Anti-staphylococcus aureus compound traditional Chinese medicine composition, compound traditional Chinese medicine preparation and preparation method thereof

Abstract

The invention relates to the technical field of traditional Chinese medicine, in particular to an anti-staphylococcus aureus compound traditional Chinese medicine composition, a compound traditional Chinese medicine preparation and a preparation method thereof, the compound traditional Chinese medicine composition comprises the following raw materials in parts by weight: 2-5 parts of sappan wood, 2-6 parts of sargentgloryvine stem, 4-6 parts of gallnut and 2-4 parts of myrobalan; 0.11 part of licorice root; the MIC of the compound traditional Chinese medicine composition is 0.49 to 0.98 mg / mL, and the MBC of the compound traditional Chinese medicine composition is 0.98 to 1.95 mg / mL. The composition has the advantages that the sappan wood, the sargentgloryvine stem, the gallnut, the medicine terminalia fruit and the liquorice root are matched for use according to a certain proportion, the synergistic antibacterial effect of the composition is the best and is better than the antibacterial effect of single traditional Chinese medicine and other proportions, and compared with a chemical synthesis bactericide, the composition has the advantages of being wide in action target spot, not prone to generating drug resistance, free of residues and the like, and has a good application prospect. Animal experiments prove that a preparation prepared from the traditional Chinese medicine composition has a remarkable curative effect, and the traditional Chinese medicine composition has a wide application prospect in practical application.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicines, and in particular to an anti-Staphylococcus aureus compound traditional Chinese medicine composition, a compound traditional Chinese medicine preparation and a preparation method thereof. Background Art

[0002] Mastitis in dairy cows is the most costly, expensive, and difficult disease to control on dairy farms. It causes significant economic losses, including excessive drug residues, increased milk waste, reduced dairy product quality, and the culling of affected cows. Staphylococcus aureus, one of the primary pathogens causing mastitis in dairy cows, is highly drug-resistant and difficult to eradicate. Despite the implementation of several mastitis prevention programs in various countries, the prevalence of S. aureus in dairy cows remains high. Furthermore, S. aureus in milk can cause pneumonia and enteritis in humans, posing a public health threat. Antibiotics are often used to treat staphylococcal infections. However, the irrational use of antibiotics in dairy farming has led to the emergence of multidrug-resistant strains and antibiotic residues, seriously impacting human health. The development and implementation of new drugs is a difficult, time-consuming, and expensive process, and the pace of new drug development often lags behind the rapid evolution of bacterial resistance. Therefore, the search for botanical antimicrobial agents is an effective measure to control the continued development of bacterial resistance. Summary of the Invention

[0003] The purpose of the present invention is to overcome the deficiencies in the prior art and provide a compound Chinese medicine composition for resisting Staphylococcus aureus, a compound Chinese medicine preparation and a preparation method thereof. The composition is mainly composed of sappan wood, safflower vine, gallnut, terminalia chebula, and licorice. Antibacterial tests have shown that sappan wood, gallnut, terminalia chebula, and safflower vine all have strong anti-Staphylococcus aureus activity and are the main ingredients of this prescription, while licorice is the auxiliary ingredient. From the perspective of traditional Chinese medicine,

[0004] The formula's ingredients, Su Mu and Sargentodoxae, promote blood circulation, remove blood stasis, promote lactation, reduce swelling, and relieve pain, alleviating symptoms of mastitis in dairy cows infected with Staphylococcus. Terminalia chebula and Gallnut have the effects of astringing the lungs and reducing internal heat, purging heat and cooling the blood, alleviating fever symptoms caused by Staphylococcus infection in dairy cows. Licorice is also added to the formula to tonify the spleen and replenish qi, harmonizing the various medicinal components. The resulting compound Chinese medicine preparation and its preparation method not only prevent bacterial resistance, toxin accumulation, and drug residues, but also reduce production costs.

[0005] The present invention is achieved through the following technical solutions: on the one hand, an anti-Staphylococcus aureus compound traditional Chinese medicine composition is provided, wherein the raw materials of the compound traditional Chinese medicine composition, by weight, include 2-5 parts of sappan wood, 2-6 parts of Sargentodoxae chinensis, 4-6 parts of Galla chinensis, 2-4 parts of Terminalia chebula, and 0.11 part of Licorice root; and the MIC of the compound traditional Chinese medicine composition is 0.49-0.98 mg / mL, and the MBC is 0.98-1.95 mg / mL.

[0006] Furthermore, the ingredients of the compound Chinese medicine composition, calculated by weight percentage, include 0.25% to 0.63% of total hematoxylin, 25.00% to 33.75% of tannic acid, and 0.13% to 3.75% of total phenolic ketones; wherein the content of the total hematoxylin is calculated based on the total amount of brazilin and hematoxylin B.

[0007] Provided is an anti-Staphylococcus aureus compound traditional Chinese medicine preparation, comprising the above-mentioned anti-Staphylococcus aureus compound traditional Chinese medicine composition, wherein the compound traditional Chinese medicine preparation is selected from liquid preparation, semi-solid preparation, solid preparation or gaseous preparation.

[0008] A method for preparing the above-mentioned anti-Staphylococcus aureus compound Chinese medicine preparation is provided. The compound Chinese medicine preparation is a liquid preparation containing 30% of the compound Chinese medicine extract. The preparation steps of the compound Chinese medicine extract are as follows:

[0009] Step S1, weighing Sappan wood, Sargentodon seu Radix, Galla chinensis, Terminalia chebula, and Glycyrrhiza uralensis according to the above weight parts, ultrafinely grinding and passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0010] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0011] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0012] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0013] Step S5: Add 0.5% polysorbate 80 to the filtrate obtained in step S4, stir at 50-55° C. to dissolve it, and then concentrate the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract with a relative density of 1.19-1.24 and a pH of 3.6-4.0.

[0014] Furthermore, in step S1, after the obtained mixed powder is ultrafinely pulverized, Chinese medicine powder with a particle size of 25 to 48 μm is collected.

[0015] Furthermore, in step S4, the impurities in the compound Chinese herbal medicine extract are precipitated by alcohol precipitation.

[0016] Beneficial effects

[0017] The present invention selects sappan wood, gallnut, terminalia chebula, and Sargentodoxae, and uses them in a certain proportion to have a good anti-Staphylococcus aureus effect. Compared with chemical synthetic fungicides, the traditional Chinese medicine ingredients have the advantages of a wide target area, low resistance to drug resistance, and no residue. Animal experiments have proved that the efficacy is significant, showing broad application prospects in practical applications. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0019] All other embodiments obtained by persons of ordinary skill in the art based on the embodiments herein without inventive effort are intended to fall within the scope of protection of the present invention. Experimental methods in the following examples, where specific conditions are not specified, were generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise indicated, all percentages, ratios, proportions, and parts are by weight.

[0020] Unless otherwise specified, the reagents and raw materials used in the examples and comparative examples of the present invention can be obtained through commercial channels.

[0021] Example 1: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0022] Step S1, weighing 5 parts of Sappan wood, 4 parts of Sargentodoxae truncatula, 5 parts of Galla chinensis, 2 parts of Terminalia chebula, and 0.11 parts of Licorice root according to the above weight ratio, ultrafinely grinding and passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0023] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0024] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0025] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0026] Step S5: Add 0.5% polysorbate 80 to the filtrate obtained in step S4, stir at 50-55° C. to dissolve it, and then concentrate the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract. The relative density is controlled at 1.20 and the pH is 3.8, wherein the total hematoxylin (brazilin, hematoxylin B) content is 0.63%, tannic acid is 25.6%, and total phenols are 2.5%.

[0027] Example 2: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0028] Step S1, weighing 2 parts of Sappan wood, 6 parts of Sargentodoxae cyrtonema, 5 parts of Galla chinensis, 3 parts of Terminalia chebula, and 0.11 parts of Licorice root according to the above weight ratio, ultrafinely grinding and passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0029] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0030] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0031] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0032] Step S5: Add 0.5% polysorbate 80 to the filtrate obtained in step S4, stir at 50-55° C. to dissolve it, and then concentrate the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract. The relative density is controlled at 1.22 and the pH is 3.7, wherein the total hematoxylin (brazilin, hematoxylin B) content is 0.25%, tannic acid is 27.51%, and total phenols are 3.75%.

[0033] Example 3: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0034] Step S1, weighing 4 parts of Sappan wood, 2 parts of Sargentodoxae truncatula, 6 parts of Galla chinensis, 4 parts of Terminalia chebula, and 0.11 parts of Licorice root according to the above weight ratio, ultrafinely grinding and passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0035] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0036] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0037] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0038] Step S5: Add 0.5% polysorbate 80 to the filtrate obtained in step S4, stir at 50-55° C. to dissolve it, and then concentrate the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract. The relative density is controlled at 1.24 and the pH is 3.6, wherein the total hematoxylin (brazilin, hematoxylin B) content is 0.50%, tannic acid is 33.75%, and total phenols are 0.13%.

[0039] Example 4: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0040] Step S1, weighing 4 parts of Sappan wood, 4 parts of Sargentodoxae chinensis, 4 parts of Galla chinensis, 4 parts of Terminalia chebula, and 0.11 parts of Licorice root according to the above weight ratio, ultrafinely grinding and passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0041] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0042] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0043] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0044] Step S5, adding 0.5% polysorbate 80 to the filtrate obtained in step S4, stirring at 50-55° C. to dissolve it, and then concentrating the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract, with a relative density controlled at 1.19 and a pH of 4.0, wherein the total hematoxylin (brazilin, hematoxylin B) content is 0.50%, tannic acid is 25.0%, and total phenols are 2.50%.

[0045] Comparative Example 1: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0046] Step S1, weighing 5 parts of Sappan wood, 5 parts of Sargentodoxae truncatula, 6 parts of Galla chinensis, and 0.11 parts of Licorice root according to the above weight ratio, ultrafinely grinding them and passing them through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0047] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0048] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0049] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0050] Step S5: Add 0.5% polysorbate 80 to the filtrate obtained in step S4, stir at 50-55° C. to dissolve it, and then concentrate the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract. The relative density is controlled at 1.22 and the pH is 3.8, wherein the total hematoxylin (brazilin, hematoxylin B) content is 0.63%, tannic acid is 26.3%, and total phenols are 0.31%.

[0051] Comparative Example 2: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0052] Step S1, weighing 5 parts of Sappan wood, 5 parts of Sargentodoxae truncatulae, 6 parts of Terminalia chebula, and 0.11 parts of Glycyrrhiza uralensis according to the above weight ratio, ultrafinely grinding and passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0053] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0054] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0055] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0056] Step S5, adding 0.5% polysorbate 80 to the filtrate obtained in step S4, stirring at 50-55° C. to dissolve it, and then concentrating the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract, controlling the relative density at 1.14 and the pH at 4.3, wherein the total hematoxylin (brazilin, hematoxylin B) content is 0.63%, tannic acid is 12.00%, and total phenols are 0.31%.

[0057] Comparative Example 3: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0058] Step S1, weighing 5 parts of Sappan wood, 5 parts of Terminalia chebula, 6 parts of Gallnut, and 0.11 parts of Licorice according to the above weight ratio, ultrafine grinding and then passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0059] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0060] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0061] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0062] Step S5: Add 0.5% polysorbate 80 to the filtrate obtained in step S4, stir at 50-55° C. to dissolve it, and then concentrate the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract. The relative density is controlled at 1.24 and the pH is 3.8, wherein the total hematoxylin (brazilin, hematoxylin B) content is 0.63% and the tannic acid content is 35.62%.

[0063] Comparative Example 4: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0064] Step S1, weighing 5 parts of Sargentodon dahurica, 5 parts of Terminalia chebula, 6 parts of Galla chinensis, and 0.11 parts of Licorice root according to the above weight ratio, ultrafinely grinding them and passing them through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0065] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0066] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0067] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0068] Step S5: Add 0.5% polysorbate 80 to the filtrate obtained in step S4, stir at 50-55° C. to dissolve it, and then concentrate the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract with a relative density of 1.23 and a pH of 3.7. The total phenol content is 3.13% and the tannic acid content is 35.62%.

[0069] Comparative Example 5: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0070] Step S1, weighing 8 parts of Sappan wood, 3 parts of Sargentodoxae chinensis, 3 parts of Galla chinensis, 2 parts of Terminalia chebula, and 0.11 parts of Licorice root according to the above weight ratio, ultrafinely grinding and passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0071] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0072] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0073] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0074] Step S5: Add 0.5% polysorbate 80 to the filtrate obtained in step S4, stir at 50-55° C. to dissolve it, and then concentrate the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract. The relative density is controlled at 1.12 and the pH is 4.5, wherein the total hematoxylin (brazilin, hematoxylin B) content is 1.01%, tannic acid is 16.88%, and total phenols are 1.88%.

[0075] Comparative Example 6: Preparation of anti-Staphylococcus aureus compound Chinese medicine composition

[0076] Step S1, weighing 6 parts of Sappan wood, 6 parts of Sargentodoxae chinensis, 2 parts of Galla chinensis, 2 parts of Terminalia chebula, and 0.11 parts of Licorice root according to the above weight ratio, ultrafinely grinding and passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm;

[0077] Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract;

[0078] Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract;

[0079] Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate;

[0080] Step S5, adding 0.5% polysorbate 80 to the filtrate obtained in step S4, stirring at 50-55° C. to dissolve it, and then concentrating the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract, with a relative density controlled at 1.11 and a pH of 4.5, wherein the total hematoxylin (brazilin, hematoxylin B) content is 0.75%, tannic acid is 12.51%, and total phenols are 3.75%.

[0081] Antibacterial performance test:

[0082] MIC (Minimum Inhibitory Concentration) / MBC (Minimum Bactericidal Concentration) Determination: Use the modified microduplicate method: Take a 48-well culture plate and add 500 μL of MHB broth containing 5% defibrinated sheep blood to wells 1-16. Add 500 μL of the Chinese herbal medicine to be tested to well 1. After repeated pipetting five times, add 500 μL to well 2, and so on to well 14. After mixing well 14, discard 500 μL. No drug was added to well 15 as a positive control, and no bacteria were added to well 16 as a negative control. Inoculate 20 μL of bacterial solution into wells 1-15, with three replicates for each drug. After incubation at 37°C for 24 hours, visually inspect the culture medium for clarity. The lowest concentration corresponding to the clarification is the MIC of the Chinese herbal medicine extract. Use a sterile cotton swab to inoculate the test bacterial solutions from wells 1 to 15 respectively onto 5% defibrinated sheep blood MHA culture medium. The minimum drug concentration of the test bacterial solution for sterile growth is the MBC of the Chinese medicine.

[0083] Table 1 Antibacterial activity test results of Examples and Comparative Examples

[0084]

[0085]

[0086] The results showed that the four Chinese herbal extracts of Sappan wood, Terminalia chebula, Sargentodoxae and Gallnut had a synergistic antibacterial effect after being compounded in a specific proportion. In the four examples, the MIC was 0.49-0.98 mg / mL and the MBC was 0.98-1.95 mg / mL, indicating that compared with the comparative example, the four examples of the present invention had a stronger anti-Staphylococcus aureus effect and were superior to single Chinese medicines and other combination ratios.

[0087] Preparation of compound Chinese medicine extracts:

[0088] The compound Chinese medicine extracts prepared in Examples 1-5 and Comparative Examples 1-7 were used to prepare compound Chinese medicine preparations according to the formula in Table 2, to obtain the compound Chinese medicine preparations of Examples 6-10 and Comparative Examples 8-15, respectively.

[0089] Table 2 Formula of compound Chinese medicine preparation for anti-Staphylococcus aureus

[0090] raw material Quality score Compound Chinese herbal extract 30％ N,N-Dimethylformamide 5％ Nepal Gold Ester 1％ Sodium benzoate 1％ α-Pyrrolidone 10％ Polyvinylpyrrolidone (film-forming agent) 10％ Deionized water margin

[0091] Preparation method of compound Chinese herbal medicine extract preparation:

[0092] A. Add deionized water to a stirring pot, add α-pyrrolidone, fructose syrup, and N,N-dimethylformamide to the stirring pot, stir until completely dispersed, heat to 60-75℃, and homogenize at high speed for 5 minutes.

[0093] B. Add Nepal gold ester, sodium benzoate, and Bentian-50 and mix evenly. Add the compound antibacterial Chinese medicine compositions prepared in Examples 1-5 and Comparative Examples 6-10, respectively, stir and heat to 40-55° C., completely dissolve, add to the mixture in step A, homogenize at high speed for 5 minutes, cool to 40° C., and stir until the material is uniform to obtain the compound Chinese medicine preparations of Examples 5-8 and Comparative Examples 7-12.

[0094] The quality standards of the compound Chinese medicine extract preparations in Examples 6-10 of this patent are as follows:

[0095] Table 3 Quality standards of compound Chinese medicine preparations

[0096] project index Appearance Reddish brown liquid PH 4.4～6.5 Total hematoxylin 0.75％～0.19％ Tannic acid 7.50％～10.13％ Total phenols 0.04％～1.13％ arsenic ≤2ppm mercury ≤1ppm lead ≤10ppm cadmium ≤5ppm heavy metal ≤20ppm Total bacteria count ≤1000 CFU / g Mold and yeast count ≤100 CFU / g Escherichia coli Not to be detected Staphylococcus aureus Not to be detected salmonella Not to be detected Pseudomonas aeruginosa Not to be detected Thermal stability Freeze at -10℃ for 24h and thaw at room temperature without stratification Freeze stability Keep at 45℃ for 24h, and dissolve at room temperature without stratification

[0097] Clinical efficacy test of compound Chinese medicine extract:

[0098] 120 dairy cows suffering from clinical mastitis were selected (the clinical symptoms of clinical mastitis dairy cows were determined as follows: the clinical symptoms of the milk region were redness, tenderness, and fever; the milk color turned yellow or was light and flocculent; the somatic cell count was more than 800,000 / mL; and the milk sample contained Staphylococcus aureus through laboratory bacterial culture separation and identification). 120 dairy cows in the medication group were each group of 10 cows and 30 mL of compound Chinese medicine extracts of Examples 5-8 and Comparative Examples 7-12 were poured into the milk region twice a day. The chemical drug group (10 cows) was poured with ceftiofur sodium, and the control group (10 cows) was poured with distilled water. The results showed that the cure rates of Examples 5-8 were between 46.6% and 66.6%, and the total effective rate was between 86.6% and 100%, both of which were better than the comparative examples.

[0099]

[0100] In summary, the present invention selects Sappan wood, Sargentodoxae chinensis, Gallnut and Terminalia chebula in a certain proportion for use, which has a good anti-Staphylococcus aureus effect. Compared with chemical synthetic fungicides, the traditional Chinese medicine ingredients have the advantages of a wide target area, low resistance to drug resistance and no residue. After being prepared into preparations, animal tests have shown significant efficacy, showing broad application prospects in practical applications.

[0101] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. An anti-Staphylococcus aureus compound Chinese medicine composition, characterized in that: The raw materials of the compound Chinese medicine composition include, by weight, 2-5 parts of sappan wood, 2-6 parts of safflower vine, 4-6 parts of gallnut, 2-4 parts of terminalia chebula, and 0.11 part of liquorice. The MIC of the compound Chinese medicine composition is 0.49-0.98 mg / mL, and the MBC is 0.98-1.95 mg / mL.

2. The anti-Staphylococcus aureus compound Chinese medicine composition according to claim 1, characterized in that: The components of the compound Chinese medicine composition include, by weight percentage, 0.25% to 0.63% of total hematoxylin, 25.00% to 33.75% of tannic acid, and 0.13% to 3.75% of total phenols; wherein the content of the total hematoxylin is calculated based on the total amount of brazilin and hematoxylin B.

3. An anti-Staphylococcus aureus compound Chinese medicine preparation, comprising the anti-Staphylococcus aureus compound Chinese medicine composition according to any one of claims 1 to 2, characterized in that: The compound Chinese medicine preparation is selected from liquid preparation, semi-solid preparation, solid preparation or gas preparation.

4. A method for preparing the anti-Staphylococcus aureus compound Chinese medicine preparation according to claim 3, characterized in that: The compound Chinese medicine preparation is a liquid preparation containing 30% of the compound Chinese medicine extract; the preparation steps of the compound Chinese medicine extract are as follows: Step S1, weighing Sappan wood, Sargentodon seu Radix, Galla chinensis, Terminalia chebula, and Glycyrrhiza uralensis according to the above weight parts, ultrafinely grinding and passing through a 300-mesh sieve to obtain a compound Chinese medicine mixed powder with a particle size of 25 to 48 μm; Step S2, adding the mixed powder and ethanol at a solid-liquid ratio of 1:10 for ultrasonic pre-extraction, wherein the ultrasonic extraction temperature is 60-65° C., the ultrasonic time is 2-4 hours, standing for precipitation, and taking the supernatant to obtain the ethanol phase of the compound Chinese medicine extract; Step S3, adding 15 times the amount of purified water to the precipitated medicinal residue and boiling, then reducing the heat to simmer and concentrating the medicinal liquid to 10 times the total weight of the mixed powder, standing and settling, and collecting the supernatant to obtain the aqueous phase of the compound Chinese medicine extract; Step S4, mixing the aqueous phase and ethanol phase of the compound Chinese medicine extract cooled to room temperature, allowing them to settle for 24 to 48 hours, and filtering the supernatant through a 0.45 μm membrane to obtain a filtrate; Step S5: Add 0.5% polysorbate 80 to the filtrate obtained in step S4, stir at 50-55° C. to dissolve it, and then concentrate the filtrate by vacuum distillation to 1 times the total weight of the traditional Chinese medicine to obtain a 1 g / mL compound traditional Chinese medicine extract with a relative density of 1.19-1.24 and a pH of 3.6-4.

0.

5. The method for preparing the anti-Staphylococcus aureus compound Chinese medicine preparation according to claim 4, wherein: In step S1, the obtained mixed powder is ultrafinely pulverized to collect Chinese medicinal powder with a particle size of 25 to 48 μm.

6. The method for preparing the anti-Staphylococcus aureus compound Chinese medicine preparation according to claim 5, characterized in that: In step S4, the impurities in the compound Chinese herbal medicine extract are precipitated by alcohol precipitation.

Images