中药决明子有效成分提取分离及结构的电喷雾串联质谱（ESI-MS～n）研究

【作者】 谢亚；

【导师】 梁勇；

【作者基本信息】 华南师范大学 ， 有机化学， 2007， 硕士

【摘要】 决明子不仅具有广泛的药用价值，而且还是一味较好的保健药品，对许多疾病如：高血压、高胆固醇、习惯性便秘、抗病毒等疗效较好，因此决明子具有较好的开发应用前景。决明子中化学成分较为复杂，要研究和应用其中的有效成分，必须将其有效成分提取并进一步加以分离和精制，得到有效单体。本论文以决明子为研究对象对其有效成分进行提取、分离和结构鉴定，主要研究内容如下：1．采用新型的高压均质提取技术提取决明子中的有效成分并与回流提取、超声提取方法进行比较，通过正交实验设计优选三种提取方法提取决明子有效成分的最佳条件。在最佳条件下高压均质提取决明子中黄酮、蒽醌、多糖的提取率分别达到1.233％，0.089％，10.55％，比回流提取和超声提取含量高约30％，得出高压均质是一种提取速率快、提取效率高的提取技术，并对高压均质提取方法进行了研究。2．建立了决明子的高效液相色谱(HPLC)指纹图谱，对三种方法提取成分用HPLC指纹图谱进行比较，进一步验证高压均质是一种高效提取技术。应用高效液相色谱-电喷雾串联质谱联用(HPLC／ESI-MS~n)技术对决明子中全成分进行了分析，检测到20多种组分，结合参考文献鉴定出了其中部分组分的结构，初步建立了液-质联用技术准确、快速分析中药决明子中有效活性成分的方法。3．参照正己烷／乙酸乙酯／甲醇／水的经典溶剂系统，通过改变溶剂系统，尝试不同的溶剂系统配比，应用高速逆流色谱(HSCCC)，采用正己烷／乙酸乙酯／甲醇／水=4：1：3：2的溶剂系统，分离出了决明子中的五个蒽醌类单体成分，经HPLC检测其纯度分别为98％，95％，96％，95％和96％。应用电喷雾串联质谱(ESI-MS~n)技术对高速逆流色谱分离出决明子中的五个单体成分结构进行了研究，结合参考文献鉴定出五个成分分别为：1，2，6-三羟基-7，8-二甲氧基-3-甲基蒽醌；1，2，6，8-四羟基-7-甲氧基-3-甲基蒽醌；2-羟基-1，6，7，8-四甲氧基-3-甲基蒽醌；2，6-二羟基-1，7，8-三甲氧基-3-甲基蒽醌；1，2-二羟基-6，7，8-三甲氧基-3-甲基蒽醌。更多还原

【Abstract】 Cassia obtusifolia have been widely used in traditional Chinesemedicine for the treatments of hypertension activity, higher cholesterin,astriction, antimicrobial, etc. It is highly valuable to study the chemicalconstituents of Cassia obtusifolia. The extraction and separation of themain functional components from Cassia obtusifolia is necessary. The maincontents are belows.1. The extraction of effective components from Cassia obtusifolia atroom temperature was carried out by High Pressure HomogenizationExtraction (HPHE) method. Compared to heat reflux extraction method andultrasonic extraction method, the best way for the extraction of effectivecomponents from Cassia obtusifolia was HPHE method. The optimumextraction conditions of HPHE method were obtained by orthogonal test.Under the optimal conditions of HPHE method, the yield of flavonoids,anthraquinones and polysaccharide were up to 1.233%, 0.089%, 10.55%,which were at least 20 to 30% higher than the figures obtained from thereflux extraction and ultrasonic extraction. The HPHE was proved to be anew process for extraction of the effective components of traditionalChinese medicine.2. HPLC fingerprint of Cassia obtusifolia was established by Highpressure liqid chromatography. The chemical components were analyzed byHPLC-electrosprary ionization-mass spectrometry (HPLC/ESI-MS~n) and thestructure were elucidated according to mass-to-charge ratio (m/z) ofmolecule ionic peak, the fragmental and reference. There were more than 20chemical components isolated, but only 10 chemical components weredetermined. The data obtained for chemical component using this methodprovide a Cassia obtusifolia profile database useful for the rapididentification of Cassia obtusifolia. The method of HPLC/ESI-MS~n is rapidand sensitive. It is suitable to application in the field of natural productsanalysis.3. The separation of Cassia obtusifolia was carried out by High-speed countercurrent chromatography (HSCCC), with the solvent systemcomposed of n-hexane-ethyl acetate-methanol-water (4:1:3:2, v/v) anda total of five well-separated peaks were obtained in the HSCCCchromatogram and their purities were determined by HPLC-UV absorptionspectrometry, the purities of five peaks were 98%, 95%, 96%, 95%, 96%,respectively, which further verified HSCCC was a versatile separationsystem for active ingredient of plant. These peaks were characterized byESI-MS~n and the data compared with the reference standards. Five peakswere identified as 1, 2, 6-trihydroxy-7, 8-dimethoxy-3-methylanthraquinone1, 2, 6, 8-tetrahydroxy-7-methoxy-3-methylanthraquinone, 2-hydroxy-1, 6,7, 8-teramethoxy-3-methylanthraquinone, 2, 6-dihydroxy-1, 7, 8-trimethoxy-3-methylanthraquinone and 1, 2-dihydroxy-6, 7, 8-trimethoxy-3-methyl-anthraquinone, respectively.更多还原