Study On Chemical Constituents And Extraction,Isolation,Purification Of Flavonoids From The Leaves Of Camellia. Nitidissima Var.longistyla

Objectives: Study on the chemical constituents of n-butanol?saturated with water?ether portion and chloroform portion about the leaves of Camellia.nitidissima var.longistyla.To establish the optimum extraction and the preparation process technology for total flavonoids from Camellia.nitidissima var.longistyla.For reference to Vc,through determining �OH?DPPH free radical scavenging capacity,which provided a scientific basis for the development and analysis of Camellia.nitidissima var.longistyla.Methods: The leaves of C.nitidissima var.longistyla were collected and dried,extracted by 95% ethanol and leached by petroleum ether,chloroform extract,ethyl acetate and n-butanol?saturated with water?respectively.Sixteen compounds were isolated from C.nitidissima var.longistyla by the different process of the isolation such as Macroporous resin chromatography,thin layer chromatography,Silica gel column chromatography,sephadex column chromatography,polyamide chromatography,ODS column chromatography,high performance liquid chromatography preparation recrystallization.On the basic of the physical constants,chemical evidence and spectrum analysis [MS,IR,NMR(including ¹H-NMR,¹³C-NMR,DEPT)] twelve compounds were characterized.The content of total flavonoids in C.nitidissima var.longistyla was determined by spectrophotometry using rutin as standard substance.The L₉?3⁴?orthogonal extraction was used to investigate the influence of ethanol concentrations?extraction time?ratio of material to solvent?extraction temperature on the extracting amount of total flavonoids from C.nitidissima var.Longistyla.The influences of different factors and levels on the total flavonoids preparation process were investigated by means of single factor test.For reference to Vc,through determining �OH?DPPH free radical scavenging capacity.Results: Twelve compounds were separated and identified from chloroform extract and butyl alcohol extract of C.nitidissima var.longistyla,which were palmitic acid?CZ-1??3?-Acetoxy-6?,13?-dihydroxyolean-7-one?CZ-2?? keampferol-3-O-?-D-glucopy rainoside?CZ-3?? ?-spinasterol?CZ-4?? ?-daucosterol?CZ-5?? 5,7,3',4'-tetramethoxyflavone?CZ-6??Trimetboxy-7,3',4'-bydroxy-5-flavone?CZ-7?? 5,7,3',4',5'-pentamethoxyflavone?CZ-8??3-O-acetyl oleanolic acid?CZ-9??(n-tetratriacontanol?CZ-10?? ?-amyrin?CZ-11???-spinasterol-3-O-?-D-glucopyranoside?CZ-12?.For reference to rutin the content of total flavonoids in C.nitidissima var.longistyla is 57.50 mg/g.The optimum extraction process including the ethanol solvent concentration was 95%,the extracting time was 2h,respectively extraction 3 times,the ratio of material to solvent was 1:40,the extraction temperature was 40?.The antioxidant assays showed that the extracted total flavonoids from C.nitidissima var.Longistyla presented a strong antioxidant acativity to scavenge DPPH and �OH radicals.Conclusion: Twelve compounds were separated and identified from C.nitidissima var.Longistyla.Among them,seven compounds were isolated from Camellia nitidissima?Hu?Tuyama for the first time,which were palmitic acid?CZ-1?? 3?-Acetoxy-6?,13?-dihydroxyolean-7-one?CZ-2???-daucosterol?CZ-5?? 5,7,3',4'-tetramethoxyflavone?CZ-6??Trimethoxy-7,3',4'-bydroxy-5-flavone?CZ-7?? 5,7,3',4',5'-pentamethoxyflavone?CZ-8??3-O-acetyl oleanolic acid?CZ-9?.The content of total flavonoids in C.nitidissima var.longistyla is 57.50 mg/g.The optimum extraction process : the ethanol solvent concentration was 95%,the ratio of material to solvent was 1:40,the extraction temperature was 40 ?,the extracting time was 2h,respectively extraction 3 times.Establish the preparation technology of total flavonoids in C.nitidissima var.longistyla.drying through vaporation to get intermediates,with purity of total flavonoids in Camellia.nitidissima var.longistyla at 62.10%.The extracted total flavonoids from C.nitidissima var.Longistyla presented a strong antioxidant acativity.