Efficient Extraction And Steady-state Preparation Of Anthocyanins From Cinnamomum Camphora Fruit

The fruit of camphor tree(Cinnamomum camphora L.Presl)is rich in anthocyanins.It is a kind of natural pigment with important utilization value.It is expected to be widely used in food,chemical industry,medicine and cosmetics.In this study,the main purpose of the research is to achieve the high-efficiency extraction and steady-state of anthocyanins extraction from C.camphora fruit.This research is mainly including the extraction of anthocyanins from C.camphora fruit with deep eutectic solvent(DES),the preliminarily determination of the main components of anthocyanins from C.camphora fruit by adopting chromatographic purification and compound property analysis,and the research on the molecular copigmentation of anthocyanins from C.camphora fruit with a variety of phenolic acids.The specific research content is as follows:In this thesis,the mature C.camphora fruits were collected,and then removed the kernel and thus freeze-dried to make the fruits peel powder as the experimental raw material.Five kinds of choline chloride based deep eutectic solvents were synthesized and used for the extraction of anthocyanins from C.camphora fruit.The results show that the extraction rate of of anthocyanins from C.camphora fruit with choline chloride-acetic acid was significantly higher than that with ethanol and the other four kinds of deep eutectic solvents.The four single factors(the molar ratio of the extraction solvent choline chloride to acetic acid,the material-to-liquid ratio,the extraction time and the extraction temperature)that affect the extraction of anthocyanins from C.camphora fruit were carried out by single factor experiments.And the value of each factor was optimized by Box-Behnken response surface method.The optimal extraction parameters were obtained as follows:choline chloride:acetic acid equals to 1:3,solid-liquid ratio is 1:20,extraction time is 30 min and extraction temperature is 30℃.Under these conditions,the one-time extraction rate of anthocyanins from C.camphora fruit was 9.12±0.2 mg/g.The separation and purification conditions and properties of anthocyanins from C.camphora fruit were studied.Among 10 kinds of macroporous resins with different polarities and pore diameters,the most suitable macroporous resin for the separation of camphor fruit anthocyanins was selected as D4020;After treated with macroporous resin and dextran gel,the camphor fruit anthocyanins were further purified.The main anthocyanins in C.camphora fruit were preliminarily determined to be cyanidin-3-O-glucoside and cyanidin-3-O-rutinoside through a ultraviolet-visible spectroscopy analysis and HPLC comparison experiment.These two kinds of anthocyanins accounted for 83.14%of the total anthocyanins content of C.camphora fruit.The results on the stability of anthocyanins in C.camphora fruit showed that the stability of camphor fruit anthocyanin solution is strong when the temperature is lower than 30℃and lower than p H 6,and different exogenous ions have different extent of influence on the stability of anthocyanins.Among them,Na~+has a minimal influence on the stability of anthocyanins.The effects of eight different phenolic acids on the molecular copigmentation of anthocyanins in C.camphora fruit were studied.And the effects of various phenolic acids on the colouring,wavelength red shift and thermodynamic properties of anthocyanins in C.camphora fruit varies at different concentrations,p H values and temperatures were investigated.The results showed that there were great differences in the molecular copigmentation effects of different phenolic acids,among which the most significant copigmentation happens when the erucic acid at 1:50 molar ration,and the hyperchromatic effect reaches to 118.45%,and the wavelength red shift 5 nm.At p H 1.0,the copigmentation between 4-coumaric acid and camphor fruit anthocyanin reaches significant level,and the hyperchromatic effect up to 116%.The wavelength red shift effect is the most significant when the erucic acid,caffeic acid and chlorogenic acid copigmented with camphor fruit anthocyanins at low p H value(1-4),and the red shift reaches 8nm.Under high temperature treatment,it is found from the thermal degradation kinetics study of anthocyanins in C.camphora fruit that the thermal degradation of anthocyanins was significantly inhibited when chlorogenic acid is at 70℃,80℃and 90℃.