联用色谱在中药及血浆氨基酸中的应用研究

【作者】 季晓燕；

【导师】 梁逸曾；

【作者基本信息】 中南大学 ， 分析化学， 2010， 硕士

【摘要】 色谱及色谱-质谱联用技术借助新的分析理论和技术,为中药现代化、生命科学和环境科学等学科分析问题的解决提供了强有力的工具和手段。本论文从实际问题出发,从色谱及色谱质谱联用仪器入手,通过有效利用仪器分析技术提供的有用信息,分析比较了八种化湿药挥发油成分异同,同时建立了一个代谢组学的新平台并成功应用于超重、肥胖与正常儿童的判别分析。本论文的研究内容主要为以下两个部分：一、采用气相色谱-质谱联用技术分离和检测八种化湿药挥发油,在色谱分离与质谱表征的条件下借助直观推导式演进特征投影法(HELP)实现了重叠色谱峰的分辨。八种化湿药共定性出182种化学成分,占挥发油总量的85.3%以上。定性出共有组分67种,八种药材挥发性成分中均含有桉叶油醇(Eucalyptol,0.01％～77.26％).环氧石竹烯(Caryophyllene oxide, 0.02％～5.5％)和和D-柠檬烯(D-Limonene,tr-8.64％).挥发油共有成分大多数为萜类化合物及其衍生物,但其含量存在较大差异。研究结果表明采用HELP大大提高了定性、定量结果的准确度,为阐明中药的化学物质组成和科学合理用药提供了参考依据,奠定了基础。二、采用液相色谱分离(HPLC)荧光检测快速测定儿童血浆中的16种氨基酸。考察方法的线性范围、精密度、稳定性和回收率。方法成功应用于儿童血浆的分析。对于获得的复杂的色谱数据,在比较了化学计量学各种不同的模式识别方法后,成功建立了一种基于HPLC结合非相关线性判别分析(ULDA)的代谢平台用以寻找儿童超重与肥胖的代谢模式紊乱,典型相关分析(CCA)的方法被用来寻找潜在生物标记物。结果表明非相关线性判别分析(ULDA)法能够反映超重、肥胖和正常儿童氨基酸代谢谱的动态变化,建立的超重、肥胖和正常儿童判别模型总判别正确率达99.71%。并且ADMA和Thr可能是儿童超重与肥胖的潜在生物标记物。本文的研究也表明TC、TG、HDL以及LDL可能是与儿童超重与肥胖代谢模式紊乱关系最密切的临床参数。更多还原

【Abstract】 With the help of the novel theories and technologies, chromatography and chromatographic-mass spectrum prvided us with powerful tools for the resolution of analytical problems, like the modernlization of Chinese traditional medicine, biology and environmental science, and so on. In this thesis, essential oils of Eight Damp Dispersing Herbs were analyzed and compared; meanwhile, a new metabolomics platform was constructed and applied successfully in discriminant analysis of the patients and the controls. The main contents of this thesis include the following parts.(Ⅰ). The similarities and differences of essential oil components in Eight Damp Dispersing Herbs were investigated by GC-MS combined with a chemometric method, Heuristic Evolving Latent Projection (HELP).182 relatively common components were identified, which represented 85.3%of their total relative contents.67 relatively common components were identified. Eucalyptol(relative content tr～82.41%), Caryophyllene oxide(0.02%～5.5%) and D-Limonene (tr-8.64%)are the common main compounds of the eight plants. There was a lot differences in the chemical compounds among the eight damp dispersing herbs, Terpenes and their derivatives are the main components, and also in percentages on many levels. The researches show that multivariate-resolution technologies of chemometrics greatly enhanced the accuracy of the qualitative and quantitative results, giving great help to analyze the chemical compositions of Traditional Chinese Medicine and provide a basis for using the Traditional Chinese Medicine in right way.(Ⅱ). Amino acids in plasma of children were determined by high performance liquid chromatography (HPLC) and fluorescence detector. Method validation was investigated by linearity range, accuracy, stability and returns-ratio. The method was applied in the analysis of children plasma successfully. Then, for the complicated chromatography data obtained, uncorrelated linear discriminate analysis (ULDA) was selected to extract the useful information and build a robust model after the comparison of different chemometrics pattern recognition methods. Finally, a new metabolism research platform based on HPLC and ULDA was established for the first time, and Canonical correlation analysis (CCA) was employed to identify potential biomarkers. This platform was successfully applied in the discriminant analysis of overweight, obese and the controls, with a correct rate of 99.71%. ADMA and threonine might be the potential bio-markers of obese. This article also indicated that TC, TG, HDL and LDL might be the closest clinical parameter relate with metabolism pattern disorder of overweight and the obese children.更多还原