用红外光谱对不同来源的仙鹤草进行分析和评估.doc

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1、2014年度本科生毕业论文（设计）英语文献翻译院 系： 理学院化学系 专 业： 化 学 年 级： 2011级 学生姓名： 何宣鹏 学 号： 201101020132 导师及职称： 王泽锋 2014年4月2014 Annual Graduation Thesis (Project) of the College Undergraduate Translation of EnglishLiteratureDepartment: Dpartment of Chemistry, School of ScienceMajor: ChemistryGrade: 2011Students Name: He

2、XuanpengStudent No.: 201101020132Tutor: Wang ZefengApril, 201423Analysis and Assessment of Agrimonia Pilosa Ledeb fromDifferent Sources Using FT-IR Spectroscopy 用红外光谱对不同来源的仙鹤草进行分析和评估Bao-qing Wang (王宝庆)，Zhe-xiong Jin(金哲雄)(College of Pharmacy，Harbin University of Commerce，Harbin 150076，China)(哈尔滨商业大学,

3、药学院，哈尔滨150076，中国)ABSTRACT摘要To get the IR spectrums of Agrimonia Pilosa Ledeb(APL) from China and Japan 从中国和日本地区得到仙鹤草的红外光谱，areas，and to find out the characters of IR spectrums through the content of diferent 通过不同化学成分的含量找出红外谱图特征，chemical constituents，to provide a fast and effective analysis method mon

4、itor the 提供一个快速和有效的分析方法，监测仙鹤草传统中医治疗inherent qualities of traditional Chinese medicine APLFourier transform Infrared 的内在品质。 用傅里叶变换红外光谱（FT-IR）Spectroscopy (FT-IR) was applied to detect sample of APL from China and Japan 技术检测中国和日本地区的仙鹤草样品。areas This study showed that the IR spectra of APL from China an

5、d Japan areas 这项研究表明，来自中国和日本地区的仙鹤草的红外光谱有其独特的红have their unique IR fingerprint featuresThe contents of tannin and calcium 外指纹特征。 从中国和日本得到的仙鹤草phosphate in APL from China is diferent APL from JapanSo FT-IR is a very 中的鞣酸和钙的含量都不相同。 所以，傅里叶变换红外光谱 quick,efective and well repetitive method for monitoring an

6、d distinguishing the 傅里叶变换红外光谱是一种很快的，有效的和重复的监测、识别传统中医的方法。traditional Chinese medicineKeywords: Agrimonia Pilosa Ledeb ;Fourier Transform Infrared Spectroscopy 关键词： 仙鹤草; 傅里叶变换红外光谱;(FT-IR)；chemical constituents 化学成分1 Introduction1 前言Agrimonia Pilosa Ledeb(APL) is a widely used medicinal plant by 仙鹤草（A

7、PL）是一种广泛使用的药用植物asian countries and has been reported to possess various biological 亚洲国家已经报道其具有有多种生物学活性。activities In Asia，APL is used in folk medicine as haemostatic，tonic for 在亚洲，仙鹤草在民间是一种作为止血，补虚，涩的腹泻，利尿的药物。asthenia，astringent in diarrhoea，and diureticFor example，APL has been 例如，仙鹤草已用于血液，心used for

8、blood，cardiovascular，gastrointestinal，genitourinary，血管，胃肠道，泌尿生殖系统，炎症，肝，呼吸道，皮肤和其他一些疾病。inflammatory，liver，respiratory tract，skin and some other conditionsIn addition, APL is also a famous medicinal plant used for cancer 此外,仙鹤草是著名的药用植物, 用于治疗癌症，therapy in Japan and the antitumor activity of the methanol

9、 extract of 在日本，用甲醇从仙鹤草根中提取抗肿瘤活性物质， the roots of this plant has been widely reported In China，this plant is 已经被普遍报道。在中国, 这种植物traditionally used to suppress diarrhoea，to reduce gastric ulcers，to 传统上是用来抑制腹泻， 治疗胃溃疡， relieve inflammation，to improve eyesight，to calm the liver，to detoxify 消炎， 明目， 平肝， 解毒的。

10、poison from bodyThe tannin is the main component of the plant that acts 单宁酸是仙鹤草的主要成分之一，against several rodent tumors and it has been suggested that the immune 研究表明，单宁酸是调整免疫系统和system adjustment is important in the antitumor effect of tannin1 抗肿瘤的主要成分。In recent years it is reported that FT-IR is used

11、in traditional Chinese 近年来，据报道， 傅里叶变换红外光谱在传统中医medicine on some aspects , such as identifying counterfeits，controlling 的一些方面有所应用， 如假冒产品的识别，质量的控制，qualities，forecasting stability , etcAlthough traditional Chinese 稳定性的预测等。 虽然中医是非常复杂的体系，medicine is very complicated systems，the fingerprint characters in F

12、T-IR 但只要其所含化学成分的种类和含量不同， spectra will show differences，as long as the categories or contents of 红外光谱指纹特征会显示出差异。their contained chemical components are differentUsing these differences of fingerprint characters can identify，compare and control the 使用这些指纹特征的差异可以快速识别，medicinal materials and their corre

13、sponding products quickly，accurately 比较和对照药材及其相应的产品，and effectively2准确和有效。Now the mainstream analysis method for monitoring the qualities of 现在的质量检测中药化学成分的chemical constituents in herb medicine is GC MS method ，which can 主流分析方法是气相色谱质谱法，offer much detailed information of chemical componentsHowever，th

14、e 此方法提供了更详细的的化学成分的信息。low reproductivity，expensive instrument and rigorous experiment 然而，低生产力，昂贵的仪器和严格的实验条件condition limit it to generalize in some degreeAnd only a few 在某种程度上受到了限制。components selected to test cannot reflect the real qualities of the 只有少数选定的成分测试不能反映复杂样品的真实性，complicated samples as trad

15、itional Chinese Medicine 不能作为中医的有效依据。efficientlyTherefore，it is the hot issue to find a fast and effective 因此，找到充分的评估样品的快速和有效的analysis method for evaluating fully the qualities of such samplesFT-IR 分析方法是一个热点问题。spectroscopy presenting the whole features and fingerprint characters of a 红外光谱呈现一个样本的整体特征

16、和指纹特征，sample can achieve this goal perfectlyThe analysis of FT-IR not only 可以很好地实现这一目标。can reveal clearly the main compositions in the sample but also can 红外光谱分析不仅可以显示样品中的主要成分，compare the differences of categories and quantities of chemical 而且还可以比较在相似样品中的的不同种类和数量的化学成分。constituents in the very simila

17、r samples3.Here we studied the differences of main constituents of APL using 本文用红外光谱指纹的方法研究仙鹤草主要成分的差异。FT-IR fingerprint method The purpose of this study is to develop a 本文研究的目的是开发一种 rapid，accurate and feasible analysis me thod to appraise integrally the 快速，准确和可行的分析方法来评价和inherent qualities of APL and

18、 compare sample of chemical constituents 比较来自中国和日本地区的仙鹤草样品内在品质和化学成分。APL from China an d Japan areas2 Experiment 2.实验部分2.1 Apparatus2.1仪器 Spectrometer：Spectrum GX FT-IR system (PerkinElmer)，equipped 光谱仪：GX红外光谱系统（原子吸收光谱仪），with a DTGS detector was usedScan range from 400 cm-1 to 4 000 配备DTGS检测仪。扫描范围：40

19、0 cm-1 -4 000 cm-1，cm-1 with a resolution of 4 cm-1 the velocity of OPD 0.2 cms，且分辨率为4 cm-1。 光程差速：0.2 cm/s，Spectra are obtained from the accumulation of a total of 16 scans扫描光谱：16个。2.2 Samples2.2样品obtained from The roots of plants were collected from different localities of Harbin district，China and

20、 Okayama district，Japan，and authenticated by Professor Zhe-xiong Jin from college of pharmacy， 哈尔滨商业大学药学院的教授金哲雄收集了哈尔滨地区，中国和冈山地区，日本不同地区的仙鹤草的根进行了实验，Harbin university of commerceVoucher specimens were deposited in 标本保存在大学药学院的实验室。college of pharmacy of this laboratory2.3 Chemical2.3化学药品Calcium phosphate

21、，Potassium bromide and Tannin were purchased 磷酸钙， 溴化钾，from Sigma-Chemical Company单宁酸是购自西格玛化工公司。2.4 Procedure2.4实验步骤The sample was directly frozen and dried into a powder，then， 样品直接冷冻干燥成粉末，然后，用KBr粉末混合，压制成片剂。blended with KBr powder，pressed into a tabletAfter that，collected the 收集的所有样品的红外光谱。infrared sp

22、ectra of all samplesAll the second derivative IR spectra were 13-point second derivative 所有的二阶导数红外光谱在光谱室温下有13个点。IR spectra taken at room temperature4.3 Result3实验结果3.1 IR spectra analysis of agrimonia Pilosa Ledeb in different areas3.1不同地区的仙鹤草的红外光谱分析Fig1 presents the IR spectra of APL in different ar

23、eas under the room 图1给出了不同地区的仙鹤草在室温下的红外光谱。temperatureFrom the spectra we can see clearly that positions and 从光谱中我们可以清楚地看到，位置和主要的特异性条带的形状shapes of the main specific bands are quite similar to each other， except 彼此是非常相似的，at 1736 cm-1 ，1438 cm-1 and 519 cm-1Fig.2 is in the secondary 除了在1736 cm-1 ，1438

24、cm-1 和519 cm-1之外。derivative IR spectra that amplify tiny differences of IR spectra at 1 736 图2是在二级导数红外光谱放大微小的红外光谱差异在1736 cm - 1和1438 cm - 1。cm-1 and 1438 cm-1 1438 cm-1 (C-H deformation)， 1736 cm-1 (C = O 1438 cm-1 (C-H变形)， 1 736 cm-1(C = O收缩)，stretching)，the band of the fingerprint region at about

25、1400 cm-1 1 在约1400 cm-1处1750 cm-1的指纹区带，750 cm-1,So this is a good fingerprint to distinguish APL from China and 所以在中国和日本，这是一个很好的指纹识别应用。Japan519 cm-1 is another special peak 519 cm-1的是另一组特殊的峰值。吸光度波长（cm-1） 图1：来自不同地区仙鹤草的红外光谱：（a）中国1；（b）中国2；（c）日本1；（d）日本2吸光度波长（cm-1） 图2：来自不同地区仙鹤草的二阶导数红外光谱：（a）中国1；（b）中国2；（c）

26、日本1；（d）日本23.2 IR spectra of Agrimonia Pilosa Ledeb in different ages3.2仙鹤草在不同生长段的红外光谱Fig3 is the IR spectra of APL from China in different agesOnly a 图3是来自中国不同年龄的仙鹤草的红外光谱。slight difference between each leak can be observed ，the peaks of 1736 可以观察到, 只有轻微的差异,1736 cm-1处的峰cm-1 strengthens gradually alon

27、g with the growth age increaseIR spectra 随着年龄的增加逐渐增强。results may indicate that the components in APL from china will 红外光谱结果表明,来自中国的仙鹤草有衰老成分的积累。accumulate with the agingThe analysis results of the peaks at 1736 cm-1 在1736 cm-1处的峰的分析结果表明，may indicate that the major changed constituents are the compoun

28、ds of 主要的改变成分的化合物是磷酸钙和单宁酸。calcium phosphate and tannin.吸光度波长（cm-1） 图3：来自中国不同年龄阶段的仙鹤草的红外光谱：（a）4年；（b）3年；（c）2年；（d）1年 Fig.4 is the IR spectra of APL from Japan in different agesFig.5 is 图4是来自日本的不同年龄的仙鹤草的红外光谱。the secondary derivative IR spectra of the plant from Japan in different 图5是日本在不同年龄的仙鹤草的二阶导数谱。ag

29、esThe intensity of absorption peak at 1620 cm-1 changes gradually 吸收峰强度在1620 cm-1处着年龄的增加逐渐增强。along with the growth age increaseThe growth age of influence on the 日本的仙鹤草年龄的增长对成分影响很小。constituents of APL from japan is very small吸光度波长（cm-1） 图4：来自日本不同年龄阶段的仙鹤草的红外光谱：（a）4年；（b）3年；（c）2年；（d）1年吸光度波长（cm-1） 图5：仙鹤

30、草在不同时期的二阶导数红外光谱：（a）4年；（b）3年；（c）2年；（d）1年3.3 IR spectra of Agrimonia Pilosa Ledeb in different Ages3.3仙鹤草在不同年龄阶段的红外光谱 According to the literature report，the main medicinal compounds of 根据文献报告，仙鹤草的主要药用化合物包括单宁酸。APL include the tanninSo the tannin has been analysed by FT-IRAs 因此，用红外光谱对仙鹤草进行分析。shown in Fi

31、g.6，Comparison of the FT-IR spectrum at 1736 cm-1，1447 如图6所示，对1736 cm-1，1447 cm-1的红外光谱进行比较。cm-1 1241 cm-1 reveals that the content of tannin in the plant from 1241 cm-1处峰显示，来自中国的仙鹤草单宁酸的含量China is higher than the plant from JapanThis conclusion is consistent 高于来自日本的。 这一结论with the result of the micros

32、copical identification of the plant与植物的显微鉴定结果一致。 吸光度波长（cm-1） 图6：仙鹤草和单宁酸红外光谱：（a）中国；（b）日本；（c）单宁酸图7：来自中国和日本的仙鹤草的显微鉴定（a）日本；（b）中国 From Fig.7，we can see clearly that the black fraction(rich tannin)from china 从图7，我们可以清楚的看到黑色分数（富含单宁酸）plant is obvious higer than Japan plant中国的仙鹤草明显比日本的仙鹤草高。3.4 IR spectra of

33、Agrimonia Pilosa Ledeb and calcium phosphate3.4仙鹤草中磷酸钙的红外光谱Fig8 shows the peak at 1034 cm-1 ，564 cm-1 ，469 cm-1 belonging 图8显示了在1034 cm-1 ，564 cm-1 ，469 cm-1处磷酸钙的特征吸收峰，to the characteristic absorption of calcium phosphate；and the intensity of peak in Japan plant was lightly stronger than that of Chi

34、na，the relative 日本仙鹤草的光强度稍微比中国强，content of calcium phosphate is higher than that of China，These results 磷酸钙的含量相对高于中国，are in agreement with that the conclusion of the microscopical 这些结果与仙鹤草的显微鉴定结论是一致的。identification of APLThe content of cluster crystal of calcium phosphate in Japan plant is slightly

35、higer the content of calcium phosphate of 日本仙鹤草的磷酸钙簇晶含量略高于中国仙鹤草的针状结晶磷酸钙含量acicular crystal in china plant(Fig9)This difference could be due to the (见图9)。 这种差异可能是由于气候的原因引起的。climate reason吸光度波长（cm-1） 图8：仙鹤草和磷酸钙的红外光谱：（a）中国；（b）日本；（c）磷酸钙图9：来自中国和日本的仙鹤草的显微结构（a）日本；（b）中国4 Discussion4 探讨Using correlation analy

36、sis technique to treat a series of IR spectrathe 采用相关分析技术来探讨一系列的红外光谱。identification in Chinese Traditional Medicine can be simplifiedThese 在中药鉴定时可以简化。analyses show that the distinctive IR feature in APL from China and 这些分析表明，从中国和日本获得的仙鹤草Japan had been obtained at 1 736 cm-1,1 438 cm -1and 519 cm-1 T

37、hese 在1 736 cm-1,1 438 cm -1， 519 cm-1处具有独特的红外光谱特征。bands are contribute to the variation of chemical components 这些特征有助于化学成分变化的分别。respectivelyThe following experiments can provide powerful evidence 以下的实验提供了强有力的证据表明，that the main differences of chemical components are tannin and calcium 化学成分的主要区别是丹宁酸和

38、磷酸钙。phosphateIn the IR spectra，the characteristic peaks of tannin and 在红外光谱中，单宁和磷酸钙的特征峰在l736 cm-1 ,1447 cm-1，calcium phosphate could be at l736 cm-1 ,1447 cm-1，1241 cm-1 1241 cm-1处。The contents of tannin and calcium phosphate contained in the China 中国和日本仙鹤草中单宁酸和磷酸钙的含量plant and Japan plant can be ide

39、ntified according to the intensities of the 可以根据其强度确定吸收峰。absorption peaks. The content of tannin reveals that APL from China have 从单宁酸的含量表明，more medicinal ingredients than the plant from Japan，the high content of 中国的仙鹤草比日本的仙鹤草有更多的药用成分， calcium phosphate is easier to reduce the content of other medic

40、inal 磷酸钙含量高，更容易降低其它药用成分的含量。ingredientsBased on these contentdifferences，the results is the same 这些内容的差异，结果是与从传统的仙鹤草显微鉴定相同。with that from conventional microscopical identification of APLThe microscopical identification of the crude plant or the plant powder is one 天然植物或植物粉末显微鉴别是一of the most important

41、 techniques to authenticate a crude plant drugBut 是一项重要的鉴定天然药物的技术 。these bands of IR spectra in the different growth age of the plant are quite 但这些波段的红外光谱在植物的不同生长年限的彼此非常相似。similar to each otherwhich may embody the complexity of the chemical 这可能体现了复杂的化学成分，composition，the secondary derivative and two-

42、dimensional IR spectra 二阶导数和二维红外光谱可能是一个不错的选择.could be a good choice5Two dimensional infrared spectroscopy is a 二维红外光谱技术是一种非线性红外光谱技术,nonlinear infrared spectroscopy technique that has the ability to correlate 必须在凝聚相系统的振动模式体现相关的能力。vibrational modes in condensed-phase systemsThis technique provides 这种技

43、术提供了information beyond linear infrared spectra，by spreading the vibrational 信息超线性的红外光谱，information along multiple axes, yielding a frequency correlation spectrum通过传播沿多轴振动信息，产生一个频率相关谱。 Generally，the secondary derivative IR spectra can obviously enhance 一般情况下，二阶导数谱能明显提高光谱分辨率和放大的红外光谱的微小the spectrum resolution an d amplify tiny differences of IR spectra， also 的差别，provide a powerful method for the quality control of APL from China and Japan6-7 也提供来自中国和日本的仙鹤草的质量控制的有效方法。 Currently，the main method for identifying various APL is the 目前，用于识别不同的仙鹤草的主要方法是micro