Sophora flavescens flavonoid active fraction with tyrosinase inhibitory activity and antibacterial activity as well as preparation method and application of sophora flavescens flavonoid active fraction

A technology with inhibitory activity and antibacterial activity, applied in the field of medicine, can solve the problems of green, sustainable development of the Sophora flavescens resource industry, large amount of solid waste and by-products, waste of resources, etc. The effect of high flavonoid content

Active Publication Date: 2019-02-15
NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Sophora flavescens is a commonly used variety of traditional Chinese medicinal materials, and the market demand is increasing year by year. At the same time, the solid waste and by-products generated in the preparation pr

Method used

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  • Sophora flavescens flavonoid active fraction with tyrosinase inhibitory activity and antibacterial activity as well as preparation method and application of sophora flavescens flavonoid active fraction
  • Sophora flavescens flavonoid active fraction with tyrosinase inhibitory activity and antibacterial activity as well as preparation method and application of sophora flavescens flavonoid active fraction
  • Sophora flavescens flavonoid active fraction with tyrosinase inhibitory activity and antibacterial activity as well as preparation method and application of sophora flavescens flavonoid active fraction

Examples

Experimental program
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Effect test

Example Embodiment

[0025] Example 1 Preparation of effective parts

[0026] 10 kg of Sophora flavescens dregs (15 times the volume concentration of 1% acetic acid aqueous solution dialysis method to extract alkaloids Sophora flavescens dregs) was ultrasonically extracted 3 times with 8 times the amount of 80% ethanol, each time for 1 hour, and concentrated to Dry, take the dried product and dissolve it with 95% ethanol by volume, filter, concentrate the filtrate under reduced pressure, dry, then add methanol to ultrasonically dissolve, and then apply the macroporous resin AB-8 chromatography column, using 8 column volumes with a volume concentration of 10 %, 20%, 30%, 40%, 50%, 60%, 70% and 80% ethanol were eluted in sequence, and each elution site was collected.

[0027] The present invention uses the following method to determine the flavonoid content of the eluate with different volume concentrations of ethanol (10% to 80%).

[0028] UPLC-TOF-MS / MS detection conditions:

[0029] Instrument: AB Sciex...

Example Embodiment

[0043] Example 2 Evaluation of tyrosinase inhibitory activity of effective parts of Sophora flavescens dregs extract

[0044] Take the 10-80% ethanol eluate prepared in Example 1, respectively, dissolve them in methanol, first prepare a 5mg / mL solution, and then dilute to 2.500, 1.250, 0.625, 0.312, 0.156, 0.078mg·mL -1 Six concentrations. Prepare 5mg·mL -1 L-tyrosine solution, 0.1mol / L phosphate buffer, 15mmol / L L-tyrosine solution, 0.25mg·mL -1 Tyrosinase and other solutions. Put each group of reaction solution in a test tube, after constant temperature in a 37℃ water bath for 10 minutes, add 0.50 mL of L-tyrosine solution, react for 15 minutes, quickly move it into a cuvette, measure the absorbance of each group at 475nm, and calculate Inhibition rate of tyrosinase by extracts of different polar parts.

[0045] Enzyme activity inhibition rate=[(A2-A1)-(B2-B1)] / (A2-A1)×100%

[0046] A1 is the absorption value without inhibitor at 0 min; A2 is the absorption value without inhibito...

Example Embodiment

[0051] Example 3 Evaluation of antibacterial activity of effective parts of Sophora flavescens medicine residue extract

[0052] Take the 10-80% ethanol eluate prepared in Example 1, respectively, dissolve them in methanol, first prepare a 5mg / mL solution, and then dilute to 2.500, 1.250, 0.625, 0.312, 0.156, 0.078mg·mL -1 Six concentrations. The qualitative filter paper is made into a 6mm diameter disc with a hole punch, and placed in a dry petri dish. After autoclaving at 121°C for 20 minutes, the filter paper is taken and placed in the sterilized petri dish. Add 5μL of liquid medicine dropwise to each piece of paper, and place the solvent in a drying box to evaporate the solvent for later use. Use a sterile cotton swab to evenly smear the diluted bacterial solution on the LB agar, take the sterilized paper slices in turn with sterilized tweezers, and dip them on the agar in turn. After placing them for 30 minutes, incubate them in a 37°C incubator for 24 hours Then observe th...

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Abstract

The invention discloses a sophora flavescens flavonoid active fraction with tyrosinase inhibitory activity and antibacterial activity and a preparation method of the sophora flavescens flavonoid active fraction. The preparation method comprises the following steps: taking sophora flavescens medicinal residues as raw materials, changing wastes into valuables, adding ethanol for reflux extraction, filtering, drying filtrate to obtain an ethanol extract, dissolving the extract with methanol, loading to AB-8 type macroporous adsorbent resin, then performing gradient elution with ethanol with the concentrations of 10 percent, 20 percent, 30 percent, 40 percent, 50 percent, 60 percent, 70 percent and 80 percent, and collecting 50-70 percent of elution part. The active fraction prepared with thepreparation method disclosed by the invention has obvious inhibitory activity to tyrosinase, has an obvious antibacterial effect on staphylococcus aureus and streptococcus agalactiae, can also well prevent and treat a human pigmentation disease caused by excessive melanin and has obvious and extensive biological activity.

Description

Technical field [0001] The invention relates to a traditional Chinese medicine extract, in particular to the effective part of flavonoids with tyrosinase inhibitory and bacteriostatic effects in the sophora flavescens dregs, belonging to the technical field of medicine. Background technique [0002] Sophora flavescens is the dried root of the leguminous plant Sophoraflavescens Ait. It has bitter taste, cold nature, clearing away heat and dampness, killing insects and diuresis. Sophora flavescens is widely distributed and grown in most parts of our country; it is mainly produced in Shanxi, Henan, Hebei, Inner Mongolia and other places. Modern pharmacological research shows that the alkaloids in Sophora flavescens have biological activities such as anti-tumor, anti-hepatitis B virus, anti-inflammatory, antibacterial and insecticidal activities, and are widely used in the development of pharmaceutical preparations, biological pesticides, veterinary drugs and other products. Sophora...

Claims

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Application Information

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IPC IPC(8): A61K36/489A61K31/352A61K8/9789A61K8/49A61Q19/02A61P17/00A61P35/00A61P31/04A23B7/154
CPCA23B7/154A23V2002/00A61K8/498A61K31/352A61K36/489A61K2236/333A61K2236/55A61K2800/782A61K2800/805A61Q19/02A61K8/9789A61P17/00A61P31/04A61P35/00A23V2200/048A23V2250/21A23V2300/14
Inventor 段金廒郭盛翁泽斌钱大玮
Owner NANJING UNIVERSITY OF TRADITIONAL CHINESE MEDICINE
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