Studies On The Chemical Constituents Of The Kernel Of Xanthoceras Sorbifolia Bunge

Xanthoceras sorbifolia Bunge（Sapindaceae）is single species genus plant,which is mainly produced in northern Inner Mongolia.It is a unique dual-use plant for medicine and food in China.Xanthoceras sorbifolia was listed as Mongolian medicine in the ethnography of China（1984 Edition）.Scholars at home and abroad mostly focus on the study of the chemical components on the shell,leaf,rhizome,flower,etc of Xanthoceras sorbifolia,However,seed kernel was the pharmaceutical raw material of clinical drug enuresis,the research on the chemical components mainly focused on more than 20 years ago.With the development of technology,there is still a lot of space deserving further study for the chemical constituents of this medicinal part.In addition,as a germplasm resource,its medicinal parts contain more bioactive substances which support biological breeding,so it has practical significance to further study.In this paper,the chemical components were separated and identified by various separation and purification technologies and spectroscopy methods,and the content determination and exploratory research on methodology of Xanthoceras sorbifolia were carried out by high performance liquid chromatography,which provides a reference for further study on material foundation of efficiency and quality control of Xanthoceras sorbifolia.In this paper,Xanthoceras sorbifolia seed kernel was treated with petroleum ether to remove oil after CO₂supercritical extraction,70%ethanol was used as the extraction solvent to further recover the concentrated maceration liquid until without ethanol taste.The ethanol extraction liquid of appropriate volume and concentration was obtained.Then,ethyl acetate,n-butanol with the saturating water were used to extract solvent,according to TLC,the ethyl acetate and n-butanol parts were separated for the study on Chemical components.A variety of chromatographic methods（thin layer chromatography,silica gel column chromatography,Sephadex LH-20 chromatography）were used to separate and purify 70%ethanol extract from the Xanthoceras sorbifolia seed kernel,and the structure of the isolated compounds was identified by physicochemical identification and spectrum method.In this paper,12 compounds isolated from Xanthoceras sorbifolia seed kernel were identified by physicochemical identification and structural identification,including 4 terpenoid saponins,2 flavonoids,2 organic acids,2 sterols,one alkaloid and 1 polyhydroxy alcohols.which were 3-O-α-L-arabinose-ivy saponin-28-O-α-L-rham-nopyranose-（1→4）-（6-O-acetyl）-β-D-glucopyranose-（1→6）-β-D-glucopyrano-side（1）,bungankasaponin D tripyridine（2）,bungankasaponin A tripyridine（3）,bungankasaponin B tripyridine（4）,adenine nucleoside（5）,5,7,4’-trihydroxy-3,6-dimethoxyflavone（6）,5,3’-dihydroxy-3,6,4’-trimethoxy-flavone-7-O-β-D-glucoside（7）,tetraol（8）,9-octade-canoic acid（9）,tripyridine（10）,β-sitosterol（11）and carotene（12）.The content of 5,7,4′-trihydroxy-3,6-dimethoxy flavone of Xanthoceras sorbifolia seed kernel from 6 batches was determined by HPLC,and the quality control evaluation method was established.Chromatographic conditions:Agilent C₁₈column（250×4.6 mm,5μm）,detection wavelength was 254 nm,column temperature was 30℃,The mobile phase was methanol-0.1%phosphoric acid water,gradient elution:0-25 min,30%-55%methanol;25-25.1min,55%-80%methanol;25.1-50 min,80%-90%methanol,injection volume was 25μL,flow rate was 1.0 m L/min.The average content of 5,7,4′-trihydroxy-3,6-dimethoxy flavone was 7.254 mg/g.The resolution between the adjacent peaks was more than 1.5,which meets the requirement of chromatographic separation.There is a good linear relationship between the mass concentration and the peak area in the range of 4.274～25.64 ug/m L.The standard curve is Y=199.3x+17.34,r>0.999.In this study,ten compounds were separated,compound 1,5,6,7,8 and 10were isolated for the first time in the Xanthoceras sorbifolia seed kernel.The method is simple and specific,which is sui Tab.for the determination of the content of Xanthoceras sorbifolia seed kernel.