Study On Chemical Constituents And Activities Of Waste Water From Levodopa Production

Objectives:To study the chemical composition of waste water from levodopa production and lay the foundation for improving the production process and improving product quality;To study the effect of petroleum ether part,ethyl acetate part,and n-butanol part of waste water on the proliferation of NG108-15 cells in vitro,and to provide scientific basis for the study of cell activity of waste water;To study the chemical constituents of petroleum ether part and ethyl acetate part from Stizolobium cochinchinensis and to study the antioxidant activity of petroleum ether part,further supplement the research data of non-levodopa components of Stizolobium cochinchinensis.Methods:We collected the waste water of the refining process in the production process of levodopa,concentrated and dried it,and broke it as concentrate sample.Stizolobium cochinchinensis was broken as sample.Two extracts samples were used 95%ethanol refluxing extraction,and achieve the ethanol extract,then respectively used system solvent separation method,extracted and achieved petroleum ether extract,ethyl acetate extract and n-butanol extract.By thin layer chromatography,silica gel column chromatographyandrecrystallizationmethod,sephadexcolumn chromatography,macroporous resin chromatography,preparation of high performance water chromatography?HPLC?technology and method for the separation and purification.Structure identification by IR,¹ H-NMR,¹³C-NMR,MS and so on,combined with the physical and chemical properties.The effects of petroleum ether extract,ethyl acetate extract and n-butanol extract on the proliferation of NG108-15 cells were studied by MTT method.We used GC-MS technology to test and analyse raw materials's petroleum ether extract,and used the clearing DPPH and ABTS⁺free radical ability experiments verify its antioxidant activity ability.Results:Twelve compounds were isolated from the waste water,respectively:Soyasapogrnol B?Octadecanoic acid?Linoleic acid?5?,8?-epi-?22E,24R?-dioxyergosta-6,22-dien-3?-ol?protocatechuic acid?6,7-dihydroxy-1-methyl-1,2,3,4-tetrahydro-isoquinoline-3-carboylic acid?6,7-dihydroxy-1-benzyl-1,2,3,4-tetrahydro-isoquinoline-3-carboylic acid?Phenylalanine.Seven compounds were isolated from Stizolobium cochinchinensis,five of them were identified,respectively:?-daucosterol?Stigmasterol?palmitic acid?1-triacontanol?Octadecanoic acid,2,3-dihydroxypropyl ester.Nineteen components that from petroleum ether extract of Stizolobium cochinchinensis were detected and identified by GC-MS.Petroleum ether extract,ethyl acetate extract and n-butyl alcohol extract of the waste water on NG108-15 has obvious inhibitory effect on cell proliferation in vitro,IC₅₀0 of petroleum ether extract is 37.14?g?mL^-1,IC₅₀0 of ethyl acetate extract is 44.89?g?mL^-1,IC₅₀0 of n-butanol extract is 49.03?g?mL^-1,IC₅₀0 of levodopa is 44.56?g?mL^-1.Petroleum ether extract of raw materials the has certain antioxidant activity ability,concentration of half clearance IC₅₀ alue concentration were46.62 mg?mL^-1,41.51 mg?mL^-1.Conclusion:Eight compounds were isolated and identified from the production waste water and five compounds were isolated and identified from the production Stizolobium cochinchinensis that all were isolated for the first time.It was first used by GC-MS to detect the presence of 19 components in petroleum ether extract of Stizolobium cochinchinensis,and all of them were identified,mainly fatty acids.Petroleum ether extract,ethyl acetate extract and n-butyl alcohol extract of waste water on NG108-15 has obvious inhibitory effect on cell proliferation in vitro,compared with the blank group?p<0.05 or p<0.01?that had statistical significance,compared with the control group,the value of IC₅₀ was arranged in order n-butyl alcohol extract>ethyl acetate extract>levodopa reference substance>petroleum ether extract.The inhibitory effect of petroleum ether extract was the strongest.By the method of purging DPPH and ABTS⁺free radical ability was used to detect the antioxidant activity of waste water's petroleum ether extract and showed a good linear relationship.