Traditional Chinese medicine disinfection composition as well as preparation method and application thereof

Abstract

The invention belongs to the field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine disinfection composition as well as a preparation method and application thereof. And 3-13 parts of pratia nummularia. The ligularia dichotoma and the pratia nummularia are matched for use for the first time, and the ligularia dichotoma and the pratia nummularia have a synergistic effect, so that a remarkable antibacterial and disinfection effect is shown. In-vitro experiments prove that the activity inhibition IC50 of the traditional Chinese medicine disinfection composition to the H1N1 virus can reach 18.4 mu g / mL, the inhibition rate of the traditional Chinese medicine disinfection composition to pathogenic bacteria such as staphylococcus aureus, candida albicans and escherichia coli is larger than or equal to 95.6%, and the traditional Chinese medicine disinfection composition shows broad-spectrum and efficient bacteriostasis and disinfection characteristics, can be widely applied to multiple fields such as medical environment disinfection, public health and epidemic prevention and daily household cleaning and has broad application prospects. The invention provides a novel method for solving the problems of poor effect and drug resistance of the existing disinfection product.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to traditional Chinese medicine disinfection compositions, their preparation methods, and applications. Background Technology

[0002] Currently, the disinfection product market is diversified, mainly divided into two categories: chemically synthesized disinfectants and some naturally derived disinfectants. Chemically synthesized disinfectants include chlorine-containing disinfectants (such as 84 disinfectant), peroxide disinfectants (such as peracetic acid), and aldehyde disinfectants (such as glutaraldehyde). These chemical disinfectants can rapidly destroy the cell structure of microorganisms and effectively kill various pathogenic microorganisms, playing a crucial role in responding to emergencies such as outbreaks of acute infectious diseases. However, with the widespread use of chemically synthesized disinfectants, a series of problems exposed in their practical application have become increasingly prominent. On the one hand, most of these disinfectants have a strong, irritating odor, and the chemicals released during disinfection can directly irritate and damage the human respiratory mucosa and skin. Long-term or frequent exposure to these chemical disinfectants may trigger allergic reactions, respiratory diseases, and other health problems, posing a potential threat to the health of personnel engaged in disinfection work and the public in disinfection environments. On the other hand, bacteria, viruses, and other microorganisms gradually develop resistance to chemical disinfectants under long-term exposure. The emergence of this drug resistance has significantly reduced the effectiveness of previously effective disinfectants in killing or inhibiting microorganisms, making it difficult to meet the increasingly complex and severe disinfection needs and posing new challenges to public health and safety.

[0003] Therefore, disinfectants from natural sources, with their significant advantages such as gentleness, safety, and environmental friendliness, are gradually gaining widespread attention from researchers and consumers. Traditional Chinese medicine, as a shining pearl of traditional medicine, contains abundant natural active ingredients. These ingredients possess various pharmacological effects, including antibacterial, antiviral, and anti-inflammatory properties, providing a rich resource for the development of new disinfection products. Through scientific formulation and modern extraction processes, many traditional Chinese medicinal materials can be processed into drugs with significant disinfection effects. These traditional Chinese medicine disinfectants not only effectively kill or inhibit pathogenic microorganisms, but also significantly reduce side effects on humans and the environment compared to chemically synthesized disinfectants, meeting modern society's pursuit of green and healthy disinfection products.

[0004] However, disinfection products developed based on traditional Chinese medicine still have some limitations. Due to the complexity of the ingredients and the diversity of their mechanisms of action, some products have a relatively narrow antibacterial spectrum, exhibiting good killing or inhibitory effects only against specific types of microorganisms, making it difficult to address diverse microbial contamination situations. Other products have insufficient disinfection effects, failing to meet the standards of highly efficient sterilization. They are ineffective when facing high concentrations of pathogen contamination or urgent disinfection needs, thus limiting their application in a wider range of disinfection scenarios. For example, the traditional Chinese medicine composition developed in patent CN113633685B includes: Scutellaria baicalensis, Lonicera japonica, Isatis indigotica, Forsythia suspensa, Crataegus pinnatifida, Mentha haplocalyx, and Galla chinensis, but it only has a significant antibacterial effect against Staphylococcus aureus, Escherichia coli, and Candida albicans. The traditional Chinese medicine air disinfectant developed in patent CN115997796A includes: Atractylodes lancea, Mentha haplocalyx, Coptis chinensis, Dryopteris crassirhizoma, Artemisia argyi, and Citrus reticulata, which has the function of purifying air and eliminating odors, but currently it only has an antibacterial effect against Staphylococcus aureus and Escherichia coli. The traditional Chinese medicine disinfectant developed in patent CN102847028A includes 10-40 parts of Cnidium monnieri, 10-40 parts of Hedyotis diffusa, 5-20 parts of Zanthoxylum bungeanum, 5-20 parts of Mentha haplocalyx, 5-20 parts of Sophora flavescens, and 5-20 parts of Stemona japonica, which has a significant inhibitory effect against Staphylococcus aureus, Escherichia coli, yeast, and Candida albicans. Current research on traditional Chinese medicine disinfectants mainly focuses on their antibacterial effects. However, their antibacterial range is narrow, and their disinfection effect is greatly reduced when faced with high concentrations of pathogens with complex compositions.

[0005] In summary, there is an urgent need for a method to prepare a traditional Chinese medicine disinfectant composition that can inhibit a broad spectrum of pathogens, so as to prepare a traditional Chinese medicine disinfectant composition that can effectively inhibit diverse microbial contamination. Summary of the Invention

[0006] The purpose of this invention is to overcome the shortcomings of the prior art, reduce chemically synthesized disinfectants, prevent damage to the human body and the environment, avoid microorganisms developing resistance to chemical disinfectants, and provide a traditional Chinese medicine disinfection composition that can inhibit a broad spectrum of pathogenic microorganisms.

[0007] The object of the present invention is achieved through the following technical solutions: Firstly, the traditional Chinese medicine disinfectant composition comprises the following components in parts by weight: Ten to twenty portions of *Ligustrum lucidum* (a type of plant); Copper Hammer Jade Belt Grass 3-13 parts.

[0008] Preferably, the traditional Chinese medicine disinfectant composition is made from the following components in parts by weight: 15 parts of the *Ligustrum lucidum* and 7.5 parts of the *Ligustrum lucidum*. Specifically, the mass ratio of *Ligusticum striatum* and *Ligusticum striatum* in the traditional Chinese medicine disinfectant composition is 2:1.

[0009] *Ligustrum reticulata* is a perennial herb belonging to the genus *Ligustrum* in the family Asteraceae, mainly distributed in northwestern Yunnan and southwestern Sichuan. In traditional folk medicine, its roots and rhizomes are often used as a substitute for the Chinese herb *Aster tataricus*, possessing effects such as clearing the lungs and regulating qi, relieving cough and expectoration, and promoting urination; it is mainly used to treat common ailments such as colds and coughs. Modern pharmacological studies have shown that plants of the *Ligustrum* genus contain various chemical components, including triterpenoids, sesquiterpenoids, phenylpropyl esters, volatile oils, acrylic acids, and alkaloids, which play important pharmacological roles in anti-inflammatory and antitussive effects.

[0010] Copper Hammer Jade Belt Grass belongs to the Campanulaceae family and is also known as Ground Eggplant Grass or Small Copper Hammer. It mostly grows in shady and damp environments such as field embankments or shady places in mountain forests. The whole plant can be used as medicine, and it has the effects of dispelling wind and dampness. At the same time, it shows good pharmacological activity in anti-inflammatory and analgesic aspects.

[0011] On the other hand, the preparation method of the traditional Chinese medicine disinfectant composition includes the following specific steps: The *Ligustrum lucidum* and *Hylocereus undatus* are mixed and pulverized to obtain a traditional Chinese medicine mixture; The herbal mixture was extracted with water under reduced pressure and reflux, then filtered. The resulting aqueous extract was concentrated and dried to obtain the herbal disinfectant composition.

[0012] Specifically, the working conditions for the reduced pressure reflux extraction include: pressure of 8 kPa to 15 kPa; temperature of 46°C to 59°C; and the number of reduced pressure reflux extractions of 1 to 3 times. Preferably, the pressure of the vacuum reflux extraction is 11 kPa, and the temperature of the vacuum reflux extraction is 53°C.

[0013] Preferably, the weight ratio of the herbal medicine mixture to the water is 1:(8~12).

[0014] On the other hand, traditional Chinese medicine disinfectant compositions can be used in the preparation of antibacterial and disinfectant products and in the preparation of anti-H1N1 virus drugs.

[0015] The beneficial effects of this invention are: (1) The present invention provides a traditional Chinese medicine disinfection composition, its preparation method and application, and disinfectant. The present invention is the first to use Ligusticum chuanxiong and Copper Hammer Jade Belt Grass in combination. The two have a synergistic effect and show significant antibacterial and disinfection effects.

[0016] (2) In vitro experiments have demonstrated that the traditional Chinese medicine disinfectant composition provided by this invention has an inhibitory effect on the H1N1 virus (IC50). 50It can reach 18.4 μg / mL, and has an inhibition rate of ≥95.6% against bacteria such as Staphylococcus aureus, Candida albicans, and Escherichia coli, exhibiting broad-spectrum and highly effective antibacterial properties. It can be widely used in multiple fields such as medical environment disinfection, public health and epidemic prevention, and daily household cleaning, providing a new method to solve the problems of poor efficacy and drug resistance of existing disinfection products. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0018] Unless otherwise specified, all raw materials, reagents, instruments, and equipment used in this invention can be purchased commercially or prepared using existing methods. Furthermore, unless otherwise specified or detailed, the steps and parameters involved can be performed according to existing processing techniques or using existing equipment; these will not be elaborated upon in detail in this invention document.

[0019] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments that do not specify specific conditions are generally determined according to national standards. If no corresponding national standard exists, then generally accepted international standards, conventional conditions, or conditions recommended by the manufacturer shall be followed.

[0020] Example 1 This example provides a traditional Chinese medicine disinfectant composition, which, by weight, comprises the following components: 15 parts of Ligusticum striatum and 7.5 parts of Clematis armandii.

[0021] The preparation method of the above-mentioned traditional Chinese medicine disinfectant composition includes the following steps: Mix and pulverize Ligustrum lucidum and Clematis armandii to obtain a traditional Chinese medicine mixture; The herbal mixture was added to water and subjected to vacuum reflux extraction at a pressure of 11 kPa and a temperature of 53°C. The weight ratio of the herbal mixture to the water was 1:10. The mixture was then filtered, and the resulting aqueous extract was concentrated and dried to constant weight. The extraction was repeated twice to obtain the herbal disinfection composition.

[0022] Example 2 This example provides a traditional Chinese medicine disinfection composition, which, by weight, comprises the following components: 10 parts of Ligusticum striatum and 3 parts of Clematis armandii.

[0023] The preparation method of the above-mentioned traditional Chinese medicine disinfectant composition includes the following steps: Mix and pulverize Ligustrum lucidum and Clematis armandii to obtain a traditional Chinese medicine mixture; The herbal mixture was added to water and subjected to vacuum reflux extraction at a pressure of 15 kPa and a temperature of 59°C. The weight ratio of the herbal mixture to the water was 1:12. The mixture was then filtered, and the resulting aqueous extract was concentrated and dried to constant weight. The extraction was repeated twice to obtain the herbal disinfection composition.

[0024] Example 3 This example provides a traditional Chinese medicine disinfectant composition, which, by weight, comprises the following components: 20 parts of Ligusticum striatum and 13 parts of Clematis armandii.

[0025] The preparation method of the above-mentioned traditional Chinese medicine disinfectant composition includes the following steps: Mix and pulverize Ligustrum lucidum and Clematis armandii to obtain a traditional Chinese medicine mixture; The herbal mixture was added to water and subjected to vacuum reflux extraction at a pressure of 8 kPa and a temperature of 46°C. The weight ratio of the herbal mixture to the water was 1:8. The mixture was then filtered, and the resulting aqueous extract was concentrated and dried to constant weight. The extraction was repeated twice to obtain the herbal disinfection composition.

[0026] Comparative Example 1 This example provides a traditional Chinese medicine disinfectant extract and its preparation method. The only difference from Example 1 is that the amount of *Ligusticum chuanxiong* is adjusted to 0 parts and the amount of *Ligusticum striatum* is adjusted to 22.5 parts (i.e., only *Ligusticum striatum* is used as the traditional Chinese medicine raw material); the other steps and parameters are the same.

[0027] Comparative Example 2 This example provides a traditional Chinese medicine disinfectant extract and its preparation method. The only difference from Example 1 is that the amount of *Ligusticum repens* is adjusted to 22.5 parts and the amount of *Euphorbia pekinensis* is adjusted to 0 parts (i.e., only *Ligusticum repens* is used as the traditional Chinese medicine raw material); the other steps and parameters are the same.

[0028] Comparative Example 3 This example provides a traditional Chinese medicine disinfectant extract and its preparation method. The only difference from Example 1 is that the dosage of *Ligusticum repens* is adjusted to 7.5 parts and the dosage of *Hedyotis diffusa* is adjusted to 15 parts (i.e., the dosage ratio of *Ligusticum repens* and *Hedyotis diffusa* is adjusted); the remaining steps and parameters are the same.

[0029] Experimental Example 1 This study examines the inhibitory effect of the traditional Chinese medicine disinfectant extracts obtained in Examples 1-3 and Comparative Examples 1-3 on the activity of H1N1 virus, with ribavirin as a control.

[0030] The testing method is as follows: 1. Experimental Materials and Grouping Cells: MDCK cells (Madin-Darby Canine Kidney cells, logarithmic growth phase, viability > 95%).

[0031] Virus: H1N1 influenza virus (TCID) 50 The MOI was determined to be 0.01.

[0032] Drugs: Traditional Chinese medicine extracts (Examples 1-3, Comparative Examples 1-3), ribavirin (positive control).

[0033] Culture medium: MEM, a synthetic culture medium for cell culture.

[0034] Group design: blank control group (MEM only), cell control group (MEM + cells only), virus control group (virus + MEM + cells), and drug treatment group (drug + virus + MEM + cells).

[0035] 2. Experimental Procedure 2.1 Cell Plating MDCK cells were loaded at a rate of 1×10 4 / wells were inoculated into 96-well plates and cultured at 37°C with 5% CO2 until a monolayer was formed.

[0036] 2.2 Drug and Virus Pretreatment Drug dilution: Dilute the drug 2-fold serially with serum-free MEM (8 concentrations, covering 1-1000 μg / mL).

[0037] Virus treatment: Dilute the virus solution to 200 TCID using serum-free MEM. 50 / 100 μL.

[0038] Simultaneous addition: Add 50 μL of drug diluent + 50 μL of virus solution to each well (total volume 100 μL) to act on the cells simultaneously.

[0039] 2.3 Incubation conditions Incubate at 37℃ and 5% CO2 for 48 hours.

[0040] 2.4 Cell viability assay (WST-1 method) Aspirate the supernatant and wash gently once with PBS (to avoid cell detachment).

[0041] Add 100 μL of serum-free MEM containing 0.25 mM WST-1 to each well and incubate at 37°C in the dark for 2 hours.

[0042] The OD value was measured at 450 nm using an ELISA reader, with a reference wavelength of 630 nm.

[0043] 3. Data Processing and Computation 3.1 Cell Viability Correction Formula Where A is the OD value.

[0044] 3.2 IC 50 calculate A four-parameter logistic regression model was used to fit the dose-response curve and plot it. The concentration corresponding to 50% cell viability was then determined as the IC50. 50 The test results are shown in Table 1.

[0045] Table 1 50% cell viability As shown in Table 1, by combining Ligusticum repens and Clematis armandii in appropriate proportions, the present invention significantly improves the inhibitory effect of the obtained traditional Chinese medicine disinfection composition on H1N1 influenza virus, and its anti-H1N1 influenza virus effect is significantly better than that of ribavirin and comparative examples 1-3.

[0046] Experiment Example 2 Based on the above test examples, this invention further investigated the inhibitory effect of the traditional Chinese medicine disinfectant extracts obtained in Examples 1-3 on Staphylococcus aureus (ATCC6538), Candida albicans (ATCC10231), and Escherichia coli (ATCC8099) after 1 minute of action, according to the WS / T 650-2019 standard "Evaluation Method for Antibacterial and Bacteriostatic Effects". The results of the antibacterial rate test are shown in Table 2.

[0047] Table 2. Antibacterial rate of the traditional Chinese medicine disinfectant extract in this application. As shown in Table 2, the traditional Chinese medicine disinfection composition provided by the present invention has an inhibition rate of ≥95.6% against bacteria such as Staphylococcus aureus, Candida albicans, and Escherichia coli, exhibiting broad-spectrum and highly effective antibacterial and disinfection properties. It can be widely used in multiple fields such as medical environment disinfection, public health and epidemic prevention, and daily household cleaning.

[0048] In summary, this invention provides a traditional Chinese medicine disinfectant composition, its preparation method, and its application as a disinfectant. This invention is the first to combine *Ligustrum lucidum* and *Ligustrum sinense*, which synergistically enhance each other's effects, exhibiting significant antibacterial and disinfectant properties. In vitro experiments have demonstrated that the traditional Chinese medicine disinfectant composition provided by this invention has an inhibitory effect (IC50) against the H1N1 virus. 50It can reach 18.4 μg / mL, and has an inhibition rate of ≥95.6% against bacteria such as Staphylococcus aureus, Candida albicans, and Escherichia coli, exhibiting broad-spectrum and highly effective antibacterial and disinfection properties. It can be widely used in multiple fields such as medical environment disinfection, public health and epidemic prevention, and daily household cleaning, providing a new method to solve the problems of poor efficacy and drug resistance of existing disinfection products.

[0049] The above description is merely a preferred embodiment of the present invention. It should be understood that the present invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the concept described herein through the above teachings or related technologies or knowledge. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present invention should be within the protection scope of the appended claims.

Claims

1. A traditional Chinese medicine disinfectant composition, characterized in that, include: Ten to twenty portions of *Ligustrum lucidum* (a type of plant); Copper Hammer Jade Belt Grass 3-13 parts.

2. The traditional Chinese medicine disinfectant composition according to claim 1, characterized in that: The traditional Chinese medicine disinfectant composition includes 15 parts of the *Ligusticum striatum* and 7.5 parts of the *Euphorbia fischeriana*.

3. The traditional Chinese medicine disinfectant composition according to claim 1, characterized in that: The mass ratio of the *Ligustrum lucidum* to the *Ligustrum sinense* is 2:

1.

4. The method for preparing the traditional Chinese medicine disinfectant composition according to any one of claims 1-3, characterized in that, The specific steps include: The *Ligustrum lucidum* and *Hylocereus undatus* are mixed and pulverized to obtain a traditional Chinese medicine mixture; The herbal mixture was extracted with water under reduced pressure and reflux, then filtered. The resulting aqueous extract was concentrated and dried to obtain the herbal disinfectant composition.

5. The preparation method according to claim 4, characterized in that, The working conditions for the reduced pressure reflux extraction include: pressure of 8 kPa to 15 kPa; temperature of 46℃ to 59℃; and the number of reduced pressure reflux extractions of 1 to 3 times.

6. The preparation method according to claim 4, characterized in that: The weight ratio of the herbal medicine mixture to the water is 1:8~12.

7. The application of the traditional Chinese medicine disinfectant composition as described in any one of claims 1-3 or the traditional Chinese medicine disinfectant composition prepared by the preparation method as described in any one of claims 4-6 in the preparation of antibacterial and disinfectant products.

8. The use of the traditional Chinese medicine disinfectant composition as described in any one of claims 1-3 or the traditional Chinese medicine disinfectant composition prepared by the preparation method as described in any one of claims 4-6 in the preparation of anti-H1N1 virus drugs.

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