Preparation method of traditional Chinese medicine compound injury treating ointment and product of traditional Chinese medicine compound injury treating ointment

Abstract

The invention provides a preparation method of a traditional Chinese medicine compound injury treating ointment. The preparation method comprises the following steps: firstly, dividing raw materials into animal raw materials, volatile oil raw materials and toxic raw materials; respectively carrying out ultrasonic extraction on the three raw material medicines by taking camellia oil as a solvent, mixing, and carrying out ultrasonic extraction to obtain an oily mixture containing protein, volatile oil and curcumin; adding beeswax to prepare an oil-wax mixture; finally, vanillyl butyl ether and azone are added, and the traditional Chinese medicine compound injury treating ointment is prepared. According to the invention, by optimizing the ultrasonic extraction conditions of each group of raw material medicines, fat-soluble active ingredients such as protein, volatile oil and curcumin, especially small-molecular fat-soluble protein, are extracted as much as possible, and the penetration enhancing effects of camellia oil, volatile oil, vanillyl butyl ether and azone are matched, so that the transdermal absorption efficiency and clinical curative effect of the injury treating ointment are improved; the prepared injury treating ointment has good effects in the aspects of quick itching relieving after treatment of soft tissue injury, burns and scalds and mosquito bites and allergy resistance, and is suitable for people with different skin types.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine external plasters, in particular to a preparation method of a traditional Chinese medicine compound wound healing plaster and a product thereof. Background Art

[0002] Sports injuries (such as muscle and ligament sprains, soft tissue injuries, joint injuries, fractures, dislocations, etc.), skin itching caused by various allergies and insect bites, wound ulceration and scarring caused by burns, etc., have seriously affected people's physical health and daily life.

[0003] Conventional Western medicine topical treatments are limited. Commonly used topical ointments for treating soft tissue injuries are mostly non-steroidal analgesics, which have little effect; topical ointments for treating skin itching caused by allergies and burns often contain hormones, which can easily cause damage to liver and kidney function and have many side effects of skin irritation.

[0004] Traditional Chinese medicine for treating soft tissue injuries, skin itching, and burns: beeswax ointment or black plaster, often has toxic and irritating reactions due to problems with the manufacturing process, which affects the drug loading capacity, limits the usage time, and also affects the efficacy.

[0005] The traditional ointment preparation method has the following defects: (1) The traditional process of preparing ointments by soaking, decocting, filtering, and exploding (collecting ointments) often uses water as a solvent. Although water-soluble active ingredients (such as alkaloids, saponins, polysaccharides, etc.) are extracted efficiently, a large amount of volatile oil contained in the drug is lost with the evaporation of water vapor during the preparation process, and a large amount of protein is precipitated due to high-temperature denaturation and coagulation during the preparation process; in addition, for fat-soluble active ingredients, the "water extraction method" is not applicable.

[0006] (2) In the preparation of topical ointments, red lead is often added to increase the adhesion of the ointment, thereby increasing the permeability of the ointment; however, due to individual differences, users often experience skin allergies and lead accumulation poisoning, which has become one of the potential risk factors for the safety of topical ointments.

[0007] (3) For animal drugs, volatile drugs, and mineral drugs, the "fine drug" addition method is usually adopted, that is, the powder made by crushing the medicinal materials (i.e., "fine drug") is added to the melted ointment and stirred continuously. Although the protein, volatile oil, metal elements, etc. in the medicinal materials can be retained as much as possible, the addition of crude drug powder interferes with the effective preservation of the external ointment and affects the skin's intuitive feeling and drug penetration and absorption during the use of the external ointment. Summary of the invention

[0008] The first object of the present invention is to provide a method for preparing a Chinese medicine compound wound healing ointment. The prepared ointment not only has little irritation to the skin, but also has good penetration and absorption effect of the active ingredients of the medicine. It has good efficacy in treating soft tissue injuries, burns and scalds, quickly relieving itching after mosquito bites and anti-allergy, and has basically no toxic side effects.

[0009] A method for preparing a Chinese medicine compound wound healing ointment comprises the following steps: (1) Divide APIs into the following four groups: Group 1: Animals - 3-5 portions of scorpions, 3-6 portions of centipedes, 3-5 portions of earth dragons; Group 2: Volatile oils - 3-5 parts of Saposhnikovia divaricata, 3-5 parts of Angelica dahurica, 3-5 parts of Angelica sinensis, 3-5 parts of Asarum, 3-5 parts of Chuanxiong, 3-5 parts of Salvia miltiorrhiza, 11-14 parts of Trigonosciadium japonicum, 10-13 parts of Curcuma zedoaria, 5-7 parts of Frankincense, 5-7 parts of Myrrh, 5-7 parts of Papaya, 3-5 parts of Rhubarb, 3-5 parts of Herba Polygoni Multiflori; Group 3: Toxic—5-7 parts of raw aconite, 3-5 parts of Arisaema consanguineum, 3-5 parts of Chuanwu, 3-5 parts of Caowu; (2) The raw material drug of group 1 is crushed, sieved, and subjected to ultrasonic extraction using camellia oil as a solvent. The ultrasonic extraction temperature is 60-80° C., the ultrasonic power is 1440-1450 W, and the extraction time is 60-70 minutes to obtain an oily substance containing protein; (3) The raw materials of group 2 are crushed, sieved, and subjected to ultrasonic extraction using camellia oil as a solvent. The ultrasonic extraction temperature is 40 to 60° C., the ultrasonic power is 350 to 360 W, and the extraction time is 40 to 50 minutes to obtain an oily substance containing volatile oil; (4) The raw materials of group 3 are crushed, sieved, and ultrasonically extracted with camellia oil as a solvent. The ultrasonic extraction temperature is 90-100° C., the ultrasonic power is 350-360 W, and the extraction time is 120-130 minutes to obtain an oily substance containing curcumin; (5) mixing the oily substance containing protein obtained in step (2), the oily substance containing volatile oil obtained in step (3), and the oily substance containing curcumin obtained in step (4), and performing ultrasonic extraction again after mixing. The ultrasonic extraction temperature in this step is 60 to 80° C., the ultrasonic power is 350 to 360 W, and the extraction time is 20 to 30 minutes to obtain an oily mixture; (6) Adding beeswax to the oily mixture obtained in step (5), melting the beeswax at 60 to 80° C. and mixing the mixture with the oily mixture to prepare an oil-wax mixture for later use; (7) Adding vanillin butyl ether and azone (N-laurylazacycloheptane-2-one) to the oil-wax mixture obtained in step (6), wherein the amount of vanillin butyl ether added is controlled to be 0.03-0.05%, and the amount of azone added is controlled to be 0.5-0.8%, stirring and mixing, packaging, cooling, and obtaining the traditional Chinese medicine compound wound healing ointment.

[0010] Preferably, the ultrasonic extraction in steps (2) to (5) is performed using a dual-frequency ultrasonic extractor.

[0011] The second object of the present invention is to provide a Chinese medicine compound injury-removing ointment prepared according to the preparation method of the Chinese medicine compound injury-removing ointment described in the first object of the present invention.

[0012] The preparation method of the Chinese medicine compound wound-healing ointment of the present invention and the product thereof have the following advantages: (1) The protein raw materials of Group 1 of the present invention use camellia oil as the extraction solvent. It is found that the obtained oily substance of Group 1 contains a large amount of protein (fat-soluble protein), and there are large differences in the molecular weight of the protein obtained under different ultrasonic extraction conditions. Therefore, in order to improve the absorption rate of protein, the inventors have optimized the ultrasonic extraction conditions to obtain more small molecular fat-soluble proteins with a molecular weight of less than 20KDa. Small molecular fat-soluble proteins, combined with the penetration-promoting effects of camellia oil, volatile oil, vanillyl alcohol butyl ether and azone (N-lauryl azacycloheptane-2-one), can be better absorbed by the skin, thereby enhancing the transdermal absorption efficiency and clinical efficacy of the wound-removing ointment; and camellia oil can also play an antibacterial, anti-corruption, anti-inflammatory and detumescent role, and reduce irritation and allergy. Therefore, the traditional Chinese medicine compound wound-removing ointment prepared by the present invention has good efficacy in treating soft tissue injuries, burns and scalds, rapid itching relief after mosquito bites, and anti-allergy, and is suitable for people with different skin types; (2) The present invention divides the raw materials into group 1 animal, group 2 volatile oil and group 3 toxicity according to the physicochemical properties of Chinese medicinal materials and their effective parts, combined with the "drug-excipient integration theory" of Chinese medicine preparations. Ultrasonic extraction is used, and different extraction parameters are used according to the different physicochemical properties of the extracted effective parts to extract as much fat-soluble effective components contained in the drug combination as possible, such as: the sum of animal proteins, the sum of plant volatile oils, non-volatile effective components (curcumin compounds), etc.; (3) Since the effective parts of the volatile oil raw materials in Group 2 are mostly volatile oils. Chinese medicine volatile oils not only have pharmacological effects such as promoting blood circulation and removing blood stasis, reducing swelling and relieving pain, and anti-inflammatory, but can also be used as penetration enhancers for external ointments to promote the transdermal absorption of proteins and active ingredients of drugs. The present invention uses camellia oil as an extraction solvent to maximize the extraction rate of fat-soluble active ingredients such as volatile oils and curcumin, while combining appropriate ultrasonic extraction conditions to minimize the volatilization of volatile oils; (4) Since the toxic components of group III toxic Chinese medicines are mostly water-soluble alkaloids, using plant oil (camellia oil) as an extraction solvent can significantly reduce the content of water-soluble alkaloids in topical ointments, thereby reducing the irritation of such components to the patient's epidermis and subcutaneous tissue; it is also beneficial for the pH value of the topical ointment to be close to the pH value of human skin; (5) In addition, the present invention also uses beeswax as a preparation matrix to adjust the consistency of the ointment and enhance the stability of the preparation, thereby avoiding the toxic and side effects of red lead on patients. At the same time, in traditional Chinese medicine, beeswax also has the effects of detoxification, sore astringence, tissue regeneration, and analgesia. It can be used externally to treat ulcers that do not heal, ecthyma erosion, trauma ulceration, and burns, and can enhance the relevant clinical efficacy of the wound-relief ointment; (6) The present invention does not need to add "fine powder of crude drug made by crushing medicinal materials", which avoids the interference of moisture, low-level fatty acids, undegraded lanolin and microorganisms in the fine powder of crude drug on the deterioration and sourness of the external ointment. At the same time, the present invention uses a plant oil solvent (camellia oil) as an extraction solvent and beeswax as a preparation matrix, so that the water content of the wound ointment is extremely low, ensuring the pressure difference between the inside and outside of the bacterial or viral cell membrane in the microenvironment of the external ointment, so that bacteria or viruses cannot survive and reproduce in the external ointment, thereby improving the antiseptic performance of the external ointment. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 It is a technical roadmap of the preparation method of the Chinese medicine compound wound healing ointment of the present invention; Figure 2 It is the SDS-PAGE band diagram of the oily substance containing protein of the present invention obtained at different temperatures. DETAILED DESCRIPTION

[0014] The preparation method of the Chinese medicine compound wound-relief ointment of the present invention and the implementation methods of the product are now specifically described: like Figure 1 As shown, a method for preparing a Chinese medicine compound wound healing ointment comprises the following steps: (1) Divide APIs into the following four groups: Group 1: Animals—3-5 parts of scorpions, 3-6 parts of centipedes, and 3-5 parts of earthworms (i.e., “raw drug 1”); Group 2: Volatile oils - 3-5 parts of Saposhnikovia divaricata, 3-5 parts of Angelica dahurica, 3-5 parts of Angelica sinensis, 3-5 parts of Asarum, 3-5 parts of Ligusticum chuanxiong, 3-5 parts of Salvia miltiorrhiza, 11-14 parts of Trigonosciadium japonicum, 10-13 parts of Curcuma zedoaria, 5-7 parts of Frankincense, 5-7 parts of Myrrh, 5-7 parts of Papaya, 3-5 parts of Rhubarb, 3-5 parts of Herba speranskiae Root (i.e. "raw drug 2"); Group 3: Toxic—5-7 parts of raw aconite, 3-5 parts of Arisaema consanguineum, 3-5 parts of Chuanwu, and 3-5 parts of Caowu (i.e., “raw drug 3”); (2) The raw material drug of group 1 was crushed, sieved, and subjected to ultrasonic extraction using camellia oil as a solvent. The ultrasonic extraction temperature was 60-80° C., the ultrasonic power was 1440-1450 W, and the extraction time was 60-70 minutes to obtain an oily substance containing protein (i.e., “oily substance 1”); (3) The raw materials of group 2 were crushed, sieved, and subjected to ultrasonic extraction using camellia oil as a solvent. The ultrasonic extraction temperature was 40-60° C., the ultrasonic power was 350-360 W, and the extraction time was 40-50 minutes to obtain an oily substance containing volatile oil (i.e., “oily substance 2”); (4) The raw material drug of group 4 was crushed, sieved, and subjected to ultrasonic extraction using camellia oil as a solvent. The ultrasonic extraction temperature was 90° C., the ultrasonic power was 350-360 W, and the extraction time was 120-130 minutes to obtain an oily substance containing curcumin (i.e., “Oily substance 3”); (5) mixing the oily substance containing protein obtained in step (2), the oily substance containing volatile oil obtained in step (3), and the oily substance containing curcumin obtained in step (4), and performing ultrasonic extraction again after mixing. The ultrasonic extraction temperature in this step is 60 to 80° C., the ultrasonic power is 350 to 360 W, and the extraction time is 20 to 360 minutes to obtain an oily mixture; (6) Adding beeswax to the oily mixture obtained in step (5), melting the beeswax at 60 to 80° C. and mixing the mixture with the oily mixture to prepare an oil-wax mixture for later use; (7) Adding vanillin butyl ether and azone (N-laurylazacycloheptane-2-one) to the oil-wax mixture obtained in step (6), wherein the amount of vanillin butyl ether added is controlled to be 0.03-0.05%, and the amount of azone added is controlled to be 0.5-0.8%, stirring and mixing, packaging, and cooling to obtain the traditional Chinese medicine compound wound healing ointment (also referred to as "compound wound healing ointment for external use").

[0015] The ultrasonic extraction in steps (2) to (5) is performed using a dual-frequency ultrasonic extractor.

[0016] The inventors conducted multiple investigations during the experimental stage, and some of the experiments are summarized as follows: 1. Effect of different extraction temperatures on the protein extraction rate and protein molecular weight of Group 1 API This experiment conducted a single factor experimental investigation on the ultrasonic extraction temperature of the raw material medicine of group 1 in the preparation method of the Chinese medicine compound wound-relief ointment of the present invention. According to the weight proportion of the raw material medicine of group 1 of the present invention and the specific operation steps of the above step (2), protein extraction was carried out, and the extraction temperatures were 60 ℃, 70 ℃, 80 ℃, 90 ℃, and 100 ℃, respectively, the extraction power was 1440-1450W, the extraction time was 60-70 min, and after the extraction was completed, the excess camellia oil was removed by a rotary evaporator to obtain the oil samples of group 1 corresponding to different ultrasonic extraction temperatures.

[0017] Protein precipitation was performed for each of the oil samples in the group 1, and the specific steps were as follows: each extract was mixed with 4 volumes of pre-cooled acetone, and after standing at -20°C for 1 hour, centrifuged, and the conditions were set as follows: 4°C, 12000 rpm, centrifuged for 10 minutes, and the protein precipitate was collected. The precipitate was then washed twice with pre-cooled acetone, and the residual oil and acetone were removed by centrifugation. The precipitate was dissolved in PBS buffer to obtain a protein sample solution.

[0018] The BCA method was then used for protein quantification. The specific steps were as follows: prepare the BCA working solution according to the instructions of the BCA kit; add the sample and standard to the 96-well plate, add the BCA working solution, mix well, and incubate at 37°C for 30 minutes; after the incubation, use an enzyme reader to measure the absorbance at 562 nm. The experimental results are shown in Table 1. The experimental results show that as the extraction temperature increases (60-90°C), the protein extraction rate gradually increases, but there is a slight decrease at 100°C. The reason may be that the high temperature causes some protein denaturation and degradation.

[0019] Table 1 Sample protein extraction rate (n=3)

[0020] SDS-PAGE analysis was performed on each protein extract, and the gel imaging system was used to capture gel images and analyze the distribution and intensity of the bands. The results are shown in Figure 2 .Depend on Figure 2The results show that the main bands of the samples extracted at 60℃ are concentrated in the range of 10-50 kDa; the bands of the samples extracted at 70℃ are mainly concentrated in the range of 10-50 kDa, and the bands are clearer; the bands of the samples extracted at 80℃ are the most abundant, and the bands are wide, concentrated in the range of 10-50 kDa; the bands of the samples extracted at 90℃ are mainly distributed in the range of 30-50kDa, and the bands are the clearest, but the bands in the range of 20kDa are obviously narrowed; the bands of the samples extracted at 100℃, overall, the remaining bands are not significantly different from those at 90℃, but the bands in the range of 20kDa are almost invisible. It can be seen that although the protein extraction rate at 60~80℃ did not reach the maximum value, it contained more small molecular proteins (≤20kDa) that are more conducive to skin absorption and effective therapeutic effects, while the extraction rate of the samples extracted at 90℃ reached the highest, but the small molecular proteins of ≤20kDa were significantly reduced, and the small molecular proteins of ≤20kDa in the samples extracted at 100℃ were further reduced. Therefore, according to Table 2 and Figure 2 According to the experimental results, in the subsequent preparation process, the ultrasonic extraction conditions of the group 1 raw materials were set to 1440-1450 W / 60-80 ℃ / 60-70 min.

[0021] (II) Effect of different extraction temperatures on the extraction rate of volatile oil from group 2 API This experiment conducted a single factor experimental investigation on the ultrasonic extraction temperature of the second raw material medicine in the preparation method of the traditional Chinese medicine compound wound-relief ointment of the present invention. According to the weight proportion of the raw material medicine of the second group of the present invention and the specific operation steps of the above step (3), volatile oil extraction was carried out, and the ultrasonic extraction temperature was 30 ℃, 40 ℃, 50 ℃, 60 ℃, 70 ℃, 80 ℃, respectively, the extraction power was 350-360 W, and the extraction time was 40-50 min, and the second oil sample corresponding to different ultrasonic extraction temperature conditions was obtained for standby use.

[0022] Each of the two oil samples in the above group was extracted three times with 50 mL of petroleum ether, the petroleum ether extraction layers were combined, placed in a rotary evaporator to evaporate the petroleum ether, and then accurately weighed to obtain the volatile oil content of each sample. The results are shown in Table 2.

[0023] Table 2 Extraction rate of volatile oil from samples ( n =3)

[0024] From the experimental results in Table 2, it can be seen that when the extraction temperature is from 30 to 50 °C, the extraction rate of volatile oil gradually increases and reaches the maximum value at 50 °C. Thereafter, as the extraction temperature further increases (60 to 80 °C), the extraction rate of volatile oil gradually decreases. In addition, the experimental data also show that when the extraction temperature is 40 °C, 50 °C, and 60 °C, there are slight differences in the average value of the volatile oil extraction rate, and the volatile oil extraction rates overlap in each actual extraction experiment, indicating that within this extraction temperature range (40 to 60 °C), the volatile oil extraction rate is not much different, so in the subsequent preparation process, the ultrasonic extraction conditions are set to 350 to 360W / 40 to 60 °C / 40 to 50min.

[0025] 3. Effect of different extraction temperatures on the clinical efficacy of preparations According to the patented technical solution of the present invention, the ultrasonic extraction temperature of step (2) is set at 60, 70, 80, 90, and 100° C., respectively, and the rest of the preparation process remains unchanged to prepare the corresponding Chinese medicine compound wound healing ointment product.

[0026] Subjects: SD rats, male, about 300 g, raised in a clean environment.

[0027] To construct an acute soft tissue injury model, the specific steps are as follows: Anesthesia: Rats were weighed and numbered and anesthetized with isoflurane breathing; Hair removal: After anesthesia takes effect, shave both lower limbs to the thigh root, use depilatory cream to remove hair from the calf (to the knee joint), and wipe with 75% ethanol to remove the residual depilatory cream; Establish the injury model: trim the hair at the gastrocnemius muscle, and use a cushion (5cm × 5cm) to pad the right lower limb to prevent it from hanging in the air. Slightly externally rotate the right lower limb to place the posterior calf muscles on the inner side of the tibia and fibula, and place the fixator (a 50mL centrifuge tube with the bottom of the conical tube facing down, and wrap the tip of the tube with a Band-Aid to prevent skin damage caused by friction) above the gastrocnemius muscle of the calf, with the tip in a fixed position. A 500g weight is freely dropped along the pipe from a height of 45cm, acting on the top of the fixator, causing a muscle contusion, which is repeated 10 times, causing blunt soft tissue injuries to the right hind limb of the rat; the left hind limb is not subjected to any injury treatment as a control; Treatment method: Weigh 2g of the wound healing ointment of the present invention, apply it evenly to the injured gastrocnemius muscle of the mouse, and then wrap it with gauze; then fix it on the outer layer with adhesive tape; change the dressing once a day (24 hours).

[0028] The scoring criteria are: (1) Swelling degree: no swelling was scored as 0 points, mild swelling was scored as 1 point, and severe swelling was scored as 2 points; (2) Subcutaneous bruising: no bruising was scored as 0 points, a small amount of bruising was scored as 1 point, and a large amount of bruising was scored as 2 points; (3) Functional impairment: no lameness was scored as 0 points, mild lameness was scored as 1 point, and severe lameness was scored as 2 points; (4) Tenderness reaction: no tenderness was scored as 0 points, mild tenderness was scored as 1 point, and obvious tenderness was scored as 2 points.

[0029] Soft tissue injury evaluation results: (1) Swelling degree: 24 hours after administration, all the treatment groups had no obvious swelling reduction effect; 72 hours after administration, all the treatment groups had a certain swelling reduction effect. When the extraction temperature was 60-80℃, the swelling reduction effect of the wound cream product was the best, as shown in Table 3.

[0030] Table 3 Swelling degree scoring results (n=5)

[0031] (2) Subcutaneous hemostasis: 72 hours after administration, the hemostasis of mice in all treatment groups improved to a certain extent. The best effect was achieved when the extraction temperature was 60-80°C. See Table 4 for details.

[0032] Table 4 Subcutaneous congestion score results (n=5)

[0033] (3) Functional impairment: 24 hours after administration, there was no significant therapeutic effect in all the drug administration groups; 48 hours after administration, the preparation prepared with an extraction temperature of 80°C had the best therapeutic effect, and the therapeutic effects of the other groups were not much different; 72 hours after administration, the preparation prepared with an extraction temperature of 60-80°C had a good therapeutic effect, as shown in Table 5 for details.

[0034] Table 5 Results of functional impairment scores (n=5)

[0035] (4) Tenderness reaction: 24 hours after administration, there was no significant therapeutic effect in all the treatment groups. At 48 hours and 72 hours after administration, the best therapeutic effect was achieved at 60-80°C, as shown in Table 6.

[0036] Table 6 Results of tenderness response scores (n=5)

[0037] (IV) Skin toxicology experiment of Chinese medicine compound wound-relieving ointment 1. Acute skin toxicity test 40 guinea pigs, half male and half female, were taken and adaptively raised for 7 days. They were divided into low, medium, high dose groups and a blank group by random number table method, with 10 in each group. All guinea pigs were depilated on both sides of the back before administration, with a depilatory area of ​​4×4 cm. Each of the low, medium and high dose groups was evenly applied with 1.5 g, 2.0 g and 2.5 g of the Chinese medicine compound wound-relief ointment of the present invention, and then wrapped with gauze and fixed with tape. The wound-relief ointment was removed after 24 hours; the blank group was not treated. Once a day, repeated for 14 days. The guinea pigs were weighed daily, and their food intake, breathing, body shape, eye and limb activities, fecal characteristics, and death were observed.

[0038] Results of acute skin toxicity experiment: After administration, the food intake of guinea pigs in each group did not decrease significantly, and there were no significant changes in breathing, posture, eyes, limb activities, mental state, and feces characteristics, and no deaths occurred. Compared with the blank group, there was no significant difference in the weight of guinea pigs in the low, medium, and high dose groups at different time points, see Table 7 for details.

[0039] Table 7 Comparison of body weight of guinea pigs in each group at different time points (x±n)

[0040] 2. Skin irritation test Eight New Zealand white rabbits, half male and half female, were adaptively fed for 7 days and randomly divided into intact skin group and damaged skin group, with 4 rabbits in each group. All New Zealand white rabbits were depilated on both sides of the spine 24 hours before administration, with a depilated area of ​​6×6 cm. After 24 hours, the depilated skin was checked for damage. After 24 hours of depilation, the depilated skin in the damaged skin group was disinfected and scratched with a surgical blade until slight bleeding occurred. The degree of damage on both sides was kept consistent as much as possible. 2 g of traditional Chinese medicine compound injury plaster was applied to the depilated area on the left side in both groups; the right side was used as a blank control group and was not treated. After 24 hours of application, the injury plaster was washed off. In the process, the skin reaction was observed 1, 24, 48, and 72 hours after the application of the injury plaster. The experimental results were scored according to the skin irritation reaction scoring standard of the "Guidelines for the Study of New Chinese Medicines" issued by the Drug Administration of the Ministry of Health, and the average score was calculated to evaluate the irritation intensity.

[0041] Results of skin irritation experiment: New Zealand rabbits in each group were observed for 3 consecutive days after administration. The intact skin group showed no erythema and edema during the whole process; the damaged skin group showed erythema 1 hour after administration, as shown in Table 8 for details.

[0042] Table 8 Skin reaction scores and irritation intensity of different groups in skin irritation test

[0043] 3. Skin allergy test Thirty guinea pigs, half male and half female, were taken and adaptively raised for 7 days. They were divided into a drug-treated group, a positive group and a blank group by a random number table method, with 10 in each group. 24 hours before drug administration, both sides of the back of the guinea pigs were depilated, with an area of ​​4×4 cm. The positive sensitizing drug was prepared with 2,4-dinitrochlorobenzene to a sensitizing concentration of 1% and a provocative concentration of 0.1% (70% ethanol as solvent).

[0044] Sensitization contact: The drug-treated group was treated with 0.2 g of wound cream on the left hair removal area, the positive group was treated with 0.2 ml of 1% 2,4-dinitrochlorobenzene solution, and the blank group was not treated. After the drug administration, the skin was fixed with gauze and tape, and the drug was washed and removed after 6 hours. The above operation was repeated on the 7th and 14th days.

[0045] Provocative contact: 14 days after the last sensitization contact with the test substance, the drug-treated group applied 0.2 g of wound cream on the left depilatory area, the positive group applied 0.2 ml of 1% 2,4-dinitrochlorobenzene solution, and the blank group was not treated. Skin allergic reactions were observed and scored.

[0046] Results of skin allergy experiment: After sensitization, no allergic reaction or sensitization was observed in the depilated skin of guinea pigs in the drug-treated group and the blank group at any time point. After the positive control group was given the sensitizer, allergic reaction and extreme sensitization were observed at any time point, as shown in Table 9 for details.

[0047] Table 9 Results of skin allergic reaction test

[0048] For ordinary technicians in the technical field to which the present invention belongs, several simple deductions or substitutions can be made without departing from the concept of the present invention, which should be regarded as falling within the protection scope of the present invention.

Claims

1. A method for preparing a Chinese medicine compound wound-healing ointment, characterized in that: The following steps are involved: (1) Divide APIs into the following four groups: Group 1: Animals - 3-5 portions of scorpions, 3-6 portions of centipedes, 3-5 portions of earth dragons; Group 2: Volatile oils - 3-5 parts of Saposhnikovia divaricata, 3-5 parts of Angelica dahurica, 3-5 parts of Angelica sinensis, 3-5 parts of Asarum, 3-5 parts of Chuanxiong, 3-5 parts of Salvia miltiorrhiza, 11-14 parts of Trigonosciadium japonicum, 10-13 parts of Curcuma zedoaria, 5-7 parts of Frankincense, 5-7 parts of Myrrh, 5-7 parts of Papaya, 3-5 parts of Rhubarb, 3-5 parts of Herba Polygoni Multiflori; Group 3: Toxic—5-7 parts of raw aconite, 3-5 parts of Arisaema consanguineum, 3-5 parts of Chuanwu, 3-5 parts of Caowu; (2) The raw material drug of group 1 is crushed, sieved, and subjected to ultrasonic extraction using camellia oil as a solvent. The ultrasonic extraction temperature is 60-80° C., the ultrasonic power is 1440-1450 W, and the extraction time is 60-70 minutes to obtain an oily substance containing protein; (3) The raw materials of group 2 are crushed, sieved, and subjected to ultrasonic extraction using camellia oil as a solvent. The ultrasonic extraction temperature is 40 to 60° C., the ultrasonic power is 350 to 360 W, and the extraction time is 40 to 50 minutes to obtain an oily substance containing volatile oil; (4) The raw materials of group 3 are crushed, sieved, and ultrasonically extracted with camellia oil as a solvent. The ultrasonic extraction temperature is 90-100° C., the ultrasonic power is 350-360 W, and the extraction time is 120-130 minutes to obtain an oily substance containing curcumin; (5) mixing the oily substance containing protein obtained in step (2), the oily substance containing volatile oil obtained in step (3), and the oily substance containing curcumin obtained in step (4), and performing ultrasonic extraction again after mixing. The ultrasonic extraction temperature in this step is 60 to 80° C., the ultrasonic power is 350 to 360 W, and the extraction time is 20 to 30 minutes to obtain an oily mixture; (6) Adding beeswax to the oily mixture obtained in step (5), melting the beeswax at 60 to 80° C. and mixing the mixture with the oily mixture to prepare an oil-wax mixture for later use; (7) Adding butyl vanillin and azone to the oil-wax mixture obtained in step (6), wherein the amount of butyl vanillin added is controlled to be 0.03-0.05%, and the amount of azone added is controlled to be 0.5-0.8%, stirring and mixing, packaging, and cooling to obtain the traditional Chinese medicine compound wound healing ointment.

2. The method for preparing the Chinese medicine compound wound-treating ointment according to claim 1, wherein: The ultrasonic extraction in steps (2) to (5) is performed using a dual-frequency ultrasonic extractor.

3. A Chinese medicine compound wound healing ointment, characterized in that: The Chinese herbal compound injury-healing ointment is a Chinese herbal compound injury-healing ointment prepared according to the preparation method of the Chinese herbal compound injury-healing ointment according to claim 1 or 2.

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