【作者】 秦海燕；

【导师】 刘文哲；

【作者基本信息】 西北大学 ， 植物学， 2007， 硕士

【摘要】 丹参（Salvia miltiorrhiza Bge）为唇形科（Labiatae）鼠尾草属多年生草本植物，传统以根入药，其应用已有悠久的历史。本文采用植物解剖学、植物化学以及色谱分析等手段，系统研究了丹参营养器官的结构、药用部位的结构发育及药用成分的积累规律，建立了3种丹参制剂中药用成分的高效液相色谱分析方法。本研究为丹参的种植、采收、品质鉴定、综合利用以及丹参制剂的质量控制提供了科学依据。全文共分四章，作者的主要贡献如下：1、采用植物解剖学方法研究了丹参营养器官的结构及常用药用部位的结构发育。结果表明：（1）丹参根的结构、发育过程与一般双子叶植物相同。根尖顶端的原分生组织衍生的细胞分化为原表皮、基本分生组织和原形成层，以后分别分化为表皮、皮层和维管柱，构成了初生结构。在次生生长中，维管形成层分裂产生次生木质部和次生韧皮部，次生韧皮部由韧皮薄壁细胞、筛管、伴胞、韧皮射线组成，次生木质部由木薄壁细胞、导管、木纤维和木射线构成，导管呈切向排列与木纤维相伴形成束状结构，导管束在次生木质部中呈放射状排列。根的木栓形成层起源于中柱鞘，向外分化出木栓层，向内分化出栓内层。周皮外侧有时可见落皮层。（2）丹参茎为四棱形，由表皮、皮层、维管束、髓、髓射线组成。茎的四个棱角处有丰富的厚角组织，维管组织由多个维管束组成，形成环状，维管束为无限外韧维管束，韧皮部较窄，木质部较宽，髓部由大量的薄壁细胞构成。（3）丹参的叶为异面叶，上下表皮均具大量的表皮毛，上表皮细胞有较厚的角质层，栅栏组织细胞2层，排列紧密，海绵组织细胞呈不规则状，细胞间隙明显。2、采用高效液相色谱—电化学检测器（HPLC—ECD）和高效液相色谱—二极管阵列检测器（HPLC—DAD），分别建立了丹参营养器官中电活性的水溶性成分和非电活性脂溶性成分的分析方法。研究了丹参水溶性成分丹参素、原儿茶醛和咖啡酸及脂溶性成分丹参酮Ⅰ和丹参酮ⅡA在营养器官中的积累规律。结果表明：（1）HPLC—ECD对丹参中电活性的水溶性成分丹参素、原儿茶醛、咖啡酸的检测灵敏度是各成分在最大吸收波长处检测灵敏度的35.9，25.3，11.6倍，解决了由于水溶性成份含量较低且紫外吸收较弱而难以用HPLC—DAD测定的问题；对于非电活性脂溶性成分丹参酮Ⅰ和丹参酮ⅡA，分别在246以及270 nm处有很强的紫外吸收，可同时得到灵敏检测。（2）丹参的根、茎、叶中均含有水溶性成分。叶中水溶性成分含量由高到低依次为丹参素＞咖啡酸＞原儿茶醛。在一个生长期中，丹参素含量在6月份最高，达到1.9％，以后逐渐减少，12月降至最低；咖啡酸在6月和9月含量最高，分别为0.14％和0.12％；原儿茶醛的含量变化不明显；在叶由嫩到老的发育过程中，总水溶性成分含量逐渐降低。茎中水溶性成份含量很低。丹参根中水溶性成份含量由高到低依次为丹参素＞原儿茶醛＞咖啡酸，在一个生长期中，丹参素含量在7月达到最高，为1.65％，且6～9月含量较为稳定，以后丹参素含量逐渐降低，10月中旬以后，丹参素含量趋于稳定；原儿茶醛、咖啡酸的含量在7、8、9月较为稳定且稍高于其它月份，以后逐渐降低，至12月降到最低。根中水溶性成分含量与其粗度无相关性，随着皮部与木质部厚度之比增大，水溶性成分含量有增加的趋势，皮部的水溶性成分含量高于木质部。（3）丹参的茎、叶中不含有脂溶性丹参酮，根中含量较高。丹参酮ⅡA和丹参酮Ⅰ在一个生长季节的含量变化规律大体呈“单峰”曲线，丹参酮ⅡA的含量高于丹参酮Ⅰ，丹参酮ⅡA在7月份含量最高，丹参酮Ⅰ在5月份含量最高。3、采用高相液相色谱-电化学检测（HPLC-ECD）以及高相液相色谱-二极管阵列检测器-电化学检测器（HPLC-DAD-ECD）联用技术，同时测定了以丹参为君药的消栓通络片、复方丹参片以及尿塞通片中电活性和非电活性药用成分的含量。结果表明：对于电活性药用成分丹参素、原儿茶酸、原儿茶醛、咖啡酸、和阿魏酸在HPLC-ECD上均可得到灵敏的检测，检出限可达1.0×10^-3μg·mL^-1；对于非电活性脂溶性成分丹参酮ⅡA，在270 nm处有很强的紫外吸收，也可得到灵敏检测；HPLC-DAD-ECD联用技术同时利用DAD和ECD各自的检测优势，不需要通过富集、衍生、分别检测等复杂操作，一次进样就可同时灵敏地对电活性和非电活性成分进行分析。更多还原

【Abstract】 Salvia miltiorrhiza Bge, a kind of perennial herb plant from labiate family, has been widely used in China for centuries, its root can be used to make traditional Chinese medicine. The structure of vegetative organs, the growth development of medicinal parts, and the accumulation of medicinal components of Salvia miltiorrhiza Bge is systematically studied with the methodology of botanical anatomy, phytochemistry and high performance liquid chromatography, and, an analytical method of high performance liquid chromatography for three kinds of Danshen preparations is established. A scientific basis for salvia miltiorrhiza planting, harvesting, quality appraisal, utilization, and quality control of Danshen preparations, therefore, is provided. The thesis consists of four chapters. The author’s contributions can be summarized as follows:1. The structure of vegetative organs of Salvia miltiorrhiza and its growth development of medicinal part were studied with the methodology of the botanical anatomy. (1) Experimental results indicated that the structure and growth development of the root of Salvia miltiorrhiza are the same to other dicotyledons. The cell derived from primordial meristem at the top of the root tip can differentiation into protoderm, ground meristem and procambium, and then epidermis, cortex and vascular cylinder respectively, which become the primary structure of the root. In secondary growth, the cambium splits into secondary xylem inside and secondary phloem outside. The former is composed of phloem parenchyma, sieve tube, companion cells and phloem ray, and the latter, xylem parenchyma, vessel, fiber and xylem ray. In addition, vessels in secondary xylem are arranged radially. The cork cambium of root derived from pericycle produces pellem outward and phelloderm inward. Rhytidome outside the periderm can be seen sometimes. (2) Salvia miltiorrhiza stem, shaped as four arris, is composed of the epidermis, the cortex, the vascular bundle, the pith and the ray. There are plentiful of collenchyma in the four arris. The vascular tissue, composed of several vascular bundles, is circularity; and the vascular bundles, which have the narrower phloem, and wider xylem, are open collateral bundle. There is a mass of parenchyma in the pith.(3)The leaf of Salvia miltiorrhiza is dorsi-ventral, and there are abundant trichomes in the surface of the leaf and the thicker cutin covers the epidermis cells of the leaf. Palisade tissue, two tiers ranks closely and spongy tissue has bigger lacuna between their cells.2、The analytical methods for determination of electroactive water-soluble and non-electroactive liposoluble components in vegetative organs of Salvia miltiorrhiza by high performance liquid chromatography with electrochemical detector (HPLC -ECD) and high performance liquid chromatography with diode array detector(HPLC-DAD) were established respectively. The accumulation of medicinal components in vegetative organs was studied. The experimental results showed that: (1) The sensitivity for electroactive water-soluble components detected by HPLC-ECD was 35.9, 25.3, 11.6 times than those obtained by HPLC-DAD at maximum absorption wavelength respectively. This method created a solution to resolve the problem that it is difficult to detect water-soluble components by HPLC-DAD because of low level and weak UV absorption. At the same time, the non-electroactive liposoluble components were detected sensitively by HPLC-DAD at 246 and 270 nm. (2)There are phenolic compounds in root, stem and leaf. The content of phenolic compounds from high to low was Danshensu, caffeic acid and protocatechuic aldehyde in leaf. The highest content of Danshensu in leaf is in June within a whole growth period, up to 1.9% and gradually drop off till the lowest in December; caffeic acid is in June and August, respectively 0.14% and 0.12%; and the change of content of protocatechuic aldehyde is inconspicuous. The content of water-soluble phenolic compounds in leaf is gradually decreases with the leaf becoming old within a growth period. The stem contains very little phenolic compounds. The content of water-soluble phenolic compounds from high to low is Danshensu, protocatechuic aldehyde and caffeic acid in root. The content of Danshensu in root arrives at the highest (1.65%) in July and consistent from June to September, and gradually reduces thereafter until a low stable level in mid-October; and for protocatechuic aldehyde and caffeic acid in root, it is little higher in July, August and September than other months. And the content of the water-soluble phenolic compounds in the root has no relevance with its diameter, but will increase with the increase of bark to xylem ratio. Water-soluble phenolic compounds were more in bark of the root than those in xylem. (3) There are no tanshinone in the stem and the leaf, and the content of tanshinone is higher in the root. Tanshinone IIA and tanshinone I roughly assumed "the single peak" within a growth period and the content of tanshinone IIA was higher than that of tanshinone I . The highest content of tanshinone IIA is in July (2.94%) and tanshinone I is in May (0.78%).3. The methods for simultaneous separation and determination of medicinal components in Xiaoshuantongluo, Fufangdanshen, Niaosaitong tablets by HPLC-ECD and high performance liquid chromatography combined electrochemical detection with diode array detection (HPLC-ECD-DAD) was established. The experimental results showed that the medicinal components were detected sensitively. The method combining ECD with DAD can not only exert their respective advantages, but also widen its applications to detect various analytes in low level directly without the complicated process of enrichment, derivation and separate detection.更多还原