| 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. |
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