Herba epimedii total flavonoid enzymatic product and preparing method and application thereof
An enzymatic hydrolysis product, the technology of Epimedium, applied in the field of medicine, can solve the problems of cumbersome separation and purification steps, the unfavorable rational application of traditional Chinese medicine resources, and the inability to fully utilize active ingredients, so as to reduce production costs, be suitable for industrial production, and facilitate recycling and repetition. The effect of reuse
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Embodiment 1
[0030] This example provides a preferred way of enzymatically hydrolyzing total flavonoids of Epimedium using the method of the present invention, and further specifically describes the enzymatic hydrolysis method of the present invention.
[0031] 1. Preparation of total flavonoids from Epimedium
[0032] Epimedium is extracted with 6-40 (m / v) times of 40%-90% ethanol by heat reflux, concentrated, and further purified by macroporous adsorption resin method to obtain total Epimedium with a mass content of not less than 60%. flavonoids.
[0033] The main components of the total flavonoids of Epimedium are icariin, epimedin A, epimedin B, epimedin C and baohuoside I, and icariin, epimedin A, epimedin B, The total mass content of Huodin C and Baohuoside I is not less than 30%.
[0034] 2. Preparation and characterization of amino mesoporous silica support
[0035] Immerse silica in toluene dissolved with 3-aminopropyltriethoxysilane (C=50mM), and stir magnetically at 800r·min -...
Embodiment 2
[0062] This example illustrates the effect of enzymatic hydrolysis of total flavonoids of Epimedium using other immobilized carriers.
[0063] This example compares the enzymatic hydrolysis effects of total flavonoids of Epimedium herba using barium alginate, microspheres and the amino mesoporous silicon-immobilized enzyme of Example 1. The results are shown in Table 2:
[0064] Table 2 Comparison of immobilized helicase prepared by different enzyme immobilization techniques
[0065]
[0066] It can be seen that the use of amino mesoporous silicon immobilized helicase has higher binding efficiency, better stability and higher conversion rate than other immobilized carriers.
Embodiment 3
[0068] This example illustrates the antitumor activity of the enzymatic hydrolyzate of the present invention.
[0069] Precisely weigh 2.65 mg of the total flavonoids of Epimedium and the free enzyme hydrolyzate and immobilized enzyme hydrolyzate obtained by the method of Example 1, respectively dissolve in 25 μL DMSO to prepare a mother liquor. Then the mother solution was diluted 100 times with RPMI-1640 incomplete medium solution, and then diluted to different dosage concentrations: 4.0, 17.0, 66.0, 265.0, 1060.0 μg·mL -1 . The MCF-7, A549, HepG2 and Hela cells in the logarithmic growth phase were inoculated in 96-well plates, and after 24 hours of culture, 20 μL of the drug solution to be tested was added. The final concentrations of the drug solution were 0.4, 1.7, 6.6, 26.5, and 106.0 μg mL -1 . Set up 6 duplicate wells for each sample, and set up a blank group of incomplete medium without drugs. After continuing to culture for 48 hours, MTT solution (5.0 mg·mL -1 ) ...
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