The Isolation, Purification And Structure Identification Of The Effective Components In Pseudostellaria Heterophylla (Miq.) Pax And The Study Of The Fingerprinting

Isolation, structure identification of the effective component in Pseudostellaria heterophylla (Miq.) Pax; and the fingerprinting of Pseudostellaria heterophylla (Miq.) Pax (Taizishen, Chinese name) based on activity constituent are studied in this dissertation. Towards these objectives, the following five topics discussed in the thesis.1. The contents of polysaccharide in Taizishen harvested at different times during the growing period were determined by phenol-sulfuric acid colorimetry. The result showed that the content of polysaccharide could be used to determine the optimum harvest time for Taizishen and provide scientific information for the rational exploiting and utilization of the herb.2. A new RP-HPLC method was established to assay the free amino acids in Taizishen. In this method, OPA and FMOC were selected as the pre-column derivatizing reagent, NVAL as the Internal Standard, and DAD UV/Vis absorption as the detection mechanism. The results indicate that the method is simple, rapid and accurate for the determination of amino acids in Taizishen.3. Pseudostellarin B (cyclic peptide) was isolated and purified from Taizishen for the first time by high-speed counter-current chromatography (HSCCC) using a two-phase solvent system. Under the optimum operation conditions, four peaks were observed and collected individually for further characterization. Pseudostellarin B is the major peak, and the UV purity of the peak is 96%. The chemical structure of the compound has been positively confirmed by electrospray ionization time of flight (TOF) MS, 1H-NMR, 13C-NMR and 1H-13C-COSY analyses. The costs of running HSCCC is cheaper compared to those of preparative HPLC chromatography. It provides an attractive alternative to HPLC for the semi-preparative scale separation and purification of bioactive components in herbal extracts.4. A high performance liquid chromatographic (HPLC) method for the determination of Pseudostellarin B (cyclic peptide) in Taizishen has been established, and a HPLC/MS method was developed for the analysis of cyclic peptides (Pseudostellarins) in Taizishen The extract...