化学计量学用于几种复杂中药体系分析及其指纹图谱的研究
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摘要
随着中药研究的不断深入，中药现代化得到不断推进，中药所含物质基础的现代化和中药质量控制的现代化成为当前中药现代化研究的两个重要方面。然而，中药是十分复杂的黑色分析体系，其药效是通过众多化学成分的协同作用的综合结果，到目前为止，人们还远没有弄清楚中药的各种化学成分，但中药行业的发展却迫切需要能够综合评价中药质量和进行中药质量控制的指标。所以，人们在充分利用各种现代化分析技术和方法进行中药化学成分研究的同时积极广泛地开展中药质量控制的现代化研究。
在中药化学的研究中，色谱是中药化学成分分离检测的重要工具，但影响色谱分离的因素很多，而中药的提取物往往又是灰色或黑色体系，最佳色谱分离条件往往难以获得。在本文的第二章，用化学计量学中的实验设计方法优化了蚕蛹虫草提取物的色谱分离条件，较快地找到了蚕蛹虫草提取物的最佳色谱分离条件，并将该色谱条件应用于蚕蛹虫草和冬虫夏草水溶性成分的分析，取得满意的分离效果。本章的研究为复杂中药分析体系的色谱分离条件的优化提供了一定的参考。
由于许多道地药材的短缺，人们开始寻求和培养人工的同属药材，如何从化学活性物质含量的角度来客观合理地判断人工培养药材与道地药材间的差异性就成为中药研究中十分重要的问题。由于药材中众多的组分未知，要找到所有组分的标准物质来进行一一对照分析几乎不可能。在标准物质不完全知道的情况下，有效地对替代药材和道地药材的化学指纹图谱中各组分进行归属分析就十分必要了。本文的第三章，基于相同检测条件下，相同物质具有相同的光谱(或质谱)，充分利用HPLC-DAD提供的包含色谱和光谱信息的二维数据，利用光谱相关色谱，对蚕蛹虫草和冬虫夏草的水溶性成分进行相对归属分析，找到了它们的共有组分和不同组分，并对部分组分进行了定性定量分析，为蚕蛹虫草和冬虫夏草进一步的药理药效研究打下了基础，同时为人工培养药材与道地药材间的比较分析提供了新的分析思路。
蚕蛹虫草和冬虫夏草中的甾醇类物质具有十分重要的药理活性，而麦角甾醇是真菌类的特征甾醇，具有抗肿瘤等作用，在第四章，比较了热水回流法和超声波法对蚕蛹虫草中麦角甾醇提取的影响，并对蚕蛹虫草和冬虫夏草的麦角甾醇进行了定性定量分析。
白术(Atractylodes macroeephala koidz)为菊科植物卜
根茎，具有健脾益气，燥湿利水，安胎，lt..汗的功效，为常用中、
挥发油是其重要的活性成分。在第五章中，以‘，.维联用色谱仪器GC一MS
和化学计量学发展的多元分辨技术对白术的挥发油进行了定性定量
分析，并比较了水蒸气蒸馏法和渗辘法对挥发油提取的影响，比较了
三个产地白术挥发油指纹图谱的异同，为白术生药的质量控制和鉴别
的指纹
在
谱建立提供了科学依据。
国，鱼腥草广泛分布于各地，而且鱼腥草注射液生产厂家众
多，大部分的省份均有鱼腥草注射液生产厂家。不同的厂家生产鱼腥
草注射液时，所用的药材部位也不尽相同，为制定出合乎客观要求的
鱼腥草注射液指纹图谱，有必要对不同产地的鱼腥草以及鱼腥草不同
部位的挥发油进行对比分析。本文第六章以二维联用色谱仪器GC一MS
和多元分辨技术对云南省不同地方产鱼腥草鲜草、相同批次鱼腥草鲜
草的不同部位的挥发油进行了定性定量分析，同时从整体上比较了它
们的指纹图谱，掌握了云南省鱼腥草挥发油较全面的化学成分信息及
指纹图谱信息，为鱼腥草注射液指纹图谱标准的制定提供了十分有用
的信息。
In recent decades, traditional Chinese medicines (TCMs) have attracted more and more people because of their advantages of low toxicity and high bioactivity. TCM is a very complicated system, which contains many constituents. There are still many constituents can't be identified clearly. The pharmacological activities of TCMs are not the action of single compound, but the interaction of many ingredients in it. However, traditional methods, by which only few components were determined, are still used for the quality control, but it is unreasonable from the view that functions of TCM emphasis it's unity. On the other hand, it is impossible to qualitatively and quantitatively determine every component accurately from a TCM on account of it's complexity. To improve and control the quality of TCM, a good and compositive method is strongly required in TCM industry. On the base of the modern and advanced instruments and techniques, the research about the modernization of TCMs was carried out deeply and widely. On o
ne side, the chemical constituents of TCMs were determined. And on other side, the chemical fingerprint of a TCM is studied, which can be used as an approach to the quality control and technologic standardization of the TCM.
In the study of TCMs, chromatography is a very important method in separating and determining the constituents in TCMs. However, it is difficult to obtain the optimum chromatographic separation conditions, especially for a complicate TCM system, because there are many parameters in the separation experiment to be concerned. A chemometric approach, called uniform design, was applied to find out the optimum chromatographic separation conditions of the extraction from Cordyceps Militaris. Satisfactory results were obtained when the optimum chromatographic conditions were applied to determine the water-soluble constituents of Cordyceps Sinensis and Cordyceps Militaris. It is easy and convenient to obtain the optimum chromatographic conditions for a TCMs system by this method.
Today many cultivated TCMs were used because of the short of natural TCMs. In fact, there is some differentiation between the two kinds of TCMs. It is very important to analyze the common and different constituents between natural and cultivated TCMs. In chapter three, a method called spectral correlative chromatography was applied to analyze water-soluble constituents between cultivated Cordyceps Militaris and natural Cordyceps Sinensis. Compared with the conventional methods, the method is easier and more convenient since it fully utilized both information from spectrum and chromatography obtained from hyphenated instruments. It will be a good method for analyzing and comparing the cultivated and natural TCMs. In this chapter, some important constituents of Cordyceps Sinensis and Cordyceps Militaris were determined qualitatively and quantitatively. A fat-soluble compound, ergosterol, which is a very important active component of cultivated Cordyceps Militaris and natural Cordyceps Sinensis, was also detected qualitatively and quantitatively by HPLC method in chapter four.
Atractylodes macrocephala koidz is an important TCM in China, in which essential oil is very important active constituents. To avoid the effects of overlapping peaks and of background on the qualitative and quantitative analysis, the GC-MS data from essential oil of Atractylodes macrocephala koidz were resolved with chemometric resolution method such as heuristic evolving latent projections (HELP) method. By using the HELP method, the components of Atractylodes macrocephala koidz from three different places, Huaihua, Pingjiang and Zhejiang, and those extracted by two different extracting methods, water distillation method and seeping method, were determined qualitatively and quantitatively. The fingerprints of three Atractylodes macrocephala koidz are studied in details. The results in this chapter will be very important in the development of the fingerprint for identification and quality control of this TCM.
To establish a fingerprint standardizat
在中药化学的研究中，色谱是中药化学成分分离检测的重要工具，但影响色谱分离的因素很多，而中药的提取物往往又是灰色或黑色体系，最佳色谱分离条件往往难以获得。在本文的第二章，用化学计量学中的实验设计方法优化了蚕蛹虫草提取物的色谱分离条件，较快地找到了蚕蛹虫草提取物的最佳色谱分离条件，并将该色谱条件应用于蚕蛹虫草和冬虫夏草水溶性成分的分析，取得满意的分离效果。本章的研究为复杂中药分析体系的色谱分离条件的优化提供了一定的参考。
由于许多道地药材的短缺，人们开始寻求和培养人工的同属药材，如何从化学活性物质含量的角度来客观合理地判断人工培养药材与道地药材间的差异性就成为中药研究中十分重要的问题。由于药材中众多的组分未知，要找到所有组分的标准物质来进行一一对照分析几乎不可能。在标准物质不完全知道的情况下，有效地对替代药材和道地药材的化学指纹图谱中各组分进行归属分析就十分必要了。本文的第三章，基于相同检测条件下，相同物质具有相同的光谱(或质谱)，充分利用HPLC-DAD提供的包含色谱和光谱信息的二维数据，利用光谱相关色谱，对蚕蛹虫草和冬虫夏草的水溶性成分进行相对归属分析，找到了它们的共有组分和不同组分，并对部分组分进行了定性定量分析，为蚕蛹虫草和冬虫夏草进一步的药理药效研究打下了基础，同时为人工培养药材与道地药材间的比较分析提供了新的分析思路。
蚕蛹虫草和冬虫夏草中的甾醇类物质具有十分重要的药理活性，而麦角甾醇是真菌类的特征甾醇，具有抗肿瘤等作用，在第四章，比较了热水回流法和超声波法对蚕蛹虫草中麦角甾醇提取的影响，并对蚕蛹虫草和冬虫夏草的麦角甾醇进行了定性定量分析。
白术(Atractylodes macroeephala koidz)为菊科植物卜
根茎，具有健脾益气，燥湿利水，安胎，lt..汗的功效，为常用中、
挥发油是其重要的活性成分。在第五章中，以‘，.维联用色谱仪器GC一MS
和化学计量学发展的多元分辨技术对白术的挥发油进行了定性定量
分析，并比较了水蒸气蒸馏法和渗辘法对挥发油提取的影响，比较了
三个产地白术挥发油指纹图谱的异同，为白术生药的质量控制和鉴别
的指纹
在
谱建立提供了科学依据。
国，鱼腥草广泛分布于各地，而且鱼腥草注射液生产厂家众
多，大部分的省份均有鱼腥草注射液生产厂家。不同的厂家生产鱼腥
草注射液时，所用的药材部位也不尽相同，为制定出合乎客观要求的
鱼腥草注射液指纹图谱，有必要对不同产地的鱼腥草以及鱼腥草不同
部位的挥发油进行对比分析。本文第六章以二维联用色谱仪器GC一MS
和多元分辨技术对云南省不同地方产鱼腥草鲜草、相同批次鱼腥草鲜
草的不同部位的挥发油进行了定性定量分析，同时从整体上比较了它
们的指纹图谱，掌握了云南省鱼腥草挥发油较全面的化学成分信息及
指纹图谱信息，为鱼腥草注射液指纹图谱标准的制定提供了十分有用
的信息。
In recent decades, traditional Chinese medicines (TCMs) have attracted more and more people because of their advantages of low toxicity and high bioactivity. TCM is a very complicated system, which contains many constituents. There are still many constituents can't be identified clearly. The pharmacological activities of TCMs are not the action of single compound, but the interaction of many ingredients in it. However, traditional methods, by which only few components were determined, are still used for the quality control, but it is unreasonable from the view that functions of TCM emphasis it's unity. On the other hand, it is impossible to qualitatively and quantitatively determine every component accurately from a TCM on account of it's complexity. To improve and control the quality of TCM, a good and compositive method is strongly required in TCM industry. On the base of the modern and advanced instruments and techniques, the research about the modernization of TCMs was carried out deeply and widely. On o
ne side, the chemical constituents of TCMs were determined. And on other side, the chemical fingerprint of a TCM is studied, which can be used as an approach to the quality control and technologic standardization of the TCM.
In the study of TCMs, chromatography is a very important method in separating and determining the constituents in TCMs. However, it is difficult to obtain the optimum chromatographic separation conditions, especially for a complicate TCM system, because there are many parameters in the separation experiment to be concerned. A chemometric approach, called uniform design, was applied to find out the optimum chromatographic separation conditions of the extraction from Cordyceps Militaris. Satisfactory results were obtained when the optimum chromatographic conditions were applied to determine the water-soluble constituents of Cordyceps Sinensis and Cordyceps Militaris. It is easy and convenient to obtain the optimum chromatographic conditions for a TCMs system by this method.
Today many cultivated TCMs were used because of the short of natural TCMs. In fact, there is some differentiation between the two kinds of TCMs. It is very important to analyze the common and different constituents between natural and cultivated TCMs. In chapter three, a method called spectral correlative chromatography was applied to analyze water-soluble constituents between cultivated Cordyceps Militaris and natural Cordyceps Sinensis. Compared with the conventional methods, the method is easier and more convenient since it fully utilized both information from spectrum and chromatography obtained from hyphenated instruments. It will be a good method for analyzing and comparing the cultivated and natural TCMs. In this chapter, some important constituents of Cordyceps Sinensis and Cordyceps Militaris were determined qualitatively and quantitatively. A fat-soluble compound, ergosterol, which is a very important active component of cultivated Cordyceps Militaris and natural Cordyceps Sinensis, was also detected qualitatively and quantitatively by HPLC method in chapter four.
Atractylodes macrocephala koidz is an important TCM in China, in which essential oil is very important active constituents. To avoid the effects of overlapping peaks and of background on the qualitative and quantitative analysis, the GC-MS data from essential oil of Atractylodes macrocephala koidz were resolved with chemometric resolution method such as heuristic evolving latent projections (HELP) method. By using the HELP method, the components of Atractylodes macrocephala koidz from three different places, Huaihua, Pingjiang and Zhejiang, and those extracted by two different extracting methods, water distillation method and seeping method, were determined qualitatively and quantitatively. The fingerprints of three Atractylodes macrocephala koidz are studied in details. The results in this chapter will be very important in the development of the fingerprint for identification and quality control of this TCM.
To establish a fingerprint standardizat
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