Preparation method of high-activity oldenlandia diffusa total flavonoid and application of high-activity oldenlandia diffusa general flavone in liver peroxidation injury
The technology of Hedyotis diffusa and Hedyotis diffusa is applied in the extraction technology of traditional Chinese medicinal materials and the field of medicine, and can solve the problems of low total flavonoid biological activity, complicated operation and high energy consumption, and achieve the effect of convenient clinical application.
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Embodiment 1
[0032] A method for enzymolysis and extraction of total flavonoids from Hedyotis diffusa with high activity
[0033] 1. Experimental method
[0034] A highly active total flavonoids of Hedyotis diffusa that can prevent and treat non-immune liver damage, which is obtained by the following preparation method: sample pretreatment: dry and pulverize the medicinal material of Hedyotis diffusa, and pass through a 60-mesh sieve (aperture 0.25mm) , ultrasonically treated with petroleum ether, the petroleum ether was discarded, the filter residue was evaporated to dry petroleum ether, and this operation was repeated once, dried and cooled for later use. This step is to remove chlorophyll and fat-soluble components in Hedyotis diffusa, so as to facilitate the dissolution of flavonoids in the extraction process.
[0035] Take the pretreated Hedyotis diffusa powder, add 0.5-1.0% cellulase in an appropriate amount, add 30-50 times the volume of 50% ethanol solution, adjust the pH to 4.0-5...
Embodiment 2
[0044] Composition analysis of total flavonoids in Hedyotis diffusa prepared by different methods
[0045] 1. Experimental method
[0046] Using UPLC-MS to analyze the composition of the total flavonoids of Hedyotis diffusa prepared by different methods in Example 1, and to measure the content of flavonoids, it is intended to find the difference between the cellulase extraction method used in the present invention and other extraction methods. Differential components.
[0047] A Waters chromatographic column (HSS T3 2.5μm, 2.1mm×150mm column) was used; the mobile phase was phase A water (0.1% formic acid): phase B acetonitrile (0.1% formic acid); the flow rate was 1mL / min; the column temperature was 4°C; The injection volume was 5 μL. The gradient elution program is shown in Table 2. Sciex 5500 QTRAP mass spectrometer was used for MRM mass spectrometry analysis in positive and negative ion switching mode. The ion source parameters in positive ion mode are as follows: Sourc...
Embodiment 3
[0066] Protective effect of total flavonoids of Hedyotis diffusa under different preparation methods on hydrogen peroxide-induced liver injury
[0067] 1. Experimental method
[0068] H 2 o 2 Act on HL-02 cells for 4 hours, measure OD value by MTT method, calculate H 2 o 2 For the inhibition rate of cells, determine the optimal modeling concentration according to the inhibition rate.
[0069] The HL-02 cells to be tested were divided into a drug group, a model group, a control group, and a positive drug group, and at least three replicate wells were set up in each group to carry out the liver protection activity screening experiment. Control group: without any treatment. Model group: use the optimal concentration of H 2 o 2 Culture medium for 4h. Drug group: add drugs in advance (the total flavonoids of Hedyotis diffusa under different preparation methods in Example 1) to act for 12h, and use the optimal concentration of H 2 o 2 The culture medium was continued for 4...
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