The Methodological Study Of One-dimensional And Two-dimensional High Performance Liquid Chromatography Fingerprint Of The Traditional Chinese Medicines

With the development of the technology of the analytical instruments, the detection and information processing, the fingerprint of traditional Chinese medicine has got great improvements in recent years. However, there are many deficiencies in the development of the fingerprint. Firstly, the correlation between the fingerprint and the chemical components in the Chinese medicines is not clear. Secondly, the dependency of the fingerprint and the pharmacologic actions is uncertain. Last but not least, it is necessary to evaluable whether the fingerprint is reliability and accuracy. Therefore, in this study, we extended a HPLC-DAD and comprehensive two dimensional high performance liquid chromatography (2D-HPLC) system to establish the fingerprint of Fructus Schisandrae and Rhizoma polygoni cuspidati. By the methodological evaluation and effect evaluation, the result of this research provided the methodological examples and technique platform for research on modernization for quality control of these two traditional Chinese medicines.First part, fingerprint analysis of Fructus Schisandrae and Rhizoma polygoni cuspidate by HPLC-DAD1. By optimization the conditions of the chromatograph, a HPLC-DAD method was established for separation and analysis of the both Schisandra chinensis and Schisandra sphenanthera. The established and validated HPLC fingerprint were further analysis with chemometric methods including similarity evaluation, principal component analysis and hierarchical clustering analysis. The obtained results indicated that the established and validated HPLC fingerprint can be successfully used to distinguish and control the quality the Schisandra chinensis and Schisandra sphenanther.2. Application of response surface methodology for optimization of the conditions of the sample extraction, a HPLC-DAD method was established for separation and analysis of the Rhizoma polygoni cuspidati from different producing areas. By the methodological validation and chemometric methods evaluation, research reveals that the established and validated the fingerprint can be suitable for quality control and geoherbalism research of Rhizoma polygoni cuspidati.Second part, fingerprint analysis of Fructus Schisandrae and Rhizoma polygoni cuspidati by comprehensive 2D-HPLC-DAD-MS1. A comprehensive two-dimensional chromatography approach was developed for the separation and identification of membrane permeable compounds in Schisandra chinensis. The first dimensional column was the immobilized liposome chromatography(ILC)column, which mimics the biological membranes and can be used to study drug-membrane interactions in liquid chromatography. More than 30 components in Schisandra chinensis were resolved using the developed separation system and among them 14 compounds were identified interacting with the ILC column based on their retention action, UV spectra and mass data. With this comprehensive two dimensional high performance liquid chromatographic system, three-dimensional chromatographic fingerprints of Schisandra chinensis were preliminarily established and processed using principal component analysis and hierarchical clustering analysis. The result demonstrated that the proposed fingerprint was a useful means to control the quality and to clarify the membrane permeability of the compounds in Schisandra chinensis.2. A comprehensive two-dimensional biochromatography method using a silica-bonded human strum albumin (HSA) column and a monolithic column was developed for the biological fingerprinting analysis of bioactive components in Rhizoma polygoni cuspidati. The biochromatography with HSA-immobilized stationary phase was applied to study the interaction of multiple components in Rhizoma polygoni cuspidati. Many compounds were interacted with the HSA column and among them 9 compounds were identified based on their retention time, UV spectra and mass data. And this technology was successfully used to the study on the biopharmaceutical analysis of Rhizoma polygoni cuspidate. The established three-dimensional chromatographic fingerprints, which included the information of chemical components and their interaction of the silica-bonded human strum albumin, were further analysis with chemometric methods. The results show that the developed comprehensive two-dimensional biochromatography system is capable of being used for biological fingerprinting analysis and the quality control of Rhizoma polygoni cuspidati.