源、结构类型、药理活性、合成途径以及绵马贯众的化学成分、药理活性、质量研究等进行了较深入的总结分析。
第二部分实验研究
1.东北贯众素ABBA对照品的制备
先后尝试了加压薄层色谱法、重结晶法、高速逆流色谱法、反相硅胶柱分离法,减压正相硅胶柱分离法、正相硅胶柱分离法以及薄层色谱制备法等多种方法分离制备东北贯众素ABBA,最后,通过薄层色谱制备的方法制备出了东北贯众素ABBA对照品,并首次从绵马贯众中分离得到东北贯众素ABBP。
2.薄层色谱鉴别研究
改进了现行版药典记载的绵马贯众项下的薄层鉴别方法,对薄层板、活化条件、展开剂、预饱和时间、显色温度和时间进行了选择,并通过正交试验考察了相对湿度、温度和预饱和时间对薄层分离效果的综合影响。最后,确定改进后的薄层条件为：将枸橼酸-磷酸氢二钠缓冲液(pH7)浸润后的GF254预制板于110℃活化30min,以正已烷-氯仿-甲醇(10：20：1)为展开剂,控制相对湿度为50%,温度为25℃,薄层板置展开缸中预饱和1h,展开,取出,立即喷以0.3%的坚牢蓝BB盐稀乙醇溶液,用吹风机热风吹至斑点清晰。实验证明,改进后的TLC鉴别方法分离效果良好,且减少了实验中制板和环境温度、湿度等不稳定因素的影响,简化了实验操作步骤,大大缩短了实验时间,提高了实验效率。
建立了绵马贯众的生物自显影TLC鉴别方法,使其质量评价与其抗氧化活性直接相关联,从而更好地控制绵马贯众的质量。
3.检查研究
对绵马贯众进行了水分、总灰分以及酸不溶性灰分的测定,为建立绵马贯众质量标准提供了参考,建议绵马贯众炭的水分限度不超过7%,总灰分含量不超过8%,酸不溶性灰分含量不超过0.6%；绵马贯众饮片的酸不溶性灰分含量不超过1%。
4.总间苯三酚的含量测定
利用比色法测定了12批绵马贯众药材、12批绵马贯众饮片以及3批绵马贯众炭中总间苯三酚的含量,以东北贯众素ABBA为对照品,建立回归方程y=19.1x-0.084,r=0.9998,结果显示,对照品的浓度在12.12μg/ml～48.48μg／ml范围内与吸收度值呈良好的线性关系。由测定结果可知,购自不同地区的绵马贯众药材、饮片所含总间苯三酚的量虽然存在一定差异,但绝大部分具有较好的一致性,仅各有一批药材和饮片的总间苯三酚含量极低,提示这2批样品可能为伪品。3批绵马贯众炭的含量测定结果显示,其总间苯三酚含量在药材含量的十分之一以下,说明炮制过程中大部分间苯三酚类成分已经遭到破坏。
5.指纹图谱研究
首次建立了绵马贯众的HPLC指纹图谱测定方法,精密度、重复性和稳定性均符合要求。测定了12批绵马贯众药材、12批绵马贯众饮片以及3批绵马贯众炭的指纹图谱,通过相似度软件分析,11批绵马贯众药材的相似度在0.908和0.987之间,均高于0.900,有l批样品相似度仅为0.182；11批绵马贯众饮片的相似度在0.936和0.992之间,均高于0.900,有1批样品相似度仅为0.183；3批绵马贯众炭的相似度分别为0.869,0.661,0.893。其中,绵马贯众药材和饮片各有一批样品与对照指纹图谱差异显著,证明本文所建立的绵马贯众HPLC指纹图谱检测方法可有效鉴别绵马贯众药材和饮片
第三部分总结与讨论
起草绵马贯众药材及饮片质量标准草案,并对本论文进行归纳总结,得出创新点：
1.改进绵马贯众TLC鉴别方法,极大简化了实验步骤,缩短了实验时间,提高了实验效率。
2.首次将TLC-生物自显影技术用于绵马贯众的鉴别,将其抗氧化活性检测与TLC鉴别完美结合。
3.首次建立了绵马贯众的指纹图谱测定方法。
Part one:Literature review
The documents about the present research situation in Chinese medicine Guan Zhong were summed up and analyzed. According to studying on the related literature domestic and overseas until2012, a review on the research progress of phloroglucinol compounds' plant origin, structure classification, pharmacological activities, synthetic routes and Dryopteris crassirhizoma's chemical constituents, pharmacological activities and quality control research was presented.
Part two:Experimentation
1. Preparation of reference substance of Dryocrassine ABBA
To prepare Dryocrassine ABBA, many methods has been tried, including overpressured-layer chromatography(OPLC), recrystallization, High speed counter-current chromatography (HSCCC), reverse phase silica gel column chromatography, vacuum silica gel column chromatography, silica gel column chromatography and preparative thin-layer chromatography. Finally, reference substance of Dryocrassine ABBA was gotten by preparative thin-layer chromatography, and Dryocrassine ABBP was isolated from Dryopteris crassirhizoma for the first time.
2. TLC identification method of Dryopteris crassirhizoma
Studying on the different type of plates, activated conditions, developing solvents, monitor conditions to improve the TLC identification method of Dryopteris crassirhizoma recording in Chinese pharmacopoeia2010edition. Further, using orthogonal experiment method to investigate the influence of temperature, relative humidity, saturating time on the TLC results. When the relative humidity was50%, the temperature was at25℃, infiltrate silica plate in pH7citric acid-disodium hydrogen phosphate buffer solution,110℃activated for30min, the developing solvent was n-hexane-chloroform-methanol(10:20:1), saturated for1h, the TLC separation for Dryopteris crassirhizoma was satisfied and with good repeatability. It has been proved by experiments that the improved method has better separation effect and it can reduce influence of environmental temperature, humidity and other factors, and remarkably save the time so that it can improve the experimental efficiency.
Establishing the TLC-bioautography identification method of Dryopteris crassirhizoma, associate its antioxidative activity with quality evaluation to control its quality well.
3. Research on content determination of moisture,total ash,acid-insoluble ash in Dryopteris crassirhizoma. According to the results, we suggest that the moisture content, total ash content and acid-insoluble ash content of Dryopteris crassirhizoma charcoal should be controlled no more than7%,8%, and0.6%; the acid-insoluble ash content of Dryopteris crassirhizoma pieces should be controlled no more than1%.
4. Content of total dryocrassins was evaluated by colorimetry
Evaluating content of total dryocrassins in12batches of Dryopteris crassirhizoma materials,12batches of Dryopteris crassirhizoma pieces, and3batches of Dryopteris crassirhizoma charcoal by colorimetry. Regression equations of Dryocrassine ABBA were y=19.1x-0.084, r=0.9998, which showed that the calibration curve was linear within the range of12.12μg/ml～48.48μg/ml for Dryocrassine ABBA. The result shows that, there were some differences among Dryopteris crassirhizoma materials and pieces, but most of them have good consistency. In addition, there were2samples'total dryocrassins content were extremely low which respectively belong to Dryopteris crassirhizoma materials and pieces, it seems like that they were counterfeits. According to the results of Dryopteris crassirhizoma charcoal, we can find that their total dryocrassins were lower than1/10of Dryopteris crassirhizoma materials and pieces, implying that most of dryocrassins in them have been destoried during processing.
5. Study of HPLC fingerprint spectrum
Establishing the determination method of HPLC fingerprint spectrum related to Dryopteris crassirhizoma for the first time. And evaluating fingerprint spectrums of12batches of Dryopteris crassirhizoma materials,12batches of Dryopteris crassirhizoma pieces, and3batches of Dryopteris crassirhizoma charcoal. Then, analyzing the fingerprint spectrums by similarity calculation software, the results showed that, the similarity of11batches of Dryopteris crassirhizoma materials were ranged from0.908to0.987, there was1sample's similarity just was0.182; the similarity of11batches of Dryopteris crassirhizoma pieces were ranged from0.936to0.992, there also was1sample's similarity just was0.183; and the similarity of3batches of Dryopteris crassirhizoma charcoal were0.869,0.661,0.893respectively. From the result above, we can get that the method was steady and reliable.
Part three:Summary and discussion
To draft the quality standard draft of Dryopteris crassirhizoma materials and pieces, and summarize the paper and carry out the Innovations as below:
1. Improved the TLC identification method recording in Chinese pharmacopoeia2010edition so that we can simplify experimental procedure, save experimental time and improve experimental efficiency.
2. First time to establish the TLC-bioautography identification method of Dryopteris crassirhizoma, associate its antioxidative activity with quality evaluation to control its quality well.
3. To establish the HPLC fingerprint spectrum of Dryopteris crassirhizoma for the first time.
第一部分文献综述
本论文通过系统查阅2012年以前国内外的相关文献,对间苯三酚类成分的植物来源、结构类型、药理活性、合成途径以及绵马贯众的化学成分、药理活性、质量研究等进行了较深入的总结分析。
第二部分实验研究
1.东北贯众素ABBA对照品的制备
先后尝试了加压薄层色谱法、重结晶法、高速逆流色谱法、反相硅胶柱分离法,减压正相硅胶柱分离法、正相硅胶柱分离法以及薄层色谱制备法等多种方法分离制备东北贯众素ABBA,最后,通过薄层色谱制备的方法制备出了东北贯众素ABBA对照品,并首次从绵马贯众中分离得到东北贯众素ABBP。
2.薄层色谱鉴别研究
改进了现行版药典记载的绵马贯众项下的薄层鉴别方法,对薄层板、活化条件、展开剂、预饱和时间、显色温度和时间进行了选择,并通过正交试验考察了相对湿度、温度和预饱和时间对薄层分离效果的综合影响。最后,确定改进后的薄层条件为：将枸橼酸-磷酸氢二钠缓冲液(pH7)浸润后的GF254预制板于110℃活化30min,以正已烷-氯仿-甲醇(10：20：1)为展开剂,控制相对湿度为50%,温度为25℃,薄层板置展开缸中预饱和1h,展开,取出,立即喷以0.3%的坚牢蓝BB盐稀乙醇溶液,用吹风机热风吹至斑点清晰。实验证明,改进后的TLC鉴别方法分离效果良好,且减少了实验中制板和环境温度、湿度等不稳定因素的影响,简化了实验操作步骤,大大缩短了实验时间,提高了实验效率。
建立了绵马贯众的生物自显影TLC鉴别方法,使其质量评价与其抗氧化活性直接相关联,从而更好地控制绵马贯众的质量。
3.检查研究
对绵马贯众进行了水分、总灰分以及酸不溶性灰分的测定,为建立绵马贯众质量标准提供了参考,建议绵马贯众炭的水分限度不超过7%,总灰分含量不超过8%,酸不溶性灰分含量不超过0.6%；绵马贯众饮片的酸不溶性灰分含量不超过1%。
4.总间苯三酚的含量测定
利用比色法测定了12批绵马贯众药材、12批绵马贯众饮片以及3批绵马贯众炭中总间苯三酚的含量,以东北贯众素ABBA为对照品,建立回归方程y=19.1x-0.084,r=0.9998,结果显示,对照品的浓度在12.12μg/ml～48.48μg／ml范围内与吸收度值呈良好的线性关系。由测定结果可知,购自不同地区的绵马贯众药材、饮片所含总间苯三酚的量虽然存在一定差异,但绝大部分具有较好的一致性,仅各有一批药材和饮片的总间苯三酚含量极低,提示这2批样品可能为伪品。3批绵马贯众炭的含量测定结果显示,其总间苯三酚含量在药材含量的十分之一以下,说明炮制过程中大部分间苯三酚类成分已经遭到破坏。
5.指纹图谱研究
首次建立了绵马贯众的HPLC指纹图谱测定方法,精密度、重复性和稳定性均符合要求。测定了12批绵马贯众药材、12批绵马贯众饮片以及3批绵马贯众炭的指纹图谱,通过相似度软件分析,11批绵马贯众药材的相似度在0.908和0.987之间,均高于0.900,有l批样品相似度仅为0.182；11批绵马贯众饮片的相似度在0.936和0.992之间,均高于0.900,有1批样品相似度仅为0.183；3批绵马贯众炭的相似度分别为0.869,0.661,0.893。其中,绵马贯众药材和饮片各有一批样品与对照指纹图谱差异显著,证明本文所建立的绵马贯众HPLC指纹图谱检测方法可有效鉴别绵马贯众药材和饮片
第三部分总结与讨论
起草绵马贯众药材及饮片质量标准草案,并对本论文进行归纳总结,得出创新点：
1.改进绵马贯众TLC鉴别方法,极大简化了实验步骤,缩短了实验时间,提高了实验效率。
2.首次将TLC-生物自显影技术用于绵马贯众的鉴别,将其抗氧化活性检测与TLC鉴别完美结合。
3.首次建立了绵马贯众的指纹图谱测定方法。
Part one:Literature review
The documents about the present research situation in Chinese medicine Guan Zhong were summed up and analyzed. According to studying on the related literature domestic and overseas until2012, a review on the research progress of phloroglucinol compounds' plant origin, structure classification, pharmacological activities, synthetic routes and Dryopteris crassirhizoma's chemical constituents, pharmacological activities and quality control research was presented.
Part two:Experimentation
1. Preparation of reference substance of Dryocrassine ABBA
To prepare Dryocrassine ABBA, many methods has been tried, including overpressured-layer chromatography(OPLC), recrystallization, High speed counter-current chromatography (HSCCC), reverse phase silica gel column chromatography, vacuum silica gel column chromatography, silica gel column chromatography and preparative thin-layer chromatography. Finally, reference substance of Dryocrassine ABBA was gotten by preparative thin-layer chromatography, and Dryocrassine ABBP was isolated from Dryopteris crassirhizoma for the first time.
2. TLC identification method of Dryopteris crassirhizoma
Studying on the different type of plates, activated conditions, developing solvents, monitor conditions to improve the TLC identification method of Dryopteris crassirhizoma recording in Chinese pharmacopoeia2010edition. Further, using orthogonal experiment method to investigate the influence of temperature, relative humidity, saturating time on the TLC results. When the relative humidity was50%, the temperature was at25℃, infiltrate silica plate in pH7citric acid-disodium hydrogen phosphate buffer solution,110℃activated for30min, the developing solvent was n-hexane-chloroform-methanol(10:20:1), saturated for1h, the TLC separation for Dryopteris crassirhizoma was satisfied and with good repeatability. It has been proved by experiments that the improved method has better separation effect and it can reduce influence of environmental temperature, humidity and other factors, and remarkably save the time so that it can improve the experimental efficiency.
Establishing the TLC-bioautography identification method of Dryopteris crassirhizoma, associate its antioxidative activity with quality evaluation to control its quality well.
3. Research on content determination of moisture,total ash,acid-insoluble ash in Dryopteris crassirhizoma. According to the results, we suggest that the moisture content, total ash content and acid-insoluble ash content of Dryopteris crassirhizoma charcoal should be controlled no more than7%,8%, and0.6%; the acid-insoluble ash content of Dryopteris crassirhizoma pieces should be controlled no more than1%.
4. Content of total dryocrassins was evaluated by colorimetry
Evaluating content of total dryocrassins in12batches of Dryopteris crassirhizoma materials,12batches of Dryopteris crassirhizoma pieces, and3batches of Dryopteris crassirhizoma charcoal by colorimetry. Regression equations of Dryocrassine ABBA were y=19.1x-0.084, r=0.9998, which showed that the calibration curve was linear within the range of12.12μg/ml～48.48μg/ml for Dryocrassine ABBA. The result shows that, there were some differences among Dryopteris crassirhizoma materials and pieces, but most of them have good consistency. In addition, there were2samples'total dryocrassins content were extremely low which respectively belong to Dryopteris crassirhizoma materials and pieces, it seems like that they were counterfeits. According to the results of Dryopteris crassirhizoma charcoal, we can find that their total dryocrassins were lower than1/10of Dryopteris crassirhizoma materials and pieces, implying that most of dryocrassins in them have been destoried during processing.
5. Study of HPLC fingerprint spectrum
Establishing the determination method of HPLC fingerprint spectrum related to Dryopteris crassirhizoma for the first time. And evaluating fingerprint spectrums of12batches of Dryopteris crassirhizoma materials,12batches of Dryopteris crassirhizoma pieces, and3batches of Dryopteris crassirhizoma charcoal. Then, analyzing the fingerprint spectrums by similarity calculation software, the results showed that, the similarity of11batches of Dryopteris crassirhizoma materials were ranged from0.908to0.987, there was1sample's similarity just was0.182; the similarity of11batches of Dryopteris crassirhizoma pieces were ranged from0.936to0.992, there also was1sample's similarity just was0.183; and the similarity of3batches of Dryopteris crassirhizoma charcoal were0.869,0.661,0.893respectively. From the result above, we can get that the method was steady and reliable.
Part three:Summary and discussion
To draft the quality standard draft of Dryopteris crassirhizoma materials and pieces, and summarize the paper and carry out the Innovations as below:
1. Improved the TLC identification method recording in Chinese pharmacopoeia2010edition so that we can simplify experimental procedure, save experimental time and improve experimental efficiency.
2. First time to establish the TLC-bioautography identification method of Dryopteris crassirhizoma, associate its antioxidative activity with quality evaluation to control its quality well.
3. To establish the HPLC fingerprint spectrum of Dryopteris crassirhizoma for the first time.
引文
[1]Inder Pal Singh, Sandip B. Bharate. Phloroglucinol compounds of natural origin[J]. The Royal Society of Chemistry,2006,23:558-591.
[2]高增平,李世文,陆蕴如,等.中药绵马贯众的化学成分研究[J].中国药学杂志,2003,38(4)：260-261.
[3]沈志滨.香鳞毛蕨药效物质基础研究和肤痒宁软膏的研制[D].黑龙江：黑龙江中医药大学中药学系,2005.
[4]李辉敏,阮金兰.鳞毛蕨科药用蕨类植物研究进展[J].九江学院学报,2004，3：78.
[5]马瑜,桂伟.宽羽鳞毛蕨化学成分研究[J].中草药,2006，37(9)：1315-1316.
[6]李辉敏.复叶耳蕨属植物的研究进展[J].九江医学,2009，24(3)：92.
[7]吴融.滇鸡血藤和复方鸡血藤膏的质量标准研究及荚果蕨贯众的化学成分研究[D].上海：复旦大学生药学系,2010.
[8]Adam K.-P., Five new phenolic compounds from fronds of Phegopteris connectilis[J]. Phytochemistry,1999,52:929.
[9]Miyake A, Yamamoto H, Takebayashi Y, et al. The novel natural product YM-26567-1[(+)-trans-4-(3-dodecanoyl-2,4,6-trihydroxyphenyl)-7-hydroxy-2-(4-hydroxy phenyl)chroman]:a competitive inhibitor of group Ⅱ phospholipase A2[J]. Pharmacol. Exp. Ther.,1992,263:1302.
[10]钟芳芳.傣药人面果树皮的化学成分与生物活性研究[D].武汉：中南民族大学生命科学学院生化与分子生物学系,2008.
[11]宋馨,祝建,吕洪飞.金丝桃属植物研究进展[J].西北植物学报,2005,25(4)：844-849.
[12]郭娜,陈宣钦,赵勤实.岷江金丝桃中一个新的间苯三酚类化合物[J].云南植物研究,2008,30(4)：515-518.
[13]曾祎含.元宝草果实部位抗多药耐药金葡菌活性成分的研究[D].上海：复旦大学药学院药物化学系,2008.
[14]李宁,张鹏,肖皖,等.间苯三酚类化合物——鹤草酚的NMR研究[J].波谱学杂志,2009,26(1)：103-109.
[15]黄相中.柳叶金叶子根和叶中活性成分的研究及具有扩张血管活性的间苯三酚类衍生物的合成[D].北京：北京协和医学院药物研究所药物化学系,2006.
[16]吴先富.仪花根、叶以及野八角果实化学成分的研究[D].北京：北京协和医学院药物研究所药物化学系,2009.
[17]Yamakoshi Y, Murata M, Shimizu A, et al. Isolation and characterization of Macro carpal s B-G antibacterial compounds from Eucalyptus macrocarpa[J]. Biosci. Biotech. Biochem.,1992,56:1570-1576.
[18]Simonson H T, Aderson A, Bremner P, et al. Antifungal constituents of Melicope borbonica[J].Phytother. Res.,2004,18:542.
[19]Satake T, Coskun M, Hori K, et al. An acetophenon and three Naphthalides from turkish Rhamnus libanoticus[J]. Phytochemistry,1991,30(12):4191-4192.
[20]Santos B V D O, Da-Cunha E V L, Chaves M C D O, et al. Phenylalkanoids from Piper marginatum[J]. Phytochemistry,1998,49:1381-1384.
[21]苏雨雷.商品贯众的品种及质量研究[D].北京：北京中医药大学中药学院中药化学系,2007.
[22]查传龙,陈兴浴.槟榔、厚朴等对肝吸虫作用的体外观察[J].南京中医学院学报,1990,(4)：30.
[23]左丽,陈若芸.鳞毛蕨属植物化学成分和药理活性研究进展[J].中草药,2005,36(9)：1427-1430.
[24]丁骁,王磊,欧阳明安.藤黄科植物化学成分的研究概况[J].化学工程与装备，2009,(1)：117.
[25]陈红云,蒋金元,王超芬,等.绵马贯众的抗菌活性研究[J].大理学院学报,2005,4(3)：22-23.
[26]常小龙.东北贯众、紫金牛和白花银背藤的化学成分研究[D].沈阳：沈阳药科大学,2006.
[27]韦四煌,方鉴,詹皓,等.贯众提取物的保肝降酶作用[J],航空军医,2004，32(3)：109-111.
[28]刘钊.绵马贯众抗感染活性成分研究[D].北京：北京协和医学院药用植物研究所生药学系,2010.
[29]徐程,于艳,孙科峰,等.绵马贯众提取物抗甲型流感病毒FM1株实验研究[J].辽宁中医药大学学报,2010,12(7)：29-31.
[30]Dona M, Dell' Aica I, Pezzato E, et al. Hyperforin inhibits cancer invasion and metastasis[J]. Cancer Res.,2004,64:6225-6232.
[31]薛惟建,王士贤,李德华.贯众抗肿瘤有效组分对P388细胞超微结构及细胞呼吸作用[J].中国药理学通报,1987，3(5)：291.
[32]Ito H, Muranaka T, Mori K, Ichthyotoxic phloroglucinol derivatives from Dryopteris fagrans and their anti-tumor promoting activity[J]. Chem Pharm Bull, 2000,48(8):1190-1195.
[33]Minoru S, Satoru K, Atsushi I. Flavon-3-ol structure-containing glycosides as histidin decarboxlase inhibitors and pharmaceutical compositions containing the inhibitors[P]. JP: 08217674A2,1996-08-27.
[34]窦昊,高志成,李薇.中药抗抑郁有效成分研究进展[J].中国药业,2010，19(21)：89.
[35]郑清明.元宝草及同属植物的生药学研究[D].上海：第二军医大学药学院生药学系,2005.
[36]陈绮.贯众抗生育有效成分的药理研究[J].天津医药,1980，(8)：188.
[37]吴先富.单刀根活性成分的研究[D].沈阳：沈阳药科大学药物化学,2006.
[38]任强.中药抗急性淋巴细胞白血病活性成分的研究[D].北京：北京协和医学院药用植物研究所生药学系,2009.
[39]高增平,陆蕴如,李国福,等.绵马贯众抗疟活性筛选[J].北京中医药大学学报,2002,25(4)：45.
[40]高增平,陆蕴如,江佩芬,等.绵马贯众部位Ⅱ的抗疟作用和急性毒性实验研究[J].北京中医药大学学报,2002,25(2)：52-53.
[41]涂小军.天然产物靛玉红及间苯三酚类衍生物的合成研究[D].西安：西北大学有机化学系,2010.
[42]中国药典委员会.中华人民共和国药典2010年版(一部).北京：化学工业出版社,2010年1月,310页.
[43]吴寿金,杨秀贤.绵马贯众化学成分的研究(Ⅰ)[J].中草药,1996,27(8)：458-459.
[44]高增平.绵马贯众有效成分的研究[D].北京：北京中医药大学中药化学系,2003.
[45]齐峰,王娥丽.常用药材绵马贯众活性成分研究[J].天津医科大学学报,2007,13(2)：191-193.
[46]陈建新,曾振灵,杨运云,等.液相色谱-串联质谱法分析绵马贯众抗禽流感病毒活性部位的化学成分[J].质谱学报,2006,27：118-120.
[47]吴寿金,杨秀贤.绵马贯众化学成分的研究Ⅱ[J].绵马贯众中间苯三酚衍生物的质谱-质谱分析[J].中草药,1997,28(12)：712-714.
[48]陈建新,曾振灵,杨运云,等.绵马贯众抗禽流感病毒药效部位的高效液相色谱-点喷雾串联质谱分析[J].分析测试学报,2005,24：73—75.
[49]陈建新,曾振灵,方炳虎,等.绵马贯众抗禽流感病毒活性部位的高效液相色谱-质谱法分析[J].分析测试学报,2008,27(6)：623—626.
[50]周荣华.东北贯众中抗菌活性成分的研究[D].哈尔滨：黑龙江大学生命科学学院,2010.
[51]高增平,马秉智,陆蕴如.绵马贯众的化学成分研究(Ⅱ)[J].北京中医药大学学报,2004,27(1)：52-53.
[52]胡昌江,叶茂,刘俊达,等.绵马贯众不同切制方法及炒炭的比较研究[J].中成药,2004,26(8)：641-643.
[53]黄勤挽,齐红艺,刘蕾,等.绵马贯众饮片“火力”与“火候”的探讨[A].中华中医药学会炮制分会.中华中医药学会第四届中药炮制学术会议论文集[C].2009：65-71.
[54]肖国君,叶利明,吴纯洁,等HPLC法测定绵马贯众中绵马贯众素的含量[J].药物分析杂志,2005,25(5)：502-504.
[55]司云珊,徐暾海,韩冬,等RP-HPLC法测定绵马贯众药材中绵马贯众素[J].中草药,2008,39(12)：1894-1895.
[56]高增平,王宝华,陆蕴如,等HPLC和TLCS测定绵马贯众抗疟有效部位中东北贯众素含量[J].中国药学杂志,2003,38(11)：878.
[57]肖国君.绵马贯众素对照品的研究及绵马贯众片的质量标准研究[D].四川：四川大学药物分析系,2004.
[58]鲁明明.中药材贯众质量标准研究[D].吉林：吉林农业大学中药,2002.
[59]张思巨,艾铁民,杨立新,等.绵马贯众贮存时间紧与药材质量相关性研究[J].中国中药杂志,1996,21(4)：206-207.
[60]侯冬岩,回瑞华,杨梅东,等.东北贯众中总黄酮的光谱分析[J].分析测试学报,2004,23(3)：110-112.
[61]柳芳.蛾眉蕨贯众的化学成分研究[D].北京：北京中医药大学,2009年.
[62]王晓丹,宗希明,吴洪斌,等.佳木斯地产白头翁、玉竹、萹蓄、长白沙参、绵马贯众、关卷术中微量元素的测定[J].佳木斯医学院学报,1997,20(4)：5.
[63]梁洪华,邓君丽.抗感颗粒中绵马贯众的鉴别和赤芍的含量测定[J].中国药事,2005,19(5)：290-292.
[64]胡海清,田宝成.抗病毒合剂的制备及质控初探[J].制剂技术,2007,16(7)：28-29.
[65]王波,胡美芳,牛胜德,等.抗感口服液的高效液相色谱鉴别[J].药物鉴定,2007,16(18)：36.
[1]高增平,王宝华,陆蕴如,等HPLC和TLCS测定绵马贯众抗疟有效部位中东北贯众素含量[J].中国药学杂志,2003,38(11)：878.
[2]吴寿金,杨秀贤,张丽,等.绵马贯众化学成分的研究(Ⅰ)[J].中草药,1996,27(8)458.
[3]高增平.绵马贯众有效成分的研究[D].北京：北京中医药大学中药化学系,2003.
[4]肖国君,叶利明,吴纯洁,等HPLC去测定绵马贯众中绵马贯众素的含量[J].药物分析杂志,2005,25(5)：502-504.
[5]司云珊,徐暾海,韩冬,等RP-HPLC法测定绵马贯众药材中绵马贯众素[J].中草药,2008,39(12)：1894-1895.
[1]中国药典委员会.中华人民共和国药典2010年版(一部).北京：化学工业出版社,2010年1月,310页.
[2]谷丽华,吴弢,张紫佳,等.应用薄层色谱-生物自显影技术评价乌药等三种中药的抗氧化活性[J].药学学报,2006,41(10)：956-962.
[3]Zijia Zhang, Liping Liao, Jeffrey Moore.Antioxidant phenolic compounds from walnut kernels (Juglans regia L.)[J].Food Chemistry,2009,113:160-165.
[4]曲建博,娄红祥,范培红.TLC-生物自显影技术在药物筛选中的应用[J].中草药,2005，36(1)：132-137.
[1]中国药典委员会.中华人民共和国药典2010年版(一部).北京：化学工业出版社,2010年1月.
[1]高增平.绵马贯众有效成分的研究[D].北京：北京中医药大学中药化学系,2003.
[2]李辉敏,阮金兰.鳞毛蕨科药用蕨类植物研究进展[J].九江学院学报,2004，3：78.
[3]左丽,陈若芸.鳞毛蕨属植物化学成分和药理活性研究进展[J].中草药,2005,36(9)：1427-1430.
[4]齐峰,王娥丽.常用药材绵马贯众活性成分研究[J].天津医科大学学报,2007,13(2)：191-193.
[5]陈红云,蒋金元,王超芬,等.绵马贯众的抗菌活性研究[J].大理学院学报,2005,4(3)：22-23.
[6]薛惟建,王士贤,李德华.贯众抗肿瘤有效组分对P388细胞超微结构及细胞呼吸作用[J].中国药理学通报,1987，3(5)：291.
[7]苏雨雷.商品贯众的品种及质量研究[D].北京：北京中医药大学中药学院中药化学系,2007.
[2]高增平,李世文,陆蕴如,等.中药绵马贯众的化学成分研究[J].中国药学杂志,2003,38(4)：260-261.
[3]沈志滨.香鳞毛蕨药效物质基础研究和肤痒宁软膏的研制[D].黑龙江：黑龙江中医药大学中药学系,2005.
[4]李辉敏,阮金兰.鳞毛蕨科药用蕨类植物研究进展[J].九江学院学报,2004，3：78.
[5]马瑜,桂伟.宽羽鳞毛蕨化学成分研究[J].中草药,2006，37(9)：1315-1316.
[6]李辉敏.复叶耳蕨属植物的研究进展[J].九江医学,2009，24(3)：92.
[7]吴融.滇鸡血藤和复方鸡血藤膏的质量标准研究及荚果蕨贯众的化学成分研究[D].上海：复旦大学生药学系,2010.
[8]Adam K.-P., Five new phenolic compounds from fronds of Phegopteris connectilis[J]. Phytochemistry,1999,52:929.
[9]Miyake A, Yamamoto H, Takebayashi Y, et al. The novel natural product YM-26567-1[(+)-trans-4-(3-dodecanoyl-2,4,6-trihydroxyphenyl)-7-hydroxy-2-(4-hydroxy phenyl)chroman]:a competitive inhibitor of group Ⅱ phospholipase A2[J]. Pharmacol. Exp. Ther.,1992,263:1302.
[10]钟芳芳.傣药人面果树皮的化学成分与生物活性研究[D].武汉：中南民族大学生命科学学院生化与分子生物学系,2008.
[11]宋馨,祝建,吕洪飞.金丝桃属植物研究进展[J].西北植物学报,2005,25(4)：844-849.
[12]郭娜,陈宣钦,赵勤实.岷江金丝桃中一个新的间苯三酚类化合物[J].云南植物研究,2008,30(4)：515-518.
[13]曾祎含.元宝草果实部位抗多药耐药金葡菌活性成分的研究[D].上海：复旦大学药学院药物化学系,2008.
[14]李宁,张鹏,肖皖,等.间苯三酚类化合物——鹤草酚的NMR研究[J].波谱学杂志,2009,26(1)：103-109.
[15]黄相中.柳叶金叶子根和叶中活性成分的研究及具有扩张血管活性的间苯三酚类衍生物的合成[D].北京：北京协和医学院药物研究所药物化学系,2006.
[16]吴先富.仪花根、叶以及野八角果实化学成分的研究[D].北京：北京协和医学院药物研究所药物化学系,2009.
[17]Yamakoshi Y, Murata M, Shimizu A, et al. Isolation and characterization of Macro carpal s B-G antibacterial compounds from Eucalyptus macrocarpa[J]. Biosci. Biotech. Biochem.,1992,56:1570-1576.
[18]Simonson H T, Aderson A, Bremner P, et al. Antifungal constituents of Melicope borbonica[J].Phytother. Res.,2004,18:542.
[19]Satake T, Coskun M, Hori K, et al. An acetophenon and three Naphthalides from turkish Rhamnus libanoticus[J]. Phytochemistry,1991,30(12):4191-4192.
[20]Santos B V D O, Da-Cunha E V L, Chaves M C D O, et al. Phenylalkanoids from Piper marginatum[J]. Phytochemistry,1998,49:1381-1384.
[21]苏雨雷.商品贯众的品种及质量研究[D].北京：北京中医药大学中药学院中药化学系,2007.
[22]查传龙,陈兴浴.槟榔、厚朴等对肝吸虫作用的体外观察[J].南京中医学院学报,1990,(4)：30.
[23]左丽,陈若芸.鳞毛蕨属植物化学成分和药理活性研究进展[J].中草药,2005,36(9)：1427-1430.
[24]丁骁,王磊,欧阳明安.藤黄科植物化学成分的研究概况[J].化学工程与装备，2009,(1)：117.
[25]陈红云,蒋金元,王超芬,等.绵马贯众的抗菌活性研究[J].大理学院学报,2005,4(3)：22-23.
[26]常小龙.东北贯众、紫金牛和白花银背藤的化学成分研究[D].沈阳：沈阳药科大学,2006.
[27]韦四煌,方鉴,詹皓,等.贯众提取物的保肝降酶作用[J],航空军医,2004，32(3)：109-111.
[28]刘钊.绵马贯众抗感染活性成分研究[D].北京：北京协和医学院药用植物研究所生药学系,2010.
[29]徐程,于艳,孙科峰,等.绵马贯众提取物抗甲型流感病毒FM1株实验研究[J].辽宁中医药大学学报,2010,12(7)：29-31.
[30]Dona M, Dell' Aica I, Pezzato E, et al. Hyperforin inhibits cancer invasion and metastasis[J]. Cancer Res.,2004,64:6225-6232.
[31]薛惟建,王士贤,李德华.贯众抗肿瘤有效组分对P388细胞超微结构及细胞呼吸作用[J].中国药理学通报,1987，3(5)：291.
[32]Ito H, Muranaka T, Mori K, Ichthyotoxic phloroglucinol derivatives from Dryopteris fagrans and their anti-tumor promoting activity[J]. Chem Pharm Bull, 2000,48(8):1190-1195.
[33]Minoru S, Satoru K, Atsushi I. Flavon-3-ol structure-containing glycosides as histidin decarboxlase inhibitors and pharmaceutical compositions containing the inhibitors[P]. JP: 08217674A2,1996-08-27.
[34]窦昊,高志成,李薇.中药抗抑郁有效成分研究进展[J].中国药业,2010，19(21)：89.
[35]郑清明.元宝草及同属植物的生药学研究[D].上海：第二军医大学药学院生药学系,2005.
[36]陈绮.贯众抗生育有效成分的药理研究[J].天津医药,1980，(8)：188.
[37]吴先富.单刀根活性成分的研究[D].沈阳：沈阳药科大学药物化学,2006.
[38]任强.中药抗急性淋巴细胞白血病活性成分的研究[D].北京：北京协和医学院药用植物研究所生药学系,2009.
[39]高增平,陆蕴如,李国福,等.绵马贯众抗疟活性筛选[J].北京中医药大学学报,2002,25(4)：45.
[40]高增平,陆蕴如,江佩芬,等.绵马贯众部位Ⅱ的抗疟作用和急性毒性实验研究[J].北京中医药大学学报,2002,25(2)：52-53.
[41]涂小军.天然产物靛玉红及间苯三酚类衍生物的合成研究[D].西安：西北大学有机化学系,2010.
[42]中国药典委员会.中华人民共和国药典2010年版(一部).北京：化学工业出版社,2010年1月,310页.
[43]吴寿金,杨秀贤.绵马贯众化学成分的研究(Ⅰ)[J].中草药,1996,27(8)：458-459.
[44]高增平.绵马贯众有效成分的研究[D].北京：北京中医药大学中药化学系,2003.
[45]齐峰,王娥丽.常用药材绵马贯众活性成分研究[J].天津医科大学学报,2007,13(2)：191-193.
[46]陈建新,曾振灵,杨运云,等.液相色谱-串联质谱法分析绵马贯众抗禽流感病毒活性部位的化学成分[J].质谱学报,2006,27：118-120.
[47]吴寿金,杨秀贤.绵马贯众化学成分的研究Ⅱ[J].绵马贯众中间苯三酚衍生物的质谱-质谱分析[J].中草药,1997,28(12)：712-714.
[48]陈建新,曾振灵,杨运云,等.绵马贯众抗禽流感病毒药效部位的高效液相色谱-点喷雾串联质谱分析[J].分析测试学报,2005,24：73—75.
[49]陈建新,曾振灵,方炳虎,等.绵马贯众抗禽流感病毒活性部位的高效液相色谱-质谱法分析[J].分析测试学报,2008,27(6)：623—626.
[50]周荣华.东北贯众中抗菌活性成分的研究[D].哈尔滨：黑龙江大学生命科学学院,2010.
[51]高增平,马秉智,陆蕴如.绵马贯众的化学成分研究(Ⅱ)[J].北京中医药大学学报,2004,27(1)：52-53.
[52]胡昌江,叶茂,刘俊达,等.绵马贯众不同切制方法及炒炭的比较研究[J].中成药,2004,26(8)：641-643.
[53]黄勤挽,齐红艺,刘蕾,等.绵马贯众饮片“火力”与“火候”的探讨[A].中华中医药学会炮制分会.中华中医药学会第四届中药炮制学术会议论文集[C].2009：65-71.
[54]肖国君,叶利明,吴纯洁,等HPLC法测定绵马贯众中绵马贯众素的含量[J].药物分析杂志,2005,25(5)：502-504.
[55]司云珊,徐暾海,韩冬,等RP-HPLC法测定绵马贯众药材中绵马贯众素[J].中草药,2008,39(12)：1894-1895.
[56]高增平,王宝华,陆蕴如,等HPLC和TLCS测定绵马贯众抗疟有效部位中东北贯众素含量[J].中国药学杂志,2003,38(11)：878.
[57]肖国君.绵马贯众素对照品的研究及绵马贯众片的质量标准研究[D].四川：四川大学药物分析系,2004.
[58]鲁明明.中药材贯众质量标准研究[D].吉林：吉林农业大学中药,2002.
[59]张思巨,艾铁民,杨立新,等.绵马贯众贮存时间紧与药材质量相关性研究[J].中国中药杂志,1996,21(4)：206-207.
[60]侯冬岩,回瑞华,杨梅东,等.东北贯众中总黄酮的光谱分析[J].分析测试学报,2004,23(3)：110-112.
[61]柳芳.蛾眉蕨贯众的化学成分研究[D].北京：北京中医药大学,2009年.
[62]王晓丹,宗希明,吴洪斌,等.佳木斯地产白头翁、玉竹、萹蓄、长白沙参、绵马贯众、关卷术中微量元素的测定[J].佳木斯医学院学报,1997,20(4)：5.
[63]梁洪华,邓君丽.抗感颗粒中绵马贯众的鉴别和赤芍的含量测定[J].中国药事,2005,19(5)：290-292.
[64]胡海清,田宝成.抗病毒合剂的制备及质控初探[J].制剂技术,2007,16(7)：28-29.
[65]王波,胡美芳,牛胜德,等.抗感口服液的高效液相色谱鉴别[J].药物鉴定,2007,16(18)：36.
[1]高增平,王宝华,陆蕴如,等HPLC和TLCS测定绵马贯众抗疟有效部位中东北贯众素含量[J].中国药学杂志,2003,38(11)：878.
[2]吴寿金,杨秀贤,张丽,等.绵马贯众化学成分的研究(Ⅰ)[J].中草药,1996,27(8)458.
[3]高增平.绵马贯众有效成分的研究[D].北京：北京中医药大学中药化学系,2003.
[4]肖国君,叶利明,吴纯洁,等HPLC去测定绵马贯众中绵马贯众素的含量[J].药物分析杂志,2005,25(5)：502-504.
[5]司云珊,徐暾海,韩冬,等RP-HPLC法测定绵马贯众药材中绵马贯众素[J].中草药,2008,39(12)：1894-1895.
[1]中国药典委员会.中华人民共和国药典2010年版(一部).北京：化学工业出版社,2010年1月,310页.
[2]谷丽华,吴弢,张紫佳,等.应用薄层色谱-生物自显影技术评价乌药等三种中药的抗氧化活性[J].药学学报,2006,41(10)：956-962.
[3]Zijia Zhang, Liping Liao, Jeffrey Moore.Antioxidant phenolic compounds from walnut kernels (Juglans regia L.)[J].Food Chemistry,2009,113:160-165.
[4]曲建博,娄红祥,范培红.TLC-生物自显影技术在药物筛选中的应用[J].中草药,2005，36(1)：132-137.
[1]中国药典委员会.中华人民共和国药典2010年版(一部).北京：化学工业出版社,2010年1月.
[1]高增平.绵马贯众有效成分的研究[D].北京：北京中医药大学中药化学系,2003.
[2]李辉敏,阮金兰.鳞毛蕨科药用蕨类植物研究进展[J].九江学院学报,2004，3：78.
[3]左丽,陈若芸.鳞毛蕨属植物化学成分和药理活性研究进展[J].中草药,2005,36(9)：1427-1430.
[4]齐峰,王娥丽.常用药材绵马贯众活性成分研究[J].天津医科大学学报,2007,13(2)：191-193.
[5]陈红云,蒋金元,王超芬,等.绵马贯众的抗菌活性研究[J].大理学院学报,2005,4(3)：22-23.
[6]薛惟建,王士贤,李德华.贯众抗肿瘤有效组分对P388细胞超微结构及细胞呼吸作用[J].中国药理学通报,1987，3(5)：291.
[7]苏雨雷.商品贯众的品种及质量研究[D].北京：北京中医药大学中药学院中药化学系,2007.