| ObjectineThe heartwood of Dalbergia odorifera T.Chen is a traditional Chinese medicine, named as Jiangxiang,planted decades before for medicinal purposes. Dalbergia odorifera T. Chen was used for curing hypertension,but not yet see the the research reports of quality evaluation and the comprehensive utilization. So, author study on the shape and microstructure, TLC identification, content determination of the active ingredient content, fingerprint, efficacy and other aspects of the leaves of the leaves of Dalbergia odorifera T. Chen, providing a scientific basis for its development and utilization of resources.MethodsSHR rats as experimental model, study the lower blood pressure efficacy of the extract of leaves of the Dalbergia odorifera chen. T Chen by determinating The systolic blood pressure before and after its administration, determinating the the content of the Ang Ⅱ、 ET-1. Used the column chromatography and slovet extraction method for the separation of chemical composition and the structures were idtentified by the analysis of the physical and chemical contants and1H-NMR、13C-NMR spectroscopy. Use the character identification method for character identicationg. Use the microscopic characteristics identificationg. Use TLC to identify the leaves of Dalbergia odorifera T. Chen. Determine the content of6-hydroxy-biochanin A and biochanin A by HPLC. Establish the fringerprint of Dalbergia odorifera T. Chen by HPLC. Optimization of the extraction technology of6-hydroxy biochanin A and biochanin A from the leaves of Dalbergia odorifera T. Chen by orthogonal experiment and the column chromatography combined with solvemt to optimize the purificationg process. Deteminate the active ingredient content and compare the fingerprint of the different parts of Dalbergia odorifera T. Chen.Result1. The study on the efficacy of blood pressure lowering of the extract of the leaves of Dalbergia odorifera T. Chen.The high dose group and low dose group of Supercritical extracts, as well an the high dose group and low dose group of volatile oil extracts of the leaves of Dalbergia odorifera T.Chen can low the systolic blood pressure of SHR rats. The high dose group and low dose group of volatile oil extract low the Systolic blood pressure obviously, and they low the pressures as18.8mmHg、25.6mmH. make T-test of the difference of volatile oil extract and the different ofthe ethanol group and the P values less than0.05,Which were significant difference.The The high dose group and low dose group of Supercritical extracts, as well an the high dose group and low dose group of volatile oil extracts of the leaves of Dalbergia odorifera T. Chen can low the congtent of AngⅡ、 ET-1of the serum of SHR rats. Make a t-test, compare the high dose group and low dose group of volatile oil extracts with the ethanol group, the different was significant.2. The study on the chemical composition of the leaves of Dalbergia odorifera T. Chen.The leaves of Dalbergia odorifera T.Chen contains aimo acids, proteins, organic-acids, phenols, carbohydrates, triterpenes, flavonoids, volat ile oil compounds. Two isoflavones were isolated from The leaves of Dalbergia odorifera T. Chen, which were identified as6-hydroxy-biochanin A and biochanin A.3.The quanlity evaluation3.1The shape and microscopic characteristics of the leaves of Dalbergia odorifera T. Chen.Describes the characteristic traits and microscopic characeteristics of the leaves of Dalbergia odorifera T. Chen as the basis of their moriphologican identification. Shape is characterized:leaves are ovate or ellipt, apex acute, blunt, base rounded or wedge-shaped, length4-7cm, width2-3cm, petiles1.5-3cm;the surface of the crude was dark green, the under surfaces green;the dried product is the pale green, leaves are nearly leathery, papery of thinly leathery and with smooth surface. The gas is slightly fragrant, slightly bitter taste. Microscopic identification of key points summarized as:(1) the cross-sectional view:the vascular is outer tough type, and the ray cells, phoem cells and veins of vascular contain more calcium oxalate column crystal or cubic;both sides of the leaf, palisade tissue and spongy are different obviously.(2) the surface view:the surface of the leaves is smooth, the pores are maily distributed in the leaf low epidermis. And there air many yellow non-glandular hairs.(3) powder view:A number of crystal fibers and fibers can be seen, some crytal fibers were branced. Calcium oxalate column crytal and cubic exist in the parenchyma cells, and bast fibers and wood fibers scattered single. Non glandular consists of two cells, near the head of the larger cells cell is shorter, darker, wall warts markedly; master catheter for screw thread.3. TLC identification of the leaves of Dalbergia odorifera T. ChenIn accordance with TLC test of the appendix VIB section of the2010editiong of Chinese Pharmacopoeia,6-hydroxy-biochanin A and biochanin A as reference,254nm UV lamp for the chromatography of the test, sample and reference substance showed the same clour spot in the corresponding position.3.3Determinationg of6-hydroxy-biochanin A and biochanin A of leaves of Dalbergia odorifera T. Chen.6-hydroxy-biochanin A in the concentration range of0.0017-0.1716u g showed a good linear relationship (r=0.9995), and the average recovery was99.04%; biochanin A in a concentration range of0.0017-0.1740μg showed a good linear relationship (r=0.9995), and the average recovery was99.02%.The content of6-hydroxy-biochanin A in A rang of0.0411%to0.6659%, and The content of biochanin in A rang of0.0411%to0.6659%, which the leaves producted in Gaozhou. The content of6-hydroxy-biochanin A of the leaves of Dalbergia odorifera T.Chen that producted in Jiangmen, Dongguan,Kaiping, Hainan is higher than the leaves that porducted in other places, and the leaves showed a content up to1.0392%which poducted in Kaiping. The content of biochanin A is generally low, the leaves which productd in Dongguan show the maximum content that is0.4382%. And the content of biochanin A of the leaves producted in Guangzhou is very low, difficult uo be detected.3.4The fingerprint of the flavonoids of the leaves of dalbergia odorifera T. Chen.Fifteen main marker peaks were selected in the standard fingerprint and two of them were identified as biochanin A and6-hydroxy-biochanin A. There were six fingerprints of samples with good similarities among the fourteen different samples of the leaves of Dalbergia odorifera T. Chen, which above0.90compared with the standard fingerprint, and there were also five of them with the similarities obove0.75. The elses were below0.75.4. The extraction and purificationg technology of the6-hydroxy-biochanin A and biochanin A of the leaves of dalbergia odorifera T.Chen.The optimal extraction technology was as follow:the concentration of ethanol was50%,the extracting time was60min, he ratio of raw materials to solvent was1:60and extracting frequency was3times.6-hydroxy-biochanin A and biochanin A were purified by the macroporous resin column an solvent.5. Study on the quality of different parts of dalbergia odorifera T.Chen.Compare the content of6-hydroxy-biochanin A and biochanin A of different parts of Dalbergia odorifera T. Chen that porducted in the same place. The contents of6-hydroxy-biochanin A and biochanin A of the flower is high than the leaves which both producted in Gaozhou. The different parts of Dalbergia odorifera T. Chen that producted in Hainan showed the content order of leaves>branches>fruits. The contents of6-hydroxy-biochanin A and biochanin A of the leaves is high than the branch, es which both producted in Jiangmen. The contents of6-hydroxy-biochanin A and biochanin A of the leaves is high than the fruits which both producted in Guangzhou. The commeercail heartwood of Dalbergia odorifera T. Chen showed the low content of the biochanin A, and the6-hydroxy-biochanin A can not be detected.The fingerprint similarity of different parts of Dalbergia odorifera T.Chen is low. And peaks of heartwood are basically concentrated in the rang of30th to50th min, and the peaks of the parts of leaves, branches, fruits are more evenly distributed. From the UV absorption spetra of the fingerprint peaks, it can be kown that the composition of heartwood is different from other parts. The common peaks of the heartwood are less found in th selection of common peaks with other peaks except the biochanin A. Excluding the heartwood, compared the other parts, it can be found that the other parts with high similarty except the fruits.ConclousionThe high dose group and low dose group of the volatile oil extracts of the leaves of Dalbergia odorifera T.Chen can low the systolic blood pressure of SHR rats, and the difference is significant. The high dose group and low dose group of volatile oil extracts of the leaves of Dalbergia odorifera T.Chen can low the congtent of AngⅡ、ET-1of the serum of SHR rats. The mechanism of lower blood pressure may be by the lower of the content of AngⅡ、ET-1of serum to achieve the effect.The leaves of Dalbergia odorifera T. Chen contains aimo acids, proteins, organic-acids, phenols, carbohydrates, triterpenes, flavonoids, volatile oil compounds. And the content of flavonoid are rich. Two isof lavones were isolated from The leaves of Dalbergia odorifera T.Chen, which were identified as6-hydroxy-biochanin A and biochanin A.6-hydroxy-biochanin A was for the first time isolated from the leaves of Dalbergia odorifera T. Chen. Optimization of the extraction technology of6-hydroxy-biochanin A and biochanin A from the leaves of Dalbergia odorifera T. Chen by orthogonal experiment, purified them by the macroporous resin column an solvent.Creat a quality evaluation system of the leaves of Dalbergia odorifera T.Chen by combinating the serval modern ainalatical tool such as the shape and microscopie identification, the TLC identification,the determination and the fingerprints. |
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