Preparation method and application of a novel external tiger balm for treating wounds

Abstract

This application discloses a method for preparing a novel topical Tiger Yellow Healing Traditional Chinese Medicine (TCM) preparation for treating wounds, comprising the following steps: S1, preparation of medicinal materials: Seven medicinal materials powders are obtained by grinding or pulverizing Bauhinia bark, Astragalus membranaceus, Angelica pubescens, Saxifraga stolonifera, Paeonia lactiflora, Acorus tatarinowii, and Angelica dahurica; S2, preparation of homologous carbon quantum dots; S3, preparation of a TCM compound; S4, mixing the homologous carbon quantum dots obtained in step S2 with the TCM compound obtained in step S3, dialysis the mixture with distilled water to remove unloaded small molecules of the TCM compound, and then drying to obtain the Tiger Yellow Healing TCM preparation. This application also provides an application of this novel topical Tiger Yellow Healing TCM preparation for treating wounds. This application is the first to propose a method for preparing a TCM compound loaded with homologous carbon quantum dots, increasing the efficacy of the TCM compound itself. It can be used to treat various acute wounds and chronic, difficult-to-heal wounds, promote rapid wound healing, and has good antibacterial, antioxidant, and wound angiogenesis effects.

Description

Technical Field

[0001] This application relates to the field of traditional Chinese medicine technology, and in particular to the preparation method and application of a novel external application of Tiger Yellow Healing Chinese medicine preparation for treating wounds. Background Technology

[0002] With the accelerating aging of the population and changes in the spectrum of human diseases, the number of patients with acute wounds caused by trauma and burns, as well as chronic, difficult-to-heal wounds caused by diseases such as diabetes, is increasing year by year, affecting people's physical and mental health and quality of life. Wound repair has become a hot topic in clinical medicine, and since any wound can potentially lead to bacterial infection, the development of new drugs that effectively combat drug-resistant bacterial infections while simultaneously promoting the healing of infected wounds is of great significance. However, the use of antibiotics has led to the emergence and spread of multidrug-resistant strains within hospitals, especially methicillin-resistant Staphylococcus aureus (MRSA). Bacterial infections can exacerbate the inflammatory response during wound repair, even leading to infection-related complications and slowing down the wound healing process. Moreover, infected wounds are difficult to treat, require long treatment times, and incur high medical expenses.

[0003] Recent research has shown that carbon quantum dots (CQDs) have emerged as promising biomedical materials, primarily due to their fabrication processes, structure, and physicochemical properties. CQDs are zero-dimensional carbon-based nanomaterials with quasi-spherical dimensions less than 10 nm. All CQDs possess sp2 or sp3 hybridized orbitals, providing stability and unique optical characteristics, including a large Stokes shift in photoluminescence and a strong dependence of emission color on the excitation wavelength, depending on their size, core structure, and composition. Due to their small size, CQDs exhibit unique photoluminescence properties. In addition to photoluminescence, CQDs possess other useful properties such as water solubility, biocompatibility, low toxicity, low cost, and environmental sustainability. They also exhibit high adsorption interactions with biological surfaces due to their controllable surface functional groups and small size. These properties make them attractive materials for biomedical applications and have been extensively studied for bioimaging, drug delivery, biosensors, cancer therapy, and antibacterial applications. Among these methods, the hydrothermal synthesis of carbon quantum dots from traditional Chinese medicine (TCM) herbs combines the inherent good water solubility and low toxicity of carbon quantum dots with the inherent properties of TCM carbon sources. This can solve the problems of poorly soluble TCM components with low bioavailability and high biotoxicity. This one-step hydrothermal synthesis of TCM-derived carbon quantum dots using advanced modern engineering technology holds promise for developing new applications of TCM. However, current domestic and international research primarily focuses on carbon quantum dots derived from single TCM herbs, with limited research on carbon quantum dots derived from TCM compound preparations. Furthermore, the low yield of TCM-derived carbon quantum dots significantly limits their clinical application. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides a method for preparing and applying a novel topical Tiger Yellow Healing Traditional Chinese Medicine preparation for treating wounds.

[0005] The technical solution provided by this invention is as follows:

[0006] A method for preparing a novel topical Tiger Yellow Healing Traditional Chinese Medicine preparation for treating wounds includes the following steps:

[0007] S1. Preparation of medicinal materials: Grind or pulverize the following seven medicinal materials into powder: Bauhinia bark, Astragalus membranaceus, Angelica pubescens, Saxifraga stolonifera, Paeonia lactiflora, Acorus tatarinowii and Angelica dahurica.

[0008] S2. Preparation of homologous carbon quantum dots: Take appropriate amounts of the seven medicinal powders from step S1, mix them evenly to obtain a mixed powder, add deionized water to the mixed powder, and disperse it by ultrasonication (ultrasonication time 10-60 min) to obtain a mixed solution; subject the mixed solution to a hydrothermal reaction, collect the supernatant after centrifugation of the reaction product, and filter it through a 0.22 μm filter membrane. The filtered product is dialyzed with ultrapure water, and the dialyzed reaction product is freeze-dried to obtain homologous carbon quantum dots. The carbon quantum dots exhibit blue fluorescence under a 365 nm ultraviolet lamp, and the particle size is <10 nm as measured by transmission electron microscopy.

[0009] S3. Preparation of Traditional Chinese Medicine Compound: Take appropriate amounts of the seven medicinal powders from step S1, mix them evenly, add the mixed powders and anhydrous ethanol to a round-bottom flask and soak overnight. Then, extract with a reflux reflux device for 1 hour. After extraction, separate the filtrate and the residue. Put the obtained residue back into the reflux reflux device, add the same volume of deionized water as anhydrous ethanol, and perform a second reflux extraction for 1 hour. Separate the filtrate and the residue. Mix the two filtrates obtained from the two refluxes to obtain a mixture. Concentrate the mixture, freeze-dry the concentrated mixture in a freeze dryer, and grind it in a mortar and pestle to obtain the Traditional Chinese Medicine Compound.

[0010] S4. Mix the homologous carbon quantum dots obtained in step S2 with the traditional Chinese medicine mixture obtained in step S3, and then stir at room temperature for 48 hours (800 rpm / min). Load the traditional Chinese medicine mixture with homologous carbon quantum dots by liquid-phase mixing. Dialyze the mixture with distilled water for 24 hours (dialysis bag 500-1000 Da) to remove unloaded small molecules of the traditional Chinese medicine mixture. After freeze-drying, the traditional Chinese medicine preparation for treating ulcers is obtained. The final product is a powder and can be stored in a dry place at room temperature.

[0011] Preferably, in step S1, the seven medicinal powders are pulverized using a high-speed pulverizer and then passed through a 40-mesh sieve.

[0012] Preferably, in step S2, the mass ratio of Bauhinia bark, Astragalus, Angelica pubescens, Saxifraga stolonifera, Paeonia lactiflora, Acorus tatarinowii, and Angelica dahurica is 5-7:3-5:3-5:1-3:1-3:1-3:1-3:1-3; and in step S3, the mass ratio of Bauhinia bark, Astragalus, Angelica pubescens, Saxifraga stolonifera, Paeonia lactiflora, Acorus tatarinowii, and Angelica dahurica is 5-7:3-5:3-5:1-3:1-3:1-3:1-3:1-3.

[0013] Preferably, in step S2, the mass ratio of the mixed powder to deionized water is 1:5 to 20.

[0014] Preferably, the hydrothermal reaction temperature in step S2 is 160–240°C, and the hydrothermal reaction time is 2–16 h.

[0015] Preferably, in step S3, the mass ratio of the mixed powder to anhydrous ethanol is 1:8 to 10.

[0016] Preferably, in step S3, the mixture is placed in a rotary evaporator for concentration until the concentrated liquid is 1 / 5 to 1 / 3 of the original volume, and then the concentration is stopped.

[0017] Preferably, in step S4, the mass ratio of the traditional Chinese medicine mixture to homologous carbon quantum dots is 1 to 150:1.

[0018] A traditional Chinese medicine preparation for treating acute wounds or chronic, non-healing wounds, prepared using the above-described method, can produce, but is not limited to, antibacterial, antioxidant, anti-inflammatory, hemostatic, and analgesic effects.

[0019] Preferably, the dosage form of the Tiger Yellow Healing Herbal Preparation is one of the following: ointment, powder, spray, emulsion, paste, smear, gel, or the Tiger Yellow Healing Herbal Preparation is an effective ingredient of a Band-Aid or other external dressing.

[0020] This application has the following advantages over the prior art:

[0021] This invention proposes for the first time a method for preparing traditional Chinese medicine compound loaded with homologous carbon quantum dots, which increases the efficacy of the compound itself. It can be used to treat various acute wounds and chronic refractory wounds, promote rapid wound healing, and has good antibacterial, antioxidant, and angiogenesis effects. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 These are TEM images of homologous carbon quantum dots in embodiments of the present invention;

[0024] Figure 2 This is a particle size analysis diagram of homologous carbon quantum dots in an embodiment of the present invention;

[0025] Figure 3 The images show the UV-Vis absorption spectrum of homologous carbon quantum dots, a general sunlight image of the carbon quantum dot solution, a general fluorescence image under a 365nm UV lamp, and a fluorescence spectrum in the embodiments of the present invention.

[0026] Figure 4 The infrared spectrum of homologous carbon quantum dots in the embodiments of the present invention;

[0027] Figure 5 This is an XPS diagram of homologous carbon quantum dots in an embodiment of the present invention;

[0028] Figure 6 The images show the ultraviolet-visible absorption spectra of the traditional Chinese medicine compound, homologous carbon quantum dots, and the final product in the embodiments of the present invention.

[0029] Figure 7 The images shown are infrared spectra of the traditional Chinese medicine compound, homologous carbon quantum dots, and the final product in the embodiments of the present invention.

[0030] Figure 8 The figures show the antibacterial experimental results of the traditional Chinese medicine compound, homologous carbon quantum dots, and the final product in the embodiments of the present invention.

[0031] Figure 9 This is a diagram showing the animal experiment results of the final product in treating MRSA infected wounds in an embodiment of the present invention. Detailed Implementation

[0032] To enable those skilled in the art to better understand the technical solutions in this application, the technical solutions in the embodiments of this application will be clearly and completely described below. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0033] 1. Preparation of homologous carbon quantum dots

[0034] Carbon quantum dots for synthesizing traditional Chinese medicine compound preparations were synthesized using a hydrothermal method. Seven dried medicinal herbs—Bauhinia bark, Astragalus membranaceus, Angelica pubescens, Saxifraga stolonifera, Paeonia lactiflora, Acorus tatarinowii, and Angelica dahurica—were pulverized into powder using a micro-grinder. The powders were then mixed in a mass ratio of 5–7:3–5:3–5:1–3:1–3:1–3:1–3, with a total powder-to-water ratio of 1:15. The aqueous solution was then thoroughly stirred and ultrasonically dispersed. The ultrasonically dispersed solution was transferred to a polytetrafluoroethylene-lined reactor and heated at 200°C for 6 hours in an oven. Subsequently, the reactor was cooled to room temperature, and the resulting mixture was centrifuged (8000 rpm, 10 minutes). The supernatant was collected and filtered through a 0.22 μm filter membrane to remove large particles, yielding a brownish-yellow solution. The solution was dialyzed with deionized water in a 1000 Da dialysis bag for 24 hours to remove unreacted components and other byproducts, and finally the homologous carbon quantum dots of the purified traditional Chinese medicine compound were obtained for characterization and other examples.

[0035] 2. Preparation of Traditional Chinese Medicine Compound Preparations

[0036] A mixture of herbal powders, including Bauhinia bark, Astragalus root, Angelica pubescens root, Saxifraga stolonifera root, Paeonia lactiflora root, Acorus tatarinowii rhizome, and Angelica dahurica root, was prepared in a mass ratio of 5–7:3–5:3–5:1–3:1–3:1–3:1–3. The mixture was soaked overnight in a round-bottom flask with anhydrous ethanol at a mass ratio of 1:8–10. The mixture was then extracted using a reflux apparatus for 1 hour, and the filtrate and residue were separated. The residue was returned to the reflux apparatus, and an equal volume of deionized water (equal to the volume of anhydrous ethanol) was added for a second reflux extraction for 1 hour. The filtrate and residue were then separated again. The two filtrates obtained from the two reflux extractions were mixed to obtain a final mixture. This mixture was then concentrated in a rotary evaporator until the concentrated liquid reached 1 / 5–1 / 3 of its original volume. The concentrated mixture was then dried in a freeze dryer and ground until a lyophilized powder of the traditional Chinese medicine mixture was obtained.

[0037] 3. Preparation of Tiger Yellow Healing Traditional Chinese Medicine Preparation

[0038] Different mass concentrations of traditional Chinese medicine (TCM) compounds and homologous carbon quantum dots were mixed at a mass ratio of 1–150:1. The mixture was then stirred at room temperature for 48 hours (800 rpm / min) to prepare a novel external TCM preparation for treating ulcers by mixing homologous carbon quantum dots with the TCM compound. The resulting product was dialyzed with distilled water for 24 hours (dialysis bag 500–1000 Da) to remove unloaded small molecules of the TCM compound. After freeze-drying, the obtained powder was stored in a dry place at room temperature.

[0039] Homologous carbon quantum dots were imaged using a JEM-2100F field emission transmission electron microscope at an accelerating voltage of 200 kV. The results are shown in Figures 1 and 2. The homologous carbon quantum dots have a uniform shape and no obvious aggregation. As shown in Figure 3, the average diameter of the carbon quantum dots is 2.47 ± 0.9 nm.

[0040] The homologous carbon quantum dots were analyzed using a UV-Vis spectrophotometer, and the results are as follows: Figure 3 The homologous carbon quantum dots of the mixture have an ultraviolet absorption peak at 276 nm; when excited at 365 nm, they emit strong fluorescence in the range of 380–700 nm, with a peak intensity of around 460 nm.

[0041] The homologous carbon quantum dots were detected by FTIR spectroscopy using an infrared spectrometer. The results are shown in Figure 4, which shows the FTIR spectra of the homologous carbon quantum dots and the corresponding group vibrations at the peaks.

[0042] The homologous carbon quantum dots were detected using photoelectron spectroscopy, and the results are shown in Figure 5. The XPS spectrum of the homologous carbon quantum dots indicates that they are mainly composed of C, N and O elements.

[0043] The UV-Vis absorption spectra of the traditional Chinese medicine compound, homologous carbon quantum dots, and the Huhuang Yuyang traditional Chinese medicine preparation were detected using a UV-Vis spectrophotometer. The results, shown in Figure 6, indicate that the novel Huhuang Yuyang traditional Chinese medicine preparation exhibits a peak at 278 nm, which differs from the peak values ​​of homologous carbon quantum dots (276 nm) and the traditional Chinese medicine compound (281 nm). Furthermore, compared to the small absorption peak at 322 nm observed in the traditional Chinese medicine compound, the Huhuang Yuyang traditional Chinese medicine preparation shows almost no absorption peak in this band. A valley appears at 265 nm, which differs from the valleys of homologous carbon quantum dots (269 nm) and the traditional Chinese medicine compound (262 nm). This indicates that the Huhuang Yuyang traditional Chinese medicine preparation was successfully prepared by combining the two components.

[0044] The traditional Chinese medicine compound, homologous carbon quantum dots, and tiger huang hemorrhoid traditional Chinese medicine preparation were analyzed by FTIR spectroscopy. The results are shown in Figure 7. The FTIR spectrum of homologous carbon quantum dots shows that the peak intensity of the tiger huang hemorrhoid traditional Chinese medicine preparation changed compared to the traditional Chinese medicine compound and homologous carbon quantum dots, and the peaks at 3336 and 1608 cm⁻¹ were significantly different. −1 A new peak appeared, at 1706 cm. −1 The peak value almost disappeared, indicating that the combination of the two ingredients successfully prepared a traditional Chinese medicine preparation for treating ulcers.

[0045] 4. In vitro antibacterial and antimicrobial experiments

[0046] The bactericidal effect of the traditional Chinese medicine preparation for treating phlegm-heat syndrome against bacteria (Staphylococcus aureus and methicillin-resistant Staphylococcus aureus) was determined by agar plate counting. 200 μL of PBS, the traditional Chinese medicine mixture solution, homologous carbon quantum dot solution, and the phlegm-heat syndrome preparation solution were added to 48-well plates, followed by 20 μL of bacterial suspension (LB, 1.5 × 10⁻⁶). 8 CFU / mL was added to each sample well. After placing the culture plate in an incubator at 37°C for 24 hours, the mixture was diluted appropriately and 100 μL was inoculated onto an agar plate. The plate was spread evenly and then placed in a constant temperature incubator (37°C), with the plate inverted to prevent contamination. After 18 hours, the culture plates were photographed, and the bacterial growth on each agar plate was recorded. Finally, the relative bacterial viability was obtained by counting the number of colonies.

[0047] Figure 8 shows photographs of homologous carbon quantum dots, traditional Chinese medicine mixtures, and tiger huang hemorrhoid preparations prepared in the embodiments of the present invention on agar plates under different bacterial treatments. Figure 8 It can be seen that, compared with traditional Chinese medicine compound preparations and homologous carbon quantum dots, the traditional Chinese medicine preparation of Tiger Yellow Healing Sore has the best antibacterial effect.

[0048] Example 5: Animal experimental results of in vivo treatment of MRSA-infected wounds. Clean-grade 8-10 week old male SD rats were selected. After acclimatization for 7 days, the rats were anesthetized via inhalation of a mixture of isoflurane and oxygen. First, the skin of the rat's back area was prepared, disinfected with 75% alcohol, and a circular full-thickness skin of approximately 1.5 cm in diameter was surgically removed to create a wound. A rubber band was fixed to the wound to prevent skin contraction during healing. Postoperatively, 100 µL of fresh MRSA bacterial suspension (bacterial density 1×10⁻⁶) was instilled into the wound area. 8 (CFU / mL). After 24 hours, rats with successful infection were selected based on whether the wound was red, swollen, and hot, and whether there was purulent exudate. The successfully infected rats were randomly divided into a control group, a traditional Chinese medicine compound group, a homologous carbon quantum dot group, and a tiger huang healer traditional Chinese medicine preparation group. Each group received the corresponding treatment, and wound healing was observed and photographed on postoperative days 0, 5, 10, and 15.

[0049] As shown in Figure 9: the first column represents the control group, which received saline treatment; the second column represents the traditional Chinese medicine compound group, which received the same compound; the third column represents the homologous carbon quantum dot group, which received the same carbon quantum dot treatment; and the fourth column represents the tiger huang healer traditional Chinese medicine preparation group, which received the same preparation. Treatment continued for 15 days, with regular photographic observation of wound changes. Results showed that compared to the control group, the traditional Chinese medicine compound, homologous carbon quantum dots, and tiger huang healer traditional Chinese medicine preparation all significantly promoted the healing of MRSA infected wounds, with the tiger huang healer traditional Chinese medicine preparation showing the best effect.

[0050] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing a novel topical Tiger Yellow Healing Traditional Chinese Medicine preparation for treating wounds, characterized in that, The following steps are included: S1. Preparation of medicinal materials: Grind or pulverize the following seven medicinal materials into powder: Bauhinia bark, Astragalus membranaceus, Angelica pubescens, Saxifraga stolonifera, Paeonia lactiflora, Acorus tatarinowii and Angelica dahurica. S2. Preparation of homologous carbon quantum dots: Take appropriate amounts of the seven medicinal powders from step S1, with the mass ratio of Bauhinia bark, Astragalus membranaceus, Angelica pubescens, Saxifraga stolonifera, Paeonia lactiflora, Acorus tatarinowii, and Angelica dahurica all being 5-7:3-5:3-5:1-3:1-3:1-3:1-3. Mix them evenly to obtain a mixed powder. Add deionized water to the mixed powder and disperse it by ultrasonication to obtain a mixed solution. Perform a hydrothermal reaction on the mixed solution. The reaction product is then centrifuged, filtered, dialyzed with ultrapure water, and dried to obtain homologous carbon quantum dots. S3. Preparation of Traditional Chinese Medicine Compound: Take appropriate amounts of the seven medicinal powders from step S1, with the mass ratio of Bauhinia bark, Astragalus membranaceus, Angelica pubescens, Saxifraga stolonifera, Paeonia lactiflora, Acorus tatarinowii, and Angelica dahurica all being 5-7:3-5:3-5:1-3:1-3:1-3:1-3. Mix them thoroughly. Add the mixed powders and anhydrous ethanol to a round-bottom flask and soak overnight. Then, extract using a reflux evaporator. After extraction, separate the filtrate and the residue. Return the residue to the reflux evaporator and add the same volume of deionized water as anhydrous ethanol for a second reflux extraction. Separate the filtrate and the residue. Mix the two filtrates obtained from the two refluxes to obtain a mixture. Concentrate the mixture, freeze-dry and grind the concentrated mixture to obtain the traditional Chinese medicine compound. S4. Mix the homologous carbon quantum dots obtained in step S2 with the traditional Chinese medicine mixture obtained in step S3. The mass ratio of the traditional Chinese medicine mixture to the homologous carbon quantum dots is 1 to 150:

1. The mixture is dialyzed with distilled water to remove unloaded small molecules of the traditional Chinese medicine mixture. After drying, the traditional Chinese medicine preparation for treating ulcers caused by tiger huang is obtained.

2. The preparation method of the novel external application of Tiger Yellow Healing Traditional Chinese Medicine preparation for treating wounds according to claim 1, characterized in that, In step S1, the seven medicinal powders are pulverized using a high-speed pulverizer and then passed through a 40-mesh sieve.

3. The preparation method of the novel external application of Tiger Yellow Healing Traditional Chinese Medicine preparation for treating wounds according to claim 1, characterized in that, In step S2, the mass ratio of the mixed powder to deionized water is 1:5 to 20.

4. The preparation method of the novel external application of Tiger Yellow Healing Traditional Chinese Medicine preparation for treating wounds according to claim 1, characterized in that, In step S2, the hydrothermal reaction temperature is 160–240°C, and the hydrothermal reaction time is 2–16 hours.

5. The preparation method of the novel external application of Tiger Yellow Healing Traditional Chinese Medicine preparation for treating wounds according to claim 1, characterized in that, In step S3, the mass ratio of the mixed powder to anhydrous ethanol is 1:8 to 10.

6. The preparation method of the novel external application of Tiger Yellow Healing Traditional Chinese Medicine preparation for treating wounds according to claim 1, characterized in that, In step S3, the mixture is placed in a rotary evaporator for concentration until the concentrated liquid is 1 / 5 to 1 / 3 of the original volume, and then the concentration is stopped.

7. A traditional Chinese medicine preparation for treating acute wounds or chronic, difficult-to-heal wounds, characterized in that, The product obtained by any one of the preparation methods of claims 1 to 6.

8. The traditional Chinese medicine preparation for treating acute wounds or chronic, difficult-to-heal wounds according to claim 7, characterized in that, The dosage form of the Tiger Yellow Healing Herbal Preparation is one of the following: ointment, powder, spray, emulsion, paste, or gel; or the Tiger Yellow Healing Herbal Preparation is the effective ingredient of a Band-Aid or other external dressing.

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