Processing method for promoting dissolution of effective substances in buffalo horn

Abstract

This invention discloses a processing method for promoting the dissolution of effective substances in buffalo horn, belonging to the field of drug extraction technology, including the following steps: S1, crushing raw buffalo horn into particles of 3-5 mm to obtain buffalo horn granules; S2, adding the buffalo horn granules to distilled water for hydrolysis for 30-60 minutes to obtain buffalo horn hydrolysate; the hydrolysis temperature is 130-230℃ and the pressure is 2-20 MPa; S3, filtering the hydrolysate and directly collecting the filtrate, or evaporating the filtrate to dryness to obtain the processed buffalo horn product. This invention uses microwave hydrolysis, which can hydrolyze the insoluble substances in buffalo horn in a short time. The average buffalo horn extract content of the processed buffalo horn product reaches 18.57%, which is nearly six times higher than the extract content of long-term decoction, greatly increasing the extract content of buffalo horn. Therefore, microwave hydrolysis of buffalo horn dissolves the sample more completely than decoction, and can significantly increase the water-soluble extract content of buffalo horn.

Description

Technical Field

[0001] This invention belongs to the field of drug extraction technology, specifically a processing method for promoting the dissolution of effective substances from buffalo horn. Background Technology

[0002] Buffalo horn, an important traditional Chinese medicine with heat-clearing and detoxifying properties, has wide applications in medicine. However, current pharmacological research on buffalo horn mainly focuses on different molecular weight fractions of buffalo horn extract or its components. The exact functional components have not yet been identified, and further research is needed to explore the functional components and mechanisms of action of active substances in buffalo horn. This will lay the foundation for subsequent quality evaluation and promote the application of buffalo horn in medicine.

[0003] Both buffalo horn and rhinoceros horn belong to the category of heat-clearing and blood-cooling medicinal herbs, used for symptoms such as fever, dizziness, rashes, and excessive bleeding. They share similar medicinal properties, structures, and components with rhinoceros horn, containing proteins, peptides, various amino acids, cholesterol, muscle fibers, and trace elements, thus exhibiting similar pharmacological effects, including anti-inflammatory, analgesic, and antipyretic properties. Since buffalo horn was included in the Chinese Pharmacopoeia, it has gradually become a substitute for rhinoceros horn. Because buffalo horn is less potent, a larger quantity and longer treatment time are required compared to rhinoceros horn. Processing can enhance the medicinal effects of buffalo horn.

[0004] Using buffalo horn as a substitute for rhinoceros horn in traditional Chinese medicine has protected the survival and population of endangered animals. However, its weaker efficacy necessitates higher dosages, increasing demand for buffalo horn and leading to a gradual decline in its numbers. The pharmacopoeia's methods for processing buffalo horn are relatively simple, involving only the use of larger quantities of slices or powder. While this increases its efficacy, the dosage is still far greater than that of rhinoceros horn. Therefore, a processing method that promotes the dissolution of effective substances from buffalo horn is urgently needed to better protect this medicinal resource. Summary of the Invention

[0005] To solve the above-mentioned technical problems, the present invention provides a processing method for promoting the dissolution of effective substances from buffalo horn, comprising the following steps:

[0006] S1. Crush the raw buffalo horn into particles with a diameter of 3-5 mm to obtain buffalo horn granules;

[0007] S2. Add the ox horn particles to distilled water and hydrolyze for 30-60 minutes to obtain ox horn hydrolysate; the hydrolysis temperature is 130-230℃ and the pressure is 2-20MPa.

[0008] S3. Filter the hydrolysate and take the filtrate directly, or evaporate the filtrate to dryness to obtain the horn-shaped firecracker product.

[0009] Furthermore, in step S2, the specific method for hydrolyzing the horn particles is as follows:

[0010] Add the ox horn particles to a microwave hydrolysis vessel and maintain the pressure at 2-10 MPa and temperature at 130-140℃ for 10-20 minutes. Then, increase the pressure to 10-20 MPa and temperature to 140-230℃ and maintain the temperature for 20-40 minutes.

[0011] Furthermore, the microwave frequency of the microwave hydrolysis vessel is periodically switched within the range of 1950-2950MHz, with a single frequency dwell time of 30-60 seconds; the microwave power of the microwave hydrolysis vessel is output intermittently with an on / off ratio of 1:1-1:3, and a single cycle duration of 2-5 minutes.

[0012] Furthermore, the microwave frequency of the microwave hydrolysis vessel alternates between a low frequency of 1950-2150MHz and a high frequency of 2450-2950MHz, with an alternation period of 1-2 minutes; the low frequency of 1950-2150MHz is used for deep penetration, and the high frequency of 2450-2950MHz is used for targeted bond breaking.

[0013] Furthermore, in step S2, the mass ratio of the horn particles to distilled water is 1:10 to 1:20.

[0014] On the other hand, the present invention provides a buffalo horn processing product prepared by a processing method that promotes the dissolution of effective substances in buffalo horn.

[0015] On the other hand, the present invention provides an application of buffalo horn processed products for the preparation of anti-inflammatory, hemostatic, and analgesic drugs.

[0016] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0017] 1. Traditional decoction methods are ineffective at extracting the active ingredients from hard medicinal materials like buffalo horn in a short time. Experiments show that the water-soluble extract content of raw buffalo horn is typically 0.8%, while that of cooked buffalo horn is 1.8%. Even the extract content of concentrated buffalo horn powder in the pharmacopoeia is only 5%. This invention uses microwave hydrolysis to hydrolyze the insoluble substances in buffalo horn in a short time. The average extract content of the processed buffalo horn product reaches 18.57%, which is nearly six times higher than that obtained by long-term decoction. This significantly increases the extract content of buffalo horn. Therefore, microwave hydrolysis of buffalo horn is more complete than decoction and can significantly increase the water-soluble extract content of buffalo horn.

[0018] 2. Compared with existing decoctions, this invention allows for the intake of more effective substances with a smaller oral dose than traditional methods, thereby reducing the oral dosage required to achieve the same therapeutic effect. Detailed Implementation

[0019] Example 1

[0020] A method for processing buffalo horn to promote the dissolution of its effective substances includes the following steps:

[0021] S1. Crush the raw buffalo horn to a particle size of 3mm to obtain buffalo horn particles;

[0022] S2. Add the ox horn particles to distilled water and hydrolyze for 30 minutes to obtain ox horn hydrolysate;

[0023] S3. Filter the hydrolysate, take the filtrate, and obtain the ox horn cannon product.

[0024] Furthermore, the specific method for hydrolyzing the horn particles in step S2 is as follows:

[0025] Add the ox horn particles to a microwave hydrolysis vessel and maintain the pressure at 2 MPa and temperature at 130°C for 10 minutes, then increase the pressure to 10 MPa and temperature at 140°C and maintain the pressure at 140°C for 20 minutes.

[0026] Furthermore, the microwave frequency of the microwave hydrolysis vessel alternates between a low frequency of 1950MHz and a high frequency of 2450MHz, with an alternation period of 1 minute; the low frequency of 1950MHz is used for deep penetration, and the high frequency of 2450MHz is used for targeted bond breaking; the residence time of a single frequency is 30 seconds; the microwave power of the microwave hydrolysis vessel is output intermittently with an on / off ratio of 1:1, and the duration of a single cycle is 2 minutes.

[0027] Moreover, in step S2, the mass ratio of the horn particles to distilled water is 1:10.

[0028] Example 2

[0029] A method for processing buffalo horn to promote the dissolution of its effective substances includes the following steps:

[0030] S1. Crush the raw buffalo horn to a particle size of 5mm to obtain buffalo horn particles;

[0031] S2. Add the ox horn particles to distilled water and hydrolyze for 60 minutes to obtain ox horn hydrolysate;

[0032] S3. Filter the hydrolysate and evaporate the filtrate to dryness to obtain the ox horn cannon product.

[0033] Furthermore, the specific method for hydrolyzing the horn particles in step S2 is as follows:

[0034] The ox horn particles were added to a microwave hydrolysis vessel and maintained at a pressure of 10 MPa and a temperature of 140°C for 20 minutes. Then the pressure was increased to 20 MPa and the temperature was increased to 230°C and maintained for 40 minutes.

[0035] Furthermore, the microwave frequency of the microwave hydrolysis vessel alternates between a low frequency of 2150MHz and a high frequency of 2950MHz, with an alternation period of 2 minutes; the low frequency of 2150MHz is used for deep penetration, and the high frequency of 2950MHz is used for targeted bond breaking; the residence time of a single frequency is 60 seconds; the microwave power of the microwave hydrolysis vessel is output intermittently with an on / off ratio of 1:3, and the duration of a single cycle is 5 minutes.

[0036] Furthermore, in step S2, the mass ratio of the horn particles to distilled water is 1:20.

[0037] Example 3

[0038] A method for processing buffalo horn to promote the dissolution of its effective substances includes the following steps:

[0039] S1. Crush the raw buffalo horn to a particle size of 4mm to obtain buffalo horn particles;

[0040] S2. Add the ox horn particles to distilled water and hydrolyze for 45 minutes to obtain ox horn hydrolysate;

[0041] S3. Filter the hydrolysate and take the filtrate directly, or evaporate the filtrate to dryness to obtain the horn-shaped firecracker product.

[0042] Furthermore, the specific method for hydrolyzing the horn particles in step S2 is as follows:

[0043] Add the ox horn particles to a microwave hydrolysis vessel and maintain the pressure at 5 MPa and temperature at 135°C for 15 minutes, then increase the pressure to 15 MPa and temperature at 200°C and maintain the temperature for 30 minutes.

[0044] Furthermore, the microwave frequency of the microwave hydrolysis vessel alternates between a low frequency of 2000MHz and a high frequency of 2700MHz, with an alternation period of 1.5 minutes; the low frequency of 2000MHz is used for deep penetration, and the high frequency of 2700MHz is used for targeted bond breaking, with a single frequency residence time of 45 seconds; the microwave power of the microwave hydrolysis vessel is output intermittently with an on / off ratio of 1:2, with a single cycle duration of 3.5 minutes.

[0045] Furthermore, in step S2, the mass ratio of the horn particles to distilled water is 1:15.

[0046] Experimental Section

[0047] 1. Experimental materials

[0048] 1.1 Medications: Buffalo horn, distilled water, aspirin, xylene (Hunan Huihong Reagent Company)

[0049] 1.2 Animals: Kunming mice, weighing 20±2g

[0050] 1.3 Instruments: DAF-6210 vacuum drying oven, reflux condenser, KDM type electric heating mantle, desiccator, HH-4 digital display constant temperature water bath, DT-500B type electronic balance, CPA225DA type 0.001% balance, microwave hydrolyzer (model EXPEC 790S, produced by Puyu Technology Co., Ltd.), 8mm ear punch (Beijing Jiandeer Technology Co., Ltd., batch number 20120614), surgical scissors, pipette, intelligent hot plate apparatus (Chengdu Taimeng Technology Co., Ltd., Sichuan Province), 5L pilot-scale automated reactor (Beijing Century Senlang Experimental Instrument Co., Ltd.).

[0051] Experimental data were statistically analyzed using SPSS software, and measurement data were analyzed using (…). Statistical description was performed, and the data between groups were analyzed using the LSD method and t-test. The test criterion was P<0.1.

[0052] 2. Determination of buffalo horn extract

[0053] Take 3.25g of raw ox horn and process it according to the method in Example 1 to obtain processed ox horn product. Then take 3.25g of raw / cooked ox horn to prepare raw ox horn decoction (raw ox horn decocted in water over medium heat for 60 minutes) / cooked ox horn decoction (cooked ox horn decocted in water over medium heat for 60 minutes). The content of ox horn extract was tested respectively, and the results are shown in Table 1.

[0054] Table 1. Content of water-soluble extracts from microwave hydrolysis of buffalo horn

[0055]

[0056] The data above shows that the water-soluble extract content of raw buffalo horn is usually 0.8%, while that of cooked buffalo horn is 1.8%. The extract content of concentrated buffalo horn powder in the pharmacopoeia is only 5%. However, the buffalo horn extract content of the processed buffalo horn product obtained in Example 1 of this invention reached an average of 18.57%, which is nearly six times higher than that of extracts obtained by long-term decoction. This greatly increases the extract content of buffalo horn, while significantly shortening the extraction time and reducing labor intensity.

[0057] 3. Mouse experiment

[0058] 3.1 Mouse ear swelling test

[0059] Eighty mice, half male and half female, were divided into eight groups of ten mice each.

[0060] (1) Control group: distilled water, once a day for 7 consecutive days;

[0061] (2) Aspirin group: Administered via ig, aspirin solution, 0.2 g / kg, once a day, for 7 consecutive days;

[0062] (3) Raw ox horn decoction group: Administered by ig, raw ox horn was decocted in water over medium heat for 60 minutes, 13mg / 20g, once a day, for 7 consecutive days;

[0063] (4) Cooked ox horn decoction group: Administered by ig, cooked ox horn was decocted in water over medium heat for 60 minutes, 13mg / 20g, once a day, for 7 consecutive days;

[0064] (5) Low-dose group of ox horn hydrolysate: Administered by gavage, 3.25 mg / 20g of the ox horn product provided in Example 1, once a day for 7 consecutive days;

[0065] (6) Medium dose group of ox horn hydrolysate: ig administration, ox horn processed product provided in Example 1, 6.5 mg / 20g, once a day, for 7 consecutive days;

[0066] (7) High-dose group of ox horn hydrolysate: Administered by gavage, 13 mg / 20 g of the ox horn product provided in Example 1, once a day for 7 consecutive days;

[0067] (8) Reactor processing group: Crush the raw buffalo horn to a particle size of 3mm, add distilled water (1:15), put it into a pilot-scale 5L automated reactor, maintain it at a pressure of 5MPa and a temperature of 135℃ for 15 minutes, then raise the pressure to 15MPa and a temperature of 200℃ and maintain it for 30 minutes, filter, and take the filtrate.

[0068] The aspirin group was given 0.2 g / kg of aspirin solution, the raw ox horn decoction group and the cooked ox horn decoction group were given 13 mg / 20 g, the buffalo horn hydrolysate (the buffalo horn processed product provided in Example 1) low, medium and high dose groups were given 3.25, 6.5 and 13 mg / 20 g of buffalo horn hydrolysate, respectively, and the control group was given distilled water. The mice in the other groups were given the above drugs by gavage once a day for 7 consecutive days. One hour after the last administration, except for the control group, 25 μL of xylene was applied to the front and back of the right ear of each group of mice to induce inflammation. The mice were sacrificed 1 hour later, and ear pieces were punched from the same location in the left and right ears. The ear pieces were accurately weighed, and the degree of swelling and the swelling inhibition rate were calculated. The results are shown in Table 2.

[0069]

[0070]

[0071] Table 2 Comparison of the degree of ear swelling in mice induced by xylene in each group

[0072]

[0073] Compared with the control group, P<0.05.

[0074] As shown in Table 2, the ear swelling was significantly reduced in the positive control group compared to the control group (P<0.05). Although there was no significant difference between the drug administration groups and the control group, they all showed an anti-inflammatory trend. The ear swelling of mice treated with horn hydrolysate was similar and at the same level, with the low-dose horn hydrolysate group showing the best effect. The inhibition rate results also indicated that the low-dose horn hydrolysate group was the most effective, proving that the horn processed products treated with the technical solution of this invention can achieve good results with low doses. Although the reaction vessel treatment group reached the level of high pressure and high temperature, it lacked microwave treatment and was less effective than the horn hydrolysate group.

[0075] 3.2 Mouse tail docking experiment

[0076] Dissolve Yunnan Baiyao capsules in distilled water to prepare a 0.02g / ml Yunnan Baiyao suspension.

[0077] Seventy mice, half male and half female, were used. The Yunnan Baiyao group was treated with 0.2 g / kg, while the other groups were treated in the same way as the mouse ear swelling experiment. The mice were administered the medication by gavage for 5 consecutive days. One hour after the last administration, the tail was cut 3 mm from the tip using surgical scissors. Timing began when blood began to flow out spontaneously. Every 30 seconds, a drop of blood was absorbed with filter paper until no more blood was absorbed, indicating natural cessation of bleeding. The bleeding time was calculated, and the results are shown in Table 3.

[0078] Table 3. Effect of tail-barre hemorrhage time in mice (s, (n=10)

[0079]

[0080] Compared with the control group, P<0.1.

[0081] Compared with the control group, the low-dose and medium-dose groups of ox horn hydrolysate significantly shortened bleeding time, and the hemostatic effect was significantly different (P<0.1), while the effects of the other groups were not obvious. Meanwhile, the low-dose group of ox horn hydrolysate and the decoction of cooked ox horn showed similar bleeding time in mice after tail amputation, indicating that the new processing method significantly enhanced efficacy, with the low-dose group of ox horn hydrolysate showing the best effect. Although the reaction vessel treatment group achieved high pressure and high temperature, it lacked microwave treatment and was less effective than the ox horn hydrolysate group.

[0082] 3.3 Mouse Hot Plate Test

[0083] Seventy female mice were selected and divided into groups, treated in the same way as the mouse ear swelling test. The female mice were placed on a (55±0.5)℃ intelligent hot plate apparatus. The pain response was measured by licking the hind paw and turning the head back. The pain threshold was recorded. Mice with an average pain threshold of 5-30 seconds between two tests without jumping were used for the experiment. To avoid damaging the skin of the mouse paws, they were removed after a maximum of 60 seconds. The average value was used as the pre-drug pain threshold. The mice were administered intragastric gavage (ig) for 5 consecutive days. After the last administration, the pain threshold of each mouse was measured at 30, 60, and 90 minutes using the same method. The rate of increase in pain threshold for each group was calculated.

[0084]

[0085] Table 4. Effects on heat-induced pain response in mice (s, (n=10)

[0086]

[0087] Although there were no significant differences among the treatment groups compared to the control group, it was noticeable that the low-dose group of bovine horn hydrolysate showed a prolonged licking latency at 30, 60, and 90 minutes after administration, while the medium-dose group showed a prolonged licking latency at 60 and 90 minutes. These effects were more pronounced than those of the raw and processed bovine horn decoction groups. The pain threshold in the reaction vessel treatment group was slightly higher than that in the control group, but significantly lower than that in the microwave hydrolysate low-dose group, indicating that microwave hydrolysis is more advantageous in releasing active ingredients. These results suggest that processing may enhance the efficacy of bovine horn, with the low-dose group of bovine horn hydrolysate showing the best effect.

Claims

1. A method for processing buffalo horn to promote the dissolution of effective substances, characterized in that, Includes the following steps: S1. Crush the raw buffalo horn into particles with a diameter of 3-5 mm to obtain buffalo horn granules; S2. Add the ox horn particles to distilled water and hydrolyze for 30-60 minutes to obtain ox horn hydrolysate; the hydrolysis temperature is 130-230℃ and the pressure is 2-20MPa. S3. Filter the hydrolysate and take the filtrate directly, or evaporate the filtrate to dryness to obtain the horn-shaped firecracker product. The specific method of hydrolysis is as follows: add the ox horn particles into a microwave hydrolysis tank, maintain the pressure of 2-10MPa and the temperature of 130-140℃ for 10-20 minutes, and then raise the pressure to 10-20MPa and the temperature of 140-230℃ and maintain it for 20-40 minutes. The microwave frequency of the microwave hydrolysis vessel switches periodically within the range of 1950-2950MHz, with a single frequency dwell time of 30-60 seconds; the microwave power of the microwave hydrolysis vessel is output intermittently with an on / off ratio of 1:1-1:3, and the duration of a single cycle is 2-5 minutes. The microwave hydrolysis vessel alternates between a low frequency of 1950-2150MHz and a high frequency of 2450-2950MHz, with an alternation period of 1-2 minutes. The low frequency of 1950-2150MHz is used for deep penetration, and the high frequency of 2450-2950MHz is used for targeted bond breaking.

2. The method for processing buffalo horn to promote the dissolution of effective substances according to claim 1, characterized in that, In step S2, the mass ratio of the ox horn particles to distilled water is 1:10 to 1:

20.

3. The buffalo horn processed product prepared by the processing method for promoting the dissolution of effective substances in buffalo horn as described in claim 2.

4. The application of the buffalo horn cannon product as described in claim 3, characterized in that, Used in the preparation of anti-inflammatory drugs.

5. The application of the buffalo horn cannon product as described in claim 3, characterized in that, Used in the preparation of hemostatic drugs.

6. The application of the buffalo horn cannon product as described in claim 3, characterized in that... Used in the preparation of analgesic drugs.

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