Method for sifting or detecting antibiotic active compound from plant extracts

A technology of antibacterial active substances and plant extracts, which is applied in the field of biological active ingredients, can solve the problems that there is no method for screening or detecting antibacterial active substances, and achieve the effect of simple methods, simple experimental equipment, and less workload

Inactive Publication Date: 2008-04-16
GUIZHOU UNIV
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  • Description
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  • Application Information

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Problems solved by technology

So far, there is no patent application for a method of screening

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Screening of 163 plant extracts

[0017] (1) Build an extraction library: get the extracts of 163 kinds of plants, mash 2 g of each plant raw material with a mortar and pestle respectively, then add 8 ml of 50% ethanol, and each extract requires a No. 6 tissue culture dish Shake for 24 hours in order to filter from the filter paper; then freeze-dry the filtrate for 16-24 hours to form a powder; the dry extract obtained by the freeze-drying method is suspended in 100 μl 50% ethanol; 163 A library of plant extracts.

[0018] (2) screen by this method, and judge which extracts have antibacterial activity according to the indicator color;

[0019] ①Extracts are screened for their activity by high-throughput methods:

[0020] Add 1 μl of each extract to a 96-well plate containing 100 μl of TSBYE medium and 10% overnight culture of S.mutans and A.actinomycetemcomitans liquid. Plates were incubated in petri dishes under anaerobic conditions for 16-18 hours. Afte...

Embodiment 2

[0022] Example 2 Antibacterial Activity Screening of Separation Fractions of Mulberry Leaf Extract:

[0023] (1) HPLC separation of crude extract:

[0024] The HPLC gradient elution method was adopted for separation, and 71 components were obtained.

[0025] (2) screen by this method, and judge which extracts have antibacterial activity according to the indicator color;

[0026] ① Antibacterial activity screening of 71 components by a high-throughput method: the method is the same as in Example 1.

[0027] ②Display the result

[0028] Components 4, 20, and 31 were blue, indicating that these three components had strong antibacterial activity.

[0029] (3) Blood agar plate to verify the antibacterial activity of component 31

[0030] Component 31 was diluted and added to the blood agar plate of the blood agar plate produced by Remel Company of the United States. After culturing, the number of clones was calculated, and the result was:

[0031] ①Inhibitory effect on Actinob...

Embodiment 3

[0035] Example 3 Antifungal Activity Screening of 86 Chinese Medicinal Materials

[0036] (1) Establishment of sample library

[0037] Collect 86 kinds of Chinese medicinal materials, each 100g, dry at 45°C, use a universal pulverizer to crush through a 40-mesh sieve, weigh 50g of each crushed medicinal material, place them in 250ml triangular flasks, and use 50% of 180ml soaked in ethanol solution, placed on a 37°C constant temperature shaker at 200 rad / min, and extracted for 12 hours. The extract solution and the solid residue are obtained, the extract solution and the solid residue are placed in a 4000rad / min centrifuge, centrifuged for 5 minutes, the supernatant is taken, and the extract of the medicinal material is obtained after suction filtration.

[0038] (2) Treatment of sample extract

[0039] The extraction solution of the medicinal material was concentrated under reduced pressure and rotation at 48°C to obtain a concentrated solution, which was frozen at -20°C ov...

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Abstract

The invention discloses a method for screening or detecting antibacterial active substance from plant extract. The method includes the following steps: (1) plant extract is prepared, or an extract bank is created based on the plant extract, or constituent or ingredient is obtained through separation and purification; (2) cultivation is completed according to the culture conditions of different bacteria and fungi; (3) each sample to be tested is added in bacterium culture solution or fungus culture solution; (4) 12 hours to 24 hours cultivation is completed under bacterium culture condition or fungus culture condition; (5) resazurin indicator is added in each culture solution; (6) according to the color change of resazurin, bacterium survival status is judged so as to judge whether the constituent or ingredient of the plant extract has bacteriostatic activity. With easy operation, simple experiment apparatus and less work load, the invention can obtain obvious results; moreover, the invention realizes fast screening of plant extract with antibacterial activity and screens out antibacterial active constituent or ingredient from the plant extract quickly, thereby providing basis for drug research; in addition, the invention can also be used in detecting plant antibacterial active constituent.

Description

technical field [0001] The present invention relates to biologically active components of natural substances, in particular to a method for screening or detecting antibacterial active substances from plant extracts. Background technique [0002] At present, due to a large number of people abusing antibiotics and various reasons, bacterial strains have developed strong drug resistance, and the use of antibiotics has also been greatly restricted. Compared with traditional antibiotics, natural antibiotics have the advantages of less toxic side effects, high antibacterial properties, and low resistance of bacterial strains to them. Therefore, searching for new antibiotics from natural resources will become a key to finding new antibiotics. way out. China has a tradition of using herbal medicines and their parts to treat diseases. Herbal medicines are rich in resources and produce many secondary metabolites, some of which can be used to treat bacterial diseases. Therefore, we c...

Claims

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Application Information

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IPC IPC(8): C12Q1/18
Inventor 周英黄赤夫钱利安
Owner GUIZHOU UNIV
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