發(fā)布時間：2018-07-02 21:31

  本文選題：三臣丸 + 高效液相色譜法　； 參考：《內(nèi)蒙古醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:建立蒙成藥三臣丸體外HPLC-DAD指紋圖譜,并應(yīng)用一測多評(quantitative analysis of multicomponents by single marker,QAMS)技術(shù)同時測定紫丁香苷,羥基紅花黃色素A,蘆丁,山奈酚-3-O-蕓香糖苷,脫水紅花黃色素B,膽酸,豬去氧膽酸等7種成分的含量,采取體外指紋圖譜結(jié)合一測多評技術(shù)的質(zhì)量評價模式,旨在為蒙成藥三臣丸體外質(zhì)量控制方法提供比較全面的科學(xué)依據(jù);基于中藥血清藥物化學(xué)理論,應(yīng)用于蒙成藥三臣丸復(fù)方當(dāng)中,建立三臣丸含藥血清HPLC-DAD指紋圖譜,分析給藥后吸收入血的成分,對蒙成藥三臣丸的血清藥物化學(xué)進(jìn)行初步研究。方法:(1)以羥基紅花黃色素A色譜峰為參照峰,對10批三臣丸樣品進(jìn)行分析,建立蒙成藥三臣丸HPLC-DAD體外指紋圖譜共有模式,進(jìn)行相似度分析,并與混合對照品溶液對比指認(rèn)共有峰。(2)以羥基紅花黃色素A為內(nèi)參物,建立其他6種成分的相對校正因子(relative correction factor,RCFs),用相對校正因子和外標(biāo)法分別計算含量;同時在3種不同類型的C18色譜柱及3臺不同品牌的色譜儀器上考察相對校正因子的重現(xiàn)性。(3)制備5批三臣丸混懸液,并灌胃(ig)給予Wistar大鼠,制備含藥血清,與空白血清樣品對比建立蒙成藥三臣丸含藥血清HPLC-DAD指紋圖譜共有模式,進(jìn)行相似度分析,并與混合對照品溶液對比指認(rèn)入血成分。(4)在獲得體內(nèi)外三臣丸指紋圖譜的基礎(chǔ)上,結(jié)合三臣丸全方、缺味藥組、單味藥組含藥血清樣品色譜圖進(jìn)行對比,對三臣丸復(fù)方中吸收入血成分及其來源進(jìn)行初步解析和說明。結(jié)果:(1)應(yīng)用HPLC-DAD法測定了10批三臣丸樣品指紋圖譜,建立了蒙成藥三臣丸特征指紋圖譜評價模式,標(biāo)定了其中36個共有峰,經(jīng)與對照品比較指認(rèn)了7個共有峰,其中4號峰為紫丁香苷,5號峰羥基紅花黃色素A,15號峰為蘆丁,16號峰為山奈酚-3-O-蕓香糖苷,18號峰為脫水紅花黃色素B,24號峰為膽酸,26號峰為豬去氧膽酸;將10批三臣丸樣品與對照圖譜進(jìn)行比對,相似度結(jié)果均在0.99以上。(2)在體外指紋圖譜基礎(chǔ)上建立了一測多評法測定蒙成藥三臣丸中7種成分含量的方法,應(yīng)用相對校正因子與外標(biāo)法計算10批三臣丸樣品中7種成分的含量,以相對誤差評價兩種方法,結(jié)果顯示無顯著性差異;相對校正因子在不同類型C18反相色譜柱及不同品牌色譜儀器上的重現(xiàn)性良好;選擇“兩點(diǎn)校正法”與相對保留時間法相結(jié)合的形式對7個待測色譜峰進(jìn)行定位。(3)建立了蒙成藥三臣丸含藥血清HPLC-DAD指紋圖譜,測定5批三臣丸含藥血清樣品,相似度在0.90以上,并指認(rèn)了其中6個共有峰,其中2號峰為紫丁香苷,3號峰為羥基紅花黃色素A,10號峰為山奈酚-3-O-蕓香糖苷,12號峰為脫水紅花黃色素B,24號峰為膽酸,25號峰為豬去氧膽酸。(4)三臣丸混懸液灌胃后吸收入血的成分有30個,其中直接以原型被吸收入血的有24個,其余有可能為代謝后新產(chǎn)生的藥源性成分或通過藥物刺激產(chǎn)生的血液內(nèi)源性成分;并對各個色譜峰的來源和歸屬進(jìn)行了初步解析和說明。結(jié)論:(1)本研究建立了蒙成藥三臣丸的HPLC-DAD指紋圖譜評價體系,所得對照圖譜與10批三臣丸樣品指紋圖譜基本類似,為完善三臣丸的質(zhì)量控制提供了新的科學(xué)依據(jù)。(2)本研究所建立的QAMS法可以作為一種簡便準(zhǔn)確的質(zhì)量評價模式用于三臣丸中多種成分的定量測定,為蒙成藥三臣丸的質(zhì)量標(biāo)準(zhǔn)提供新的科學(xué)依據(jù)。(3)本研究建立了三臣丸含藥血清HPLC-DAD指紋圖譜評價體系,所得對照圖譜與5批三臣丸灌胃所得指紋圖譜基本類似,反映了三臣丸口服給藥后的成分吸收入血情況,為三臣丸的藥效物質(zhì)基礎(chǔ)研究提供科學(xué)依據(jù)。(4)本研究中蒙成藥三臣丸吸收入血的成分有30個,其中人工牛黃貢獻(xiàn)3個;其余成分均由紅花以直接或者間接的方式貢獻(xiàn),其中15個成分為單味紅花貢獻(xiàn),9個成分是在紅花與人工牛黃配伍的情況下提供的,1個成分是在紅花與天竺黃配伍的情況下提供的,另外2個是在三臣丸全方配伍的條件下提供的成分。
[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.
【學(xué)位授予單位】：內(nèi)蒙古醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R29

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1 白玉琴;蒙成藥三臣丸質(zhì)量控制與血清藥物化學(xué)初步研究[D];內(nèi)蒙古醫(yī)科大學(xué);2017年

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本文編號：2090988