The Separationand Identification Of Chemicals From Puffball And The Inhibition Of Them To Cathepsin L

This article mainly studies the separation and identifying chemicals of puffball and the inhibition effect of puffball on Cathepsin L.The major medical applications of puffballs are for hematischesis, antibiosis, anticancer, anti-inflammatory and others. It was found that puffball was capable of inhibiting the activity of Cathepsin L in this study.Cathepsin L is acidophilia protease, which is regulated strictly on expression. If its expression is abnormal, it will cause disease such as cancer, cardiovascular. So, we must inhibit the activity of cathepsin L. Puffball has good inhibition on cathepsin L, which means some active compounds in puffball could be next generation drugs for anticancer and anticardiovascular. If we can find the active compound, it will benefit the human and society.Under this circumstances, we mainly studies the separation and identifying chemicals of puffball and the inhibition effect of the separated componets on Cathepsin L.The analytical HPLC is to optimize the separation condition. We introduce two preparative scheme namly routine repeatedly preparative HPLC and 2D Sepbox high throughput preparation, then the separated components are used to test bioactivity with Cathepsin L. The result indicats the 2D Sepbox high throughput preparation is better than routine preparative HPLC.LC-IT-TOFMS was applied to identify bioactive compounds. We inferred the structure of one compound,1,2-bihydro-2-hydroxy-5,10-dioxobenzo[g] quinoline-9-carboxylic acid. We researched it with molecular docking technology and the active spots was same with report.To analyze the compositons of puffball, different pretreatment methods, including headspace solid phase microextraction (HS-SPME), supercritical CO2 fluid (SFE-CO?), and traditional solvent ultrasonication, were applied. The extractants obtained from these methods were identified and their relative contents were determined by GC-MS.19 compounds, mainly alkanes and morpholine derivatives, identified by HS-SPME-GCMS and 24 chemicals with unusual high level of amides by SFE-CO2.17-26 components were obtained by traditional solvent extractions depending upon the solvent used. In this study, a total of 62 compounds were identified in puffball. The results showed that a combination of multiple methods should be a reasonable choice for the analysis.Because of the different inhibition effect from Inner Mongolia and Jilin, we analyzed the differences of them from microstructure, main class and chemical compositions by SEM, FTIR and Py-GCMS.In a word, we separated chemicals of puffball applying HPLC, GC-MS and identified the active compound. We tested the inhibition effect with puffball extractants from different places of origin and by different pretreatment for Cathepsin L. All the results provided great references for futher studing of chemicals puffball.