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1、成都中医药大学 博士学位论文 基于桑叶桑椹功效的多元光谱色谱信息研究及桑叶品质评价 姓名：游元元 申请学位级别：博士 专业：中药学 指导教师：万德光 2008-05 摘 要 摘 要 中华人民共和国药典2005 年版第一部规定，桑叶 Folium Mori 与桑椹 Fructus Mori 的基原仅一个，即桑科植物桑Morus alba L 。通过文献查考及 资源、品种调查发现，本草记载供药用的桑历来不止一种，而药材流通领域的 桑类药材基原也不单一。 桑叶与桑椹两味中药主治功能不同。通过免疫调节、镇咳祛痰、对胰岛 素受体的影响及体外抗氧化等药效学实验发现，两味药材在在上述药效学方面 既有区别，又
2、有共通之处，体现了同属植物多器官入药的特点。 中药的光谱与色谱图具有物种特异性与个体相似性特征，其图谱面貌可反 映中药的化学成分群体状况。首次采用多元光谱色谱法对桑叶桑椹药材进行多 品种多极性部位研究及两味药材的横向比较。采用红外光谱法、薄层色谱法、 高效液相色谱法分别建立了桑叶、桑椹药材的整体性化学成分特征表达图谱， 再对两味桑类药材的不同极性部位进行了紫外光谱、薄层色谱及高效液相色谱 的比对研究。在此基础上将相同极性部位的桑叶与桑椹药材图谱进行对比。从 实验结果看，仅采用一种光谱或色谱方法评价中药材的整体品质有其局限性。 将药材化学组分按极性分为几个部位，分别研究不同极性部位的特征，所获取
3、 的信息量明显增多。 除 TLC 法外， 其余方法都进行了不同样品间的相似度评价。 将某一样品不同极性部位所有的相似度值作为样本特征进行聚类分析，结果更 能代表样品的总体相似情况。 历代本草强调霜桑叶、冬桑叶的品质佳。首次采用高效液相色谱法对不同 采收时间、不同品种来源的桑叶进行了全面品质研究。实验结果显示，嫩桑叶 化学成分种类与数量均较多，而晚桑叶某些成分的峰面积更大。不同月份采收 的桑叶样品的总峰面积、 绿原酸与 4 种槲皮素苷的峰面积均处于动态变化之中。 从 10 月起，上述峰面积开始增加，至 11 或 12 月达到最大值，随后快速下降。 所以采收于深秋季节的市售桑叶药材能较好地保证药用
4、质量。同一产地不同品 种的冬桑叶 HPLC 图谱有较高的相似度，而相同品种不同产地的桑叶却有较明 显的差异。说明外界因素对冬桑叶化学成分影响的十分重要。以上实验结果丰 富了冬桑叶的科学内涵。 关键词：桑叶；桑椹；光谱；色谱；冬桑叶；品质评价 Abstract In Pharmacopoeia of the Peoples Republic of China (Edition 2005), both mulberry leaf (Folium Mori) and mulberry fruit (Fructus Mori) have exclusive original plant, Morus
5、alba Linn. Literatures indicated pharmaceutical mulberries were manifold in all ages. Medicinal materials of mulberry in markets are also sourcing from many kinds of plants of Moracease family. Mulberry leaf and mulberry fruit, having different therapeutic range, are two traditional Chinese medicine
6、s rooting from different organs of mulberries. Differentiate as well as commonness between both was shown through following experiments: immunoregulatory, relieving cough and expelling phlegm, effect on insulin receptor and antioxidant activity in vitro. Spectrum and chromatogram prints of plant med
7、icines, which embody species distinguishability and individual comparability, reflect chemical ingredients in whole. We obtained some mulberry leaves and mulberry fruits in different breeds, the two medicines were compared severally in 5 polarity fractions, they were also contrasted with each other.
8、 Infrared spectrum(IR), thin layer chromatography(TLC) and high performance liquid chromatography(HPLC) were used to establish unitary chemical token figures of mulberry leaf and mulberry fruit respectively. Afterwards, ultraviolet spectrophotometry (UV), TLC, HPLC were applied to draw comparisons a
9、mong disparate polarity fractions of each medicine. Comparisons also occurred between two medicines extract in same polarity fraction. Results illuminated single spectrum or chromatogram method was deficient to estimate holistic quality of traditional Chinese drug. While being divided into several p
10、arts according to polarity, the medicinal materials provided more information by means of studying the character of every polar portion. Similarities of samples were gained in all methods except TLC. Cluster analysis based on all similarities of every sample represented the likeness more logically.
11、Ancient medicament literatures pointed out that mulberry leaf should be collected in winter for assuring the quality. HPLC was used to research mulberry leaf samples with different gather time and dissimilar breed. From the results, we discovered mulberry leaves gathered in spring have more ingredie
12、nt sorts and quantity than that gathered in other seasons, yet leaves collected in winter had larger peak area in certain component. Area of total peaks, chlorogenic acid peak and four quercitin glycosides peaks were all in dynamic changes. Area of peaks above-mentioned increased from October and re
13、ached maximum on November or December, then descended rapidly. Thus it can be seen that merchandise of mulberry leaf gathered in late autumn were reassuring. Similarities of HPLC prints were relative high among Winter Folium Mori samples with different breed from same agriculture area, while lower a
14、mong that with same breed but different producing place, which reflected that the environment was significant for ingredients accumulation of mulberry leaf. Key words: Mulberry leaf; Mulberry fruit; Spectrum; Chromatogram; Winter Folium Mori; Quality evaluation 140 声 明 声 明 博士学位论文 基于桑叶桑椹功效的多元光谱色谱信息研
15、究及桑叶品质评价 基于桑叶桑椹功效的多元光谱色谱信息研 究及桑叶品质评价 是本人在成都中医药大学攻读博士学位期间 在导师指导下完成的，该论文涉及的有关知识产权归成都中医药大 学所有。有关该学位论文成果的开发、转让、论文发表等，均需征 得导师和成都中医药大学的同意。 博士生签名： 导 师签名： 前 言 1 前 言 前 言 栽桑养蚕是中国古代先民重要的生产活动，五六千年前的新石器时期中国 人已经种桑养蚕。殷商时代甲骨文中的僲（桑）字，战国青铜器上提筐采桑的 图纹均证明中国栽种和利用桑树之早。小农经济时代，富国以农桑为本，劝农 课桑是历朝历代极为重视的国家基本政策。在夏商西周时期栽桑养蚕业已初具 规
16、模，经过多年发展，人们累积经验并创造了许多提高桑叶质量与产量的技术， 繁殖与改良出了若干桑树品种，使桑叶的品质不断提高，为蚕业的发展奠定了 基础，至今我国的蚕丝产业仍居世界领先地位。 尽管养蚕是栽桑最主要的用途，但并非唯一用途。桑树除采叶养蚕外，桑 根皮、桑枝、桑椹及多余的桑叶都有独特的药用价值，是中医学中的常用药物。 桑叶与桑椹药用历史悠久，桑叶的药用始载于神农本草经桑白皮项下，桑 椹入药始载于唐代新修本草 。后世历代本草对两味药材均有记载。中国是桑 资源大国，桑树的自然分布和人工栽培几乎遍及全国。对桑叶与桑椹的深入研 究可以丰富桑类药材的科学内涵，增加桑资源的综合利用效率。 2005 年版中华人民共和国药典规定，桑叶、桑椹均来源于桑科桑属植 物桑 Morus alba L.。 中国植物志桑科桑属下记载了 11 个种，而目前人工培 育的桑树品种多达上
