Preparation process and application of tobacco extract skin bacteriostatic spray

Abstract

The invention discloses a preparation process and application of a tobacco extract skin bacteriostatic spray. The skin bacteriostatic spray comprises tobacco, a gastrodia elata extract, modified chitosan, a humectant and a stabilizer. The preparation process comprises the steps of extraction of nicotine and a scutellaria baicalensis extract, modification of chitosan, preparation of the skin antibacterial spray and the like. Nicotine in tobacco and a radix scutellariae extract are used as main antibacterial components of the antibacterial spray, modified chitosan is modified and then grafted with carboxyl containing a carbon-carbon double bond, the modified chitosan and the nicotine can be subjected to an addition reaction, chitosan is used for film forming, the antibacterial time is prolonged, the nicotine is fixed, stimulation is reduced, and the antibacterial effect is improved. The spray has the advantages that the problems that the traditional spray is short in time efficiency and nicotine is irritant to human bodies are solved, a good antibacterial effect on common pathogenic bacteria such as staphylococcus aureus, escherichia coli and candida albicans is achieved, the antibacterial time is long, the irritation to skin is little, and the spray can be widely applied to daily skin care and bacteriostasis.

Description

Technical Field

[0001] This invention relates to the field of antibacterial skin sprays and their production technology, and particularly to the preparation process and application of a tobacco extract antibacterial skin spray. Background Technology

[0002] As the body's largest defense organ, the skin is susceptible to microbial invasion and infection. Therefore, highly effective and safe skin antibacterial agents have become a research hotspot in the pharmaceutical and daily chemical industries. While traditional chemically synthesized antibacterial agents such as benzalkonium chloride have significant antibacterial effects, long-term use can easily lead to skin barrier damage, allergic reactions, and bacterial resistance, failing to meet consumers' demand for gentle products. Natural plant extracts, due to their high biocompatibility and broad antibacterial spectrum, are gradually becoming the preferred alternative to chemical antibacterial agents.

[0003] Tobacco, an economic crop rich in alkaloids, polyphenols, and fiber, contains extracts whose alkaloids and polyphenols exhibit potent inhibitory effects against common pathogenic bacteria such as Staphylococcus aureus and Escherichia coli. The antibacterial mechanism is clearly defined—it achieves its bactericidal effect by disrupting the integrity of bacterial cell membranes. However, nicotine's high volatility, poor thermal stability, and irritant properties to the skin and mucous membranes when applied directly severely limit its application in topical formulations. Chlorogenic acid, a polyphenol, is easily photodegraded, degrading by 30% after 10 days at 60°C. Therefore, improving the stability and reducing the irritation of nicotine and chlorogenic acid through formulation technology has become a core challenge in the development of tobacco extracts.

[0004] In existing technologies, natural antibacterial agents generally suffer from drawbacks such as insufficient component stability, short antibacterial duration, and high irritation. For example, simple tobacco extract solutions are prone to decreased antibacterial efficacy due to nicotine volatilization; they also lack targeted soothing ingredients, making it difficult to alleviate the potential skin irritation caused by nicotine. Therefore, developing a composite antibacterial spray with tobacco extract as the main component, synergistic effects of Scutellaria baicalensis extract and chlorhexidine acetate, combined with modified carrier materials, soothing ingredients, and a stabilizing system to achieve a highly effective antibacterial-soothing synergistic effect has become an urgent technical problem to be solved in this field. Summary of the Invention

[0005] The purpose of this invention is to address the shortcomings of existing technologies by proposing a preparation process and application of a tobacco extract skin antibacterial spray.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] A tobacco extract skin antibacterial spray, comprising the following components in parts by weight:

[0008] 1-5 parts of tobacco extract A: an aqueous solution of alkaloids, including 10% nicotine, 1% nornicotine, 0.25% neonicotine and the balance water;

[0009] 15-20 parts of tobacco extract B: an aqueous solution of polyphenols, including 20-25% chlorogenic acid, 3-5% quercetin, 0.1-0.5% ascorbic acid and the balance water;

[0010] 10-25 parts Scutellaria baicalensis extract: a mixture of Scutellaria baicalensis including 25% baicalin, 5% baicalein, 5% quercetin and the balance water;

[0011] 1-5 parts modified chitosan: chitosan with surface-grafted carboxyl groups;

[0012] 1-5 parts moisturizer: glycerin;

[0013] 0.1-0.5 parts of excipient: chlorhexidine acetate;

[0014] 0.01-0.5 parts stabilizer: disodium ethylenediaminetetraacetate and citric acid;

[0015] 40-70 parts solvent: deionized water.

[0016] Preferably, the degree of deacetylation of the chitosan is ≥85%.

[0017] This invention also proposes a preparation process for the aforementioned tobacco extract antibacterial skin spray, comprising the following steps:

[0018] S1, extraction of tobacco and Scutellaria baicalensis extracts:

[0019] S101. Mix tobacco powder with 60%-80% ethanol solution at a volume ratio of 1:10-1:20, extract ultrasonically at 40-50℃ for 30-60 min, centrifuge for 10-15 min, and purify the supernatant by passing it through a macroporous adsorption resin column. First, wash the macroporous adsorption resin with deionized water 2-3 times, then elute with 0.08 mol / L hydrochloric acid. Collect the eluent and concentrate it by vacuum evaporation at 40-50℃ to obtain tobacco extract A.

[0020] S102. The resin effluent from S101 is purified by LX-8 macroporous adsorption resin, eluted with 50% ethanol (v / v), concentrated by vacuum distillation at 40°C, and 0.1-0.5 wt% ascorbic acid is added. The solution is then stored in the dark to obtain tobacco extract B.

[0021] S103. Take Scutellaria baicalensis slices, add water at a liquid-to-solid ratio of 13:1, extract at 60°C in a reflux device for 2 hours, filter out the filtrate, and repeat the extraction of the residue at the same liquid-to-solid ratio. Combine the two filtrates, centrifuge, and concentrate the supernatant to obtain Scutellaria baicalensis extract.

[0022] S2, Modification of chitosan:

[0023] S201. Freeze-dry tobacco extract A, dissolve it in dichloromethane at a ratio of 1:10-15, gradually add trans-2-butenoyl chloride and triethylamine in a molar ratio of 1:1.1:1.1 with tobacco, react at room temperature for 2-4 hours, add 3 times the volume of ice water, transfer to a separatory funnel and let stand for separation, and keep the organic phase for later use.

[0024]

[0025] S202. Take the organic phase from S201, add triethylamine to adjust the pH to 8.0-10.0, and gradually add an equal amount of chitosan powder to the tobacco extract at 30℃ while stirring. After the addition is complete, continue stirring and react for 1-2 hours. Filter, take the supernatant and distill it under reduced pressure at 40-60℃, wash it 2-3 times with anhydrous diethyl ether, and dry it under vacuum at 60℃ to obtain tobacco-chitosan powder.

[0026]

[0027] S3. Preparation of antibacterial skin spray:

[0028] Add 0.01-0.5% of disodium ethylenediaminetetraacetate and citric acid 1:1 compounding agent to deionized water, 1-5% of tobacco-chitosan powder, and stir for 10-20 minutes. Add 15-20% of tobacco extract B, 10-25% of Scutellaria baicalensis extract, and 0.1-0.5% of chlorhexidine acetate, and stir for 15-20 minutes. Then add 1-5% of glycerol, and stir for 20-30 minutes. Filter the solution and dispense it into brown spray bottles, sealing them with caps.

[0029] Preferably, the D-101 type macroporous resin in S101 performs highly selective adsorption of alkaloids through a synergistic effect of nonpolar hydrophobic interactions and pore sieving effect. Among the adsorbed alkaloids, 95% are nicotine, with less nornicotine and neonicotine.

[0030] Preferably, the reduced pressure evaporation operation in S101 serves two purposes: firstly, it lowers the boiling point of the ethanol-water mixed solvent, allowing ethanol and hydrochloric acid to fully evaporate; secondly, it prevents nicotine from being degraded due to high temperatures.

[0031] Preferably, after purification by LX-8 macroporous adsorption resin in S102, most of the polyphenolic substances are adsorbed, of which 80-95% are chlorogenic acid, 5%-10% are quercetin, and a small amount of flavonoid glycosides are also adsorbed. Chlorogenic acid will generate oxygen free radicals after being exposed to light. These oxygen free radicals will attack the double bonds and phenolic hydroxyl groups in chlorogenic acid, causing chlorogenic acid degradation. A small amount of ascorbic acid can be added to the extract. The enol structure in ascorbic acid has high activity and will react with free radicals to inhibit the degradation of chlorogenic acid. Furthermore, the subsequent product is packaged in a light-proof container.

[0032] Preferably, in step S103, distillation is used to extract volatile flavonoids from Scutellaria baicalensis tablets, mainly baicalin, baicalein, and quercetin.

[0033] Preferably, in S201, the tobacco extract is freeze-dried to remove water to obtain tobacco extract powder. The more active N in the nicotine molecule reacts with trans-2-butenoyl chloride under anhydrous conditions, and the hydrogen atom is replaced by an acyl group. Triethylamine acts as an acid-binding agent to neutralize the generated HCl and prevent product interference.

[0034] Preferably, the purpose of adding ice water in S201 is to dissolve excess acyl chloride and HCl, and to separate the organic phase and aqueous phase by allowing the mixture to stand in a separatory funnel.

[0035] Preferably, the purpose of adding triethylamine to S202 is to adjust the pH to ensure the alkalinity required for the nucleophilic addition reaction. Ethanol and most of the triethylamine are removed in the subsequent vacuum distillation, residual triethylamine is removed during vacuum drying, and free nicotine solution is removed by anhydrous diethyl ether, further reducing the irritation of the spray to the human body.

[0036] Preferably, the purpose of the 1:1 compounding agent of disodium ethylenediaminetetraacetate and citric acid in the deionized water system at a total amount of 0.01-0.5% in S3 is to adjust the pH between 5.0 and 6.0. On the one hand, it maintains the acidity and alkalinity of the spray, making it suitable for the skin, and on the other hand, it acts as a chitosan stabilizer to dissolve carboxylic acid chitosan.

[0037] Preferably, after the components in S302 are stirred evenly, they need to be filtered to remove undissolved carboxyl-containing chitosan powder to prevent clogging of the spray nozzle. When dispensing, brown bottles are used for storage to prevent nicotine and rose essential oil from degrading and losing their activity.

[0038] The present invention also proposes a tobacco extract skin antibacterial spray prepared by the aforementioned preparation method. First, natural antibacterial components are extracted, and then chitosan is modified. The tobacco extract and chitosan are subjected to nucleophilic addition, and the effective components in the tobacco extract are grafted onto the surface of chitosan to form a macromolecular structure that is not easily absorbed by the human body. Then, other auxiliary components and stabilizers are mixed and stirred evenly to obtain the tobacco extract skin antibacterial spray.

[0039] In addition, this invention also applies the aforementioned tobacco extract antibacterial skin spray to daily skin surface antibacterial care. Before use, a facial and local skin allergy test is required. Shake the spray bottle well before use, spray evenly from a distance of 10-15cm from the skin, pressing 2-3 times each time, and allow to air dry. It is suitable for daily skin surface antibacterial care, outdoor skin protection, post-exercise skin cleaning and antibacterial care, and gentle antibacterial care around wounds.

[0040] Compared with the prior art, the beneficial effects of the present invention are:

[0041] 1. Existing technologies for the direct application of tobacco extracts suffer from problems such as strong irritation, short duration of action, and weak antibacterial effect. This invention utilizes D-101 and LX-8 macroporous resins to purify and efficiently extract the active ingredients, combined with the soothing and moisturizing effects of glycerin, reducing the risk of skin irritation. Simultaneously, a carboxyl-containing chitosan carrier forms a stable compound with nicotine through nucleophilic addition, achieving controlled release of the antibacterial components, overcoming the short duration of antibacterial action in traditional natural extract preparations, and reducing the harm of free nicotine to the human body, significantly reducing skin irritation. Alkaloids in tobacco extracts interfere with the synthesis of peptidoglycans in cell walls, thereby disrupting cell wall integrity. Chlorogenic acid, quercetin, and baicalin in baicalein extract can insert their phenolic hydroxyl groups into the phospholipid bilayer of bacterial cell membranes via hydrophobic interactions, binding to lipids or proteins on the membrane, disrupting membrane integrity and fluidity, restricting cell metabolism, and inhibiting nucleic acid synthesis to suppress bacterial growth, showing significant inhibitory effects against Staphylococcus aureus. Chlorhexidine acetate tightly adsorbs to the bacterial surface through electrostatic attraction and inserts into the bacterial cell membrane via hydrophobic interactions. The lipid bilayer of the cell membrane enhances membrane permeability, assisting the extract in entering the bacterial cell and exhibiting highly efficient and broad-spectrum antibacterial properties. Chitosan, as a film-forming agent, forms a thin film on the body surface, which on the one hand isolates some bacteria, and on the other hand covalently binds with nicotine in the extract to form a macromolecule, reducing the entry of nicotine into the human body, reducing toxicity, and prolonging the antibacterial duration of the spray. These four components form a synergistic effect: alkaloids destroy cell walls, flavonoids inhibit metabolism, chlorhexidine acetate penetrates cells, assists the components in entering cells, and chitosan prolongs the antibacterial duration.

[0042] 2. Existing chitosan-based formulations generally suffer from poor water solubility and sticky film formation. This invention addresses these issues by combining disodium EDTA with a citric acid buffer system, which effectively chelates metal ions and maintains pH stability (5.0-6.0), enhancing the stability of the spray solution and extending its shelf life. The large-molecule chitosan fixes nicotinic acid molecules, preventing free nicotinic acid from entering the body. Chitosan itself forms a film quickly and exhibits excellent breathability after film formation. The addition of glycerin moisturizing ingredients avoids the skin dryness problem caused by traditional antibacterial agents, improving user comfort. Detailed Implementation

[0043] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments.

[0044] Example 1:

[0045] A preparation process for a tobacco extract skin antibacterial spray includes the following steps:

[0046] S1, extraction of tobacco and Scutellaria baicalensis extracts:

[0047] S101. Mix tobacco powder with 75% ethanol solution at a volume ratio of 1:15, extract ultrasonically at 45℃ for 45 min, centrifuge for 10 min, and purify the supernatant by passing it through a macroporous adsorption resin column. First, wash the macroporous adsorption resin with deionized water 2-3 times, then elute with 0.08 mol / L hydrochloric acid. Collect the eluent and concentrate it by vacuum evaporation at 50℃ to obtain tobacco extract A.

[0048] S102. The resin effluent from S101 is purified by LX-8 macroporous adsorption resin, eluted with 50% ethanol (v / v), concentrated by vacuum distillation at 40°C, and 0.3 wt% ascorbic acid is added. The solution is then stored in the dark to obtain tobacco extract B.

[0049] S103. Take Scutellaria baicalensis slices, add water at a liquid-to-solid ratio of 13:1, extract at 60°C in a reflux device for 2 hours, filter out the filtrate, and repeat the extraction of the residue at the same liquid-to-solid ratio. Combine the two filtrates, centrifuge, and concentrate the supernatant to obtain Scutellaria baicalensis extract.

[0050] S2, Modification of chitosan:

[0051] S201. The tobacco extract A is freeze-dried and dissolved in dichloromethane at a ratio of 1:10. Trans-2-butenoyl chloride and triethylamine are gradually added in a molar ratio of 1:1.1:1.1 to tobacco. After reacting at room temperature for 3 hours, 3 times the volume of ice water is added. The mixture is then transferred to a separatory funnel and allowed to stand for separation. The organic phase is reserved for later use.

[0052] S202. Take the organic phase from S201, add triethylamine to adjust the pH to 9.0, and gradually add an equal amount of chitosan powder to the tobacco extract at 30°C while stirring. After the addition is complete, continue stirring and react for 1.5 hours. Filter, take the supernatant and distill it under reduced pressure at 50°C. Wash it 2-3 times with anhydrous ether and dry it under vacuum at 60°C to obtain tobacco-chitosan powder.

[0053] S3. Preparation of antibacterial skin spray:

[0054] Add 0.3% of disodium ethylenediaminetetraacetate and citric acid 1:1 compounding agent to deionized water, 1% of tobacco-chitosan powder, and stir for 20 min. Add 18% of tobacco extract B, 15% of Scutellaria baicalensis extract, and 0.3% of chlorhexidine acetate, and stir for 15 min. Then add 1-5% of glycerol, stir for 20 min, filter, and dispense the solution into brown spray bottles and seal with caps.

[0055] Example 2:

[0056] A preparation process for a tobacco extract skin antibacterial spray includes the following steps:

[0057] S1, extraction of tobacco and Scutellaria baicalensis extracts:

[0058] S101. Mix tobacco powder with 75% ethanol solution at a volume ratio of 1:15, extract ultrasonically at 45℃ for 45 min, centrifuge for 10 min, and purify the supernatant by passing it through a macroporous adsorption resin column. First, wash the macroporous adsorption resin with deionized water 2-3 times, then elute with 0.08 mol / L hydrochloric acid. Collect the eluent and concentrate it by vacuum evaporation at 50℃ to obtain tobacco extract A.

[0059] S102. The resin effluent from S101 is purified by LX-8 macroporous adsorption resin, eluted with 50% ethanol (v / v), concentrated by vacuum distillation at 40°C, and 0.3 wt% ascorbic acid is added. The solution is then stored in the dark to obtain tobacco extract B.

[0060] S103. Take Scutellaria baicalensis slices, add water at a liquid-to-solid ratio of 13:1, extract at 60°C in a reflux device for 2 hours, filter out the filtrate, and repeat the extraction of the residue at the same liquid-to-solid ratio. Combine the two filtrates, centrifuge, and concentrate the supernatant to obtain Scutellaria baicalensis extract.

[0061] S2, Modification of chitosan:

[0062] S201. The tobacco extract A is freeze-dried and dissolved in dichloromethane at a ratio of 1:10. Trans-2-butenoyl chloride and triethylamine are gradually added in a molar ratio of 1:1.1:1.1 to tobacco. After reacting at room temperature for 3 hours, 3 times the volume of ice water is added. The mixture is then transferred to a separatory funnel and allowed to stand for separation. The organic phase is reserved for later use.

[0063] S202. Take the organic phase from S201, add triethylamine to adjust the pH to 9.0, and gradually add an equal amount of chitosan powder to the tobacco extract at 30°C while stirring. After the addition is complete, continue stirring and react for 1.5 hours. Filter, take the supernatant and distill it under reduced pressure at 50°C. Wash it 2-3 times with anhydrous ether and dry it under vacuum at 60°C to obtain tobacco-chitosan powder.

[0064] S3. Preparation of antibacterial skin spray:

[0065] Add 0.3% of the total amount of disodium ethylenediaminetetraacetate and citric acid 1:1 compounding agent to deionized water, add 5% of the total amount of tobacco-chitosan powder, stir for 20 min, add 18% of the total amount of tobacco extract B, 15% of scutellaria baicalensis extract and 0.3% of the total amount of chlorhexidine acetate, stir for 15 min, then add 1-5% of the total amount of glycerol to the system, stir for 20 min, filter, dispense the solution into brown spray bottles, and seal with caps.

[0066] Example 3:

[0067] A preparation process for a tobacco extract skin antibacterial spray includes the following steps:

[0068] S1, extraction of tobacco and Scutellaria baicalensis extracts:

[0069] S101. Mix tobacco powder with 75% ethanol solution at a volume ratio of 1:15, extract ultrasonically at 45℃ for 45 min, centrifuge for 10 min, and purify the supernatant by passing it through a macroporous adsorption resin column. First, wash the macroporous adsorption resin with deionized water 2-3 times, then elute with 0.08 mol / L hydrochloric acid. Collect the eluent and concentrate it by vacuum evaporation at 50℃ to obtain tobacco extract A.

[0070] S102. The resin effluent from S101 is purified by LX-8 macroporous adsorption resin, eluted with 50% ethanol (v / v), concentrated by vacuum distillation at 40°C, and 0.3 wt% ascorbic acid is added. The solution is then stored in the dark to obtain tobacco extract B.

[0071] S103. Take Scutellaria baicalensis slices, add water at a liquid-to-solid ratio of 13:1, extract at 60°C in a reflux device for 2 hours, filter out the filtrate, and repeat the extraction of the residue at the same liquid-to-solid ratio. Combine the two filtrates, centrifuge, and concentrate the supernatant to obtain Scutellaria baicalensis extract.

[0072] S2, Modification of chitosan:

[0073] S201. The tobacco extract A is freeze-dried and dissolved in dichloromethane at a ratio of 1:10. Trans-2-butenoyl chloride and triethylamine are gradually added in a molar ratio of 1:1.1:1.1 to tobacco. After reacting at room temperature for 3 hours, 3 times the volume of ice water is added. The mixture is then transferred to a separatory funnel and allowed to stand for separation. The organic phase is reserved for later use.

[0074] S202. Take the organic phase from S201, add triethylamine to adjust the pH to 9.0, and gradually add an equal amount of chitosan powder to the tobacco extract at 30°C while stirring. After the addition is complete, continue stirring and react for 1.5 hours. Filter, take the supernatant and distill it under reduced pressure at 50°C. Wash it 2-3 times with anhydrous ether and dry it under vacuum at 60°C to obtain tobacco-chitosan powder.

[0075] S3. Preparation of antibacterial skin spray:

[0076] Add 0.3% of the total amount of disodium ethylenediaminetetraacetate and citric acid 1:1 compounding agent to deionized water, add 3% of the total amount of tobacco-chitosan powder, stir for 20 min, add 18% of the total amount of tobacco extract B, 15% of scutellaria baicalensis extract and 0.1% of the total amount of chlorhexidine acetate, stir for 15 min, then add 1-5% of the total amount of glycerol to the system, stir for 20 min, filter, dispense the solution into brown spray bottles, and seal with caps.

[0077] Example 4:

[0078] A preparation process for a tobacco extract skin antibacterial spray includes the following steps:

[0079] S1, extraction of tobacco and Scutellaria baicalensis extracts:

[0080] S101. Mix tobacco powder with 75% ethanol solution at a volume ratio of 1:15, extract ultrasonically at 45℃ for 45 min, centrifuge for 10 min, and purify the supernatant by passing it through a macroporous adsorption resin column. First, wash the macroporous adsorption resin with deionized water 2-3 times, then elute with 0.08 mol / L hydrochloric acid. Collect the eluent and concentrate it by vacuum evaporation at 50℃ to obtain tobacco extract A.

[0081] S102. The resin effluent from S101 is purified by LX-8 macroporous adsorption resin, eluted with 50% ethanol (v / v), concentrated by vacuum distillation at 40°C, and 0.3 wt% ascorbic acid is added. The solution is then stored in the dark to obtain tobacco extract B.

[0082] S103. Take Scutellaria baicalensis slices, add water at a liquid-to-solid ratio of 13:1, extract at 60°C in a reflux device for 2 hours, filter out the filtrate, and repeat the extraction of the residue at the same liquid-to-solid ratio. Combine the two filtrates, centrifuge, and concentrate the supernatant to obtain Scutellaria baicalensis extract.

[0083] S2, Modification of chitosan:

[0084] S201. The tobacco extract A is freeze-dried and dissolved in dichloromethane at a ratio of 1:10. Trans-2-butenoyl chloride and triethylamine are gradually added in a molar ratio of 1:1.1:1.1 to tobacco. After reacting at room temperature for 3 hours, 3 times the volume of ice water is added. The mixture is then transferred to a separatory funnel and allowed to stand for separation. The organic phase is reserved for later use.

[0085] S202. Take the organic phase from S201, add triethylamine to adjust the pH to 9.0, and gradually add an equal amount of chitosan powder to the tobacco extract at 30°C while stirring. After the addition is complete, continue stirring and react for 1.5 hours. Filter, take the supernatant and distill it under reduced pressure at 50°C. Wash it 2-3 times with anhydrous ether and dry it under vacuum at 60°C to obtain tobacco-chitosan powder.

[0086] S3. Preparation of antibacterial skin spray:

[0087] Add 0.3% of the total amount of disodium ethylenediaminetetraacetate and citric acid 1:1 compounding agent to deionized water, add 3% of the total amount of tobacco-chitosan powder, stir for 20 min, add 18% of the total amount of tobacco extract B, 15% of scutellaria baicalensis extract and 0.5% of the total amount of chlorhexidine acetate, stir for 15 min, then add 1-5% of the total amount of glycerol to the system, stir for 20 min, filter, dispense the solution into brown spray bottles, and seal with caps.

[0088] Example 5:

[0089] A preparation process for a tobacco extract skin antibacterial spray includes the following steps:

[0090] S1, extraction of tobacco and Scutellaria baicalensis extracts:

[0091] S101. Mix tobacco powder with 75% ethanol solution at a volume ratio of 1:15, extract ultrasonically at 45℃ for 45 min, centrifuge for 10 min, and purify the supernatant by passing it through a macroporous adsorption resin column. First, wash the macroporous adsorption resin with deionized water 2-3 times, then elute with 0.08 mol / L hydrochloric acid. Collect the eluent and concentrate it by vacuum evaporation at 50℃ to obtain tobacco extract A.

[0092] S102. The resin effluent from S101 is purified by LX-8 macroporous adsorption resin, eluted with 50% ethanol (v / v), concentrated by vacuum distillation at 40°C, and 0.3 wt% ascorbic acid is added. The solution is then stored in the dark to obtain tobacco extract B.

[0093] S103. Take Scutellaria baicalensis slices, add water at a liquid-to-solid ratio of 13:1, extract at 60°C in a reflux device for 2 hours, filter out the filtrate, and repeat the extraction of the residue at the same liquid-to-solid ratio. Combine the two filtrates, centrifuge, and concentrate the supernatant to obtain Scutellaria baicalensis extract.

[0094] S2, Modification of chitosan:

[0095] S201. The tobacco extract A is freeze-dried and dissolved in dichloromethane at a ratio of 1:10. Trans-2-butenoyl chloride and triethylamine are gradually added in a molar ratio of 1:1.1:1.1 to tobacco. After reacting at room temperature for 3 hours, 3 times the volume of ice water is added. The mixture is then transferred to a separatory funnel and allowed to stand for separation. The organic phase is reserved for later use.

[0096] S202. Take the organic phase from S201, add triethylamine to adjust the pH to 9.0, and gradually add an equal amount of chitosan powder to the tobacco extract at 30°C while stirring. After the addition is complete, continue stirring and react for 1.5 hours. Filter, take the supernatant and distill it under reduced pressure at 50°C. Wash it 2-3 times with anhydrous ether and dry it under vacuum at 60°C to obtain tobacco-chitosan powder.

[0097] S3. Preparation of antibacterial skin spray:

[0098] Add 0.3% of the total amount of disodium ethylenediaminetetraacetate and citric acid 1:1 compounding agent to deionized water, add 3% of the total amount of tobacco-chitosan powder, stir for 20 min, add 18% of the total amount of tobacco extract B, 15% of Scutellaria baicalensis extract and 0.3% of the total amount of chlorhexidine acetate, stir for 15 min, then add 1-5% of the total amount of glycerol to the system, stir for 20 min, filter, dispense the solution into brown spray bottles, and seal with caps.

[0099] Comparative Example 1:

[0100] A preparation process for a tobacco extract skin antibacterial spray includes the following steps:

[0101] S1, extraction of tobacco and Scutellaria baicalensis extracts:

[0102] S101. Mix tobacco powder with 75% ethanol solution at a volume ratio of 1:15, extract ultrasonically at 45℃ for 45 min, centrifuge for 10 min, and purify the supernatant by passing it through a macroporous adsorption resin column. First, wash the macroporous adsorption resin with deionized water 2-3 times, then elute with 0.08 mol / L hydrochloric acid. Collect the eluent and concentrate it by vacuum evaporation at 50℃ to obtain tobacco extract A.

[0103] S102. The resin effluent from S101 is purified by LX-8 macroporous adsorption resin, eluted with 50% ethanol (v / v), concentrated by vacuum distillation at 40°C, and 0.3 wt% ascorbic acid is added. The solution is then stored in the dark to obtain tobacco extract B.

[0104] S103. Take Scutellaria baicalensis slices, add water at a liquid-to-solid ratio of 13:1, extract at 60°C in a reflux device for 2 hours, filter out the filtrate, and repeat the extraction of the residue at the same liquid-to-solid ratio. Combine the two filtrates, centrifuge, and concentrate the supernatant to obtain Scutellaria baicalensis extract.

[0105] S2, Modification of chitosan:

[0106] S201. The tobacco extract A is freeze-dried and dissolved in dichloromethane at a ratio of 1:10. Trans-2-butenoyl chloride and triethylamine are gradually added in a molar ratio of 1:1.1:1.1 to tobacco. After reacting at room temperature for 3 hours, 3 times the volume of ice water is added. The mixture is then transferred to a separatory funnel and allowed to stand for separation. The organic phase is reserved for later use.

[0107] S202. Take the organic phase from S201 and gradually add an equal amount of chitosan powder to the tobacco extract at 30°C while stirring. After the addition is complete, continue stirring and react for 1.5 hours. Filter and take the supernatant for vacuum distillation at 50°C. Wash with anhydrous ether 2-3 times and vacuum dry at 60°C to obtain tobacco-chitosan powder.

[0108] S3. Preparation of antibacterial skin spray:

[0109] Add 0.3% of the total amount of disodium ethylenediaminetetraacetate and citric acid 1:1 compounding agent to deionized water, add 3% of the total amount of tobacco-chitosan powder, stir for 20 min, add 18% of the total amount of tobacco extract B, 15% of Scutellaria baicalensis extract and 0.3% of the total amount of chlorhexidine acetate, stir for 15 min, then add 1-5% of the total amount of glycerol to the system, stir for 20 min, filter, dispense the solution into brown spray bottles, and seal with caps.

[0110] Comparative Example 2:

[0111] Compared to Example 5, Comparative Example 2 added 7% of the total amount of tobacco-chitosan powder to S302.

[0112] Comparative Example 3:

[0113] Compared to Example 5, Comparative Example 3 added 0.7% chlorhexidine acetate to S302, which is the total amount of the system.

[0114] Performance testing:

[0115] 1. Antibacterial zone experiment of tobacco extract skin antibacterial spray

[0116] Sterile filter paper discs soaked in the spray were placed on agar medium inoculated with Staphylococcus aureus, Escherichia coli, and Candida albicans, respectively, and incubated at 36.5℃ for 36 hours. The distance from the edge of the filter paper disc to the edge of the inhibition zone was measured.

[0117] 2. Antibacterial duration experiment of tobacco extract skin antibacterial spray

[0118] After cleaning the skin, spray the product and allow it to air dry. Take samples every 12 hours using a sterile cotton swab, dilute and culture the samples, and count the number of colonies. The control group did not use the antibacterial spray and samples were taken.

[0119] 3. Skin irritation test

[0120] A three-dimensional skin model constructed from human keratinocytes was exposed to the spray for 36 hours, and the activity of mitochondrial dehydrogenase in live cells was measured at 12-hour intervals. The control group was not treated with the spray.

[0121] Relative activity (%) = (Experimental group activity / Control group activity) × 100%

[0122] Table 1. Experimental data on the antibacterial zone of tobacco extract skin antibacterial spray.

[0123]

[0124] Table 2. Experimental data on the antibacterial duration of tobacco extract skin antibacterial spray.

[0125]

[0126] Table 3. Data from skin irritation tests

[0127] Data Analysis:

[0128] After ultrasonic extraction, most of the ethanolic substances are extracted. Following adsorption with macroporous resin, the vast majority of alkaloids are adsorbed onto the resin column. After elution with hydrochloric acid, the hydrochloric acid is removed and the extract is concentrated, resulting in a tobacco extract with a high alkaloid concentration. Alkaloids account for 2.2% of the total tobacco content, with nicotine accounting for 95%. Therefore, the majority of alkaloids are nicotine. Polyphenols account for 2-8% of the total tobacco content, with chlorogenic acid accounting for 80-95% of the polyphenols. After distillation extraction at 60℃, the volatile flavonoids in Scutellaria baicalensis tablets dissolve in water, accounting for approximately 8-15% of the tablets' mass. Although flavonoids are polyphenols, they are virtually harmless to the human body and can have antiviral, anti-inflammatory, and anti-allergic effects. Chlorhexidine acetate is a common antibacterial drug; it has significant antibacterial effects in trace amounts, but excessive amounts may cause irritation to the human body.

[0129] Compared to Example 5, Example 1 reduced the amount of tobacco extract added, while Example 2 increased the content of tobacco extract compared to Example 5. The tobacco extract, in the antibacterial spray, exhibits a synergistic antibacterial effect with Scutellaria baicalensis extract, chlorhexidine acetate, and chitosan. Table 1 shows that the diameter of the inhibition zone in the control group was smaller than that in Example 1, which was smaller than that in Example 5, which was smaller than that in Example 2, indicating that the tobacco extract has an inhibitory effect on Staphylococcus aureus, Escherichia coli, and Candida albicans; the higher the content, the more significant the inhibitory effect. Table 2 shows the duration of antibacterial activity of the spray. Within the same time frame, the colony count in the control group was higher than in other groups. After 48 hours, the colony count in Example 1 was higher than that in Example 5, which was higher than that in Example 2, indicating that the colony count is inversely proportional to the content of tobacco extract. Table 3 shows the relative cell activity within 36 hours, used to determine the degree of irritation of the spray to cells. According to the data in Table 3, the relative cell activity in Example 1 was greater than that in Example 2, which was greater than that in Example 3, but the difference was not significant, indicating that the components in the tobacco extract have a certain irritant effect on cells, but this effect is not obvious during the application period. Nicotine, a major component of tobacco extracts, can enter the human body in its free state. A single intake exceeding 40-60 mg (approximately the total nicotine content of one cigarette) can cause nausea, vomiting, headache, and convulsions. In severe cases, it can suppress the respiratory center, leading to death. It can also bind to nicotine acetylcholine receptors in brain neurons, resulting in chronic addiction and dependence. This invention utilizes a modified chitosan-nicotine addition reaction to immobilize nicotine on chitosan, and removes free nicotine using ether, significantly reducing the damage caused by free nicotine to the human body. Long-term continuous use is prohibited.

[0130] Compared to Example 5, Example 3 contained 0.1% chlorhexidine acetate, and Example 4 contained 0.5% chlorhexidine acetate. Data from Tables 1-3 show that the antibacterial effect of the control group was significantly lower than that of Example 1, which was lower than that of Example 5, which was lower than that of Example 4. This indicates that chlorhexidine acetate exerts an antibacterial effect in the spray. The antibacterial performance and irritation of Examples 3, 4, and 5 also showed a gradient increase at the same time point. Chlorhexidine acetate is a broad-spectrum cationic antibacterial agent. Due to its strong antibacterial efficacy and long-lasting effect, direct contact of high-concentration (>1%) solutions with the skin may cause erythema and itching. Long-term use of chlorhexidine-containing sprays may lead to skin barrier damage. The slow-release effect of modified chitosan regulates the concentration of chlorhexidine acetate, preventing a sudden increase and reducing skin irritation.

[0131] Compared to Example 5, Comparative Example 1 did not add additional triethylamine solution in S202. Data from Tables 1, 2, and 3 show that the antibacterial effect was moderate with minimal irritation. In S202, triethylamine acts as a pH adjuster, ensuring the smooth nucleophilic reaction between the double bond and the primary amine. Without additional triethylamine, the pH was maintained by the remaining triethylamine in S201. In the early stages of the reaction, the pH was within a suitable range, allowing the nucleophilic reaction to proceed smoothly. However, in the middle and later stages, the pH exceeded the suitable range, hindering the nucleophilic reaction and resulting in a low nicotine grafting rate in chitosan, thus affecting the antibacterial effect. Comparative Example 2 increased the content of tobacco extract to 25%, and Comparative Example 3 increased the content of chlorhexidine acetate to 0.7% compared to Example 5. According to Table 1, both antibacterial sprays showed excellent antibacterial effects, maintaining strong antibacterial performance even after 48 hours of use; however, they also exhibited relatively high skin irritation. The content of active ingredients in the spray is directly proportional to the antibacterial effect. However, high concentrations can also cause strong irritation. Therefore, it is necessary to control the amount of tobacco extract and chlorhexidine acetate added within a reasonable range to achieve a balance between high antibacterial activity and low irritation.

[0132] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A skin antiseptic spray of tobacco extract, characterized in that, The following components by weight are included: 1-5 parts of tobacco extract A: an aqueous alkaloid solution including 10% nicotine, 1% nornicotine, 0.25% anabasine, and the balance water; 15-20 parts of tobacco extract B: an aqueous polyphenol solution including 20-25% chlorogenic acid, 3-5% quercetin, 0.1-0.5% ascorbic acid, and the balance water; 10-25 parts of scutellaria extract: a scutellaria mixture including 25% baicalin, 5% baicalein, 5% quercetin, and the balance water; 1-5 parts of modified chitosan: chitosan with carboxyl groups grafted on the surface; 1-5 parts of humectant: glycerol; 0.1-0.5 parts of auxiliary agent: chlorhexidine acetate; 0.01-0.5 parts of stabilizer: disodium ethylenediaminetetraacetate and citric acid; 40-70 parts of solvent: deionized water.

2. The skin bacteriostatic spray of claim 1, wherein, The degree of deacetylation of the chitosan is ≥ 85%.

3. The process for preparing a tobacco extract skin antiseptic spray as claimed in claim 1, wherein, The following steps are included: S1, extraction of tobacco and scutellaria extract: S101, tobacco powder is mixed with an ethanol solution having a volume fraction of 60%-80% at a volume ratio of 1:10-1:20, ultrasonic extraction is performed at a temperature of 40-50°C for 30-60 min, centrifugation is performed for 10-15 min, the supernatant is taken and purified by a macroporous adsorption resin column, the macroporous adsorption resin is first washed with deionized water for 2-3 times, then eluted with 0.08 mol / l hydrochloric acid, the eluate is collected, concentrated by evaporation under reduced pressure at 40-50°C, and tobacco extract A is obtained; S102, the resin effluent in S101 is purified by a polyphenol adsorption resin, eluted with 50% ethanol by volume, concentrated by evaporation under reduced pressure at 40°C, 0.1-0.5 wt% ascorbic acid is added, and stored in the dark to obtain tobacco extract B; S103, scutellaria tablets are taken, water is added at a liquid-solid ratio of 13:1, extraction is performed at 60°C in a reflux device for 2 h, the filtrate is filtered out, the residue is re-extracted at the same liquid-solid ratio, the two filtrates are combined, centrifuged, and the supernatant is concentrated to prepare scutellaria extract; S2, modification of chitosan: S201, tobacco extract A is freeze-dried, dissolved in dichloromethane at a solid-liquid ratio of 1:10-15, trans-2-butene chloride, triethylamine, and water are gradually added at a molar ratio of 1:1.1:1.1, the reaction is performed at room temperature for 2-4 h, 3 times the volume of ice water is added, transferred to a separatory funnel and allowed to stand to separate into layers, and the organic phase is reserved for later use; S202, the organic phase in S201 is taken, triethylamine is added to adjust the pH to 8.0-10.0, an equal amount of chitosan powder to the tobacco extract is gradually added at 30°C, stirring is performed while adding, after the addition is completed, stirring is continued to react for 1-2 h, filtered, and the supernatant is distilled under reduced pressure at 40-60°C, washed with anhydrous ether for 2-3 times, and dried at 60°C under vacuum to obtain tobacco-chitosan powder; S3, preparation of a skin bacteriostatic spray: In deionized water, add 0.01-0.5% of ethylenediaminetetraacetic acid disodium salt, citric acid 1:1 complexing agent, add 1-5% of tobacco-chitosan powder, stir for 10-20 min, add 15-20% of tobacco extract B, 10-25% of scutellaria extract and 0.1-0.5% of chlorhexidine acetate, stir for 15-20 min, add 1-5% of glycerol, stir for 20-30 min, filter, and then divide the solution into brown spray bottles and seal.

4. The process for preparing a tobacco extract skin antiseptic spray according to claim 3, characterized in that, In S101, the macroporous resin is D-101, the column size is D3 cm x H30 cm, the sample flow rate is 0.5 mL / min, after saturation, the resin column is washed with deionized water for 2-3 times, and then eluted with 6-10 resin column volumes of 0.08 mol / l hydrochloric acid; in S102, the polyphenol adsorption resin is LX-8 macroporous resin, and eluted with 6-10 resin column volumes of 50% ethanol solution.

5. The process for preparing a tobacco extract skin antiseptic spray according to claim 3, characterized in that, In S101-S103, the solution is concentrated to 1 / 4-1 / 8 of the original volume.

6. The process for preparing a tobacco extract skin antiseptic spray according to claim 3, characterized in that, In S201, the trans-2-butenoyl chloride is slowly added and stirred.

7. The process for preparing a tobacco extract skin antiseptic spray according to claim 3, characterized in that, In S202, the molar ratio of chitosan powder to tobacco is 1:1, and the amount of triethanolamine added is 10%-15% of the system.

8. The tobacco extract skin bacteriostatic spray prepared by the preparation process of claim 3 is applied to daily skin bacteriostatic care.

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