砂仁药材质量标准研究

【作者】 李晓光；

【导师】 徐鸿华； 林励；

【作者基本信息】 广州中医药大学 ， 中药学， 2000， 硕士

【摘要】 砂仁具有化湿止痛、温脾止泻、理气安胎等作用，在临床上有着广泛的应用，仅《中国药典》（九五版，一部）记载以砂仁为主要原料的中成药就有21方。因此对于砂仁药材的质量及其标准的研究成为一项重要的基础工作，对生产和临床具有指导意义。70年代以来，国内外学者对砂仁药材的内外在质量作了大量的研究工作，取得了一定的成果，特别是徐国钧等对砂仁药材作了品种整理，确定了砂仁的正品来源，并从生药学方面对常见混伪品进行了鉴别方法研究。余竞光等不但对砂仁药材的挥发油部位进行分析，还对其非挥发性成分进行了研究。这些工作为砂仁药材质量的进一步深入研究提供了思路和线索，进而为药材质量标准研究和确立提供了理论和实验基础。 本文在上述研究基础上，并为配合国家“九五”重点科技攻关子专题——砂仁药材质量标准规范化研究，主要通过对砂仁药材内在质量的较深入、系统地研究，结合本课题其他老师的研究结果，加以系统和整理，尝试从定性和定量两个方面对砂仁药材的质量标准制定提供科学资料。 伪品和混乱品种的大量存在长期以来一直是困扰砂仁药材质量的一大问题，通过对近二十年来砂仁及其常见混伪品的鉴别研究概况总结，本文提出了利用特征性成分和DNA多态性进行定性鉴别研究的思路。因此首先对砂仁药材的化学成分进行了研究。先用少量阳春砂药材进行预试验，按极性大小分为4个提取部位，研究了各部位所含化学成分的特征，并在乙酸乙酯部位分离得到一单体化合物。然后改进提取方法，加大药材量，并进行进一步分离，得到了4个单体化合物。通过对MS、IR、¹³C-NMR、¹H-NMR等光谱解析及与已知物薄层对照等鉴定方法，确定了得到的5种单体化合物的结构。分别为：香草酸、原儿茶酸、（±）-儿茶素、槲皮甙、胡萝卜甙。接下来通过对砂仁药材正品及7种常见混伪品的薄层色谱以半定量方法进行考察，发现槲皮甙斑点为砂仁正品定性鉴别的特征斑点。又采用随机扩增多态DNA（RAPD）技术对砂仁正品及其常见的几种伪品进行鉴别研究，结果显示：PCR所得指纹图清晰，砂仁正品与伪品间呈现明显指纹差异，呈现多态性的引物占所选的 12．17％，根据这些多态性可进行鉴别分析。我们的研究结果表明，以药材中特征性成分为对照品采用簿层色谱法进行鉴定，结合分子生物学技术识别药材DNA，这样就从药材的遗传物质及由此产生的化学成分特征两方面来控制药材真伪。如果再将试验条件和方法标准化，可以作为药材快速鉴定的方法，其指标可以为药材定性鉴别指标。 目前药典对于砂仁药材的定量控制是以挥发油含量为指标的，挥发油是由多种成分组成的混合物，且性质不稳定，所以直接以此为定量指标不是、十分科学。在研究中发现，砂仁药材挥发油中乙酸龙脑酯含量很大，可考虑以乙酸龙脑酯代替挥发油作为砂仁药材的定量指标，于是对乙酸龙脑酯进行了药理作用研究和含量测定。结果显示，乙酸龙脑酯具有显著的镇痛、止泻和抑制离体家兔小肠平滑肌运动的作用，与砂仁药材的临床功效相吻合，且与挥发油作用有相关性，即证明砂仁挥发油主要是通过乙酸龙脑酯来发挥其功效的，乙酸龙脑酯应为砂仁药材的主要有效成分，应以其作为定量指标。对不同产地的三种正品砂仁和一些市售品中乙酸龙脑酯含量进行了测定，结合其药理作用的最小有效浓度对其含量限度标准提出建议。 实验中还发现，砂仁药材去挥发油的水提液对麻醉大鼠十二指肠运动有抑制作用，是有效部位之一。在阳春砂水提部位中我们分离到了单体化合物儿茶素，文献报道儿茶素有止泻作用，本文的实验研究证明儿茶素对离体兔肠运动有抑制作用，应为砂仁药材中的水提部位有效成分。用IIPLC法测定不同产地正品砂仁药材中儿茶素含量，发现其含量较低，可能不是水提部位中主要有效成分，对于水提部位主要有效成分的研究还需进一步深人。 本文运用多学科手段对砂仁药材的定性和定量质量标准进行研究，成功地分离出5个单体化合物，其中4个为首次分离；首次对砂仁中乙酸龙脑酯药理作用进行了研究；首次对砂仁中栅皮贰进行了特异性考察；首次对砂仁中儿茶 4素药理作用和含量进行研究；首次运用RAPD技术对砂仁及其混伪品进行了鉴别。对实验中出现的一些方法学问题进行了讨论，对中药材质量控制的思路提出一些思考。更多还原

【Abstract】 Villous amomum fruit have many functions in clinic such as removing moist and relieving pain, warming spleen and checking diarrhea, administering gas and preventing miscarriage. There are 21 patent medicine made from villous amomum fruit, which was reported by Chinese Medicine Law. So It’s an important base work to study the quality and quality catierion of vollous amomum fruit. This work has guiding effect to production and clinical. Scholars have made plenty of studies to internal and external quality of villous amomum fruit since 70s. Xu guojun et al. arranged species of villous amomum fruit and definited its really origin. Yu jingguang et al. analyzed volate part of villous amomum fruit and studied components separated from other parts. These works provided roads to advanced research and theory and experimental foundation for quality caterion of villous amomum fruit. This article is on the base of above-mentioned works. Its task is to supply research foundations to the establishment of quality caterion of villous amomum fruit from two aspects: quality and quantity. It’s a big problem that so many false species and chaos species have been existing for such a long time. After summarizing articles about differentiation of villous arnomum fruit, this text suggested to inditify villous amomum fruit by using characteristic component and polymorphism of DNA. Firstly, we studied the chemistry components of villous amomum fruit. The first step is pretest. We studied the character of 4 parts that were getting out from villous amomum fruit and separated a monosomie compound. Then we bettered the method and separated other four monosomic compounds. The third step is defining structure. By means of analyzing spectrum and comparing thin layer, the structures of 5 monosomic compounds were definited: vanillic acid, protocatechin acid, catechin, quercitroside, 62 and daucosterol. At last we inspected the Thin Layer Chromatography of villous amomum fruit and its common false species and discovered that the spot of quercitroside is characteristic spot. Secondly, we studied identification of villous amomum fruit by using RAPD technology. The result showed that fingerprint was darity, obvious fingerprint differences appeared between villous amomum fruit and its false species. We could identify them on the basis of these differences. At present, the fixed quantity control of villous amomum fruit is regulated by the content of its volatile oil. It is not filly credible, for volatile oil is composed of many kinds of components and its nature is not stable. We found that bomyl acetate were main components of volatile oil. Experiment showed that bornyl acetate had obvious effects as relieving pain, checking diarrhea and refraining movement of small intestine smooth muscle of rabbits. These effects coincided with the clinical function of villous amomum fruit and had correlation with effects of volatile oil. So we can draw a conclusion that borncyl acetate is the chief effective matter in villous amomum fruit, and we regarded borncyl acetate as the quantity target of villous amomum fruit. Finally we raised a piece of suggestion about content limit of borncyl acetate on the base of its content and effect data. Moreover, we found that the water extract (volatile oil removed) could inhibition the movement of duodenum of anesthetized rats. Catechin was separa更多还原