Studies On Chemical Constituents And Bioactivity Of The Disporopsis Pernyi(HUA)Diels

Disporopsis pernyi (Hua) Diels which belongs to Disporopsis genus of Liliaceae family, mainly grows in south Asia up to an altitude of500-2500m. Disporopsis pernyi (Hua) Diels has been widely used in folk medicine for the treatment of abnormal sweating, chronic cough, women postpartum weakness and irregular menstrual etc. The chemical constituents, extraction process, the quality control, antimicrobial and antioxidant ability of Disporopsis pernyi (Hua) Diels were investigated in this study.Nine chemical constituents were isolated from70%ethanol extraction of the root of Disporopsis pernyi (Hua) Diels, by using silica gel column chromatography, Sephadex-LH20column chromatography. The structures of following7compounds (Stearic acid, Palmitic acid, Rutin, Luteolin, Betulin, Salicylic acid, Glucose) were fully elucidated by chemical, physical and spectroscopic methods (MS, UV, IR and NMR). Among them, Stearic acid, Palmitic acid and Betulin were isolated from Disporopsis pernyi (Hua) Diels for the first time.High performance liquid chromatography quadrupole of flight time mass spectrometry (HPLC-Q-TOF-MS) was applied to study the phenols components of Disporopsis pernyi (Hua) Diels. The analysis was performed on a Diamonsil C18column using gradient elution with the mobile phase consisting of0.1%formic acid in acetonitrile and water (pH2.4) and the flow rate was1.0mL·min-1. The detection was performed under anion pattern detection mode. Standards, references and other methods were used to identify the5phenols compounds of Disporopsis pernyi (Hua) Diels. Gas chromatography tandem mass spectrometry (GC-MS) method was developed to study the volatile compounds of Disporopsis pernyi (Hua) Diels. The essential oil analysis was carried out on SHIMADZU QP5050A series instrument coupled with HP-5MS capillary column (30m×0.25mm; film thickness0.25μm). Twelve volatile compositions of Disporopsis pernyi were identified for the first time.The content of10elements were determined by Flame Atomic Absorption Spectrometry (FAAS). The samples needed to be digested in nitric acid-perchloric acid (4:1, v/v) on the heating pad before measured. The result shows that Na, Mg, K, Cu, Zn and Ca are widely existed in Disporopsis pernyi (Hua) Diels, while the content of As, Cr, Mo and Pb are all lower than limit content. This method was found to be simple, rapid, accurate and good for the determination of Na, Mg, K, Cu, Zn, Ca, As, Cr, Mo and Pb in Disporopsis pernyi.Polyphenols were quantified by ultra-performance liquid chromatography (UPLC). A total of4polyphenolic compounds found in extract were identified and presented:rutin, luteolin, quercetin, betulinic acid.Disk diffusion method and gradually tube dilution method were used to investigate the antibacterial ability of essential oil, total saponins, total polyphenols and total polysaccharide. Through the determing the size of inhibition zone and MIC, the antibacterial ability of essential oil, total saponins, total polyphenols and total polysaccharide of Disporopsis pernyi (Hua) Diels was evaluated systematically. The antimicrobial activity of the four extracts was evaluated against4microorganisms. The total polyphenols of Disporopsis pernyi showed antimicrobial activities comparable with standard antibiotics, while the other extracts did not showed antimicrobial activity.The antioxidant potential was evaluated using following in vitro methods:FRAP and DPPH radical. The antioxidant activity of samples was compared with the activity of BHT. Results showed that all extracts possessed concentration-dependent antioxidant activity. The antioxidant activities were in descending order:BHT> total polyphenols> total saponins> total polysaccharide> essential oil.To indicates the differences between composition compounds of essential oil, in vitro antimicrobial and antioxidant activity of Disporopsis pernyi and those of its adulterants(Polygonatum odoratum, Panax japonicus, Recineckea carnea and Tupistra chinensis). Although these herbs have similarities in composition, there were remarkable differences between them. The methanol extract of Tupistra chinensis showed antimicrobial activities against the tested microorganisms in the disc diffusion test. The antioxidant potential was evaluated using following in vitro methods:Ferric reducing antioxidant power (FRAP) and2,2-diphenyl-l-picrylhydrazil (DPPH) radical. Results showed that all extracts possessed concentration-dependent antioxidant activity. The abilities of antioxidant activities were in descending order:BHT>Tupistra chinensis>Recineckea carnea> Panax japonicus>Polygonatum odoratum>Disporopsis pernyi.In conclusion, the constituents of the Disporopsis pernyi (Hua) Diels were investigated by HPLC-Q-TOF-MS, GC-MS, FAAS, UV and various analytical techniques. The the antioxidant and antimicrobial activities of Disporopsis pernyi and those of its adulterants were determined. This study provided useful information for research of chemical constituents study, quality control and biological activity of Disporopsis pernyi (Hua) Diels.