阅读说明：本技术 一种茶叶提取物及其工业化制备方法和用途 (Tea extract and industrial preparation method and application thereof ) 是由 王昭日 刘明川 杨胜杰 崔玉梅 于 2019-10-06 设计创作，主要内容包括：本发明涉及一种茶叶提取物及其工业化制备方法和用途,所述茶叶提取物中总儿茶素含量不低于85wt％,表没食子儿茶素没食子酸酯含量在65wt％以上,咖啡因含量不高于0.3％。本发明将茶叶提取、纯化、浓缩、干燥后,得到茶叶提取物,收率比现有工艺提高近3倍。未使用任何有毒有机溶剂,较为适合大规模生产。所制备的茶叶提取物具有较好的抗雾霾、保护肺功能,并且对神经细胞具有保护作用。(The invention relates to a tea extract, an industrial preparation method and application thereof, wherein the content of total catechins in the tea extract is not less than 85 wt%, the content of epigallocatechin gallate is more than 65 wt%, and the content of caffeine is not more than 0.3%. The invention extracts, purifies, concentrates and dries the tea to obtain the tea extract, and the yield is improved by nearly 3 times compared with the prior art. Does not use any toxic organic solvent, and is suitable for large-scale production. The prepared tea extract has good haze resistance, lung protection function and nerve cell protection function.)

1. A tea extract characterized by: the total catechin content is not less than 85 wt%, epigallocatechin gallate content is above 65 wt%, and caffeine content is not higher than 0.3%.

2. The tea extract of claim 1, wherein: said tea is selected from green tea, preferably Duyun Maojian, Xinyang Maojian, Biluochun and Huangshan Maofeng; the tea leaf is selected from above-ground or below-ground part, wherein the above-ground part is preferably selected from leaf, stem or seed.

3. The method for producing a tea leaf extract according to claim 1 or 2, characterized by being produced by the following method: mixing fresh or dried tea with alcohol-water solution, sequentially extracting, purifying, and drying to obtain tea extract.

4. A process for the preparation of a tea extract according to claim 3, characterized in that it is prepared by the following process:

(1) cleaning tea leaves, air drying, or pulverizing to obtain tea leaf extract material.

(2) Mixing a certain amount of tea leaf extraction raw materials with an alcohol water solution (the volume concentration of alcohol is 20-50%) according to a mass ratio of at least 1:5, and extracting at a constant temperature of 70-90 ℃ for at least 30min for at least 2 times;

(3) after extraction is finished, cooling the medicinal material extracting solution in the step (2) to room temperature, filtering, and concentrating the filtrate until no alcohol smell exists;

(4) and (3) performing No. 1 column chromatography separation on the filtrate in the step (3), concentrating the obtained eluent until no alcohol smell exists, performing No. 2 column chromatography separation, concentrating the obtained eluent until the relative specific gravity of the eluent is 1.05-1.20 at about 50 ℃, and drying to obtain the tea extract powder.

5. The method of claim 4, wherein: the filler of the No. 1 column chromatography is polyamide resin, macroporous resin such as LXD-200, HP-20, LX-8, D101, AB-8 and the like, preferably polyamide resin; the filler of the No. 2 column chromatography is macroporous resin such as LXD-200, HP-20, LX-8, D101, AB-8 and the like, and preferably LXD-200 or HP-20 macroporous resin is selected.

6. The method of claim 4, wherein: the No. 1 column chromatography separation method comprises the following steps: adding the extractive solution into a chromatographic column filled with polyamide resin, eluting with at least 2BV (column volume) of water, eluting with at least 2BV of alcohol solution with concentration of at least 60%, and collecting the alcohol eluate; the No. 2 column chromatography separation method comprises the following steps: concentrating the No. 1 column chromatography alcohol-water eluent until no alcohol smell exists, performing No. 2 column chromatography separation, eluting with at least 2BV water, eluting with at least 2BV alcohol solution with volume concentration of at least 20%, and collecting the alcohol eluent.

7. The method of claim 4, wherein: the alcohol is methanol, ethanol, n-butanol or isopropanol, preferably ethanol; the concentration method is vacuum concentration, normal pressure concentration, membrane concentration, preferably vacuum concentration; the drying method comprises vacuum drying, heating drying, airing, air drying, freeze drying and spray drying; preferably, the yield of the tea extract is not less than 6 wt%.

8. A composition comprising the tea leaf extract of 1 or 2, and a pharmaceutically or food acceptable auxiliary; preferably, the dosage form of the composition is selected from plain tablets, film-coated tablets, sugar-coated tablets, enteric-coated tablets, dispersible tablets, capsules, granules, oral solutions or oral suspensions, and cosmetic dosage forms such as liquid, emulsion, cream, powder, block and the like.

9. Use of the tea leaf extract according to claim 1, the composition according to claim 8 for the preparation of a food, health food or pharmaceutical product for preventing, ameliorating or treating a nerve-related disease; the application of the compound can be used for preparing food, health-care food or medicines for treating or preventing symptoms caused by PM 2.5 particles.

Technical Field

The invention relates to a tea extract product, which has high content of epigallocatechin gallate and chemical components of total catechins. The invention also relates to a preparation process of the tea extract, which is prepared by extracting, purifying, concentrating and drying, and the extract can be used as food, health-care food or medicine.

Background

Tea has a long history of thousands of years in China, and the modern drinking of tea is beneficial to health by the introduction that 'hundreds of grass are tasted by Shennong and the tea is solved by seventy-two toxins every day' from ancient times, so that the common general knowledge is provided. Tea varieties are numerous in China, wherein green tea has the advantages of monopoly and large head, the coverage is the widest, and the green tea is popular with the public. The tea polyphenols account for about 20-30% of the dry weight of green tea, wherein the main active ingredient is catechin compound. The catechins mainly comprise the following components: catechin (C), Epicatechin (EC), Gallocatechin (GC), Epigallocatechin (EGC), epicatechin gallate (ECG), epigallocatechin gallate (EGCG), Catechin Gallate (CG), gallocatechin gallate (GCG), etc. A large number of scientific researches at home and abroad show that the catechin compounds have various pharmacological effects of inhibiting bacteria, diminishing inflammation, resisting tumors, resisting viruses, resisting oxidation, reducing blood fat and blood sugar, inhibiting obesity, resisting senile dementia, regulating immunity and the like. Wherein EGCG is main component of green tea polyphenols, and is also main component of green tea catechins. Many studies have shown that EGCG has the effects of protecting against free radical DNA damage, against radiation and ultraviolet radiation, preventing lipid peroxidation, reducing serum low-density cholesterol, ultra-low-density cholesterol and triglyceride levels, interfering with signaling required for cancer cell survival, inhibiting carcinogens in the diet, preventing the viability of certain carcinogens in conjunction with other enzymes and antioxidants in the intestine, liver, and lung, scavenging free radicals, combating pollution, sun exposure and smoking, and preventing skin aging and wrinkling.

Because of the complex components of green tea, the preparation of a tea extract containing high-purity catechins and high-content chemical components of EGCG from the green tea has great technical difficulty. US patent No. US7763291 extracts dried green tea with hot water, extracts the aqueous extract with ethyl acetate, concentrates the ethyl acetate and then passes through macroporous resin, collects the eluent with 40% methanol solution by volume concentration, concentrates and dries to obtain tea extract with total catechins and EGCG contents respectively accounting for 85% and 55% or more, with a yield of 2.2 wt%. Although the catechin and EGCG components with higher content are prepared, a large amount of ethyl acetate and methanol are used in the process, the organic solvent is toxic and harmful, flammable and explosive, causes harm to human health, brings risks to the production environment, is neither safe nor environment-friendly, is not green in production, and in addition, the yield of the tea extract obtained in the process is lower, so that the resource utilization rate is low, the cost is increased, and the green production cannot be met.

The ethanol is a common organic reagent, has the advantages of no toxicity, low price, easy recovery and the like, and compared with expensive instruments and toxic chemical reagents, the ethanol saves the cost and reduces the pollution of waste liquid discharge to the environment. Aiming at the problem, the tea is extracted by using a low-alcohol-content ethanol water solution, and then purified by using column chromatography, so that the tea extract with high content of catechins and EGCG is obtained, and the yield is improved by nearly 3 times. The process has the advantages of high resource utilization rate, simple and convenient operation, low energy consumption, reduction of production cost, environmental protection and suitability for large-scale production.

Disclosure of Invention

The invention aims to provide a tea extract with higher content of gallocatechin gallate and total catechins.

Another object of the present invention is to provide a method for preparing a tea extract having a high content of gallocatechin gallate and total catechins.

The invention also provides application of the tea extract in preparing food, health-care food or medicine for preventing, improving or treating nerve-related diseases.

The invention also provides application of the tea extract in preparing food, health-care food or medicine for improving or treating memory decline.

The invention also provides the application of the tea extract in preparing food, health-care food or medicine for treating or preventing Parkinson's disease and Alzheimer's disease.

The invention also provides application of the tea extract in preparing food, health-care food or medicines for treating or preventing symptoms caused by PM 2.5 particles.

Another object of the present invention is to provide the use of the tea extract of the present invention for the preparation of food, health food or pharmaceutical products for protecting or improving lung function.

The invention also provides the application of the tea extract in preparing food, health food or medicine for treating or preventing lung diseases.

The purpose of the invention is realized by the following technical scheme:

a tea extract is characterized by being prepared by the following method: mixing fresh or dried tea with alcohol-water solution, sequentially extracting, purifying, and drying to obtain tea extract.

Preferably, the tea extract is characterized in that the tea is selected from above-ground or below-ground parts, wherein the above-ground parts are preferably selected from leaves, stems or seeds.

Preferably, the tea extract is characterized by being prepared by the following method:

(1) cleaning tea leaves, air drying, or pulverizing to obtain tea leaf extract material.

(2) Mixing a certain amount of tea leaf extraction raw materials with an alcohol water solution (the volume concentration of alcohol is 20-50%) according to a mass ratio of at least 1:5, and extracting at a constant temperature of 70-90 ℃ for at least 30min for at least 2 times;

(3) after extraction is finished, cooling the medicinal material extracting solution in the step (2) to room temperature, filtering, and concentrating the filtrate until no alcohol smell exists;

(4) and (3) performing No. 1 column chromatography separation on the filtrate in the step (3), concentrating the obtained eluent until no alcohol smell exists, performing No. 2 column chromatography separation, concentrating the obtained eluent until the relative specific gravity of the eluent at 50 ℃ is 1.05-1.20, and drying to obtain tea extract powder.

Preferably, the filler of the No. 1 column chromatography is polyamide resin and macroporous resin such as LXD-200, HP-20, LX-8, D101, AB-8 and the like, and the polyamide resin is more preferably selected.

Preferably, the filler of the No. 2 column chromatography is macroporous resin such as LXD-200, HP-20, LXD-8, D101, AB-8 and the like, and more preferably LXD-200 or HP-20 macroporous resin.

Preferably, the No. 1 column chromatography separation method comprises the following steps: adding the extractive solution into a chromatographic column filled with polyamide resin, eluting with at least 2BV (column volume) of water, eluting with at least 2BV of alcohol solution with concentration of at least 60%, and collecting the alcohol eluate.

Preferably, the No. 2 column chromatography separation method comprises the following steps: concentrating the No. 1 column chromatography alcohol-water eluent until no alcohol smell exists, performing No. 2 column chromatography separation, eluting with at least 2BV water, eluting with at least 2BV alcohol solution with volume concentration of at least 20%, and collecting the alcohol eluent.

Preferably, the alcohol concerned may be methanol, ethanol, n-butanol, isopropanol, etc., more preferably ethanol.

Preferably, the yield of the tea extract is not less than 6 wt%.

Preferably, the tea extract is characterized in that: the total catechin content is not less than 85 wt%, the gallocatechin gallate content is above 65 wt%, and the caffeine content is not higher than 0.3%.

Preferably, the tea extract is characterized in that the concentration method in the preparation method can be vacuum concentration, atmospheric concentration and membrane concentration, and preferably the vacuum concentration method is used.

Preferably, the tea extract is characterized in that the drying method in the preparation method can be vacuum drying, heating drying, airing, air drying, freeze drying and spray drying.

A composition comprises the above tea leaf extract and pharmaceutically or dietetically acceptable adjuvants.

Preferably, the dosage form of the composition is selected from plain tablets, film-coated tablets, sugar-coated tablets, enteric-coated tablets, dispersible tablets, capsules, granules, oral solutions or oral suspensions, and cosmetic dosage forms such as liquid, emulsion, cream, powder, block and the like.

The tea extract and the composition thereof are applied to preparing foods, health-care foods or medicines for preventing, improving or treating nerve-related diseases; the application of the compound can be used for preparing food, health-care food or medicines for treating or preventing symptoms caused by PM 2.5 particles.

Compared with the prior art, the invention has the following advantages:

(1) in the preparation process of the tea extract, only ethanol and water are used as solvents, and any other toxic, harmful, flammable and explosive chemical substances or solvents are not used, so that the preparation process is harmless to human bodies, high in process safety coefficient, green, environment-friendly, pollution-free and residue-free.

(2) The yield of the preparation process of the tea extract is 3 times of that of the prior art, the resource utilization rate is high, and the production cost is saved.

(3) The preparation process of the tea extract ensures the content of effective components and reduces the content of caffeine to be below 0.3 percent.

Examples

The invention is further illustrated by the following examples. It should be understood that the method described in the examples is only for illustrating the present invention and not for limiting the present invention, and that simple modifications of the preparation method of the present invention based on the concept of the present invention are within the scope of the claimed invention. All the raw materials and solvents used in the examples are commercially available products unless otherwise specified.

Preparation of example 1

Taking 100kg of dried green tea (all hair tips are mixed), adding 1000L of 20% ethanol water solution, heating to 80 deg.C, extracting for 30min, extracting for 2 times, mixing the two extractive solutions, cooling to room temperature, sieving with 60 mesh sieve, and concentrating until there is no alcohol smell.

And (3) sampling the extracted concentrated solution at the flow rate of 1BV/h to a No. 1 column chromatography filled with polyamide resin, eluting with 2BV of purified water after sampling, eluting with 80% ethanol water solution with the volume concentration of 2BV, collecting the eluent, and concentrating until no alcohol smell exists.

Loading the concentrated solution of the No. 1 column chromatography eluent to No. 2 column chromatography filled with LXD-200 resin, eluting with 2BV purified water, eluting with 25% ethanol water solution with 2BV volume concentration, collecting eluent, concentrating at 50 deg.C until the relative specific gravity is 1.20, and spray drying to obtain 6.1kg of tea extract (C-1), total catechin content is 90.3%, EGCG content is 68.2%, caffeine content is 0.14%, and yield is 6.1 wt%.

Preparation of example 2

Taking 100kg of dried green tea (Xinyang Maojian tea), adding 1000L of 20% ethanol water solution, heating to 70 deg.C, extracting for 30min, extracting for 3 times, mixing the three extractive solutions, cooling to room temperature, sieving with 80 mesh sieve, and concentrating until there is no alcohol smell.

And (3) sampling the extracted concentrated solution at the flow rate of 1BV/h to a No. 1 column chromatography filled with polyamide resin, eluting with 2BV of purified water after sampling, eluting with 75% ethanol water solution with the volume concentration of 2BV, collecting the eluent, and concentrating until no alcohol smell exists.

Loading the concentrated solution of the No. 1 column chromatography eluent to No. 2 column chromatography filled with LXD-200 resin, eluting with 2BV purified water, eluting with 2BV ethanol water solution with volume concentration of 40%, collecting eluent, concentrating at 50 deg.C until the relative specific gravity is 1.10, and vacuum drying to obtain 6.4kg of tea extract (C-2), total catechin content is 86.9%, EGCG content is 67.1%, caffeine content is 0.16%, and yield is 6.4 wt%.

Preparation of example 3

Taking 100kg of dried green tea (Biluochun tea), adding 1000L of 20% ethanol water solution, heating to 90 deg.C, extracting for 30min, extracting for 2 times, mixing the two extractive solutions, cooling to room temperature, filtering with plate frame, and concentrating until no alcohol smell is generated.

And (3) sampling the extracted concentrated solution at the flow rate of 1BV/h to a No. 1 column chromatography filled with polyamide resin, eluting with 2BV of purified water after sampling, eluting with 70% ethanol water solution with the volume concentration of 2BV, collecting the eluent, and concentrating until no alcohol smell exists.

Loading the concentrated solution of the No. 1 column chromatography eluent to No. 2 column chromatography filled with LXD-200 resin, eluting with 2BV purified water, eluting with 2BV ethanol water solution with volume concentration of 20%, collecting eluent, concentrating at 50 deg.C until the relative specific gravity is 1.20, and freeze-drying to obtain 6.4kg of tea extract (C-3), with total catechin content of 89.0%, EGCG content of 66.4%, caffeine content of 0.26%, and yield of 6.4 wt%.

Preparation of example 4

Taking 100kg of dried green tea (all hair tips are mixed), adding 1000L of 50% ethanol water solution, heating to 80 deg.C, extracting for 30min, extracting for 2 times, mixing the two extractive solutions, cooling to room temperature, filtering with plate frame, and concentrating until there is no alcohol smell.

And (3) sampling the extracted concentrated solution at the flow rate of 1BV/h to a No. 1 column chromatography filled with polyamide resin, eluting with 2BV of purified water after sampling, eluting with 70% ethanol water solution with the volume concentration of 2BV, collecting the eluent, and concentrating until no alcohol smell exists.

Loading the concentrated solution of the No. 1 column chromatography eluent to No. 2 column chromatography filled with HP-20 resin, eluting with 2BV purified water, eluting with 2BV ethanol water solution with volume concentration of 30%, collecting eluent, concentrating at 50 deg.C until the relative specific gravity is 1.20, and freeze-drying to obtain 6.5kg of tea extract (C-4), total catechin content of 87.1.0%, EGCG content of 66.7%, caffeine content of 0.23%, and yield of 6.5 wt%.

Preparation of example 5

Taking 100kg of dried green tea (all hair tips are mixed), adding 1000L of 20% ethanol water solution, heating to 70 deg.C, extracting for 30min, extracting for 2 times, mixing the two extractive solutions, cooling to room temperature, filtering with plate frame, and concentrating until there is no alcohol smell.

And (3) sampling the extracted concentrated solution at the flow rate of 1BV/h to a No. 1 column chromatography filled with polyamide resin, eluting with 2BV of purified water after sampling, eluting with 85% ethanol water solution with the volume concentration of 2BV, collecting the eluent, and concentrating until no alcohol smell exists.

Loading the concentrated solution of the No. 1 column chromatography eluent to No. 2 column chromatography filled with HP-20 resin, eluting with 2BV purified water, eluting with 2BV ethanol water solution with volume concentration of 30%, collecting eluent, concentrating at 50 deg.C until the relative specific gravity is 1.20, and freeze-drying to obtain 6.4kg of tea extract (C-5), total catechin content is 86.6%, EGCG content is 68.4%, caffeine content is 0.19%, and yield is 6.4 wt%.

Preparation of example 6

Taking 100kg of dried green tea (Duyun Maojian tea), adding 1000L of 30% ethanol water solution, heating to 85 deg.C, extracting for 30min, extracting for 2 times, mixing the two extractive solutions, cooling to room temperature, filtering with plate frame, and concentrating until no alcohol smell is produced.

And (3) sampling the extracted concentrated solution at the flow rate of 1BV/h to a No. 1 column chromatography filled with polyamide resin, eluting with 2BV of purified water after sampling, eluting with 65% ethanol water solution in volume concentration of 2BV, collecting the eluent, and concentrating until no alcohol smell exists.

Loading the concentrated solution of the No. 1 column chromatography eluent to No. 2 column chromatography filled with HP-20 resin, eluting with 2BV purified water, eluting with 2BV ethanol water solution with volume concentration of 20%, collecting eluent, concentrating at 50 deg.C until the relative specific gravity is 1.20, and spray drying to obtain 6.1kg of tea extract (C-6), total catechin content is 86.6%, EGCG content is 65.9%, caffeine content is 0.25%, and yield is 6.1 wt%.

Biological Activity example 1 (evaluation of protective Effect of nerve cells)

PC12 cells were cultured in high-glucose DMEM medium containing 10% fetal bovine serum, the cells were passaged by digesting with 0.125% trypsin for about 50s, digestion was stopped in DMEM medium containing 10% serum, and fresh medium was added to blow the cells uniformly.At 10⁵Cell density passages per mL. 4mL of cell-containing medium was added to each flask. At 37 ℃ 5% CO₂Culturing under the condition. PC12 cells were grown to confluency in a flask, digested with 0.125% trypsin solution, repeatedly pipetted into cell suspension, diluted to 1.0X 10 with 10% FBS-containing high-sugar DMEM medium⁵one/mL, 100. mu.L per well, were inoculated into 96-well plates, 5-6 wells per group, at 37 ℃ with 5% CO₂Culturing for 24h under the condition to obtain a fusion state.

The 96-well plate is respectively administered with 100 mu L of the drug in each well according to a certain concentration gradient, and after 24 hours of culture, the MTT method is used for detecting the cell viability. 50mg of MTT was dissolved in 10mL of PBS and filtered through a 0.22 μm microporous membrane. It was diluted to 0.5mg/mL immediately before use. The medium was discarded from each group of cells and washed twice with PBS, each time with 0.5mg/mL MTT, 37 ℃ and 5% CO₂After 3 hours of incubation under the conditions, MTT working solution was removed, 150. mu.L of DMSO was added to each well to dissolve crystals, and the crystals were shaken for 10 minutes to measure the OD value of each well (measurement wavelength 570nm, reference wavelength 650 nm). The cell viability of the model group and the administration group was calculated by taking the average value of the OD values of the control group as 100% cell viability.

Grouping experiments: 1) blank (high glucose DMEM); 2) model group (high-sugar DMEM cultured for 3H, then added with H)₂O₂Stimulating for 1h to make the final concentration be 100 mu M); 3) positive drug (NAC) group: adding high-glucose DMEM containing NAC (NAC) as positive drug at a certain concentration (500 μ M) and culturing for 3 hr, and adding 100 μ M H₂O₂Stimulating for 1 h; 4) administration group: adding high-sugar DMEM containing tea leaf extract (C-1) with various concentration gradients, culturing for 3 hr, and adding 100 μ M H₂O₂Stimulating for 1 h. The above groups were cultured under the same conditions, followed by experiments, and cell viability was measured by the MTT method, and the measurement results are shown in tables 1 and 2.

TABLE 1 Effect of tea extract on the viability of Normal PC12 cells

Compared with the control group, the compound of the formula,^***p<0.001

TABLE 2 tea extract pairs H₂O₂Effect of induced oxidative damage of PC12 cells

Compared with the control group, the compound of the formula,^###p<0.001; in comparison with the set of models,^***p<0.001

as shown in Table 1, the tea extract showed no inhibitory effect on the activity of normal PC12 cells at a concentration of 1 to 3. mu.g/mL, and showed strong inhibitory effect on the activity of PC12 cells at a concentration of 10 to 500. mu.g/mL, and showed concentration dependence. The PC12 cell is a tumor neuron cell, so that the tea extract has potential anti-tumor activity.

As shown in table 2, the cell viability of PC12 in the model group was 50.45%, and p was <0.001 compared to the control group (cell viability of 100%), indicating that the model was successfully constructed. When 500 μ M of NAC, a positive control, was added to the cells, the viability of the cells increased to 100.80%, indicating significant protection. When the concentration of the tea extract is 10 mug/mL and 30 mug/mL, the protection effect is respectively improved from 50.45% to 87.76% and 94.48%, which shows that the tea extract has stronger protection effect on nerve cells.

Biological Activity example 2 (evaluation of anti-PM 2.5 Effect)

Transgenic neutrophils and macrophage green fluorescent zebra fish 2 days (2dpf) after 90-tail fertilization are randomly selected to be placed in a six-hole plate, 30 zebra fish are treated in each hole (experimental group), and red fluorescent dye is injected into the venous sinus to establish a zebra fish macrophage phagocytic function model. The concentration of the tea extract (C-1) in the model zebra fish was 31.25. mu.g/mL by water dissolution, while the model control group was set to have a volume of 3mL per well (experimental group). After finishing, randomly taking 10 zebra fish in each group, collecting pictures under a fluorescence microscope, and calculating the number (N) of the remaining fluorescent microspheres phagocytosed and removed by macrophages by using image processing software; evaluating tea extract according to residual quantity of fluorescent microspheresThe function of promoting the phagocytic function of macrophages by fetching the materials is calculated by the following formula: macrophage phagocytosis promoting effect (%) ═ N_{Model control group}-N_{Test article group})/N_{Model control group}X 100%, statistical treatment results are expressed as mean + -SE, using ANOVA and Dunnett's T-test, p<The difference was significant at 0.05, and the results are shown in table 3.

TABLE 3 Zebra fish macrophage phagocytosis data (n ═ 10)

Compared with the model control group,^**p<0.01

as can be seen from table 3, the number of the fluorescent microspheres in the tea extract group of zebra fish is 27, and compared with the model control group (41), p is less than 0.01, and the phagocytosis promotion rate of macrophages is 34%, which indicates that the tea extract has a promotion effect on the phagocytosis function of the zebra fish macrophages.