Extraction, Purification, Antioxidation Activity and Stability of Red Pigment from Mulberries
|Course||Agricultural Products Processing and Storage|
|Keywords||Mulberry red pigment Extract Purification Qualitative Analysis Quantitative Analysis Antioxidant Stability|
Mulberry (Mulberry) commonly known as mulberry jujube, mulberry, Sang Real, mulberries, mature the Sangke Sang the genus Sang (Morusalba L.) FRUIT, mulberry contains large amounts of anthocyanin pigments, is a good The anthocyanin pigments resources. Mulberry water-soluble under alkaline conditions was blue, dark purple-red in acidic solution, with blood, Run brain, liver Lee, diuretic, anti-oxidation, anti-mutation, anti-tumor, a variety of pharmacological activity mulberry red pigment as a natural pigment, more and more the concern of the majority of scholars. Foreign mulberry red pigment reported fewer domestic mulberry red pigment research has focused on the extraction and purification process of the preparation of high purity by Mulberry has not been reported, this paper focuses on the red mulberry Purification and antioxidant activity of the pigment to provide a theoretical basis for the application of Mulberry Red Pigment. In this thesis, frozen mulberry raw materials, Mulberry Red Pigment extraction process, qualitative and quantitative analysis methods, purification processes, antioxidant activity and stability of the system, the main contents and results are as follows: studies of mulberry red pigment extracted processes. Solvent extraction method mulberry mulberry red pigment, affecting the yield of mulberry red pigment of the main factors temperature, time, liquid ratio and extraction concentration, the maximum absorption wavelength of 513 nm absorbance value of the indexes single factor experiment to study the impact of individual factors on the yield of pigment; selected on the basis of single factor experiment extraction temperature, extraction time, extraction concentration, liquid ratio for the study factors, the absorbance value of the indexes orthogonal test The results showed that mulberry red pigment extracted optimum conditions: temperature 50 ℃, extraction time 90min, the liquid ratio 15:1, the concentration of the extracts was 40% (80% ethanol: 0.1% HCl = 1:1) , mulberry red pigment extracted material was 102.6mg / g, validation tests showed that this process can be used for the extraction of mulberry red pigment. Of mulberry anthocyanins analysis methods. In this paper, four qualitative methods - color reaction, paper chromatography, thin layer chromatography and UV - visible spectroscopy main component of mulberry red pigment - mulberry anthocyanin qualitative analysis of four qualitative methods to determine the initial red mulberry The main component of the pigment cornflower -3 - glucosidase. Quantitative analysis method of mulberry red pigment colored price method, the extinction coefficient method, pH shows the difference method and nano 2 -Al (NO 3 ) 3 colorimetric method. Four quantitative methods were used for quantitative analysis of mulberry red pigment. The results show that, the purified color value is 14.8 times before purification, the extinction coefficient measured mulberry red pigment anthocyanin content to 0.015%, pH shows difference method measured obtaining content of 0.017%, Nano < / sub>-Al (NO 3 ) 3 colorimetric measured anthocyanin content of 0.0076%. pH differential method is more mature a method of previous studies, relatively accurate, simple, better than other methods. Study macroporous best conditions. To filter through static adsorption and parsing experimental four macroporous resin-NKA, D101, AB-8 and D1300, respectively, to calculate the rate of adsorption rate and resolution, the results show that, D101 macroporous resin highest rate of 72.1%, analytical rate of 68.5%, and therefore determine the best resin D101-oriented experiment. The main factors affecting the macroporous resin feed concentration, feed solution pH on the column speed, resolution effect of the main factors eluent concentration, eluent volume and elution speed, dynamic adsorption and dynamic resolution adsorption on the basis of single factor experiments, and single factor experiment and orthogonal experiment were resolved to the amount of adsorption and a resolution rate of the indexes to examine the effect of adsorption and resolve. By macroporous resin single factor and orthogonal experiment shows the best adsorption conditions: the feed concentration 1.071Abs (absorbance), the pH of the feed liquid column speed 1mL/min; through dynamic resolution single factor and is cross-experimental macroporous resin optimum analytical conditions: eluent concentration of 80% (ethanol), elution volume 2BV (column volume), elution rate 1mL/min this condition a resolution rate of 86.1% . Eluted with 4BV 0.01% HCl macroporous resin column, removal of mulberry red pigment crude was sugars and other soluble impurities; with 3BV of ethyl acetate three times to remove mulberry red pigment in the fat-soluble polyphenols impurities, carbohydrate impurities removal rate of 100%, fat-soluble polyphenols impurity removal rate was 39.65%, mulberry red pigment purified liquid. Mulberry red pigment purified liquid HPLC-DAD-ESI-MS analysis shows that the pigment contains at least 14 different anthocyanins, geranium -3 - glucose, geranium -3 - rhamnosidase, geranium -3 - rutin glucoside, cyanidin 3 - glucoside, cyanidin -3 - rutin Huai glucosidase, peony flower Su -3 - arabinoside, cornflower -3 - rutin glycosides, cornflower -3 - Bone wooden two-glucoside, cyanidin -3 - glycosides of xylose rutin, Mallow prime 3 - glucoside, cyanidin -3 arabinoside, petunia -3 - glycosides of glucose rutin, larkspur -3, 5 - Arab two glycosides, petunia -3 - glucosidase. Using Sephadex LH-20 macroporous mulberry red pigment and then purified. Preparation of different concentrations of acidic methanol solution (20%, 40%, 60%, 80%) as eluent Sephadex LH-20 column, and segment collection, the elution curve drawn three components: Component 1, Component 2 and the component 3; component 1 and component 2 of the HPLC analysis, the component 1 of cornflower -3 - glucoside-based (normalized results of the analysis showed that 85.8% of the dye component), the group sub-2 is cornflower -3 - elderberry two glycoside-based (normalized analysis showed that 78.5% of the component 2 pigment). Mulberry red pigment antioxidant experiments show that crude extracts of mulberry red pigment free radical scavenging effect is not obvious; mulberry red pigment purified liquid on superoxide anion radicals, hydroxyl radicals and peroxide hydrogen radical scavenging, when the concentration 40μg/mL, purified liquid on DPPH · scavenging rate has reached 93.5%; purified liquid · OH clearance rate increased with the increase of the concentration of the solution, and the mulberry red pigment solution · OH scavenging effect Vc. , mulberry red pigment concentration of the solution 600μg/mL · OH clearance rate reached 88.21%, the Vc and Mulberry Red Pigment IC 50 values ??were of 123.25,132.25 microg / mL; the purified liquid O 2 - sup> Clear clear effect was weaker than Vc, Vc and mulberry red pigment the IC 50 values ??respectively mulberry red pigment with oil for 304.94μg/mL and 514.02μg/mL; antioxidant effect, the effect is compared with reference BHT. Mulberry red pigment stability experiments show that: mulberry red pigment stability factor is the pH of the pigment is more stable under acidic conditions, bright colors, heating cause the pigment degradation, decreased stability; mulberry red pigment oxidant comparison sensitive to lower concentrations will make a lot of pigment degradation, even faded to colorless; metal ions Na , sup>, the Mg 2 sup> little impact on the stability of the pigment, Fe is 3 sup>, Cu 2 sup> mulberry red pigment, and the mulberry red pigment into a blue-purple.