High-fidelity bezoar and preparation method thereof

Abstract

The invention provides a high-fidelity calculus bovis and a preparation method thereof, and belongs to the technical field of traditional Chinese medicines. According to the technical scheme provided by the invention, the high-fidelity calculus bovis is accurately prepared by taking the mean value of the contents of various components of the natural calculus bovis as a reference and accurately measuring the contents of various components in the raw materials, fully utilizing the original components of the ox bile or ox gall powder, not adding other non-calculus bovis components and combining the existence forms of the components. The problems that existing artificial calculus bovis is greatly different from natural calculus bovis in efficacy, and toxic and side effects are relatively high are solved. The shape, the forming process and the components of the product are highly similar to those of natural calculus bovis; the medicine effect of the calculus bovis is improved, and the production cost is reduced.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of traditional Chinese medicines, and particularly relates to high-fidelity ox gall and a preparation method thereof. BACKGROUND

[0002] Due to the scarcity of natural ox gall, various cultivation and preparation methods of ox gall have appeared in recent decades. At present, medical ox gall is mostly artificial ox gall or in-vitro cultivated ox gall. The synthesis method of artificial ox gall is disclosed in the methods of the patents with announcement numbers CN101804195A, CN103690563A, CN115837035A and CN116077529A. The in-vitro cultivation method of ox gall is disclosed in the methods of the patents with announcement numbers CN117562927A and CN111658671A. The in-vitro cultivated ox gall is produced by simulating the pathological environment of the bile duct infected by bacteria, but the bacterial fermentation method is relatively complex and the process is tedious. The synthesis method of artificial ox gall is simple and direct preparation, but compared with the natural ox gall, the main differences of the synthetic artificial ox gall are as follows: Component difference: Compared with the components of natural ox gall, the high is not high and the low is not low. Although the main components of the substitutes are roughly the same as those of natural ox gall, a large amount of components such as starch and chitosan which are not originally in natural ox gall are added; the synergistic components are lacking; the relative content and proportion of each component are quite different; and the content and proportion of inorganic components such as calcium, zinc, potassium and sodium are also not matched with those of natural ox gall. Formation process difference: The principle of the existing ox gall preparation technology is completely different from the formation process of natural ox gall. Natural ox gall is the product of cholecystic pathological metabolism, and its formation process is a slow dynamic process. The deposition of the pathological metabolism components secreted by the upstream (liver) is continuously in the gallbladder. The healthy bovine bile taken from the gallbladder has no supply of the pathological components secreted by the upstream (liver), and the healthy bovine bile only contains several ten-thousandths of bilirubin. Even if the formation process of natural ox gall is simulated by adding escherichia coli for fermentation, the purpose of increasing bilirubin cannot be achieved due to the limitation of the upstream components.

[0003] Morphology difference: Natural ox gall is a natural compound form, while the prepared artificial ox gall is simply added with related components, and the state or form is quite different from that of natural ox gall. The material basis of the pharmacological action is also not completely the same.

[0004] Unreasonable formula and artificial difference: In the previous related inventions, only several main components of ox gall are considered, the bile components are not fully utilized, and other components with synergistic effects in ox gall are ignored. Only the products meeting the pharmacopoeia standard are regarded as qualified products, which leads to many problems such as low utilization rate, low drug efficacy and the like compared with natural ox gall. SUMMARY

[0005] Therefore, the present application provides a high-fidelity bezoar and a preparation method thereof, which realizes a product similar to natural bezoar in form, formation process and ingredients, and achieves the purpose of narrowing the pharmacodynamic gap between artificial bezoar and natural bezoar.

[0006] To achieve the above-mentioned purpose, the present application provides a high-fidelity bezoar, which comprises the following components by weight percentage: bilirubin 15-39%, cholic acid 2.5-6%, glycocholic acid 3.5-4.5%, chenodeoxycholic acid 0.5-2%, chenodeoxycholic acid 0.25-2%, glycochenodeoxycholic acid 0.2-0.5%, glycodeoxycholic acid 0.6-2%, cholic acid 0.6-2.5%, chenodeoxycholic acid 0.1-0.3%, deoxycholic acid 0.2-0.8%, lithocholic acid 0.05-0.1%, total cholesterol 0.5-2%, calcium 2.3-4%, zinc 1.4-2.4%, magnesium 0.01-0.03%, iron 0.4-0.65%, potassium 0.04-0.09%, sodium 0.6-1%, carotenoids and fatty acids 1-2.1%, taurine 0.5-0.9%, total protein amino acids 15-25%, calcium / bilirubin ratio 1:7-20, calcium / zinc ratio 1:0.1-0.5, calcium / magnesium ratio 1:0.008-0.01, and sodium / potassium ratio 1:0.1-0.12.

[0007] To achieve the above-mentioned purpose, the present application further provides a preparation method of the high-fidelity bezoar, which comprises the following steps: using bovine bile or bovine gall powder and bovine gallbladder inner wall to prepare protein bilirubin calcium zinc sodium multi-component complex, and using bovine bile or bovine gall powder to prepare total bile acid, carotenoids and fatty acids; and then mixing the protein bilirubin calcium zinc sodium multi-component complex, the total bile acid, the carotenoids and the fatty acids, and performing rotary extrusion to obtain the high-fidelity bezoar.

[0008] Optionally, when bovine bile is used as the raw material, the following preparation steps are included: S1, after taking out the bile from fresh bovine gallbladder, the bovine gall is cut open, and the intramural and mucosal layers are peeled off and taken out, to obtain the bovine gallbladder inner wall; the bovine gallbladder inner wall is cut into small pieces, and then is high-speed beaten after adding water, and centrifuged to obtain the gallbladder inner wall slurry; the bovine bile is mixed with dissolved cholesterols, the gallbladder inner wall slurry, calcium chloride, taurine, and is fully stirred, and then is adjusted in pH with an alkali solution, and is fully stirred after adding bilirubin, and is added with zinc sulfate after being kept warm in stages, and is centrifuged after being cooled, to obtain the centrifugal supernatant and the centrifugal precipitate; the centrifugal precipitate is added with water and is centrifuged again, to obtain the protein bilirubin calcium zinc sodium multi-component complex and the centrifugal supernatant of the second centrifugation; S2, combine the secondary centrifugal supernatant with the centrifugal supernatant, adjust the pH with an acid, stir thoroughly, then stand, and then centrifuge to obtain a tertiary centrifugal supernatant and a tertiary centrifugal precipitate; the tertiary centrifugal supernatant is extracted with ethyl acetate, and the upper liquid phase is taken, and after recovery of the ethyl acetate, carotenoids and fatty acids are obtained; the tertiary centrifugal precipitate is dissolved and extracted with a mixed solution, and after recovery of the mixed solution, total bile acids are obtained; the mixed solution is obtained by mixing ethyl acetate and ethanol; S3, mix the protein bilirubin calcium zinc sodium multiple complex with carotenoids and fatty acids, mix the total bile acids and potassium dihydrogen phosphate uniformly, dry to obtain refined bovine bezoar, and after grinding, mix with edible alcohol, and obtain high-fidelity bovine bezoar by rotary extrusion.

[0009] Optionally, in S1, the mass ratio of the inner wall of the bovine gallbladder to water is 1:1; the centrifugation and secondary centrifugation conditions are a rotation speed of 3500-4500 rpm and a time of 5-10 min; the alkali solution adjusts the pH to 8-8.5; the stepwise incubation is incubated at 38-42°C for 1.5-2.5 h, and then the temperature is increased to 790-810°C at a rate of 2-3°C / min and incubated for 1.5-2.5 h; the volume ratio of water to precipitate when the secondary precipitate is added with water is 3-5.

[0010] Optionally, in S2, the acid adjusts the pH to 1-2, and the standing time after thorough stirring is 2-4 h; the rotation speed of the tertiary centrifugation is 5500-6500 rpm, and the time is 15-25 min; the mass ratio of ethyl acetate to ethanol in the mixed solution is 3:1, and the ethanol is 95% ethanol.

[0011] Optionally, in S3, the powder is ground to 80-100 mesh; when the powder is mixed with edible alcohol after grinding, the weight-volume ratio of the powder to edible alcohol is 8:1, the concentration of the edible alcohol is 65%, and the rotation speed of the rotary extrusion is 40-400 rpm.

[0012] Optionally, when bovine gall powder is used as the raw material, the following preparation steps are included: Step 1, mix the bovine gall powder with an alcohol ester solution, slowly stir while incubating, and separate the insoluble material and the solution to obtain the insoluble material and the solution, repeat the above operation with the insoluble material and the alcohol ester solution to obtain a solid and a secondary solution, combine the solution and the secondary solution, and recover under reduced pressure to a syrup to obtain total bile acids, carotenoids and fatty acids; Step 2, after taking out the bile from fresh bovine gallbladder, the bovine gall is opened, the inner muscle layer and mucous membrane layer are peeled off and taken, and the bovine gall inner wall is obtained; the bovine gall inner wall is cut into small pieces, and after adding water, high-speed beating is carried out, and after centrifugation, the gallbladder inner wall slurry is obtained; after the solid alcohol ester solution is removed, water is added, and then the dissolved cholesterols, the gallbladder inner wall slurry are mixed, calcium chloride and taurine are added, and fully stirred, the pH is adjusted with an alkali solution, bilirubin is added and fully stirred, and after staged incubation, zinc sulfate is added, and after cooling, centrifugation is carried out, and the centrifugal precipitate is obtained, and the centrifugal precipitate is added with water, and secondary centrifugation is carried out, and the protein bilirubin calcium zinc sodium multi-component complex is obtained; Step 3, the total bile acid, carotenoid and fatty acid are mixed with potassium dihydrogen phosphate uniformly, and then the protein bilirubin calcium zinc sodium multi-component complex is added and mixed, and after drying, the refined bovine bezoar is obtained, and after being ground into powder, the powder is mixed with edible alcohol, and high-fidelity bovine bezoar is obtained through rotary extrusion.

[0013] Optionally, in the step 1, the mass-volume ratio of the bovine gall powder and the alcohol ester solution is 1: (5-6), the alcohol ester solution is obtained by mixing ethanol and ethyl acetate, the volume ratio of the ethanol and the ethyl acetate is 1:3; the incubation temperature is 40-50℃, the slow stirring time is 20-30min, and the temperature of the reduced pressure recovery is 55-60℃.

[0014] Optionally, in the step 2, the mass ratio of the bovine gall inner wall and water is 1:1, the centrifugation and secondary centrifugation conditions are that the rotation speed is 3500-4500rpm, and the time is 5-10min, the pH of the alkali solution is adjusted to 8-8.5; the staged incubation is that the temperature is incubated at 38-42℃ for 1.5-2.5h, then the temperature is increased to 79-81℃ at a rate of 2-3℃ / min, and incubated for 1.5-2.5h; when the secondary precipitate is added with water, the volume ratio of the water and the precipitate is 3-5.

[0015] Optionally, in the step 3, the powder is ground to 80-100 meshes; when the powder is mixed with edible alcohol after being ground, the weight-volume ratio of the powder and the edible alcohol is 8:1, the concentration of the edible alcohol is 65%, and the rotation speed of the rotary extrusion is 40-400rpm.

[0016] The above technical scheme of the present application at least has the following beneficial effects: The technical scheme provided by the application takes into account the content and proportion of main components and secondary components; makes full use of the original components of bovine gall or bovine gall powder and bovine gallbladder membrane, so that the bovine gall components are fully utilized, and the addition of other unnecessary non-ursine components is avoided; the main component is a protein bilirubin calcium zinc sodium multi-component compound; the chemical existence form of the main component is more similar to the chemical existence form of natural ursine; the bacterial fermentation process and the separation and purification process are abandoned; the average value of the content of various components of natural ursine is taken as a reference, the accurate determination of the content of various components in the raw material is taken as a basis, and the component index of high-fidelity ursine is determined according to the modern pharmacological research results of natural ursine; the high-fidelity ursine has the basic characteristics and activity of natural ursine, and has no difference in color, taste, quality and nature from natural ursine; the drug efficacy is better than that of other artificial ursine or in-vitro cultured ursine; the production process is simple, the quality is controllable, the investment is small, and the production is suitable for large-scale or industrial production. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 SEM micro-morphology diagram of high-fidelity ursine in Example 1 of the application; Figure 2 SEM micro-morphology diagram of natural ursine in Comparative Example 1 of the application. DETAILED DESCRIPTION

[0018] In order to make the purpose, technical scheme and advantages of the embodiments of the application more clear, the technical scheme of the embodiments of the application will be described clearly and completely in combination with the embodiments of the application. Obviously, the described embodiments are part of the embodiments of the application, rather than all the embodiments. Based on the described embodiments of the application, all other embodiments obtained by those skilled in the art belong to the scope of protection of the application.

[0019] Example 1 The application provides a preparation method of high-fidelity ursine, comprising the following steps: The method using bovine gall as raw material is as follows: Preparation of complex bilirubin calcium protein 1. After the fresh bovine gallbladder is taken out of the gall, the bovine gall is opened, the gallbladder muscle layer and mucous membrane layer are peeled off and reserved, and the bovine gallbladder inner wall is obtained; the bovine gallbladder inner wall is cut into small pieces, 6 kg of the bovine gallbladder inner wall is added with 6 kg of water, and then high-speed beating is performed, and 4000 rpm centrifugation is performed for 8 minutes, and the liquid is reserved; 2. 50 kg of fresh bovine gall is added with 10 kg of gallbladder inner wall slurry, 25 g of cholesterol (dissolved in 200 ml of ethanol) is mixed in a jacketed reaction kettle; 3. 0.3 kg of calcium chloride and 0.7 kg of taurine are added, and fully stirred; 4. The system PH is adjusted to 8.5 by using 20% sodium hydroxide; 5. Add 0.7 kg of bilirubin and mix well; 6. Keep the water-jacketed reactor 400C for 2 hours, then increase to 800C at a rate of 20C / min and keep for 2 hours; 7. Add 0.25 kg of zinc sulfate (ZnSO4.7H2O) and cool; 8. Centrifuge at 4000 rpm for 5-10 minutes; 9. Separate the supernatant and the precipitate; 10. Stir the precipitate with 4 times the volume of water and centrifuge at 4000 rpm for 8 minutes; 11. Obtain 2.8 kg of protein bilirubin calcium zinc sodium multi-complex and the supernatant of the second centrifugation. Combine the supernatant of the second centrifugation with the supernatant of the first centrifugation to obtain a combined supernatant; Total bile acid extraction 12. Adjust the pH of the combined supernatant obtained in step 11 to 1-2 with hydrochloric acid, mix well, and stand for 2-4 hours; 13. Centrifuge the combined supernatant at 6000 rpm for 20 minutes to obtain the supernatant of the third centrifugation and the precipitate of the third centrifugation, and retain the precipitate of the third centrifugation; 14. Extract the supernatant of the third centrifugation with 10 L of ethyl acetate once. After standing and separating the layers, take the upper liquid phase, and recover the ethyl acetate at 65C under reduced pressure to obtain 0.25 kg of carotenoids and fatty acids; 15. Dissolve and extract the precipitate of the third centrifugation with 10 L of ethyl acetate / 95% ethanol (3:1) at 45C, recover the ethyl acetate and ethanol at 65C under reduced pressure, and obtain 1.65 kg of total bile acid; 16. Quantitatively determine the content of each bile acid component in the total bile acid for later use.

[0020] Product fine blending 17. Mix the 2.8 kg of protein bilirubin calcium zinc sodium multi-complex obtained in step 11 and the 0.25 kg of carotenoids and fatty acids obtained in step 14 according to the intended metering; 18. Mix 0.56 kg of the total bile acid obtained in step 15 and 9 g of potassium dihydrogen phosphate well; after drying, obtain 3.6 kg of fine blended ox gall; dry, grind to 90 mesh; 19. Quantitatively determine each index component; 20. Use the solid molding method to spray the fine blended powder into 65% edible alcohol at a weight to volume ratio of 8:1, and place it in a modified tangyuan machine or similar equipment to produce high-fidelity ox gall of the desired diameter at a rotation speed of 200 rpm. Dry, package, label, and store in the warehouse.

[0021] The application provides a high-fidelity ox gall, which comprises the following components in percentage by weight: 21.7% of bilirubin, 4.746% of cholic acid, 3.948% of glycocholic acid, 1.631% of oxogallate, 1.822% of chenodeoxycholic acid, 0.220% of glycochenodeoxycholic acid, 1.049% of glycodeoxycholic acid, 1.907% of cholic acid, 0.141% of chenodeoxycholic acid, 0.267% of deoxycholic acid, 0.083% of lithocholic acid, 0.9% of total cholesterol, 2.75% of calcium, 1.4% of zinc, 0.03% of magnesium, 0.55% of iron, 0.087% of potassium, 0.76% of sodium, 1.5% of carotenoids and fatty acids, 0.8% of taurine, 21.4% of total protein amino acids, the ratio of calcium / bilirubin is 1:7.9, the ratio of calcium / zinc is 1:0.5, the ratio of calcium / magnesium is 1:0.01, and the ratio of sodium / potassium is 1:0.11.

[0022] Example 2 The application provides a preparation method of high-fidelity ox gall, which comprises the following steps: The method using ox gall powder as raw material is as follows: Total bile acid extraction 1. 5 kg of ox gall powder is added into 30 L of ethanol / ethyl acetate (1:3, V / V), and the mixture is kept at 45 DEG C and stirred slowly for 20-30 min; 2. The alcohol-ester solution and insoluble substances are separated, the insoluble substances are repeated once according to step 1 to obtain solid and secondary solution, and the solid is reserved after being cooled to room temperature, and the alcohol-ester solutions of the two times are combined; 3. The solvent is recovered to syrup at 55-60 DEG C under reduced pressure, and 1.45 kg of total bile acid, carotenoids and fatty acids are obtained; 4. The content of each bile acid is determined.

[0023] Preparation of compound bilirubin calcium protein 5. After the bile is taken out from fresh ox gall bladder, the ox gall is opened, and the inner muscle layer and mucous membrane layer are peeled and reserved to obtain the inner wall of ox gall bladder; the inner wall of ox gall bladder is cut into small pieces, 3.5 kg of the inner wall of ox gall bladder is added into 3.5 kg of water, and then the mixture is high-speed beaten and centrifuged at 4000 rpm for 10 min, and the liquid is reserved; 6. After the insoluble substances in step 2 are added into 10 L of water; 7. The materials in steps 5 and 6 are combined in a jacketed reaction kettle, 20 g of cholesterols (dissolved in 200 ml of ethanol) and 0.3 kg of calcium chloride, 0.7 kg of taurine are added, and the mixture is fully stirred; 8. The PH of the system is adjusted to 8.5 by using 20% sodium hydroxide; 9. 0.7 kg of bilirubin is added, and the mixture is fully stirred; 10. The circulating water jacketed reactor is kept at 400℃ for 2 hours, then increased to 80℃ at a rate of 2℃ / min and kept for 2 hours; 11. Add 0.24 kg of zinc sulfate (ZnSO4.7H2O) and cool; 12. Centrifuge at 4000 rpm for 5 minutes; 13. Obtain the centrifugal precipitate; 14. Stir the centrifugal precipitate with 5 times the volume of water, and centrifuge again at 4000 rpm for 10 minutes; dry the precipitate to obtain 2.6 kg of the protein bilirubin calcium zinc sodium multi-complex; Product fine blending 15. Mix 0.5 kg of the total bile acid, carotenoid and fatty acid obtained in step 3 with 9 g of potassium dihydrogen phosphate, and then mix with the 2.6 kg of the protein bilirubin calcium complex obtained in step 14; 16. Dry to obtain 3.1 kg of fine blending ox gall, and grind to 100 mesh; 17. Quantitatively determine each index component; 18. Adopt the solid molding method, spray the fine blended powder into 65% edible alcohol at a weight to volume ratio of 8:1, and place in a modified tangyuan machine or similar equipment to produce high fidelity ox gall of the required diameter at a rotation speed of 400 rpm. Dry, detect, package, label and store.

[0024] The present application provides a kind of high fidelity ox gall, including the following weight percentage components: bilirubin 21.7%, cholic acid 4.746%, glycocholic acid 3.948%, ox gall deoxycholic acid 1.631%, ox gall chenodeoxycholic acid 1.822%, glycochenodeoxycholic acid 0.220%, glycodeoxycholic acid 1.049%, cholic acid 1.907%, chenodeoxycholic acid 0.141%, deoxycholic acid 0.267%, lithocholic acid 0.083%, total cholesterol 0.9%, calcium 2.75%, zinc 1.4%, magnesium 0.03%, iron 0.57%, potassium 0.083%, sodium 0.76%, carotenoid and fatty acid 1.5%, taurine 0.8%, total protein amino acid 21.4%, calcium / bilirubin ratio 1:7.9, calcium / zinc ratio 1:0.5, calcium / magnesium ratio 1:0.01, sodium / potassium ratio 1:0.11.

[0025] Comparative Example 1 Natural ox gall.

[0026] The ox gall prepared in Examples 1 and 2 is subjected to component testing, and the results are compared with the component index of the present application, as shown in Table 1. The testing method is as follows: The method for determining the protein bilirubin calcium zinc sodium multi-complex is high performance liquid chromatography.

[0027] HPLC conditions: Column: Kromasil C18 (250 mm x 4.6 mm, 5 μm) Mobile phase: acetonitrile:methanol:0.1 M ammonium acetate-acetic acid pH 4.7 (10:70:20, V / V / V), isocratic elution; flow rate: 1.5 mL / min; UV detection wavelength: 450 nm. Sample preparation: Each 10 mg was weighed and dissolved in a 100 mL brown volumetric flask, and then chloroform was added to constant volume. After filtration, the sample was determined by HPLC. The content of total bile acid and various bile acids was determined by HPLC-ELSD.

[0028] HPLC-ELSD determination of bile acids: The column was Ultimate AQ-C, the mobile phase was acetonitrile 0.2% formic acid aqueous solution, gradient elution, and the flow rate was 1 mL / min; the parameters of the evaporative light scattering detector were: drift tube temperature 100°C, carrier gas (N) flow rate 1.9 L / min.

[0029] The inorganic elements in the intermediate product were determined by inductively coupled plasma mass spectrometry (ICP-MS / MS).

[0030] ICP-MS / MS determination of inorganic elements: The determination was carried out by He collision mode, the plasma mode was HMI-8, the sampling depth was 10.0 mm, the radio frequency power was 1 600 W, the carrier gas flow rate was 0.8 L / min, the atomizing gas flow rate was 0.8 L / min, the auxiliary gas flow rate was 0.8 L / min, the plasma gas flow rate was 15.0 L / min, the peristaltic pump speed was 0.10 r / min, and the atomizing chamber temperature was 2°C.

[0031] Table 1. The proposed indicators of the present application and the content detection table of each component of the bile of Example 1 and Example 2

[0032] The present application provides a high-fidelity calculus bovis and a preparation method thereof. Since there is a big difference between healthy bovine bile or bovine gall powder and the main components in natural calculus bovis, such as insufficient protein content in bile, only a few parts per hundred thousand of bilirubin, low total cholesterol, too high sugar content, total bile acid content is several times higher than that of natural calculus bovis, and the ratio of calcium and zinc is out of balance, etc., all of which cannot reach the best proportion for preparing calculus bovis. Therefore, bovine bile or bovine gall powder cannot be directly used to prepare calculus bovis. In order to reach the component content range of natural calculus bovis, the present application uses part of the sugar to reduce the proportion of sugar, uses the gallbladder wall to increase the proportion of mucin, extracts total bile acid and determines the components of each bile acid, and then adds the corresponding components according to the measurement to optimize the content ratio of bile acid. At the same time, it is also considered that the original bile acid is easily metabolized into secondary bile acid during transportation, storage and production, and finally all degraded into lithocholic acid. Although lithocholic acid has physiological activities such as inhibiting tumor growth and killing cancer cells, it can also cause liver damage. Therefore, the present application limits the amount of such components such as lithocholic acid, cholic acid and deoxycholic acid in the raw material formula.

[0033] Studies have shown that calculus bovis containing 0.7% bilirubin and calculus bovis containing 18% bilirubin have no difference in reducing the effect of elevated serum glutathione transhydrogenase. This shows that the content of bilirubin is not the main factor determining the efficacy. Therefore, the content of bilirubin is not a strict index in the formulation.

[0034] The formation of natural calculus bovis is that organic and inorganic components exist and gradually interweave together in a complex and symbiotic environment, and finally form unique complex components and structures. Natural calculus bovis is often formed with the core of exfoliated epithelium, parasite eggs, mucous blocks, bacterial blocks or other foreign matters, and the factors promoting the formation of calculus bovis are not independent of each other, but are the result of pathological balance. Bilirubin calcium stones are not single precipitates, but protein-calcium-bilirubin complexes. Glycoprotein, as a linear macromolecule, combines with calcium ions and a small amount of other elements such as zinc and sodium ions through its negatively charged part, and its hydrophobic region combines with bilirubin, forming a bilirubin-metal salt-mucin high molecular polymer complex, and the three are deposited together to form a stone embryo. Bilirubin can react with many metal ions, but the reaction rates are different. In acidic or slightly alkaline medium, the reaction of bilirubin with metal ions is not obvious, and with the increase of alkalinity of the medium, the reaction is accelerated. Therefore, when formulating calculus bovis, not only the chemical components of natural drugs should be restored, but also the component content and ratio should be consistent as much as possible, and attention should also be paid to the form, structure and configuration or conformation of the components. Calcium ions slowly form protein calcium flocculates in a weak alkaline environment, and after adding bilirubin, protein bilirubin calcium complexes are formed, and the addition of zinc sulfate can make the flocculates densely aggregated into a group.

[0035] The high-fidelity calculus bovis prepared in Example 1 and the natural calculus bovis of Comparative Example 1 are micro-characterized, and the results are as follows: Figure 1、 Figure 2 .

[0036] In combination with Table 1 Figure 1 、 Figure 2 It can be known that the protein-calcium-bilirubin multiple complex can be formed by controlling the adding amount and ratio of calcium, zinc and sodium, reaction time, adding sequence and reaction temperature and the like conditions, and the high-fidelity bovine bezoar can be prepared by accurately measuring each component in the processed intermediate raw material, and then accurately preparing through calculation, which is not only consistent with the chemical components, component content and ratio of natural bovine bezoar, but also closer to the natural bovine bezoar in micro-morphology. The product tends to approach the indexes and performances of the natural bovine bezoar as much as possible.

[0037] The above is the preferred embodiment of the present application, it should be pointed out that, for those skilled in the art, without departing from the principles of the present application, can make a number of improvements and refinements, these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A high-fidelity calculus bovis, characterized in that, Comprise the following weight percentage of components: bilirubin 15-39%, cholic acid 2.5-6%, glycocholic acid 3.5-4.5%, chenodeoxycholic acid 0.5-2%, chenodeoxycholic acid 0.25-2%, glycochenodeoxycholic acid 0.2-0.5%, glycodeoxycholic acid 0.6-2%, cholic acid 0.6-2.5%, chenodeoxycholic acid 0.1-0.3%, deoxycholic acid 0.2-0.8%, lithocholic acid 0.05-0.1%, total cholesterol 0.5-2%, calcium 2.3-4%, zinc 1.4-2.4%, magnesium 0.01-0.03%, iron 0.4-0.65%, potassium 0.04-0.09%, sodium 0.6-1%, carotenoids and fatty acids 1-2.1%, taurine 0.5-0.9%, total amino acids 15-25%, calcium / bilirubin ratio 1:7-20, calcium / zinc ratio 1:0.1-0.5, calcium / magnesium ratio 1:0.008-0.01, sodium / potassium ratio 1:0.1-0.

12.

2. The method of preparing high-fidelity calculus bovis according to claim 1, characterized in that, Comprise the following steps: The protein bilirubin calcium zinc sodium multiple complex is prepared from bovine bile or bovine gall powder and the inner wall of bovine gallbladder, and total bile acid and carotenoids and fatty acids are prepared from bovine bile or bovine gall powder; then the protein bilirubin calcium zinc sodium multiple complex, total bile acid and carotenoids and fatty acids are mixed and high-fidelity bovine bezoar is obtained by rotary extrusion.

3. The method of preparing high-fidelity calculus bovis according to claim 2, characterized in that, When bovine bile is used as raw material, the following preparation steps are included: S1, after taking out the bile from fresh bovine gallbladder, the bovine gall is cut open, the inner muscle layer and mucous membrane layer are peeled off and taken out, and the inner wall of the bovine gallbladder is obtained; the inner wall of the bovine gallbladder is cut into small pieces, water is added and high-speed beating is carried out, and after centrifugation, the gallbladder wall slurry is obtained; the bovine bile is mixed with dissolved cholesterols, gallbladder wall slurry, calcium chloride, taurine is added, stirred fully, the pH is adjusted with alkali solution, bilirubin is added and stirred fully, and after segmental preservation, zinc sulfate is added, cooled and centrifuged to obtain centrifugal supernatant and centrifugal precipitate; the centrifugal precipitate is added with water and subjected to secondary centrifugation to obtain protein bilirubin calcium zinc sodium multiple complex and secondary centrifugal supernatant; S2, the secondary centrifugal supernatant and the centrifugal supernatant are combined, the pH is adjusted with acid, stirred fully, and then centrifuged to obtain tertiary centrifugal supernatant and tertiary centrifugal precipitate; the tertiary centrifugal supernatant is extracted with ethyl acetate, the upper liquid phase is taken, and after recovery of ethyl acetate, carotenoids and fatty acids are obtained; the tertiary centrifugal precipitate is dissolved and extracted with a mixed solution, and after recovery of the mixed solution, total bile acid is obtained; the mixed solution is obtained by mixing ethyl acetate and ethanol; S3, the protein bilirubin calcium zinc sodium multiple complex and carotenoids and fatty acids are mixed, the total bile acid and potassium dihydrogen phosphate are mixed uniformly, dried to obtain refined bovine bezoar, and then ground and mixed with edible alcohol to obtain high-fidelity bovine bezoar by rotary extrusion.

4. The method of preparing high-fidelity calculus bovis according to claim 3, characterized in that, The mass ratio of the bovine gallbladder inner wall to water in S1 is 1:1; the centrifugation and secondary centrifugation conditions are 3500-4500 rpm for 5-10 min; the alkaline solution is adjusted to pH 8-8.5; the segmented incubation is 38-42℃ for 1.5-2.5 h, then increased to 790-810℃ at a rate of 2-3℃ / min for 1.5-2.5 h; the secondary precipitation is added with water in a volume ratio of 3-5.

5. The method of preparing high-fidelity calculus bovis according to claim 3, characterized in that, In S2, the acid is adjusted to pH 1-2, and the standing time after sufficient stirring is 2-4 h; the speed of the third centrifugation is 5500-6500 rpm for 15-25 min; the mass ratio of ethyl acetate to ethanol in the mixed solution is 3:1, and the ethanol is 95% ethanol.

6. The method of preparing high-fidelity calculus bovis according to claim 3, characterized in that, In S3, the powder is ground to 80-100 mesh; the powder is mixed with edible alcohol in a weight-volume ratio of 8:1, the concentration of the edible alcohol is 65%, and the rotation extrusion speed is 40-400 rpm.

7. The method for preparing high-fidelity bezoar according to claim 2, characterized in that, When bovine gall powder is used as the raw material, the following preparation steps are included: Step 1, the bovine gall powder is mixed with an alcohol-ester solution, slowly stirred while incubating, and then the insoluble matter and the solution are separated, the insoluble matter is repeatedly mixed with the alcohol-ester solution, and the solid and the secondary solution are obtained, the solution and the secondary solution are combined, and the total bile acid, carotenoids and fatty acids are obtained by recovering the solution to a syrup under reduced pressure; Step 2, the fresh bovine gallbladder is opened after removing the bile, the inner muscle layer and mucous membrane layer are peeled off and retained, and the bovine gallbladder inner wall is obtained; the bovine gallbladder inner wall is cut into small pieces, added with water, and then high-speed beaten, and the gallbladder inner wall slurry is obtained after centrifugation; the solid is added with water after removing the alcohol-ester solution, then mixed with dissolved cholesterols and gallbladder inner wall slurry, added with calcium chloride and taurine, stirred, adjusted to pH with an alkaline solution, added with bilirubin, stirred, incubated in stages, added with zinc sulfate, cooled, centrifuged, and the centrifugal precipitate is obtained; the centrifugal precipitate is added with water, and then the protein bilirubin calcium zinc sodium multi-component complex is obtained after secondary centrifugation; Step 3, the total bile acid, carotenoids and fatty acids are mixed with potassium dihydrogen phosphate, then mixed with the protein bilirubin calcium zinc sodium multi-component complex, dried, and then the refined bovine bezoar is obtained, which is ground and mixed with edible alcohol, and then the high-fidelity bovine bezoar is obtained by rotation extrusion.

8. The method of preparing high-fidelity calculus bovis according to claim 7, characterized in that, In step 1, the mass-volume ratio of the bovine gall powder to the alcohol-ester solution is 1:(5-6), the alcohol-ester solution is obtained by mixing ethanol and ethyl acetate in a volume ratio of 1:3; the incubation temperature is 40-50℃, the slow stirring time is 20-30 min, and the temperature for recovering under reduced pressure is 55-60℃.

9. The method for preparing high-fidelity bezoar according to claim 7, characterized in that, The mass ratio of the inner wall of the bovine gallbladder to water in step 2 is 1:1, the centrifugal and secondary centrifugal conditions are 3500-4500 rpm for 5-10 min, the pH of the alkaline solution is adjusted to 8-8.5; the segmented incubation is incubated at 38-42℃ for 1.5-2.5 h, then increased to 790-810C at a rate of 2-3℃ / min and incubated for 1.5-2.5 h; the volume ratio of water to precipitate in the secondary precipitation is 3-5.

10. The method for preparing high-fidelity bezoar according to claim 7, characterized in that, In step 3, the powder is ground to 80-100 mesh; the weight / volume ratio of the powder to edible alcohol after mixing is 8:1, the concentration of the edible alcohol is 65%, and the rotation speed of the rotary extrusion is 40-400 rpm.

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