蒙成药三臣丸质量控制与血清药物化学初步研究

发布时间：2018-07-02 21:31

本文选题：三臣丸 + 高效液相色谱法　；参考：《内蒙古医科大学》2017年硕士论文

【摘要】：目的:建立蒙成药三臣丸体外HPLC-DAD指纹图谱,并应用一测多评(quantitative analysis of multicomponents by single marker,QAMS)技术同时测定紫丁香苷,羟基红花黄色素A,芦丁,山奈酚-3-O-芸香糖苷,脱水红花黄色素B,胆酸,猪去氧胆酸等7种成分的含量,采取体外指纹图谱结合一测多评技术的质量评价模式,旨在为蒙成药三臣丸体外质量控制方法提供比较全面的科学依据;基于中药血清药物化学理论,应用于蒙成药三臣丸复方当中,建立三臣丸含药血清HPLC-DAD指纹图谱,分析给药后吸收入血的成分,对蒙成药三臣丸的血清药物化学进行初步研究。方法:(1)以羟基红花黄色素A色谱峰为参照峰,对10批三臣丸样品进行分析,建立蒙成药三臣丸HPLC-DAD体外指纹图谱共有模式,进行相似度分析,并与混合对照品溶液对比指认共有峰。(2)以羟基红花黄色素A为内参物,建立其他6种成分的相对校正因子(relative correction factor,RCFs),用相对校正因子和外标法分别计算含量;同时在3种不同类型的C18色谱柱及3台不同品牌的色谱仪器上考察相对校正因子的重现性。(3)制备5批三臣丸混悬液,并灌胃(ig)给予Wistar大鼠,制备含药血清,与空白血清样品对比建立蒙成药三臣丸含药血清HPLC-DAD指纹图谱共有模式,进行相似度分析,并与混合对照品溶液对比指认入血成分。(4)在获得体内外三臣丸指纹图谱的基础上,结合三臣丸全方、缺味药组、单味药组含药血清样品色谱图进行对比,对三臣丸复方中吸收入血成分及其来源进行初步解析和说明。结果:(1)应用HPLC-DAD法测定了10批三臣丸样品指纹图谱,建立了蒙成药三臣丸特征指纹图谱评价模式,标定了其中36个共有峰,经与对照品比较指认了7个共有峰,其中4号峰为紫丁香苷,5号峰羟基红花黄色素A,15号峰为芦丁,16号峰为山奈酚-3-O-芸香糖苷,18号峰为脱水红花黄色素B,24号峰为胆酸,26号峰为猪去氧胆酸;将10批三臣丸样品与对照图谱进行比对,相似度结果均在0.99以上。(2)在体外指纹图谱基础上建立了一测多评法测定蒙成药三臣丸中7种成分含量的方法,应用相对校正因子与外标法计算10批三臣丸样品中7种成分的含量,以相对误差评价两种方法,结果显示无显著性差异;相对校正因子在不同类型C18反相色谱柱及不同品牌色谱仪器上的重现性良好;选择“两点校正法”与相对保留时间法相结合的形式对7个待测色谱峰进行定位。(3)建立了蒙成药三臣丸含药血清HPLC-DAD指纹图谱,测定5批三臣丸含药血清样品,相似度在0.90以上,并指认了其中6个共有峰,其中2号峰为紫丁香苷,3号峰为羟基红花黄色素A,10号峰为山奈酚-3-O-芸香糖苷,12号峰为脱水红花黄色素B,24号峰为胆酸,25号峰为猪去氧胆酸。(4)三臣丸混悬液灌胃后吸收入血的成分有30个,其中直接以原型被吸收入血的有24个,其余有可能为代谢后新产生的药源性成分或通过药物刺激产生的血液内源性成分;并对各个色谱峰的来源和归属进行了初步解析和说明。结论:(1)本研究建立了蒙成药三臣丸的HPLC-DAD指纹图谱评价体系,所得对照图谱与10批三臣丸样品指纹图谱基本类似,为完善三臣丸的质量控制提供了新的科学依据。(2)本研究所建立的QAMS法可以作为一种简便准确的质量评价模式用于三臣丸中多种成分的定量测定,为蒙成药三臣丸的质量标准提供新的科学依据。(3)本研究建立了三臣丸含药血清HPLC-DAD指纹图谱评价体系,所得对照图谱与5批三臣丸灌胃所得指纹图谱基本类似,反映了三臣丸口服给药后的成分吸收入血情况,为三臣丸的药效物质基础研究提供科学依据。(4)本研究中蒙成药三臣丸吸收入血的成分有30个,其中人工牛黄贡献3个;其余成分均由红花以直接或者间接的方式贡献,其中15个成分为单味红花贡献,9个成分是在红花与人工牛黄配伍的情况下提供的,1个成分是在红花与天竺黄配伍的情况下提供的,另外2个是在三臣丸全方配伍的条件下提供的成分。
[Abstract]:Objective: to establish the HPLC-DAD fingerprint of Mongolian medicine three Chen pill in vitro, and to determine the content of 7 components, such as syringin, hydroxysafflor yellow A, rutin, kaempferol -3-O- rutin, dehydrated safflower yellow B, cholic acid and porcine deoxycholic acid, by one test and multiple evaluation (quantitative analysis of multicomponents by single marker, QAMS). In order to provide a comprehensive scientific basis for the in vitro quality control method of the three minister pills of Mongolian medicine, the quality evaluation model combined with the one measurement and multi evaluation technique is designed to provide a comprehensive scientific basis for the quality control of the three minister pills of Mongolian medicine. Based on the theory of serum drug chemistry of Chinese medicine, it is applied to the three minister pill compound of Mongolian medicine, to establish the HPLC-DAD fingerprint of the serum of the three minister pills, and to analyze the after drug absorption. The composition of the income blood was preliminarily studied on the serum drug chemistry of the Mongolian medicine three minister pills. Methods: (1) the samples of 10 batches of three Chen pills were analyzed with the peak of the A chromatographic peak of the hydroxyl safflower yellow as the reference peak, and the common pattern of the HPLC-DAD in vitro fingerprint of the Mongolian medicine three Chen pills was established, and the similarity analysis was carried out and compared with the mixed control solution. (2) the relative correction factor (relative correction factor, RCFs) was established with the hydroxyl safflower yellow A as the internal parameter. The relative correction factor and the external standard method were used to calculate the content, and the reproducibility of the relative correction factor was examined on 3 different types of C18 columns and 3 different brands of chromatographic instruments. (3) Preparation of 5 batches of three Chen pills suspension, and gavage (Ig) to Wistar rats, preparation of drug containing serum, compared with the blank serum samples to establish the Mongolian medicine three Chen pills serum HPLC-DAD fingerprint pattern, the similarity analysis, and compared with the mixed control solution to identify the blood components. (4) to obtain the basis of the body and outside the three Chen pills fingerprint of the base On the basis of the three minister pill, the whole prescription, the lack of taste medicine group and the serum sample chromatogram of the single drug group were compared. The composition and origin of the absorbed blood in the three minister pills were analyzed and explained. Results: (1) the fingerprint of 10 batches of three Chen pills was measured by HPLC-DAD method, and the evaluation model of the characteristic fingerprint of three Chen pills of Mongolian patent medicine was established. 36 common peaks were calibrated, and 7 common peaks were identified by comparison with the control products, of which No. 4 peak was syringin, No. 5 peak hydroxy safflower yellow A, 15 peak rutin, 16 peak of kaempferol -3-O- rutin, 18 peak of dehydrated safflower yellow B, 24 peak as cholic acid, and 26 peak of porcine deoxycholic acid; 10 batch of three Chen pills samples and pairs were used. The similarity results were all above 0.99. (2) on the basis of the in vitro fingerprint, a method was established to determine the content of 7 components in the three Chen pills of Mongolian medicine, and the content of 7 components in the 10 batches of three Chen pills was calculated with the relative correction factor and the external standard method, and the relative error was evaluated by two methods. The results showed that there was no one. Significant differences; the relative correction factors were reproducible on different types of C18 reversed phase chromatography columns and different brand chromatographic instruments; selected the "two point correction method" and the relative retention time method to locate the 7 pending chromatographic peaks. (3) the HPLC-DAD fingerprint of the serum containing the Mongolian medicine three Chen pills was established, and the 5 batches of three ministers were determined. The similarity degree is above 0.90, and 6 common peaks are identified, among which 2 peak is syringin, 3 peak is hydroxy safflor yellow A, 10 peak is -3-O- rutin, 12 peak is dehydrated safflower yellow B, 24 is cholic acid, and 25 is porcine deoxycholic acid. (4) three courtier pills are absorbed into blood after gastric lavage. There are 30 components, of which 24 are directly absorbed into the blood by prototyping, the rest may be derived from the new metabolites or the endogenous components of the blood produced by drug stimulation, and the origin and attribution of the chromatographic peaks are preliminarily analyzed and explained. Conclusion: (1) this study established the HPLC-DAD of three minister pills of Mongolian medicine. The fingerprint atlas is similar to the fingerprint of the 10 batches of three Chen pills, which provides a new scientific basis for the improvement of the quality control of the three minister pills. (2) the QAMS method established in this study can be used as a simple and accurate quality evaluation model for quantitative determination of various components in the three minister pills and three ministers of the Mongolian medicine. The quality standard of the pill provides a new scientific basis. (3) this study established the evaluation system of the HPLC-DAD fingerprint of the serum containing three Chen pills. The obtained control map is similar to the fingerprint of the 5 batches of three Chen pills. It reflects the absorption of the ingredients into the blood after oral administration of three Chen pills and provides the basis for the study of the medicinal material of the three Chen pills. (4) in this study, there were 30 components of the Chinese Mongolian medicine three Chen pills to absorb the blood, of which 3 were made by artificial bezoar; the other components were contributed directly or indirectly by safflower, of which 15 components were single saffron, and 9 were provided in the case of safflower and artificial Niu Huangpei, and 1 in safflower and guinea pigs. The other 2 were supplied under the condition of compatibility of three Chen Wan.
【学位授予单位】：内蒙古医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R29

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