The Preparation Of Ziyin Tiaojing Granule And Its Effect On Uterine Contraction Of Rats In Vitro

BackgroundZiyin Tiaojing medicinal broth is a clinical empirical recipe of professor Jianling Huang in Guangdong Provincial Hospital. It is composed by9kinds of traditional Chinese medicine and shows good effect on climacteric dysfunctional uterine bleeding in clinic. But medicinal broths are inconvenient for decoction, carrying, and storing, and its quality is difficult to control, which results to poor compliance of patients. Therefore, this paper will use the modern technology to change the Ziyin Tiaojing medicinal broth into a kind of granule entrusted by Yongchuan Chinese Medicinal hospital, so that to overcome the disadvantage of medicinal broth and more extensively use its scientific and medical value. This paper was funded by the Chongqing Municipal Health Bureau (Item number:2012-2-36).Objective:This paper aims to change the Ziyin Tiaojing medicinal broth into Ziyin Tiaojing granule, then detect the quality of the Ziyin Tiaojing granule, and formulate a quality standard for it. The stability of the Ziyin Tiaojing granule will also be detected. Finally, the effect of Ziyin Tiaojing granule on uterine contraction of rats in vitro is designed to research in this paper.Methods: 1. The preparation of Ziyin Tiaojing granuleIt was researched from three aspects:(1) The extraction:the decoction method with water was used to extract the active components. With the extraction yield, and the contents of Asperosaponin Ⅵ, Stilbene glycosides, total tannins, total saponins and total flavonoids as indicators, the extraction technology was optimized by orthogonal design.(2) The purification:After the optimization, the extract was purified by alcohol. The purification technology was optimized with the ablation rates of foreign matter and the remaining rates of Asperosaponin Ⅵ, Stilbene glycosides, total tannins, total saponins and total flavonoids as indicators.(3) The formation of granules:suitable filler was selected and optimal volume of it was screened out. The formation technology was optimized by the colour, the moldability, the uniformity, the dissolve ability, and the flavor of the granule.2. The quality and quality standards research of Ziyin Tiaojing granuleIt was researched from the following four aspects:(1) Characteristics:the appearance, color and flavor were described.(2) Identification:TLC was used to identify the medical materials (including Medicinal material A, Chinese Teasel Toot, Medicinal material D, Medicinal material B, Medicinal material C, and Tangshen).(3) Inspection:the Ziyin Tiaojing granule was inspected on the particle size, the moisture content, the dissolve ability, the content uniformity, etc. based on the appendix ⅠC in2010th edition of Chinese Pharmacopeia.(4) Determination:HPLC was used to determine the content of Asperosaponin Ⅵ and Stilbene glycosides. UV was used to determine the content of total tannins, total saponins and total flavonoids.3. The stability study of Ziyin Tiaojing granuleThe stability of Ziyin Tiaojing granule was detected by influential factor test and accelerated test.(1) Influential factor test:three groups of Ziyin Tiaojing granule were set at high temperature (60℃±2℃), high humidity (relative humidity is92.5%), and highlight (4500Lx±500Lx), respectively, to investigate the stability of Ziyin Tiaojing granule under severe environment.(2) Accelerated test:Ziyin Tiaojing granule was set at environment of40℃±2℃with the relative humidity of92.5%for6months to investigate its stability.4. The effect of Ziyin Tiaojing granule on uterine contraction of rats in vitroFirst, the effective doses of Ziyin Tiaojing granule were evaluated.10healthy female rats was selected. Diethylstilbestrol was hypodermicly injected to cause estrus in rats. A patch of uterine was cut to detect the change of uterine contraction curve after adminstered with different doses of Ziyin Tiaojing granule by BL-420biological functional experimental system. Second, the effect of different doses of Ziyin Tiaojing granule on uterine constraction of rats in vitro was assayed by the same way.Results:1. The preparation technology of Ziyin Tiaojing granuleThe9kinds of medical materials were taken based on the prescription. Medicinal material A, Medicinal material D, and Medicinal material H were crushed to the extent that could pass through10mesh sieves. After that, the crushed three materials and the other materials were mixed in n times amount of water (g:mL) to soak. After30minutes, the mixture was decoct for a times, and1h for each. Then the extract was obtained after filtration. The extract was condensed to1g.mL-1at60℃, and then added with ethanol to make the alcohol content achieved at50%. After setting for12h at room temperature, the mixture was filtrated and the filtrate was got rid of the ethanol, then condensed to a relative density of1.10-1.15at60℃. The condensed filtrate was added with65.0g.mL-1Amidulin and0.2g.mL-1Steviosin to spray drying at100℃, f m3·h-1,155mL.min-1and0.2MPa so that to obtain the medicamental pulverata. The medicamental pulverata was prepared into soft materials with70%ethanol, then granulated by crushing and sieving. The wet granules were dried at60℃for1h. After resieving, sealing and packaging, the Ziyin Tiaojing granule was successfully obtained.2. The quality of Ziyin Tiaojing granule (1) The Ziyin Tiaojing granule presented as dilute brow or brown. It tasted slightly sweet and astringent. It smelled fragrant.(2) The TLC for Ziyin Tiaojing granule showed an identical spot with the TLC for medical material controls at the same level.(3) All of the particle size, the moisture content, the dissolve ability and the content uniformity of Ziyin Tiaojing granule were in accordance with the Chinese Pharmacopeia.(4) The Ziyin Tiaojing granule contained≥2.00mg-g-1of Asperosaponin VI,>0.30mg-g-1of Stilbene glycosides,≥5.00mg·g-1of total tannins,≥1.20mg·g-1of total saponins and≥9.00mg·g-1of total flavonoids.3. The stability of Ziyin Tiaojing granuleBoth of the influential factor test and the accelerated test showed satisfactory stability of Ziyin Tiaojing granule.4. The effect of Ziyin Tiaojing granule on uterine constraction of rats in vitroThe Ziyin Tiaojing granule had good effect on the uterine constraction of rats in vitro. It could intensify the amplitude of uterine contraction and increase the motoricity of uterine smooth muscle.Conclusions:The Ziyin Tiaojing granule could be prepared as follows:9kinds of medical materials were taken based on the prescription. Medicinal material A, Medicinal material D, and Medicinal material H were crushed to the extent that could pass through10mesh sieves. After that, the crushed three materials and the other materials were mixed in n times amount of water (g:mL) to soak. After30minutes, the mixture was decoct for a times, and1h for each. Then the extract was obtained after filtration. The extract was condensed, and then added with ethanol to purify the extract. After setting for12h, the mixture was filtrated and the filtrate was got rid of the ethanol, then condensed to a relative density of1.10-1.15. The condensed filtrate was added with65.0g.mL-1Amidulin and0.2g.mL-1Steviosin to spray drying so that to obtain the medicamental pulverata. The medicamental pulverata was prepared into soft materials with70%ethanol, then granulated by crushing and sieving. The wet granules were dried, resieved, sealed and packaged.The obtained Ziyin Tiaojing grabule was controllable on quality and stable. It could promote the uterine contraction of rats in vitro.