| Imperial chrysanthemum, belong to morifolium genus and perennial herb, is one of Chinese traditional herbal medicine, possess extraordinary high edible and medicinal value. In this paper, the imperial chrysanthemum as the investigate object, through extraction, separation and purification of the imperial chrysanthemum pigment, The qualitative analysis of water-soluble pigment components was performed by means of solubility and color reaction, then the pigment components were separated and purity tested by silica gel column chromatography and HPLC. and then utilizing its biological function,in order to evaluate its hypoglycemic activity, the stability of water-soluble pigment and the ?-glucosidase inhibitory activity test in vitro were studied.Test results are as follows: 1?Through the comparison of purification effect of 8 kinds of macroporous resins, DM-28 macroporous resin, possessing the best purifying effect on the water-soluble pigments of chrysanthemum as the investigate index, Static adsorption and desorption conditions of DM-28 resin to the water-soluble pigments of chrysanthemum were optimize, as well as the dynamic, The results showed that the optimal static adsorption conditions were as follow: adsorption equilibrium time was 3 hours, the concentration of sample was A390=0.361(namely diluted 12 times), sample p H was 2.5, adsorption temperature was 30?, p H 3.0 50% ethanol as eluent. The optimal dynamic adsorption conditions were that: sample concentration was A390=0.361, the sample flow rate was 2.0ml/min, the flow rate of 1.0ml/min, 80% ethanol was collected. The color value of the pigment was 22.5, which was 3.65 times that of the former. DM-28 macroporous resin show better comprehensive adsorption property, and satisfy the requirements of large-scale production.2?The results stability of the water-soluble pigment showed that: light and commonly used food additives such as saccharose?citric-acid?tartaric-acid have scarcely effect on the stability of pigment, but the glucose can enhance the colour and lustre of pigment, on the contrary as the lactose; the pigment has good acid and alkali resistance; 90 ? or above will cause the degradation of the pigment, therefore in order to prevent the pigment changing in the structure of that degradation, the production personnel should pay attention to high temperature in the production process; deoxidizers could accelerate the degradation of pigment, and the more pigment concentration is high, the more degradation degree of pigment is serious; most metal ions have little effect on the stability of the pigment, but non-metal ions such as Fe3+?Cu2+ will have a great impact on the stability of pigment, so the pigment should avoid contacting with these metal ions in production and storage process. The pigment is relatively stable, can be considered as a natural pigment to be added to the food industry.3?The ?-glucosidase inhibitory activity test results of chrysanthemum water-soluble pigment inhibitor in vitro showed that the chrysanthemum water-soluble pigment inhibitors have a good dose-effect relationship on ?-glucosidase inhibitory effect, and have a significant inhibitory effect. Through the SPSS statistics analysis, it shows that: the inhibitory effect of the acarbose?non-purified and purified samples on IC50 values respective were 0.521,0.307, 0.779 mg/m L; and the samples combine with the enzyme rapidly, its role is non-competitive inhibition. The water-solubility extractive of chrysanthemum water-soluble pigment was considered to be developed into a ?-glucosidase inhibitor.4?The solubility of the sample observation, chromogenic reaction were used to makes a qualitative analysis of the water-soluble pigment composition, found that preliminary judgment of main component as flavonols of two hydrogen with two phenolic hydroxyl groupa; then by silica gel column chromatography, 11 compounds were separated, and the HPLC is used for the purity check of 11 compounds. The results showed that 11 compounds with purity higher than 95%, can be considered as monomer compound; by melting-point test, the melting point of these 11 monomer compounds were determined; Using high resolution mass spectrometry(HRMS)?IR(infrared spectroscopy)?1H NMR? 13 C NMR and other means to determine its chemical structure need to be further studied. |
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