The Research Of Preparation And Medical Mechanism Analysis Of Active Ingredient In Artemisia Selengensis Turcz

As a plant resource with good development prospect, Artemisia selengensis Turcz has been used in several applications, such as stop bleeding, relieve cough, diminish inflammation, and stimulate appetite. This paper focused on Artemisia selengensis Turcz,using the novel sample pretreatment techniques for extraction and isolation of flavonoid compounds (such as rutin) from Artemisia selengensis Turcz. Meanwhile the fingerprint of Artemisia selengensis Turcz was established to conduct the quality control of this Traditional Chinese Medicine and then the molecularly imprinted solid-phase extraction (MISPE) procedure for the selective pre-concentration of rutin from complicated traditional Chinese medicine samples was proposed. At last the interaction between troxerutin and human serum albumin (HAS) was studied by the UV and fluorescence spectroscopy. The concrete contents are expressed as follows:1. Ultrasonically Assisted Extraction of Rutin from Artemisia selengensis TurczUltrasonically assisted extraction (UAE) followed by high performance liquid chromatography (HPLC) analysis method has been developed for the fast extraction and determination of rutin in Artemisia selengensis Turcz. The extraction parameters were respectively optimized and the optimal conditions were 90:10 (v/v) methanol–ethanol as solvent, 30:1 liquid–solid ratio, and 40 min extraction time. In order to show the superiority of UAE, other extraction techniques were investigated, including microwave-assisted extraction, reflux extraction and marinated extraction. The results showed that UAE was the most suitable technique for the extraction of rutin from Artemisia selengensis Turcz because of its high extraction efficiency. Reversed phase-HPLC with ultraviolet detection was employed for the analysis of rutin in Artemisia selengensis Turcz. Under the optimum conditions, the calibration curve for the analyte was linear in the range of 0.34–20.7μg/mL. The mean recovery of rutin was 100.77%, and its relative standard deviation was 0.37% (n=5). Three kinds of Artemisia selengensis Turcz from different habitats were investigated. The total content of rutin was 9.90, 6.23, 5.56 mg/g, respectively.2. Molecularly Imprinted Polymer for Solid-Phase Extraction of Rutin from Complicated Traditional Chinese Medicines An improved approach for the preparation of molecularly imprinted polymer (MIP) was proposed. Based on ultrasonic eluting template, the molecularly imprinted polymers (MIPs) were prepared by bulk polymerization and water-bath heating, using rutin as the template, acrylamide as the functional monomer and AIBN as the cross linker. The MIPs prepared by other elution method were used for the comparison. The synthesized MIPs were characterized by Fourier transform infrared (FT-IR) and scanning electron microscope (SEM). Then using high performance liquid chromatography evaluated the polymer recognition mechanism and adsorption ability. As a sorbent of solid phase extraction, the selective adsorption performance and the selective pre-concentration of rutin from complicated traditional Chinese medicine (TCM) samples were studied. The selectivity coefficient (k) of the MIPs is 2.80 and 5.46 for quercetin and genistein, the recoveries ranged between 85% and 91% for TCMs demonstrated that the MISPE could provide a new approach for pre-concentrating and determinating rutin from complicated TCM samples.3. Fingerprint profile of active components for Artemisia selengensis Turcz by HPLC-PAD combined with chemometricsAn approach was proposed to develop enhanced fingerprint by means of HPLC with photodiode array detection. In comparison with common fingerprint at a fixed wavelength, enhanced fingerprint compiled additional spectral data and it was more informative. So it could be used to conduct the quality control of this TCM comprehensively. Thereafter, the chromatographic fingerprint data set was submitted for classification with a suite of chemometrics methods viz. similarity analysis (SA), hierarchical clustering analysis (HCA) and principal component analysis (PCA). The correlation coefficients of them are 0.9839-0.9883, 0.8597-0.8907 respectively, which are consistent with their collecting locations. These results were corresponding with the PCA and same to HCA. These methods provided comprehensive information about matching and discrimination of the fingerprints, and appeared to be suited for quality assurance purposes for these similar types of sample.4. Spectroscopic studies on the interaction between troxerutin and human serum albuminTroxerutin was synthesized with rutin and ethylene oxide by Williamson etherification nucleophilic substitution reaction. Then spectroscopy was applied to study the interaction mechanism of troxerutin and human serum albumin (HAS). Results: The quenching mechanism of troxerutin acted on human serum albumin was static quenching; there are two binding sites in HSA; the thermodynamic parameters wereΔH>0,ΔS>0. Conclusion: The interaction between troxerutin and HSA is driven mainly by hydrophobic force, and troxerutin could change the structure of human serum albumin. This work illustrates its binding mechanism at a molecular level and it has special significance in pharmacology and clinical medicine field as well as methodology field.