Novel polymer-based hemostatic formulations with antimicrobial, antiviral and anti-inflammatory properties and methods for their manufacture

Abstract

The present invention relates to a method for producing a gel formulation containing lavender (Lavandula spp.) oil that provides antimicrobial, antiviral, and anti-inflammatory activity and has hemostatic (bleeding-stopping) effects. The objective of the present invention is to develop a formulation that can be safely used in the treatment of minor bleeding injuries, emergency response situations, surgical incisions, and military injuries and burns.

Description

[Technical Field]

[0001] The present invention relates to polymer-based formulations (clotting gels and hemostats) that provide antimicrobial (antibacterial, antifungal, anticandida), antiviral, anti-inflammatory activity and contain lavender oil (Lavandula spp.) and have hemostatic (bleeding-stopping) effects. [Background technology]

[0002] Uncontrolled bleeding and wound infection are among the leading causes of death in medical wound care. Hemostasis is the cessation of bleeding by clotting or physical pressure. Due to its clotting properties, hemostasis is also considered the first step in wound healing. To prevent the formation of a blood clot, endothelial cells in intact blood vessels prevent blood clotting with heparin-like molecules and thrombomodulin and platelet aggregation with nitric oxide and prostacyclin. When the endothelium of a blood vessel is damaged, endothelial cells stop secreting coagulation and aggregation inhibitors and instead secrete von Willebrand factor, which initiates the maintenance of hemostasis after injury. Hemostasis is achieved through the coordinated action of three distinct mechanisms: vasoconstriction, platelet plug formation, and blood coagulation (fibrin formation from inactive fibrinogen protein) [1].

[0003] When the body's own mechanisms are not effective enough to stop bleeding, additional hemostatic agents must be applied. Currently, surgeons use different methods to achieve rapid hemostasis. Physical methods, such as manual pressure and suturing, may not be completely effective. Thermal methods, such as lasers, bipolar coagulation, and ultrasonic dissectors, may cause serious complications related to the development of infection and necrosis formation [2].

[0004] Hemostatic agents, sealants, and adhesives are used during surgical procedures to achieve hemostasis. Hemostatic agents vary widely in terms of their mechanism of action, composition, ease of application, adhesion to tissue, immunogenicity, and cost. These agents contribute to rapid hemostasis, better visualization of the surgical area, shorter surgical times, reduced need for blood transfusions, shorter wound healing times, and an overall improvement in patient recovery time. While some antihemorrhagic systemic drugs used in medicine inhibit fibrinolysis or promote coagulation, locally acting hemostatic agents act by causing vasoconstriction or promoting platelet aggregation [3].

[0005] A wide range of traditional materials, usually based on cellulose, collagen, and gelatin, are used for hemostatic applications [4]. However, in recent years, activated coagulants (chitosan or kaolin) have been incorporated into hemostatic materials to accelerate the blood clotting process and reduce bleeding [5]. In particular, many new fabrics loaded with modified sodium starch glycolate, chitosan hydroquinone-based gauzes, or other hemostatic agents have been introduced [2, 6]. The use of inorganic and organic nanofiber materials carrying inorganic components or metal ion-chelating tannic acid coatings, as well as other materials with antimicrobial and anti-inflammatory agents, has become widespread [7]. Essential oils derived from certain plants, which possess hydrophobic, pain-relieving, anti-inflammatory, antimicrobial, and antiviral properties, have been used to treat pain and bleeding caused by dental infections in humans [8].

[0006] Essential oils are widely used in traditional and alternative medicine for their anesthetic, antiseptic, astrengenic, hemostatic, deodorizing, diaphoretic, antiseptic, expectorant, antipyretic, fumigant, inhalant, insect repellent / repellent, sedative and stimulating properties, as well as anti-scar, anti-abscess, anti-arthritic, anti-asthmatic, anti-boil, anti-bronchitis, anti-burn, anti-cancer, anti-diabetic, anti-diarrheal, anti-diphtheria, anti-dysentery, anti-encephalitis, anti-enteritis, anti-erysipelas, anti-pyrexia, anti-influenza, anti-inflammatory, anti-sore throat, anti-laryngitis, anti-leprosy, anti-malarial, anti-mastitis, anti-miasma, anti-pharyngitis, anti-phytitis, anti-rhinitis, anti-wound, anti-sore throat, anti-spasmodic and anti-bronchial pain properties. In particular, recent research has scientifically proven the effectiveness of essential oils, making them highly sought after in many fields, from cleaning living spaces to the medical and beauty sectors.[9] Among these oils, lavender oil has been scientifically shown to be effective in many different areas over the years and is used in many different fields. In particular, lavender oil is used topically, orally, and via inhalation in many areas, including depression, anxiety, dementia, cardiovascular activity, carcinogenesis prevention, antimicrobial activity, and aromatherapy. In vitro studies have demonstrated the antibacterial effects of lavender oil, particularly against gram-negative microorganisms.

[10] Inhibition of the growth of methicillin-susceptible and methicillin-resistant Staphylococcus aureus (MSSA and MRSA) has been observed with various lavender oils and other essential oils (e.g., spike lavender, Lavandula latifolia, and French lavender, Lavandula stoechas) in combination

[11] . Other studies have reported potent and rapid antiparasitic activity against Giardia duodenalis and Trichomonas vaginalis, as well as both fungistatic and fungicidal activity against Candida albicans [12, 13]. After bacterial infection, aphthous ulcers take time to heal and the area repairs itself.In a rabbit study, 90% of animals with recurrent aphthous ulcerations treated with lavender oil showed complete healing by day 3, whereas redness, ulcers, and inflammation persisted in the placebo group. Histological examination in the same study showed that edematous and severe infiltration of acute and chronic inflammatory cells was observed in the placebo group, whereas tissues had a nearly normal appearance in the group treated with topical lavender oil. In a double-blind clinical trial of 115 patients with recurrent aphthous ulcerations, 100% of patients treated with topical 2% lavender gel showed improvement by day 4, whereas the placebo group continued to have ulcerations and pain.

[14]

[0007] The mechanism of antiviral activity of essential oils has been reported in the literature as the ability of essential oils and their components to inhibit capsid fragmentation, viral proliferation, and viral entry into host cells by cleaving the hemagglutinin protein in some viruses, or to inhibit later stages of the viral life cycle by targeting redox signaling pathways

[15] . When evaluating lavender oil for its antiviral activity, studies have shown its activity against herpes simplex types 1 and 2, influenza H1N1, and H5N1 viruses [16, 17]. Due to its biological activity demonstrated by in vitro, in vivo, and clinical studies, lavender oil has become a well-known candidate in the pharmaceutical and medical fields. For this reason, a wide range of products containing lavender oil, water, and active phenols have been introduced to the market in recent years. However, scientific research conducted in the literature has published many scientific papers on the anti-inflammatory, antimicrobial, and antiviral activities of plant essential oils derived from lavender species. However, the hemostatic properties of lavender seed oil have not been known until now. Within the scope of this study, it was determined for the first time that lavender oil is a hemostatic agent, and it was planned to develop lavender oil-containing coagulant gels and hemostatic preparations and / or wound dressing materials with antimicrobial (antibacterial, antifungal, anticandida), antiviral, and anti-inflammatory properties.

[0008] Lavandula oil is used topically, orally, and by inhalation for various purposes, but has not been utilized for its clotting properties.In the prior art, clotting materials can contain organic and inorganic polymers, nanoparticles, or nanofibers.Therefore, they may cause infection and allergic reactions in the area where they are used.Depending on the severity of the infection and the area of ​​application, the use of hemostatic agents may cause septic shock.In addition, in the prior art, different materials are used for surgical injuries and surface injuries, which is economically disadvantageous.

[0009] CN103446619, DE102009023269, RU2002121797 are some patent applications known in the art. [Prior art documents] [Patent documents]

[0010] [Patent Document 1] Chinese Patent Application Publication No. 103446619 [Patent Document 2] German Patent Application Publication No. 102009023269 [Patent Document 3] Russian Patent Application Publication No. 2002121797 [Non-patent literature]

[0011] [Non-Patent Document 1] Gil, Morayma Reyes, Transfusion Medicine and Hemostasis. Elsevier, 2019. 559-564 [Non-patent document 2] Suchy, Pavel et al., Materials 13.7(2020):1627 [Non-patent document 3] Vyas, Krishna S., and Sibu P. Saha, Expert opinion on biological therapy 13.12(2013):1663-1672 [Non-patent document 4] Chiara, Osvaldo et al., BMC surgery 18.1(2018):1-20 [Non-Patent Document 5] Hu, Zhang et al., Marine drugs 16.8(2018):273 [Non-patent document 6] Panwar, Vineeta et al., Carbohydrate Polymers 235(2020):115975 [Non-Patent Document 7] Koopmann, Ann-Kathrin et al., “Tannin-Based Hybrid Materials and Their Applications: A Review.” Molecules 25.21(2020):4910 [Non-patent document 8] Edris, Amr E., Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives 21.4(2007):308-323 [Non-Patent Document 9] Bakkali, Fadil et al., Food and chemical toxicology 46.2(2008):446-475.

[10] . Lodhia MH, Bhatt KR, Thaker VS. Antibacterial activity of essential oils from palmarosa, evening primrose, lavender and tuberose. Indian J Pharm Sci. 2009;71(2):134-136.20336210 [Non-Patent Document 11] Roller S, Ernest N, Buckle J., J Altern Complement Med. 2009;15(3):275-279.19249919 Rouzet M. Lab Pharma Probl Tech. 1984;32:462-466 [Non-Patent Document 12] Moon T, Wilkinson JM, Cavanagh HM., Parasitol Res. 2006;99(6):722-728.16741725 [Non-Patent Document 13] D'Auria FD, Tecca M, Strippoli V, Salvatore G, Battinelli L, Mazzanti G., Med Mycol. 2005;43(5):391-396.16178366 [Non-Patent Document 14] Altaei DT., Am J Dent. 2012;25(1):39-43.22558691 [Non-Patent Document 15] Wani, Abdul Rouf et al., Microbial Pathogenesis 152(2021):104620 [Non-Patent Document 16] Abou Baker, Doha H. et al., Journal of Agriculture and Food Research 4(2021):100135 [Non-Patent Document 17] Winska, Katarzyna et al., Molecules 24.11(2019):2130 Summary of the Invention [Problem to be solved by the invention]

[0012] The objective of the present invention is to develop a polymer-based hemostatic formulation containing lavender oil that has antimicrobial (antibacterial, antifungal, anticandida), antiviral and anti-inflammatory activity.

[0013] Another object of the present invention is to develop a formulation in the form of a gel and / or chitosan powder containing lavender oil.

[0014] A further object of the present invention is to develop a formulation that can be safely used in the treatment of minor bleeding injuries, emergency response situations, surgical incisions, and military injuries and burns. [Means for solving the problem]

[0015] The present invention relates to the development of a hemostatic formulation containing lavender oil, which has antimicrobial (antibacterial, antifungal, anticandida), antiviral, and anti-inflammatory activity, in combination with a gelling polymer to obtain a gel form and / or a chitosan component that enhances the aforementioned biological activity to obtain a powder form. The formulations developed within the scope of the present invention exhibit the hemostatic effect of lavender oil, and, unlike commercially available hemostatic agents, exhibit antibacterial, antifungal, anticandida, antiviral, and anti-inflammatory activity. The formulations of the present invention contain all-natural ingredients, and their biological activity can prevent bacterial, fungal, and viral infections that may occur in open bleeding. In addition, thanks to their anti-inflammatory properties, these formulations have the potential to be used in the treatment of many autoimmune and allergic diseases. Thanks to their biological activity, hemostatic properties, and hydrophobicity, these formulations developed within the scope of the present invention can be safely used in the treatment of minor bleeding injuries, emergency response situations, surgical incisions, and military injuries and burns. Thanks to these properties, the product exhibits antimicrobial and anti-inflammatory properties while also exhibiting bleeding-stopping properties due to its hydrophobic and hemostatic structure. DETAILED DESCRIPTION OF THE INVENTION

[0016] The method for producing the gel preparation having a hemostatic effect containing lavender oil of the present invention comprises the following steps: - forming a coagulant gel, Slowly add lipogel to lavender oil in a 1:1 weight ratio. The resulting mixture was stirred at a stirring speed of 250 rpm until it was completely gelled. Obtaining coagulant gel (lipogel) The process and - producing a hemostatic agent formulation, Polymerized chitosan (Sigma, 48165) was dissolved in 1% (v / v) acetic acid using a stirrer to form a 4% (w / v) solution, which was then transformed into a gel form. Mixing the above coagulant gel formulation and chitosan solution in a ratio of 1:2 to convert it into a liquid form; The prepared mixture is allowed to stand at a temperature of -80 to -96°C for 24 hours, and then all water therein is removed using a freeze dryer at a temperature of -82 to -96°C under a vacuum output of 0.016 mBar (1.6 Pa); Obtaining the hemostatic agent preparation of the present invention Process and Includes.

[0017] The ingredients (contents) of these preparations are as follows: 1. Oil gelling agent and lavender oil, 2. Chitosan, oil gelling agent and lavender oil

[0018] In one embodiment of the present invention, a gel formulation having a hemostatic effect containing lavender oil contains 1% to 10% chitosan by volume, 20% to 50% lavender oil, and 20% to 50% oil gelling agent.

[0019] The prepared formulations have antiseptic and hemostatic properties, as well as anti-inflammatory, antibacterial, antifungal, anticandida, and antiviral activity. These formulations can be in the form of lotions, creams, emulsions, sprays, ampoules, foams, gelatins, pastes, powders, and biomedical material applications containing them, or combinations thereof. In addition, hemostatic formulations with anti-inflammatory, antibacterial, antifungal, anticandida, and antiviral activity can also be used by being loaded onto different carrier materials. Within these product groups, product types such as gauze, bandages, sponges, tapes, wound dressings, wound closures, or any combination thereof can be manufactured for the purpose of stopping bleeding and healing burns and wounds.

[0020] Both formulations, made with lipogel and chitosan, are completely different from existing patents in terms of their components and offer new biological properties. The formulations are not only anti-inflammatory and antibacterial, but also exhibit antimicrobial and antiviral activity. The experimental results obtained are shown in Tables 2, 3, and 4. Due to their ability to rapidly clot and form a gel layer, the formulations do not pose a risk of embolism and can be successfully used in all types of bleeding. Therefore, the formulations can be used not only for specific injuries and wounds in certain parts of the body, but also for all internal and external injuries, wounds, and bleeding, such as surgical interventions, battle wounds, oral trauma, and surgery. [Example]

[0021] Experimental Research Manufacturing of coagulant gel (lipogel) formulations To produce 1000 mg of solidifying gel, slowly add 500 mg of Lipogelac to 500 mg of lavender oil. Stir the mixture at 250 rpm until completely gelled.

[0022] Manufacturing of hemostatic preparations Preparation of polymer solution: The polymer (chitosan) powder was dissolved at a ratio of 4% (w / v) in distilled water containing 1% (v / v) acetic acid to form a gel. The coagulant gel formulation and chitosan solution were mixed with acetic acid and dH2O in a 1:2 ratio to form a liquid. Due to the carrier properties of these substances, mixing should be performed at a 1:2 ratio for maximum efficiency. The prepared mixture was left at -80°C for 24 hours, and then all water in it was removed using a freeze dryer (Ilshine Biobase) at -82°C with a vacuum output of 0.016 mBar (1.6 Pa).

[0023] Antimicrobial experiments The prepared formulations were analyzed for antibacterial and antifungal tests according to international standard methods. EN 13727 (Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of bactericidal activity in the medical area) was used for the antibacterial test, and EN 13624 (Chemical disinfectants and antiseptics - Quantitative suspension test for the evaluation of fungicidal or yeasticidal activity in the medical area) was used for the antifungal test. Briefly, after dilution of the test sample, the product is introduced into a specific bacterial / fungal suspension. After a 2-minute contact time, the mixture is neutralized and inoculated into an appropriate medium to calculate the microbial reduction. After an incubation period, the viability is evaluated.

[0024] Antiviral experiments Cytotoxicity Assay: The effect of the prepared compound on cell viability was measured using a substance called MTS, which is based on mitochondrial dehydrogenase enzyme activity. In this method, HaCaT (immortalized human keratinocyte cells) were prepared in culture medium and seeded at 5,000 cells / well in a 96-well culture plate. After an incubation period (24 hours), the medium on the cells was removed, and a specific concentration of the compound was diluted in the medium and added to the cells. The cellular response to the molecule's toxicity was detected by measuring cell viability after 72 hours. After the incubation period, the MTS substance, added to the cells along with the culture medium, causes the formation of colored formazan crystals as an indicator of cell viability. This color change was evaluated based on absorbance measurement using an ELISA plate reader. The results obtained were analyzed.

[0025] Calculation of infectious titer by TCID50 The cells were removed from the flask, transferred to a 15 ml Falcon tube, and centrifuged at 500 × g for 5 minutes. The medium on the cells that had settled to the bottom of the Falcon tube was discarded, and 1 ml of fresh medium was added and dissolved with a pipette. The cells were then seeded into a 96-well plate to form a monolayer within 24 ± 2 hours and incubated at 37°C in a 5% CO2 incubator. When the cells were observed to form a monolayer under an inverted microscope, they were treated. The Vero cell line was used for adenovirus and poliovirus, and the Raw cell line was used for murine norovirus. Substances for which nontoxic doses were determined were prepared using virus medium according to the volume used.

[0026] 225 μl of virus medium was added to a new 96-well plate, and 25 μl of virus was added to the first 6 wells of the 96-well plate, and serial dilutions were performed on a logarithmic scale (Log10). The medium from the prepared monolayer cells was discarded and washed twice with virus medium. The serial dilutions prepared in the new 96-well plate were transferred to the cells, and the medium volume was brought to 200 μl with the prepared non-toxic dose of substance. The plate was incubated for 72 hours in a 37°C, 5% CO2 incubator. At the end of the incubation period, the cytopathic effect (CPE) of the virus suspension was evaluated under an inverted microscope. The results were evaluated by calculating the TCID50 using the Spearman-Karber method according to the following formula:

number

number

[0027] The antiviral activities in Table 3 were calculated using Equations 1 and 2.

[0028] European Union standards for antiviral activity studies (EN14476, EN17272, etc.) state that a product must achieve a 4-log reduction against the virus used in the experiment in order to be declared to have antiviral activity. In this regard, antiviral activity of 4 logs or more was detected in both formulations prepared by the present inventors against adenovirus type 5, poliovirus type 3, murine norovirus, vaccinia virus, bovine coronavirus, and herpes simplex virus type 1 (Tables 2 and 4).

[0029] Clotting experiments 1000cc of human venous blood was placed in a petri dish and the following amounts of substances were added to it: Separate time measurements were taken for each substance experiment.

[0030] [Table 1]

[0031] In vivo testing of hemostatic formulations Albino rats (9 weeks old) were incised at the femoral artery, femoral vein, renal artery, and liver to ensure blood flow. Immediately after the initiation of blood flow, a hemostatic agent was applied, and the duration of the application was monitored.

[0032] The results obtained from the analysis of the formulation are as follows:

[0033] Coagulant gel antiviral activity results The antiviral activity of the resulting formulation was evaluated against various DNA-RNA viruses, and the results are shown in Table 2.

[0034] [Table 2]

[0035] In vitro results of chitosan-based formulations antimicrobial activity [Table 3]

[0036] Antiviral activity [Table 4]

[0037] Assessment of clot formation Control group [Table 5]

[0038] Experimental group [Table 6]

[0039] In this study, the maximum carrier ratio of chitosan was determined to be 4%.

[0040] In vivo results of chitosan-based formulations The results obtained in the femoral artery, femoral vein, renal artery and liver sections of rats are as follows:

[0041] [Table 7]

[0042] References [1]. Gil, Morayma Reyes. “Overview of the coagulation system”, Transfusion Medicine and Hemostasis. Elsevier, 2019. 559-564. [2]. Suchy, Pavel et al., “Composite hemostatic nonwoven textiles based on hyaluronic acid, cellulose, and etamsylate”, Materials 13.7(2020):1627. [3]. Vyas, Krishna S., and Sibu P. Saha. "Comparison of hemostatic agents used in vascular surgery", Expert opinion on biological therapy 13.12(2013):1663-1672. [4]. Chiara, Osvaldo et al., "A systematic review on the use of topical hemostats in trauma and emergency surgery", BMC surgery 18.1(2018):1-20. [5]. Hu, Zhang et al., "Chitosan-based composite materials for prospective hemostatic applications", Marine drugs 16.8(2018):273. [6]. Panwar, Vineeta et al., "In-vitro and in-vivo evaluation of modified sodium starch glycolate for exploring its haemostatic potential.", Carbohydrate Polymers 235(2020):115975. [7]. Koopmann, Ann-Kathrin et al., "Tannin-Based Hybrid Materials and Their Applications: A Review.", Molecules 25.21(2020):4910. [8]. Edris,Amr E. ”Pharmaceutical and therapeutic potentials of essential oils and their individual volatile constituents: a review.” Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives 21.4(2007):308-323. [9]. Bakkali,Fadilら、”Biological effects of essential oils-a review.” Food and chemical toxicology 46.2(2008):446-475.

[10] . Lodhia MH、Bhatt KR、Thaker VS. Antibacterial activity of essential oils from palmarosa, evening primrose, lavender and tuberose. Indian J Pharm Sci. 2009;71(2):134-136.20336210

[11] . Roller S、Ernest N、Buckle J. The antimicrobial activity of high-necrodane and other lavender oils on methicillin-sensitive and -resistant Staphylococcus aureus (MSSA and MRSA). J Altern Complement Med. 2009;15(3):275-279.19249919 Rouzet M. Lab Pharma Probl Tech. 1984;32:462-466.

[12] . Moon T、Wilkinson JM、Cavanagh HM. Antiparasitic activity of two lavandula essential oils against Giardia duodenalis, Trichomonas vaginalis and Hexamita inflata. Parasitol Res. 2006;99(6):722-728.16741725

[13] . D’Auria FD、Tecca M、Strippoli V、Salvatore G、Battinelli L、Mazzanti G. Antifungal activity of Lavandula angustifolia essential oil against Candida albicans yeast and mycelial form. Med Mycol. 2005;43(5):391-396.16178366

[14] . Altaei DT. Topical lavender oil for the treatment of recurrent aphthous ulceration. Am J Dent. 2012;25(1):39-43.22558691

[15] . Wani,Abdul Roufら、”An updated and comprehensive review of the antiviral potential of essential oils and their chemical constituents with special focus on their mechanism of action against various influenza and coronaviruses.” Microbial Pathogenesis 152(2021):104620.

[16] . Abou Baker,Doha H.ら、”Antiviral activity of Lavandula angustifolia L. and Salvia officinalis L. essential oils against avian influenza H5N1 virus.” Journal of Agriculture and Food Research 4(2021):100135.

[17] . Winska,Katarzynaら、”Essential oils as antimicrobial agents-myth or real alternative?.” Molecules 24.11(2019):2130.

Claims

1. A gel formulation containing an oil gelling agent and lavender oil, which has hemostatic effects due to the presence of lavender oil components.

2. A gel formulation containing chitosan and lavender oil components, which has hemostatic effects.

3. A gel formulation having a hemostatic effect containing the lavender oil component according to claim 2, comprising 1% to 10% by volume of chitosan, 20% to 50% of lavender oil, and 20% to 50% of an oil gelling agent.

4. A gel formulation having a hemostatic effect containing the lavender oil component described in claim 2, having the form of a lotion, cream, emulsion, spray, ampoule, foam, gelatin, paste, powder, or a biomedical material imparting material containing any one or a combination thereof.

5. A gel formulation having a hemostatic effect, comprising the lavender oil component according to claim 2, which is used by being loaded into at least one carrier material.

6. A gel formulation having a hemostatic effect, containing the lavender oil component described in claim 5, which is used for the purpose of stopping bleeding, healing burns, and healing wounds by being loaded onto a carrier material that is gauze, a bandage, a sponge, tape, a wound dressing, a wound occlusive dressing, or a combination thereof.

7. A method for producing a gel preparation having a hemostatic effect containing a lavender oil component according to any one of claims 2 to 6, A process for producing a coagulant gel, Add lipogel to lavender oil, Stir the resulting mixture until it is completely gelled. Obtain a coagulant gel (lipogel). The process, A process for producing hemostatic agent preparations, Polymerized chitosan (Sigma, 48165) is dissolved in acetic acid using a stirrer to form a solution, which is then converted into a gel. The coagulant gel preparation is mixed with the chitosan solution, and the mixture is converted into a liquid form. After the prepared mixture has been allowed to stand, all the water in it is removed using a freeze-dryer. To obtain the hemostatic agent formulation of the present invention Process and A method for producing a gel preparation containing lavender oil components that has hemostatic effects.

8. A method for producing a hemostatic gel formulation containing a lavender oil component according to claim 7, wherein the lipogel is slowly added to the lavender oil in a 1:1 weight ratio.

9. A method for producing a hemostatic gel formulation containing a lavender oil component according to claim 7, wherein the mixture of lavender oil and lipogel is stirred at a stirring speed of 250 rpm until it is completely gelled.

10. A method for producing a hemostatic gel formulation containing a lavender oil component according to claim 7, wherein the chitosan is dissolved in 1% (v / v) acetic acid to form a 4% (w / v) solution.

11. A method for producing a hemostatic gel preparation containing a lavender oil component according to claim 7, wherein the coagulant gel preparation and the chitosan solution are mixed in a ratio of 1:

2.

12. A method for producing a hemostatic gel preparation containing a lavender oil component according to claim 7, wherein the mixture prepared by mixing the coagulant gel preparation and the chitosan solution is left to stand for 24 hours at a temperature of -80 to -96°C, and then all the water therein is removed using a freeze dryer at a temperature of -82 to -96°C with a vacuum output of 0.016 mBar (1.6 Pa).