Separation By Counter-Current Chromatography And Antioxidant Ability Of The Primary Chemical Composition From Caulis Lonicerae Japonica And Flower Of Forsythia Suspense

Caulis Lonicerae japonica Thunb. can contribute to clearing heat and detoxification, dispelling wind and removing obstruction in channels. Flower of Forsythia Suspense is the flower of Oleaceae plant Forsythia Suspense (Thunb.)Vahl, early spring blossoms, quantity is considerable, but its chemical composition was not still clear. Not only this has wasted the resources, also causes our development use to be blocked.High speed counter-current chromatography (HSCCC) is a unique liquid-liquid partition chromatography that uses no solid support matrix. Therefore, it eliminates irreversible adsorptive loss of samples onto the solid support matrix used in the conventional chromatographic column and the metamorphic change of samples. It is simple, economical, quick and with good repeatability. The method has been successfully applied to the analysis and separation of several natural products.1. Regarding the C. Lonicerae japonica Thunb., this experiment begins from the C. Lonicerae japonica Thunb. and Flos Lonicerae's antioxidant ingredient comparison, further carried on to the C. Lonicerae japonica Thunb. chemical composition has separated and purification.The primary coverage includes: The different collecting time C. Lonicerae japonica Thunb. and the Flos Lonicerae antioxidant ingredient's appraisal and their ethanol extract and Water extract the antioxidant analysis, C. Lonicerae japonica Thunb. high speed current-current chromatography research. Main result: (1) Their ethanol extraction's active constituent content and the antioxidant must be higher than the water extraction;In the ABTS system, in September, the C. Lonicerae japonica Thunb. antioxidant has surpassed the Flos Lonicerae;But in the FRAP system, the C. Lonicerae japonica Thunb. antioxidant must be higher than the Flos Lonicerae throughout.(2)The separation and purification of three compounds from the crude extract of C. Lonicerae japonica Thunb. was achieved by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of ethyl acetate-ethanol-water (4:1:5,v/v). Then, there are three compounds were obtained from the C. Lonicerae japonica Thunb. for the first time, including 7.5 mg of coffeic acid, 13.5 mg of chlorogenic acid, 21 mg of luteolin from 110 mg crude extracts and their purities were 95.55%, 97.24% and 98.11%, respectively.2. To the Flower of Forsythia Suspense, this experiment begins from Flower of Forsythia Suspense antioxidant ingredient content determination. Have carried on the inspection to its extraction process and the best collecting time,Then used the high-speed counter-current chromatography to carry on the separation and purification to its main chemical composition。Main research content:(1) Flower of Forsythia Suspense extraction process(2) HPLC determination different collecting time Flower of Forsythia Suspense main chemical composition content(3) Flower of Forsythia Suspense main chemical composition HSCCC analysis(4) Flower of Forsythia Suspense antioxidant ingredient content determination and ABTS and FRAP to its antioxidant analysisMain result:(1) The optimum extraction Technology is: Expects the material fluid ratio 1:30, ethyl ethanol density 95%, extraction temperature 100℃.(2) The best collecting time is: At the end of March。(3)HSCCC was applied to the separation and purification of Flower of Forsythia Suspense (Thunb.)Vahl.,140mg of crude extracts were separated by using ethyl acetate-ethanol-acetate acid- water (4:1:0.25:6,v/v) as the two-phase solvent system the purity of the three compounds was 96.39%,97.48%,95.22%, compoundⅢwas rutin, compoundⅠandⅡare identifying.(4) What the Flower of Forsythia Suspense total flavanone content the highest is on April 6; the total phenol content the highest is on March 26; In the ABTS system, what the antioxidant is strongest is on April 6; Influencing factor's primary and secondary relations are: The solvent polarity (ethyl ethanol density) > withdraws the temperature > the material fluid ratio; In the FRAP system, what the antioxidant is strongest is on March 26; Influencing factor's primary and secondary relations are: The solvent polarity (ethyl ethanol density) > the material fluid compares > withdraws the temperature.The experiment proved that in August and September recovery's C. Lonicerae japonica Thunb., its antioxidant ingredient content and antioxidant ability and Flos Lonicerae is almost the same, particularly raises the antioxidant ability has surpassed Flos Lonicerae. Through to Flower of Forsythia Suspense chemical composition's antioxidant ability appraisal, we may know that the Flower of Forsythia Suspense has the very strong antioxidant ability, particularly the end of March to the beginning of April recovery. This from now on fully will provide the important theory basis for us using the C. Lonicerae japonica Thunb. and the Flower of Forsythia Suspense resources.This experiment applies the high speed current-current chromatography to analyze the C. Lonicerae japonica Thunb. and the Flower of Forsythia Suspense for the first time, and distinguished a step separation to obtain three kind of compound. This method highly effective, fast, the specimen handling quantity big, eliminated the sample adsorption which, the pollution, the peak shape distortion brings as a result of the use carrier and so on bad situation, is a very good analysis traditional Chinese medicine plant ingredient method.