CN114052144A - Traditional Chinese medicine feed additive for improving growth performance of animals and preparation method thereof - Google Patents

Abstract

The invention provides a traditional Chinese medicine feed additive for improving the growth performance of animals and a preparation method thereof, belonging to the technical field of feed preparation. After the feed additive is used, the villus length of duodenum of broiler chickens can be increased, the crypt depth of duodenum can be reduced, the increased intestinal tract area can promote the absorption of nutrient substances, the feed intake of broiler chickens is effectively increased, and the weight gain of broiler chickens is increased. Moreover, the feed additive can improve the contents of cell factors such as IgA, IgM, IL-2, IL-4, IL-8 and TNF-alpha and the like and anti-inflammatory factors in the blood serum of the broiler chicken, thereby promoting the humoral immune response, stimulating the complement components by increasing the concentration of the immunoglobulin, enhancing the specific immune mechanism of the broiler chicken and protecting the broiler chicken from infection.

Description

Traditional Chinese medicine feed additive for improving growth performance of animals and preparation method thereof

Technical Field

The invention belongs to the technical field of feed preparation, and particularly relates to a traditional Chinese medicine feed additive for improving animal growth performance and a preparation method thereof.

Background

Bacterial drug resistance caused by feed antibiotic abuse threatens human health, and with increasing urgent demands of the people on meat quality and health, all growth-promoting drug feed additive varieties except traditional Chinese medicines are quitted according to requirements of relevant regulations in agricultural rural areas. Antibiotics are forbidden in the feed, the risk of livestock and poultry infected diseases is increased, and natural plants become hot spots for replacing the research and application of the antibiotics due to the advantages of safety, effectiveness and easy obtainment. A plurality of test data results show that the natural plants can improve the growth performance, the oxidation resistance, the intestinal morphology, the flora structure or the immune function of livestock and poultry on the premise of playing the role of substitution resistance.

Research and development of substitution technologies and products for effectively substituting antibiotics for growth promotion and disease prevention in livestock breeding become research hotspots in the livestock industry at present. The traditional Chinese medicine feed product based on natural plant medicine resources is a hotspot for research on antibiotic substitutes of feed products due to the effects of immunity enhancement, microorganism resistance, digestion assistance, growth promotion and the like. The traditional Chinese medicine resources are used for preventing and treating the diseases of the livestock and poultry with the characteristics of definite curative effect, natural sources and the like, have a long history, are widely applied to feeding products such as traditional Chinese veterinary medicines, feed additives and the like, and become an important component for supporting the healthy development of the livestock and poultry industry in China at present. According to investigation, under the driving of the national policy of banning antibiotics for feeding products, the traditional Chinese medicine feeding products become the dominant force of banning antibiotics and reducing antibiotics, among various 'substitute antibiotics', the recognition rate of farmers for the traditional Chinese medicine as an antibiotic substitute is up to 90%, and the use popularization rate of the traditional Chinese medicine feeding products in meat poultry farms is up to 95%.

The Chinese holly leaves are also named as the ilex purpurea hassk, the oil-leaf trees and the tree tops, and have the effects of clearing away heat and toxic materials, cooling blood and stopping bleeding, and healing sores. Can be used for treating scald due to hot water and fire, eczema, pyocutaneous disease, cough due to lung heat, sore throat, pyretic stranguria, dysentery, and traumatic hemorrhage. At present, no report is found on the research of the ilex purpurea Hassk extract on the aspect of promoting the growth performance of animals.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: provides a traditional Chinese medicine feed additive for improving the growth performance of animals and a preparation method thereof.

In order to solve the technical problems, the invention adopts the following technical scheme:

firstly, the invention provides an application of a Chinese holly leaf extract in preparing a chicken growth promoting feed, and a preparation method of the Chinese holly leaf extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

Preferably, the ilex purpurea Hassk extract can promote the growth of chicken duodenal villus.

Preferably, the ilex purpurea Hassk extract can reduce the depth of chicken duodenal crypts.

Secondly, the invention provides an application of the ilex purpurea Hassk extract in preparing a promoter for promoting the growth of chicken duodenal villi, and the preparation method of the ilex purpurea Hassk extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

Secondly, the invention provides an application of a Chinese holly leaf extract in preparing an inhibitor for inhibiting the depth of a chicken duodenal crypt, and a preparation method of the Chinese holly leaf extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

Secondly, the invention provides an application of the ilex purpurea Hassk extract in preparing an accelerant for promoting the expression of Cyclin-D1 and CDK-1 protein in chicken duodenum, and is characterized in that the preparation method of the ilex purpurea Hassk extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

Secondly, the invention provides an application of the ilex purpurea Hassk extract in preparing a chicken immunity enhancing medicine, which is characterized in that the preparation method of the ilex purpurea Hassk extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

Finally, the invention provides an application of the ilex purpurea Hassk extract in preparing anti-inflammatory drugs for chickens, and is characterized in that the preparation method of the ilex purpurea Hassk extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

The invention has the beneficial effects that:

1. compared with a blank group, the daily gain is obviously improved when the Chinese holly leaf extract with medium dosage is added in the whole growth period of the broilers, the experiment is taken as an example, the cost of the Chinese holly leaf extract additive is 100 yuan/kg, the Chinese holly leaf extract additive is added into feed of each ton of broilers according to 1000g/kg, the cost is 373 yuan, the cost of the formula basic feed is 3.58 yuan/kg, the sale price of the broilers is calculated according to 7.80 yuan/kg, the economic benefit of the method can be averagely increased by 0.88 yuan for one broilers, and the economic benefit of the method can be 17.72 ten thousand yuan for 20 ten thousand for broilers which are sold in batches in large-scale farms.

2. The experimental results show that the visceral indexes of the medium-dose group and the blank group are not different, which indicates that the influence of the medium-dose ilex purpurea Hassk extract on the visceral organs of animals is small and the safety of the addition is high.

3. According to the experimental results, the Chinese holly leaf extract in the medium-dose group can improve the villus length of duodenum and reduce the crypt depth of duodenum, and the expression level of proliferation-related proteins Cyclin-D1 and CDK-1 in duodenal tissues of the medium-dose group is obviously higher than that of a blank control group.

4. The experimental results show that the immunity and the anti-inflammatory capability of the broiler chicken can be improved to a certain degree by adding the ilex purpurea Hassk extract.

Drawings

FIG. 1 expression of proliferation-associated proteins Cyclin-D1 and CDK-1 in duodenal tissues in the blank and middle dose groups (two replicates).

Detailed Description

In order to clearly illustrate the technical features of the present solution, the present solution is explained below by way of specific embodiments.

Example 1

Preparation of ilex purpurea Hassk extract

(1) Drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

Example 2

Growth performance testing

(1) The henhouse, the playground, the used crib and the used basin are thoroughly disinfected one week before the test, and the empty fence is kept for 1 week. The test starts from 1 day of age, 540 healthy AA white feather broilers with similar body weight and 1 day of age are selected, the treatment is randomly divided into 6 treatments, 9 repetitions are set for each treatment, 10 repetitions are set for each treatment (male and female are grouped according to a ratio of 1: 2), and the test period is 0-42 days.

(2) The experimental groups were as follows:

TABLE 1 Experimental groups

Grouping	Treatment group	Adding amount of
			A Low dose	Low dose plant extracts	500 g/ton compound feed
Dosage in B	Medium dose plant extract	1000 g/ton compound feed
			C high dose	High dose plant extracts	1500 g/ton compound feed
F blank	Blank control group	Without adding

(3) Feeding management

The test chickens are raised in three layers on the net in cages and freely drink water and eat. 24 hours of light exposure, average temperature (26.2 + -1.2) deg.C during the test. The relative humidity of the henhouse is 60% -65%. Inoculating a chicken flock according to a conventional immunization program, namely injecting newcastle disease inactivated vaccine and new-branch bigeminy vaccine at 1 day age, injecting bursa of fabricius vaccine at a drip, and injecting avian influenza vaccine at 14 days age;

(4) test daily ration

Feeding 510 crushed materials at 1-21 days of age, and feeding 511 granular materials at 21-42 days of age;

(5) the basic ration comprises the following components:

TABLE 2 basic diet composition

Composition of raw materials	Front stage material (%)	Late stage material (%)
			Chinese corn (II)	53.14	55.29
Flour (Biaodi)	5.00	5.00
			Poultry oil	1.80	4.00
Bean pulp 46	30.82	25.02
			United states DDGS	6.00	8.00
Stone powder	1.10	1.10
			Calcium hydrogen phosphate	0.80	0.40
Salt	0.30	0.30
			Lysine, 70.0%	0.25	0.25
Methionine, 98%	0.19	0.13
			Threonine, 98%	0.03	0.01
DV32 vitamin	0.25	0.22
			M-P multiore	0.20	0.20
Choline chloride, 60%	0.10	0.08
			Total up to	100	100

(6) The nutritional levels were as follows:

TABLE 3 nutritional level composition

Weight (D)	%	1	1
				Moisture content	%	12.55	12.26
Dried substance	%	87.45	87.74
				Crude protein	%	22.00	20.01
Crude fat	%	4.83	7.12
				Coarse ash content	%	5.48	4.91
Coarse fiber	%	3.48	3.19
				Middle washing fiber	%	11.40	11.60
Dry basis crude protein	%	23.75	19.97
				Calcium carbonate	%	0.79	0.69
Total phosphorus	%	0.63	0.54
				Can utilize phosphorus-poultry	%	0.36	0.29
Salt (salt)	%	0.29	0.29
				Electrolyte balance (EB）	mEq/kg	257	222

(7) Production performance testing

During the test, the daily feeding amount, the residual material amount and the death and elutriation number are strictly recorded in the test period by taking the repetition as a unit. The remaining feed weight of each replicate group was weighed simultaneously on an empty stomach (fasted for 12h, drinking water only) in duplicate units at 08:00 am on days 1, 21 and 42 of the experiment, respectively. And (3) recording the repeated feed consumption, the animal growth condition and the death number in stages in the test period, and calculating the final-stage weight, the average daily gain, the average feed intake and the feed-weight ratio of each stage of each group.

The results obtained are shown in Table 4:

TABLE 4 results of growth Performance index at early and late stages of broiler chickens

TABLE 5 results of growth Performance index of broiler chickens at all stages

Group of	Daily food intake (g/d)	Daily gain (g/d)	Meat ratio of materials
				A Low dose	86.25±2.41	51.36±1.87	1.680±0.023
Dosage in B	88.92±3.35	53.07±2.74*	1.677±0.035
				C high dose	87.82±1.43	52.01±1.96	1.690±0.051
F blank	84.24±1.19	49.26±0.72	1.710±0.031

The experimental results show that compared with the blank group, the medium-dose group has obvious difference on the daily feed intake and daily weight gain improvement of the broiler chickens at the later stage, and the Chinese holly leaf extract with the medium dose can improve the feed intake of the broiler chickens, so that the weight gain of the broiler chickens is improved.

Example 3

Visceral index detection

(1) Rapidly taking out the heart, the liver and the spleen of the medium-dose group and the blank group, sucking the blood on the surfaces of the organs by using filter paper, weighing and recording, and then calculating the organ index according to the following calculation formula: organ index = (organ weight/body weight) × 100.

(2) The results obtained are shown in table 6:

TABLE 6 organ indices

	Heart with heart-shaped	Liver disease	Spleen
				Middle dose	0.456±0.015	2.104±0.086	0.113±0.015
Blank space	0.453±0.034	2.146±0.121	0.107±0.010

The experimental results show that compared with the control group, the visceral index of the medium-dose group has no significant difference (P > 0.05), which indicates that the Chinese holly leaf of the medium-dose group has no damage to the viscera of the broiler chicken.

Example 4

(1) Randomly selecting 6 healthy broilers with approximate average weight from the medium-dose group and the blank control group, and dissecting intestinal tract segments of which the positions are unified with duodenum, jejunum and ileum by 2cm respectively after the broiler fasts for 12 h;

(2) immediately placing the sample in 4% paraformaldehyde solution for fixing for 24 h, slicing by conventional paraffin, and staining by hematoxylin-eosin (HE);

(3) HE stained sections were visualized under a light microscope to make paraffin sections, 5 representative fields (full villi and straight trend) were selected on each section, and Villi Height (VH), Crypt Depth (CD) and villi height/crypt depth (VH/CD) were calculated using OLYMPUS cell Sens Standard 1.18 (Build 16686) software measurement analysis.

(4) The results obtained are shown in table 7:

influence of the Chinese holly leaf extract powder dose in Table 7 on intestinal tract morphology of white feather broiler

Item	Middle dose	Blank space
			Duodenum
VM，mm	1.6231±0.0183*	1.2881±0.0532
			CD，mm	0.2395±0.0147b*	0.2637±0.0197
VH/CD	6.8686±0.3815*	4.9910±0.4276

The experimental results show that compared with a blank group, the villus height of duodenum of a medium-dose group is obviously higher than that of a control group, and the crypt depth is obviously lower than that of the blank control group, so that the ilex purpurea Hassk extract of the medium-dose group can effectively promote the villus height of duodenum of broiler chickens, and can effectively inhibit the crypt depth of duodenum of broiler chickens.

Example 4

(1) Randomly selecting 6 healthy broilers with approximate average weight from the medium-dose group and the blank control group, and dissecting to take 2cm of duodenum after the broilers are killed in an empty stomach for 12 hours;

(2) grinding into powder under liquid nitrogen, and adding protein extract to extract protein;

(3) detecting the expression of proliferation-related proteins Cyclin-D1 and CDK1 by referring to the Western Blot experiment step, and obtaining the result as shown in figure 1;

from the results, it can be seen that the Chinese holly leaf extract of the medium-dose group can effectively promote the protein expression of cell proliferation-related proteins Cyclin-D1 and CDK-1, thereby effectively promoting the growth of villus.

Example 5

(1) After the test is finished, randomly selecting 10 broilers from the medium dose group and the blank control group, collecting blood by a wing vein by 10ml, placing the blood in a test tube without anticoagulant, placing the test tube for 30min, centrifuging the test tube for 10min at 4000r/min, separating serum, subpackaging the separated blood into 2 parts, and storing the blood at the temperature of minus 20 ℃;

(2) detecting Total Protein (TP), Albumin (ALB), Globulin (GLB), alanine amino converting enzyme (ALT), aspartic acid amino converting enzyme (AST), Total Cholesterol (TC), High Density Lipoprotein (HDL), Triglyceride (TG), and Low Density Lipoprotein (LDL) by using a Hitachi 7600 full-automatic biochemical analyzer;

(3) measuring the contents of IgG, IgA, IgM, IL-2, IL-4, IL-8 and TNF-alpha in serum by adopting an ELISA kit method, and detecting by adopting a Bio-Rad model-680 multifunctional microplate reader;

(4) the results obtained were as follows:

influence of the amount of the ilex purpurea Hassk extract on biochemical indexes of broiler serum in Table 8

Item	Middle dose	Blank space
			ALT(U/L)	1.0900±0.2433	1.1400±1.3915
AST(U/L)	321.6700±20.1506	402.1200±68.3855
			TP(g/L)	36.0000±5.8689*	51.1000±7.3386
ALB(g/L)	12.8000±0.4422	17.0000±3.5151
			LDL(mmol/L)	0.7860±0.1610*	1.1160±0.1903
HDL(mmol/L)	1.6710±0.1158*	1.8050±0.1905
			TC(mmol/L)	2.7670±0.2147	3.0530±0.2827
TG(mmol/L)	0.9410±0.2060*	0.7180±0.1346
			IgG(mg/ml)	17.2465±1.6544	18.4067±1.8855
IgA(mg/ml)	4.5072±0.4966*	3.0888±0.5184
			IgM(mg/ml)	26.0858±5.4692*	21.1834±3.7882

TABLE 9 influence of ilex purpurea Hassk extract on secretion of serum cytokines in broiler chickens

Item	Middle dose	Blank space
			Interleukin-2 (IL-2)	12.0080±2.5334*	8.6176±2.7938
Interleukin-4 (IL-4)	84.4516±7.5706*	53.5736±15.2424
			Interleukin-8 (IL-8)	626.6689±91.9571	566.0093±49.6149
Tumor necrosis factor-alpha (TNF-alpha)	55.8074±6.9869*	39.0042±13.0385

From the experimental results, it can be seen that the ilex purpurea Hassk extract can improve partial immunity and anti-inflammatory factors, so that the immunity and anti-inflammatory capability of the broiler chicken can be improved to a certain extent.

The technical features of the present invention which are not described in the above embodiments may be implemented by or using the prior art, and are not described herein again, of course, the above description is not intended to limit the present invention, and the present invention is not limited to the above examples, and variations, modifications, additions or substitutions which may be made by those skilled in the art within the spirit and scope of the present invention should also fall within the protection scope of the present invention.

Claims (8)

1. The application of the ilex purpurea Hassk extract in preparing the chicken growth promoting feed is characterized in that the preparation method of the ilex purpurea Hassk extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

2. The use of claim 1, wherein the ilex purpurea Hassk extract can promote the growth of chicken duodenal villus.

3. The use of claim 1, wherein the ilex purpurea Hassk extract can reduce the depth of duodenal crypts in chickens.

4. The application of the ilex purpurea Hassk extract in preparing the accelerant for promoting the growth of chicken duodenal villi is characterized in that the preparation method of the ilex purpurea Hassk extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

5. The application of the ilex purpurea Hassk extract in preparing the inhibitor for inhibiting the depth of the chicken duodenal crypt is characterized in that the preparation method of the ilex purpurea Hassk extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

6. The application of the Chinese holly leaf extract in preparing the promoter for promoting the expression of Cyclin-D1 and CDK-1 protein in chicken duodenum is characterized in that the preparation method of the Chinese holly leaf extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

7. The application of the ilex purpurea Hassk extract in preparing the chicken immunity enhancing medicine is characterized in that the preparation method of the ilex purpurea Hassk extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

8. The application of the ilex purpurea Hassk extract in preparing the anti-inflammatory drugs for the chickens is characterized in that the preparation method of the ilex purpurea Hassk extract comprises the following steps:

(1) drying folium Ilicis Purpureae, and sieving with 20 mesh sieve to obtain folium Ilicis Purpureae powder;

(2) adding 8 times volume of 40% ethanol containing 0.3% glacial acetic acid into the obtained fructus Ilicis Purpureae powder, reflux extracting for 2 hr, filtering, and collecting residue and filtrate;

(3) heating the obtained filtrate to 60 ℃, and concentrating under reduced pressure to obtain an extract with the relative density of 1.03;

(4) and carrying out spray drying on the obtained extract to obtain the Chinese holly leaf extract.

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