| Stability and homogeneity of the quality of Chinese medicinal materials and preparation process is the key to decide the quality of TCM preparation homogeneity. While the Chinese herbal medicine in the different conditions of plant species, locality of growth, harvest time and storage conditions and storage period can affect the quality of preparations stable. The research of Scutellariae baicalensis, using tri-step fourier transform infrared spectroscopy and principal component analysis were used to analyze the different varieties, origin, harvest time and storage period of baicalin, change of the infrared spectrum that can react in different samples of the material basis of difference characteristics and overall material basis. Now using lc-ms to analyse the Radix Scutellariae of different varieties, origin, harvest time and storage period, to determine the relationship between the metabonomics in the chemical composition and its content and to establish the standard library of LC-MS, using various means of analysis. The establishment standard of specific quality of Scutellariae Radix, lay the foundation for revealing the relationship between clinical medicine and curative effect standard. It can provide a theoretical basis for the quality of Chinese medicinal materials in standardization. The first partStudy on flavonoids of Scutellariae baicalensis Georgi in different harvest periodObjective:To analyse the flavonoids of Scutellaria baicalensis Georgi harvested in different harvest periods by using high performance liquid chromatographymass spectrometry, providing reference for the establishment of harvest period of baicalin.Methods:The Agilent Zorbax XDB-C18 column(2.1 mm×150 mm,3.5 μm) was used with the gradient mobile system, the flow rate was 0.2 m L·min-1, the injection volume of standards and samples was 2 μL; the temperature of column was 25 ℃, the detection wavelength was 276 nm, the mobile phase was 0.1% formic acid-acetonitrile. Electrospray ionization source(ESI)was applied and operated in positive ionization multiple reaction monitoring model, the nebulizer was 25 psi with the flow of dry gas 10 L·min-1, the temperature of dry temp was 350 ℃.Results:The content of flavonoids in different harvesting period of Scutellariae baicalensis is different. The content of baicalin which was picked from the spring of chifeng decreased from 12.89% to 1.93%, the content of wogonoside decreased from 3.48% to 0.68%, the content of baicalein decreased from 0.79% to 0.26%, the content of wogonin decreased from 3.03% to 1.46%, the content of chrysin decreased from 0.019% to 0.010%, the content of oroxylin A decreased from 0.057% to 0.029%. The content of baicalin which was picked from the autumn of chifeng decreased from 12.48% to 11.76%, the content of wogonoside decreased from 3.25% to 0.55%, the content of baica lein increased from 0.34% to 2.73%, the content of wogonin increased from 1.70% to 6.16%, the content of chrysin increased from 0.009% to 0.062%, the content of oroxylin A decreased from 0.037% to 0.175%. The content of baicalin which was picked from the autumn of shandong increased from 10.00% to 12.98%, the content of wogonoside increased from 1.25% to 2.57%, the content of baicalein decreased from 1.18% to 0.82%, the content of wogonin decreased from 4.72% to 3.34%, the content of chrysin increased from 0.015% to 0.011%, the content of oroxylin A decreased from 0.035% to 0.030%.Conclusion:The content of 6 flavonoids of Scutellariae baicalensis which was picked from chifeng in the overall downward trend, the content of 6 flavonoids of Scutellariae baicalensis which was picked from shandong in the overall upward trend. The quality of Scutellariae baicalensis from different harvest period can be reflected by this study, providing reference for the formulation of the best harvest period of Scutellariae baicalensis. The second partStudy on flavonoids of Scutellaria baicalensis Georgi in different storage periodObjective:To analyse the storage stability of Scutellaria baicalensis Georgi at room temperature by high performance liquid chromatography-mass spectrometry(HPLC-MS), provides reference for the establishment of storage of Scutellaria baicalensis Georgi.Methods:The method is the same with the first part.Results:The content of baicalin which was storage for 18 months decreased from 8.86% to 8.03% by HPLC-MS, the content of wogonoside decreased from 0.50% to 0.39%, the content of baicalein decreased from 2.08% to 1.60%, the content of wogonin decreased from 0.52% to 0.40%, the content of chrysin and oroxylin A which were storage for 3 months decreased from 0.04% to 0.03%, there is no obvious change from 3 months to 18 months.Conclusion:With the extension of storage time,the contents of the main flavonoids of Scutellaria baicalensis Georgi are in the overall downward trend. The quality of Scutellaria baicalensis Georgi from different storage period can be reflected well by this study, and it can provide a reference for the formulation of the storage period of Scutellariae baicalensis. The third partComparison of the adaptability of ultraviolet detector and mass spectrometry detectorObjective:The detector of ultraviolet and mass spectrometry were compared with the results of adaptability by HPLC, judge the peak purity of liquid chromato-graphic peaks, further accurate quantification of compounds, in order to improve the quality standard of Radix Scutellariae, and provide the reference for the further high quality standard of other traditional Chinese medicine.Methods:The Agilent Zorbax XDB-C18 column(2.1 mm×150 mm,3.5 μm) was used with the gradient mobile system, the flow rate was 0.2 m L·min-1, the injection volume of standards and samples was 2 μL; the temperature of column was 25 ℃, the detection wavelength was 276 nm, the mobile phase was 0.1% formic acid-acetonitrile. Electrospray ionization source(ESI)was applied and operated in positive ionization multiple reaction monitoring model, the nebulizer was 25 psi with the flow of dry gas 10 L·min-1, the temperature of dry temp was 350 ℃.Results:LC-UV and LC-MS were used to determinate the content of main flavonoids in Scutellaria baicalensis Georgi. The two detectors had high precision and stability through the method of comparison, but the sensitivity of LC-MS is higher than LC-UV more than 10 times. In addition, the unknown compounds were determinated by LC-MS with the retention time t R and mass-to-charge ratio, the mass spectrometry detector has higher selectivity and specificity for some ingredients that can not get good separation or have similar retention time with others by LC-UV.Conclusion:Mass spectrometry detector and ultraviolet detector have its advantages and disadvantages in determination of the content of traditional Chinese medicine. The different compounds should choose different detector, to determine the most suitable method for the determination of the content. This experiment can provide reference establishing the determination method for the content of other compounds in the traditional Chinese medicine. The forth partThe establishment,searching and application of libraryObjective:To establish the Liquid Chromatography-Mass Spectrometer-Data base(LC-MS-DS) with more than 30 natural compounds that was constructed with HPLC-ESI-IT-MS/MS. To verify the credibility of the searching results of the LC-MS-DS.Methods:The analysis was performed in alternative ion mode, Agilent Zorbax XDB-C18 column(2.1 mm×150 mm, 3.5 μm) was performed with the mobile phase of 0.1% formic acid-acetonitrile in gradient mobile system, the flow rate was 0.2 m L·min-1, the temperature of column was 25 ℃, the detection wavelength was 200~400 nm; the target mass was 300, the scanning range of compound stability was m/z 100~1200.Results:The establishment of the natural compound such as baicalin, wogonoside, wogonin, astragaloside, emodin and other 30 kinds of traditional Chinese medicine were set in LC-MS-DS, and the control library search results matching degree P, F, R were 995, 996, 1000; 999, 998, 1000; 993, 993, 1000. The matching degree of database and its searching results of each control sample spectrum are very high.Conclusion:The searching results established in this study by liquid chromatography-mass spectrometry-database has high credibility, which provides an effective method for the identification of the compouds of Chinese herbal medicine. |
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