Extraction method and application of ailanthus altissima general flavone

Abstract

The invention relates to the technical field of chemical treatment of natural medicines, in particular to an extraction method and application of total flavonoids of ailanthus altissima L. The extraction method comprises the following steps: taking ailanthus altissima L. as a raw material, air-drying and crushing the raw material to obtain a crushed raw material medicine; performing a single-factor test according to five factors and six levels, and respectively investigating the influence of the extraction temperature X1, the ultrasonic power X2, the ethanol concentration X3, the liquid-material ratio X4 and the ultrasonic time X5 on the yield of the total flavonoids in the ailanthus altissima; on the basis of a single factor level, factors having significant influence on total flavonoids of ailanthus altissima are selected, Box-Behnken response surface design is carried out, and a test is carried out according to a factor level combination; and carrying out analysis on the test data by adopting Design-Expert 8.0. 6 statistical analysis software. Compared with the prior art, the method has the advantages that the technical conditions are simple, the safety is realized, the cost is low, the technical support and theoretical basis are provided for the industrial production of the new ailanthus altissima calf diarrhea preparation, and the foundation is laid for effectively preventing and controlling the calf diarrhea.

Description

Technical Field

[0001] The invention relates to the technical field of chemical processing of natural medicines, and in particular to a method for extracting total flavonoids from water hyacinth and application thereof. Background Art

[0002] Water hyacinth is the fruit of Ailanthus altissima (Mill.) Swingle of the family Simaroubaceae, and is a commonly used Chinese herbal medicine in my country. Water hyacinth is white and yellowish in color, bitter in taste and cold in nature, and has the effects of clearing heat, stopping bleeding, promoting dampness, killing insects, and resisting bacteria. It has good efficacy in treating diarrhea and dysentery, but its application in veterinary clinic is relatively small, which restricts its promotion and application, and seriously affects the clinical advantages of water hyacinth in traditional Chinese veterinary medicine.

[0003] Calf diarrhea, also known as calf loose stools, can occur all year round and is a common gastrointestinal disease in calves. Calf diarrhea is a comprehensive disease characterized by digestive dysfunction. The main symptom is diarrhea. Due to hyperperistalsis, incomplete or difficult intestinal absorption, intestinal contents and a large amount of water are excreted from the body, and the feces are thin soup or watery. Dehydration and acidosis occur quickly, and death is relatively fast. The cause of calf diarrhea is complex, and antibiotics are often used for treatment. As a result, drug-resistant strains appear from time to time, resulting in a decrease in efficacy. Chinese veterinary medicine has the characteristics of no pollution, no residue, not easy to produce drug resistance, and no pollution to animal-derived food. With the strengthening of food safety and industry supervision, some antibiotics are restricted or banned. When looking for alternative products, Chinese veterinary medicine has become the first choice. Due to the low level of development of calves, they have not yet formed a mature disease resistance mechanism, and their immunity is low, so they are easily affected by external factors and develop diseases. Therefore, it is urgent to develop effective drugs for the prevention and treatment of calf diarrhea to promote the healthy development of my country's cattle industry.

[0004] At present, the treatment of calf diarrhea is still mainly based on antibiotics. There is an over-reliance on antibiotics, excessive or abuse of antibiotics, which increases the risk of pathogen resistance and causes public food safety issues. However, curbing animal-derived bacterial resistance and demonstrating the application of antibiotic reduction are currently tasks that my country attaches great importance to.

[0005] The Chinese herbal medicine "Eichhornia crenata" is still a blank in the research of Chinese veterinary medicine, and there are no relevant reports. In view of the good efficacy of Eichhornia crenata in the treatment of diarrhea and dysentery, according to the characteristics of calf diarrhea and the drug demand characteristics of modern large-scale ranches, modern process extraction technology, pharmacology, pharmacodynamics and traditional veterinary medicine and other related technologies are applied to develop a new preparation of Eichhornia crenata with clear clinical positioning and outstanding therapeutic effect, effectively give play to the unique advantages of Chinese veterinary medicine in the prevention and treatment of calf diarrhea, conduct in-depth research on calf diarrhea, and lay the foundation for more effective prevention and control of calf diarrhea. Summary of the invention

[0006] The purpose of the present invention is to provide a method for extracting total flavonoids from Eichhornia crassipes, which is simple to operate, safe, low in cost, and high in effective ingredient content and yield.

[0007] In order to achieve the above purpose, the technical solutions provided are as follows:

[0008] A method for extracting total flavonoids from Eichhornia crassipes, comprising the following steps:

[0009] Step 1: Take water hyacinth as raw material, air-dry and crush the raw material to obtain crushed raw material medicine;

[0010] Step 2: using an ethanol solution of corresponding concentration as a solvent, adding the crushed raw material obtained in step 1 to the solvent, setting the corresponding extraction temperature, ultrasonic power, liquid-to-solid ratio, and ultrasonic time for ultrasonic extraction, and then filtering and concentrating to obtain total flavonoids;

[0011] Step 3, a single factor experiment was conducted according to 5 factors and 6 levels to investigate the effects of extraction temperature X1, ultrasonic power X2, ethanol concentration X3, liquid-to-solid ratio X4, and ultrasonic time X5 on the yield of total flavonoids in water hyacinth;

[0012] Step 4: Based on the single factor level of step 3, select factors that have a significant impact on the total flavonoids of water hyacinth, conduct Box-Behnken response surface design, and conduct experiments according to the combination of factor levels;

[0013] Step 5: Analyze the experimental data using Design-Expert 8.0.6 statistical analysis software; the formula for extracting total flavonoids from Eichhornia crassipes is:

[0014] Y(%)=10.56+0.09X1+0.32X2-0.77X3+0.31X1X2+0.36X1X3-0.24X2X3-1.19X1 2 -0.64X2 2 -0.78X3 2

[0015] Wherein, Y represents the total flavonoids content of Herba Lycopodii, X1 represents the extraction temperature, X2 represents the ultrasonic power, and X3 represents the ethanol concentration.

[0016] Preferably, in step three, the extraction temperature is 30-80° C., the ultrasonic power is 50-100 W, the ethanol concentration is 20-70%, the liquid-to-solid ratio is 20:1-70:1, and the ultrasonic time is 20-120 min.

[0017] Preferably, in step three, each group of experimental ultrasonic extractions is repeated three times, combined, filtered, and the filtrate is concentrated under reduced pressure in a water bath at 50° C. and a vacuum degree of -0.07 MPa to obtain total flavonoids from Eichhornia crassipes.

[0018] Preferably, in step 4, the factors that have a significant impact on the total flavonoids of water hyacinth are extraction temperature, ultrasonic power and ethanol concentration.

[0019] Preferably, in step five, analysis shows that the best extraction process has an ultrasonic temperature of 50° C., an ultrasonic power of 85 W, and an ethanol concentration of 45%.

[0020] Preferably, the total flavonoids of water hyacinth are used in the treatment of calf diarrhea.

[0021] Preferably, the total flavonoids of water hyacinth are used as a special drug for calf diarrhea in the treatment of multiple mixed infectious diarrhea caused by Clostridium perfringens, Salmonella and Cryptosporidium.

[0022] The beneficial effects of the present invention are as follows: the present invention provides a method for extracting total flavonoids from water hyacinth and application thereof, the content of total flavonoids from the extracted water hyacinth reaches 10.41%, which is significantly improved compared with the prior art, and the method has simple and safe process conditions and low cost, which provides technical support and theoretical basis for the industrial production of a new preparation of water hyacinth for calf diarrhea, lays a foundation for more effective prevention and control of calf diarrhea, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is the single factor experimental result of the present invention;

[0024] Among them, A. The effect of extraction temperature on the extraction rate of total flavonoids from water hyacinth; B. The effect of ultrasonic power on the extraction rate of total flavonoids from water hyacinth; C. The effect of ethanol concentration on the extraction rate of total flavonoids from water hyacinth; D. The effect of liquid-to-solid ratio on the extraction rate of total flavonoids from water hyacinth; E. The effect of ultrasonic time on the extraction rate of total flavonoids from water hyacinth;

[0025] Figure 2 The response surface and contour lines of the effects of extraction temperature and ultrasonic power on the extraction rate of total flavonoids from Eichhornia crassipes in the present invention;

[0026] Figure 3 The response surface and contour lines of the effects of extraction temperature and ethanol concentration on the extraction rate of total flavonoids from Eichhornia crassipes in the present invention;

[0027] Figure 4 The response surface and contour lines of the effects of ultrasonic power and ethanol concentration on the extraction rate of total flavonoids from Eichhornia crassipes in the present invention are shown. DETAILED DESCRIPTION

[0028] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] Example 1: Determination of the content of total flavonoids in Eichhornia crassipes

[0030] 1. Preparation of test solution:

[0031] Weigh 1.000g of crushed water hyacinth accurately, put it into a 100mL conical flask, add ethanol of corresponding concentration according to Table 1, soak for 30min, set the corresponding extraction time, extraction temperature, and ultrasonic power in the ultrasonic equipment for ultrasonic extraction, and filter out the extract after the extraction. Repeat the ultrasonic extraction 3 times under the same conditions, combine the filtrates of the 3 times, concentrate to dryness under reduced pressure using a rotary evaporator, dissolve it with 70% ethanol solution, dilute to a 50mL volumetric flask, and let it stand for testing.

[0032] 2. Preparation of reference solution

[0033] Accurately weigh 21.93 mg of rutin standard dried to constant weight, dissolve it in 50% ethanol, and dilute to 100 mL volumetric flask for later use.

[0034] 3. Drawing of standard curve

[0035] Accurately measure 0.0, 2.5, 3.0, 3.5, 4.0, 4.5, and 5.0 mL of the reference solution and place them in 10 mL volumetric flasks, add 50% ethanol to make up to 5 mL, and shake well. Add 0.3 mL of 5% sodium nitrite solution to each, let stand for 5 min; add 0.3 mL of 10% aluminum nitrate solution, let stand for 5 min; add 4 mL of 4% sodium hydroxide solution, let stand for 1 min; make up to 50% ethanol, shake well, and let stand for 15 min. Use the first group as a blank control, measure the absorbance at 510 nm, use the concentration x (mg / mL) as the horizontal coordinate, and the absorbance y as the vertical coordinate to make a standard curve, perform regression analysis, and get the regression equation y=6.5256x+0.0032, R 2 =0.9999, indicating that the rutin reference substance has a good linear relationship in the concentration range of 0.0548-0.1097 mg / mL.

[0036] 4. Sample determination

[0037] Accurately measure 0.5 mL of the test solution, place it in a 10 mL volumetric flask, add 50% ethanol to make up to 10 mL, shake well, and measure its absorbance at 510 nm.

[0038] 5. Determination of total flavonoids extraction

[0039] Weigh one tenth of the extract of Eichhornia crassipes accurately, prepare the test solution according to the 3 preparation method, pipette 0.2 mL into a 96-well plate, and repeat three times. Then use an ELISA reader to measure the absorbance of the solution.

[0040] Example 2: Single Factor Experiment

[0041] Taking the total flavonoids yield of Eichhornia crassipes as the evaluation index, the effects of five factors, including extraction temperature (factor X1), ultrasonic power (X2), ethanol concentration (factor X3), liquid-to-solid ratio (factor X4) and ultrasonic time (factor X5), on the total flavonoids yield of Eichhornia crassipes were investigated respectively, and 6 levels were set for each factor for a single factor experiment.

[0042] Table 1 Single factor test level table

[0043]

[0044] The experimental design is as follows:

[0045] ① Comparison of extraction temperature: ultrasonic power was 80W, ethanol concentration was 70%, liquid-to-solid ratio was 20:1, ultrasonic time was 60min, and extraction temperature was set at 30, 40, 50, 60, 70 and 80℃ respectively;

[0046] ②Comparison of ultrasonic power: extraction temperature was 50℃, ethanol concentration was 70%, liquid-to-solid ratio was 20:1, ultrasonic time was 60min, and ultrasonic power was set to 50, 60, 70, 80, 90 and 100W respectively;

[0047] ③Comparison of ethanol concentration: extraction temperature was 50℃, ultrasonic power was 80W, liquid-to-solid ratio was 20:1, ultrasonic time was 60min, and ethanol concentration was set at 20, 30, 40, 50, 60 and 70℃ respectively;

[0048] ④Comparison of liquid-to-solid ratio: extraction temperature was 50°C, ultrasonic power was 80W, ethanol concentration was 70%, ultrasonic time was 60min, and liquid-to-solid ratio was set to 20:1, 35:1, 40:1, 50:1, 60:1 and 70:1 respectively;

[0049] ⑤ Comparison of ultrasonic time: the extraction temperature was 50°C, the ultrasonic power was 80W, the ethanol concentration was 70%, the liquid-to-solid ratio was 20:1, and the ultrasonic time was set to 20, 40, 60, 80, 100 and 120 min respectively.

[0050] Each group of the above experiments was repeated 3 times, and the average value was taken. The extract was processed by the method in Example 1, and the total flavonoids content of Eichhornia crassipes was determined.

[0051] Under the condition that other factors remain unchanged, the effect of extraction temperature on the extraction rate of total flavonoids from Eichhornia crassipes is as follows: Figure 1 As shown in A, it can be seen that the total flavonoids content of Eichhornia crassipes is the highest when the extraction temperature is 50°C; the effect of ultrasonic power on the extraction rate of total flavonoids of Eichhornia crassipes is shown in Figure 1 As shown in B, it can be seen that the total flavonoids extraction rate of Eichhornia crassipes is the highest when the ultrasonic power is 80W; the effect of ethanol concentration on the total flavonoids extraction rate of Eichhornia crassipes is as follows Figure 1 As shown in C, it can be seen that the extraction rate of total flavonoids from water hyacinth is the highest when the ethanol concentration is 50%; the effect of liquid-to-solid ratio on the extraction rate of total flavonoids from water hyacinth is shown in Figure 1 As shown in D, it can be seen that the extraction rate of total flavonoids from water hyacinth tends to be stable when the liquid-to-solid ratio is ≥50; the effect of ultrasonic time on the extraction rate of total flavonoids from water hyacinth is shown in Figure 1 As shown in E, it can be seen that the extraction rate of total flavonoids from Eichhornia crassipes tends to be stable when the ultrasonic time is ≥40 min.

[0052] Example 3: Model building

[0053] 1. Response surface design

[0054] According to the results of the single-factor experiment, three factors, extraction temperature (X1), ultrasonic power (X2), and ethanol concentration (X3), were selected as variables. The Box-Behnken response surface design was performed using Design-Expert 8.0.6 software, with -1, 0, and 1 representing the variable levels. The true values ​​and coding levels of the independent variables are shown in Table 2.

[0055] Table 2 Box-Behnken test design factors and levels

[0056]

[0057] Accurately weigh 5 samples of 1.0 g each, extract the total flavonoids from Eichhornia crassipes according to the process conditions, treat the test solution of the extract according to the experimental steps in Example 1, determine the total flavonoids content of Eichhornia crassipes according to the method in Example 2, and calculate the total flavonoids extraction rate of Eichhornia crassipes.

[0058] 2. Box-Behnken test and results

[0059] The experimental scheme obtained by Box-Behnken software is shown in Table 3, with a total of 17 groups. Each group was repeated 3 times and the total flavonoids content in the water hyacinth extract was determined. The average value was calculated and filled in Table 3, and analyzed using Box-Behnken software.

[0060] 3. Model building and variance analysis

[0061] Design-Expert 8.0.6 software was used to perform multivariate linear regression fitting analysis on the experimental scheme and experimental results, and the regression equation for the relationship between the total flavonoids content of Eichhornia crassipes and various factors was obtained as follows. The results of variance analysis are shown in Table 4.

[0062] Y(%)=10.56+0.09X1+0.32X2-0.77X3+0.31X1X2+0.36X1X3-0.24X2X3-1.19X1 2 -0.64X2 2 -0.78X3 2

[0063] In the formula, Y represents the total flavonoids content of Herba Lycopodii, X1 represents the extraction temperature, X2 represents the ultrasonic power, and X3 represents the ethanol concentration.

[0064] Table 3 Box-Behnken test results

[0065]

[0066]

[0067] Table 4 Response surface quadratic simulation variance analysis

[0068]

[0069] Note: P≤0.01 indicates that the factor has a very significant effect on the response value; P≥0.05 indicates that the factor has a significant effect on the response value.

[0070] As shown in Table 3, the model is extremely significant (P = 0.0001 < 0.01), and the lack of fit term is not significant (P = 0.2021 > 0.05), indicating that non-experimental factors have little effect on the results and the model has good experimental stability. 2 =0.9771, indicating that the model fits the actual experiment well and the results can be explained by the fitted model. Corrected determination coefficient R 2 Adj is 0.9477, which is consistent with R 2 The results are close, indicating that the model has sufficient accuracy and universality. Therefore, this model can be used to analyze and predict the influence of various factors on the extraction rate in the extraction experiment of water hyacinth. The order of factors affecting the total flavonoids yield in water hyacinth is: ethanol concentration > ultrasonic power > extraction time.

[0071] 4. Response surface interaction analysis

[0072] The response surface and contour lines of the model are shown in Figure 2 to Figure 4 The three groups of graphs intuitively reflect the impact of various factors on the response value.

[0073] The shape of the contour line reflects the interaction between the factors. The elliptical contour line indicates that the interaction between the two is significant. The ethanol concentration, ultrasonic power, and extraction time are fixed in sequence to form a response surface diagram. The contour line diagram is elliptical and the 3D diagram is steep, indicating that the interaction is more significant. Figure 2 It can be seen that the response surface curve of ethanol volume fraction is significantly steeper than the curves of ultrasonic power and extraction temperature, indicating that the ethanol volume fraction has a more significant effect on the response value of total flavonoid extraction.

[0074] Through response surface software Box-Behnken analysis and mathematical regression model optimization of data, the optimal extraction process for ultrasonic-assisted extraction of total flavonoids from Eichhornia crassipes was obtained as follows: ultrasonic temperature 50°C, ultrasonic power 85W, ethanol concentration 45%, and the predicted total flavonoid extraction rate was 10.8217%.

[0075] 5. Process verification test

[0076] Extraction was performed under the optimized conditions (ultrasonic temperature 50°C, ultrasonic power 85W, ethanol concentration 45%), and the determination was repeated 3 times under this condition. The content of total flavonoids in Eichhornia crassipes was determined, and the average value was 10.4146% (see Table 5), which was close to the theoretical calculated value, indicating that the optimization scheme was feasible.

[0077] Table 5 Confirmatory results analysis

[0078]

[0079] During the ultrasonic extraction process, extraction temperature, ultrasonic power, ethanol concentration, liquid-to-solid ratio, ultrasonic time, etc. will affect the amount of extract obtained. The single factor results showed that with the increase of extraction temperature, ultrasonic power and ethanol concentration, the total flavonoids extraction amount in water hyacinth first increased and then decreased, while with the increase of liquid-to-solid ratio and ultrasonic time, the total flavonoids extraction amount in water hyacinth first increased and then tended to stabilize. However, when the factors interacted with each other, the total flavonoids extract content in water hyacinth was determined by the extraction process optimized by response surface methodology, and the total flavonoids content in water hyacinth extract was 10.4146% in the validation test of the optimal extraction process, which was not much different from the simulation prediction value of 10.8217%, indicating that the optimized extraction process parameters obtained based on response surface methodology are accurate and reliable, and have good stability.

[0080] Example 4: Clinical efficacy experiment of total flavonoids extract of water hyacinth on calf diarrhea

[0081] 1. Symptoms of calf diarrhea

[0082] In a cattle farm in Honggu District, Gansu Province, calves aged 10-30 days had yellow porridge-like or watery feces, and were emaciated and dehydrated. In some cases, they died quickly. The fecal samples of the calves were cultured and separated in the laboratory, and it was determined that the calves had multiple mixed infectious diarrhea, including Clostridium perfringens, Salmonella and Cryptosporidium.

[0083] 2. Efficacy standards

[0084] Recovery: The calf’s diarrhea symptoms disappear, and the calf actively eats and drinks milk, and its physical and mental state recovers.

[0085] Ineffective: The diarrhea symptoms of calves are aggravated, and the calves lose weight and die faster.

[0086] 3. Method of use

[0087] The dosage for each use is 30.0 g of the total flavonoids extract of water hyacinth prepared in Example 1 per calf; 30.0 g of the total flavonoids extract of water hyacinth prepared in Example 1 is diluted with 100 ml of pure water and then administered to the calf orally.

[0088] 4. Treatment

[0089] Taking the medicine twice in a row has 100% efficacy for yellow stool and mild loose stools. If it is watery diarrhea, the medicine needs to be used 4 times, twice a day (once in the morning and once in the evening), which has a significant effect.

[0090] The drugs were used together to treat 6 calves with diarrhea, and 6 of them were cured, with a cure rate of 100%. The specific drug usage is shown in Table 6.

[0091] Table 6 Clinical efficacy experiment of total flavonoids extract of Eichhornia crassipes on calf diarrhea

[0092]

[0093] In summary, the present invention provides a method for extracting total flavonoids from water hyacinth and its application. The content of total flavonoids from water hyacinth extracted reaches 10.41%, which is significantly improved compared with the prior art. The method has simple and safe process conditions, low cost and is easy for industrial production. It provides a theoretical basis and optimizes process parameters for the development and application of water hyacinth in veterinary products, and has certain guiding significance for the comprehensive utilization of water hyacinth.

[0094] It will be apparent to those skilled in the art that the invention is not limited to the details of the exemplary embodiments described above and that the invention can be implemented in other specific forms without departing from the spirit or essential features of the invention. Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations falling within the meaning and scope of the equivalent elements of the claims be included in the invention. Any reference numeral in a claim should not be considered as limiting the claim to which it relates.

Claims

1. A method for extracting total flavonoids from Eichhornia crassipes, characterized in that: The steps include: Step 1: Take water hyacinth as raw material, air-dry and crush the raw material to obtain crushed raw material medicine; Step 2: using an ethanol solution of corresponding concentration as a solvent, adding the crushed raw material obtained in step 1 to the solvent, setting the corresponding extraction temperature, ultrasonic power, liquid-to-solid ratio, and ultrasonic time for ultrasonic extraction, and then filtering and concentrating to obtain total flavonoids; Step 3, a single factor experiment was conducted according to 5 factors and 6 levels to investigate the effects of extraction temperature X1, ultrasonic power X2, ethanol concentration X3, liquid-to-solid ratio X4, and ultrasonic time X5 on the yield of total flavonoids in water hyacinth; Step 4: Based on the single factor level of step 3, select factors that have a significant impact on the total flavonoids of water hyacinth, conduct Box-Behnken response surface design, and conduct experiments according to the combination of factor levels; Step 5: Analyze the experimental data using Design-Expert 8.0.6 statistical analysis software; the formula for extracting total flavonoids from Eichhornia crassipes is: Y(%)=10.56+0.09X1+0.32X2-0.77X3+0.31X1X2+0.36X1X3-0.24X2X3-1.19X1 2 -0.64X2 2 -0.78X3 2 Wherein, Y represents the total flavonoids content of Herba Lycopodii, X1 represents the extraction temperature, X2 represents the ultrasonic power, and X3 represents the ethanol concentration.

2. The method for extracting total flavonoids from Eichhornia crassipes according to claim 1, characterized in that: In step three, the extraction temperature is 30-80° C., the ultrasonic power is 50-100 W, the ethanol concentration is 20-70%, the liquid-to-solid ratio is 20:1-70:1, and the ultrasonic time is 20-120 min.

3. The method for extracting total flavonoids from Eichhornia crassipes according to claim 1, characterized in that: In step three, the ultrasonic extraction was repeated three times for each test, the mixture was combined, filtered, and the filtrate was concentrated under reduced pressure in a water bath at 50° C. and a vacuum degree of -0.07 MPa to obtain total flavonoids from Eichhornia crassipes.

4. The method for extracting total flavonoids from Eichhornia crassipes according to claim 1, characterized in that: In step 4, the factors that have a significant impact on the total flavonoids of water hyacinth are extraction temperature, ultrasonic power and ethanol concentration.

5. The method for extracting total flavonoids from Eichhornia crassipes according to claim 1, characterized in that: In step five, analysis shows that the best extraction process has an ultrasonic temperature of 50°C, an ultrasonic power of 85W, and an ethanol concentration of 45%.

6. Use of the total flavonoids from Eichhornia crassipes obtained by the method for extracting total flavonoids from Eichhornia crassipes as described in any one of claims 1 to 5 in treating calf diarrhea.

7. The use according to claim 6, characterized in that: Application of total flavonoids of water hyacinth as a special drug for calf diarrhea in the treatment of multiple mixed infectious diarrhea caused by Clostridium perfringens, Salmonella and Cryptosporidium.

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