Study On High Performance Liquid Chromatographic Analysis Theory For Traditional Chinese Medicine

Chinese medicine is a typical complicated unknown sample, to illustrate itschemical constitution is one of the key scientific subjects of Traditional ChineseMedicine (TCM) modernization. Liquid chromatography (LC) is the most commontechnology to be used in the analysis of TCM, as well as the essential separatingmethod combined with MS or NMR. In this paper, after summarizing thecharacteristic of the complicated system of Chinese medicine and the problemsneeded to be solved during the analysis, an analysis thought for Chinese medicine isbrought forward, the regular rule of LC retention values and the shape of peak underlinear gradient are investigated, a method is developed to acquire quickly the LCretention parameters and peak shape parameters, consequently, the unknown samplecan be converted into LC-known sample. Also in this paper, actual samples ofSwertia franchetiana and Astragalus are taken as examples, the LC parametersdatabase is established accordingly, the information base of the Chinese medicinefingerprint chromatogram is provided. Simultaneously, the target optimizationmethod of the Chinese medicine ingredient is developed focusing on the practicalapplication.In this dissertation, the LC retention value equation is obtained correctly andquickly by using five linear gradients, the method of estimating the retention timeunder any gradient based on the retention parameter a, c is developed, as well as therevisionary method based on the internal standard peak. The respective errors are lessthan 0.74%. The c-a relationship is inspected by applying the acquirement of theretention parameter of the unknown samples of Swertia franchetiana and Astragalus.Based on EMG model and the technology of automatic curve fitted, a method isfirstly developed to quickly obtain the peak shape parameters and peak shape rule byusing linear gradients on LC. The peak shape parameters under any gradientcondition are estimated by the rule of linear gradient peak shape, which shows theestimated values are almost identical to the resolved result, this method can beapplied to the whole chromatogram fit and chromatogram emulation. The chloroformextract of Swertia franchetiana is taken as example, resolving of authenticchromatograms under different gradient condition shows that quantification bycurve-fitting method has high quantification precision. The method of LC target optimization is developed by using MovedOverlapping Separation Ranging Map (OSRM) and simulated chromatograms. Theprocess of LC target optimization is elaborated for the analysis of total extract ofSwertia franchetiana and ethyl estate extract of Astragalus. The validation by usingseveral objects shows that optimization separation can be achieved. LC-MS analysis has been done to the astragalosides of Astragalus, themolecular weight and structure information of astragalosides are obtained, possiblestructure is deduced by using selective ion monitor (SIM) and secondary MS (CID).The preparative LC separation method of ingredients of Astragalus extract isestablished, as well as the active screening on the leukemia cell model, based onwhich an exploring research was done on the effective substantial foundation.