Application Of Modern Analytical Techniques On Quality Control And Material Base Research Of Traditional Chinese Medicines Formula

Traditional Chinese medicines (TCMs) which combined mixtures of herbal medicines each with multiple constituents are utility to prevent and cure human diseases for thousands of years. The multiplicity formulas attract more and more attentions for their mysterious curative effects. In order to ensure its maximal therapeutic effect with few or no side effect, strict quality standards and validation of suitable analytical assays should be established. But the analysis of such a complex mixture is a challenging task.The chromatographic fingerprinting method has become one of the most frequently applied approaches, which can provide the whole profile of not only the marker compounds but also other active components.In this dissertation, three Traditional Chinese Medicines (TCMs), Zuo Gui Pill (ZGP), Niu Huang Jie Du Pill (NHJDP) and Zuo Jin Pill (ZJP), are selected as the object of research. The quality evaluation system was investigated systematically, on the basic of the modem quality control trend. The method was developed for chromatographic fingerprints and simultaneous quantification of the major constituents of TCMs. The chemical constituents were analysis by HPLC-MS technique. In addition, the fingerprints were qualitative evaluated by chemometrics. This method laid a foundation for quality control of TCMs. The results were summarized as followed:1) A HPLC coupled with DAD and MS spectrometry method was developed for the quality assessment of ZGP by having a combination of chromatographic fingerprint and simultaneous quantification of ten major constituents. These compounds were identified by ESI/MS. In addition, the samples according to their manufacture corporation were classified, and ten major ingredients were screened out to be the most discriminant variables which could be regarded as the constituents for quality assessment of ZGP by using Similarity analysis and partial least squares-discrimination analysis (PLS-DA). This proposed method was applied for the original discrimination and quality control of this TCM successfully.2) An UPLC coupled with Tunable UV detector and RRLC-Q-TOF method was developed for the quality assessment of NHJDP by having a combination of chromatographic fingerprint and simultaneous determination of ten major constituents. Twelve compounds were simultaneously identified by ESI/MS. In addition, the principal component analysis (PCA) coupled with the UPLC fingerprint was applied to classify the NHJDP samples according to their manufacture corporation. The results demonstrate that this analytical method is simple and suitable for the original discrimination and quality control of this TCM.3) A rapid method based on RRLC coupled with DAD has been developed for having a combination of chromatographic fingerprint and simultaneous determination of six major constituents in ZJP. Moreover, a comparative study has been made between the performance of RRLC and HPLC techniques. Then based on the fingerprint data, hierarchical cluster analysis (HCA) was utilized for qualitative evaluation on the resemblance and difference of tested samples. The results demonstrate that this analytical method was helpful to comprehensively evaluate the quality of this TCM.