Cola flavor essence

Abstract

The invention discloses cola-flavor essence which is prepared from lemon oil, white lemon oil, sweet orange oil, cinnamon oil, clove bud oil, terpilenol, spearmint oil, borneol and menthol. The essence prepared from all natural raw materials has cola flavor fragrance, and has a good inhibition effect on staphylococcus aureus, escherichia coli, pseudomonas aeruginosa, aspergillus niger and streptococcus mutans.

Description

Technical Field

[0001] The invention relates to the field of flavor additives, in particular to a cola flavor essence. Background Art

[0002] Spices have a long history and have enriched people's lives, cultivated people's sentiments, and given people a wonderful life enjoyment through their sense of smell and taste. Spices are the fragrant raw materials used to prepare essences, and essences are a mixture of various spices.

[0003] The flavors used in toothpaste are made by blending natural and synthetic flavors with reference to the fragrance of natural plants. They have natural flavors and various flavors. Toothpaste flavors are part of food flavors and are generally compatible with chewing gum flavors and mouthwash flavors. Flavors are the most expensive component in toothpaste, and the style of toothpaste is basically determined by the flavors used. Toothpaste flavors are an important factor for consumers to distinguish toothpaste products based on their actual experience. Cooling agents are an indispensable raw material in the formula of toothpaste flavors, giving toothpaste flavors a cooling feeling, thereby ensuring the freshness and cleanliness of toothpaste, mouthwash and other oral products.

[0004] At present, the flavors used in toothpaste, mouthwash, chewing gum and other oral cleaning products are mainly cooling agents, which only bring a cool taste. The most common cooling agent is menthol (especially L-menthol), but due to its strong smell and strong irritation to the skin, mucous membrane tissue and eyes, it is not suitable for large-scale use. In addition, it has no inhibitory effect on bacteria such as pathogens, and some cooling agents can also damage the oral micro-ecological system.

[0005] Domestic and foreign studies have shown that plant spices have a good inhibitory effect on pathogenic bacteria in daily production and life. Tasson and Nych as reported in 1995 that peppermint essential oil spices have an inhibitory effect on Salmonella enteritidis and Listeria monocytogenes. Later, through the determination of the antibacterial range, it was found that the cool substances extracted from long bean kernels and the essential oil spices extracted from mastic gum can play an antibacterial role in food.

[0006] This study is based on the "Antimicrobial Susceptibility Test Operation Methods and Judgment Criteria" (9th edition) issued by the Clinical and Laboratory Standards Institute (CLSI) of the United States. A cola-flavored toothpaste flavor with all-plant flavors was formulated. It has a good antibacterial effect on 4 indicator bacteria and 1 oral anaerobic pathogen, and will not destroy the dynamic balance of microorganisms in the oral cavity. Its application in toothpaste was systematically studied to provide a basis for its comprehensive development and utilization. Summary of the invention

[0007] The problem to be solved by the present invention is to propose an innovative solution to the shortcomings of the above-mentioned prior art, especially a cola-flavored toothpaste flavor that is generally acceptable to the public and has a good inhibitory effect on Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, and Streptococcus mutans without destroying the oral microecological system.

[0008] Escherichia coli, also known as Escherichia coli, can live in the intestines of normal people. When the body's resistance is reduced due to cold or malnutrition, the E. coli living in the intestines will grow and multiply in large quantities, causing local intestinal inflammation, which in turn affects the peristaltic function of the intestines, resulting in the inability to digest and absorb food in time, and continuous fermentation or decomposition to produce unpleasant odors such as nitrogen dioxide and ammonia, causing bad breath. Some patients may also experience discomfort symptoms such as nausea and vomiting, which will also aggravate the bad breath phenomenon.

[0009] Oral infection with Staphylococcus aureus may cause symptoms such as oral pain and fever. Staphylococcus aureus infection can cause damage to the oral mucosa, trigger an inflammatory response, and stimulate nerve endings, causing pain. This symptom usually appears in the oral mucosa, gums, or tongue. Staphylococcus aureus infection can activate the body's immune system and produce inflammatory mediators and cytokines, which stimulate the body's temperature regulation center and cause an increase in body temperature. Fever may involve multiple parts of the body, but it is common in areas rich in lymph nodes such as the armpits and groin.

[0010] Pseudomonas aeruginosa is a common bacterium that can cause an inflammatory reaction in the oral mucosa after infection, leading to oral leukoplakia. It may cause damage to the oral mucosa, leading to symptoms of oral ulcers, and an inflammatory reaction, leading to symptoms of red, swollen and painful gums. If food residues in the mouth are not cleaned in time and ferment in the mouth, Pseudomonas aeruginosa can also cause symptoms of bad breath. If the patient's condition is more serious, it may also cause abnormalities in the body's temperature regulation center, leading to symptoms of fever.

[0011] When Aspergillus enters the body, the pigment it produces turns the tongue coating black, causing inflammation and tissue damage, which can further lead to abnormal tongue color. If not treated promptly, Aspergillus infection can spread to other parts of the body and cause serious health problems.

[0012] Mutans Streptococcus infection may cause symptoms such as oral pain, gingival swelling, burning sensation in the throat, fever, and headache. Mutans Streptococcus infection may cause inflammation and ulceration of the oral mucosa, which can expose nerve endings and stimulate pain. Pain usually occurs inside the mouth and may be concentrated in the teeth, gums, or tongue. The immune system will activate the defense mechanism, at which time a large number of inflammatory factors will be produced, thereby triggering an inflammatory response in the local tissue. This inflammatory response will cause capillary dilation and increased permeability, allowing water and white blood cells in the blood to leak into the interstitial space, thereby causing swelling of the gums. Swelling usually occurs at or below the gums, sometimes accompanied by symptoms such as erythema and tenderness. Because toxins or other metabolites produced by bacteria irritate the mucosa of the throat, local inflammatory reactions may occur, causing a burning sensation in the throat. Patients may feel a burning discomfort in the back of the throat, sometimes accompanied by coughing or difficulty swallowing. Infection will cause the body's immune system to respond, including the release of endogenous pyrogens, which act on the body's temperature regulation center to move the body temperature set point upward and cause fever. Fever is usually accompanied by symptoms such as malaise and fatigue. The body temperature may be higher than the normal range. The systemic inflammatory response caused by the infection may lead to increased intracranial pressure and compression of the cranial nerves, thus causing headaches. Headaches can be dull, throbbing or stabbing, with varying locations and severity.

[0013] It can be seen that Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger, and Streptococcus mutans have a great impact on oral health. Although the use of certain chemical products will have an inhibitory effect on certain bacteria, it will also have the same effect on probiotics. In severe cases, it will also destroy the dynamic balance of microorganisms in the cavity, as well as the unknown hazards of the chemical products themselves. Therefore, it is very necessary to use only natural materials to produce a flavor that has a full taste and is popular with the public without using chemical products. At the same time, this flavor also has a good antibacterial effect on 4 indicator bacteria and 1 oral anaerobic pathogens, and will not destroy the dynamic balance of microorganisms in the mouth.

[0014] To solve the above problems, the solution adopted by the present invention is as follows: a cola flavor essence, characterized in that the essence is composed of lemon oil, white lime oil, sweet orange oil, cinnamon oil, clove bud oil, pinene alcohol, spearmint oil, borneol and menthol.

[0015] Furthermore, the cola flavor is characterized in that, by mass percentage, lemon oil accounts for 5%-13%, white lime oil accounts for 20%-50%, sweet orange oil accounts for 2%-5%, cinnamon oil accounts for 0.2%-0.5%, clove bud oil accounts for 0.2%-0.5%, pineneol accounts for 1.5%-4%, spearmint oil accounts for 2.5%-7%, borneol accounts for 0.05%-0.1%, and menthol accounts for 6%-20%, with a total amount of 100%.

[0016] Furthermore, the flavor also includes spearmint oil, which is 2.5%-7% by mass. Adding 2.5%-7% of the spearmint oil component can enhance the fullness and thickness of the cola flavor.

[0017] A toothpaste using the cola flavor essence, characterized in that the cola flavor essence is applied to the toothpaste.

[0018] An oral cleaning product using the cola flavor essence is characterized in that the cola flavor essence is used in the oral cleaning product.

[0019] The technical effects of the present invention are as follows: the flavor composed of natural raw materials of lemon oil, white lime oil, sweet orange oil, cinnamon oil, clove bud oil, pine oil, spearmint oil, borneol and menthol has a cola flavor that is generally acceptable to the public, and has a softer taste than the currently commonly used menthol cooling agent, and has strong fragrance, color and stability. White lime oil can enhance the fullness and thickness of the cola flavor, and can induce cola flavor when combined with other components.

[0020] The cola flavor of the present application can effectively inhibit the growth of cariogenic bacteria, and the natural plant raw materials used in the present application have the advantages of rich sources, good antibacterial effect and small toxic side effects. The lemon oil, white lime oil and other plant flavors contained in this flavor; lemon oil is mainly found in the oil cells of the outer skin of lemons, with an average content of about 0.7% to 0.8%. Lemon oil is rich in limonene, citral and other special ingredients, which can be used for skin care and health care to promote metabolism, antibacterial and antiseptic effects, and can also be made into food flavor additives. Studies have found that the natural plant flavor mixture of the present application, this toothpaste flavor, can inhibit the growth of Streptococcus mutans, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Aspergillus niger, and has an inhibitory effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Antibacterial test of the cola flavor essence of the present application on Escherichia coli.

[0022] Figure 2 The cola flavor essence of the present application is tested for the inhibition of Staphylococcus aureus.

[0023] Figure 3 The cola flavor essence of the present application is tested for its antibacterial effect on Aspergillus niger.

[0024] Figure 4 Antibacterial test of the cola flavor essence of the present application on Streptococcus mutans.

[0025] Figure 5 Antibacterial test of the cola flavor essence of the present application on Pseudomonas aeruginosa.

[0026] Figure 6The cola flavor of the present application is tested for MIC of Escherichia coli.

[0027] Figure 7 The cola flavor of the present application is tested for MIC of Staphylococcus aureus.

[0028] Figure 8 The cola flavor of the present application is tested for MIC of Aspergillus niger.

[0029] Fig. 9 The cola flavor of the present application is used for MIC determination of Streptococcus mutans.

[0030] Fig.10 The cola flavor of the present application is used for MIC determination of Pseudomonas aeruginosa. DETAILED DESCRIPTION

[0031] Embodiment: A cola flavor essence is composed of lemon oil, white lime oil, sweet orange oil, cinnamon oil, clove bud oil, pineneol, spearmint oil, borneol and menthol.

[0032] Antibacterial test Strains: Escherichia coli ATCC 8739, Staphylococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC9027, Aspergillus brasiliensis ATCC 16404 were from Shanghai University of Technology, and Streptococcus mutans ATCC 35668 was purchased from Shanghai Xuanke Biotechnology Co., Ltd.

[0033] Experimental culture media: Aerobic bacteria use tryptone soy agar (TSA), tryptone soy broth (TSB) for anaerobic bacteria use brain heart infusion agar (BHI) agar plates, glucose agar (SDA), brain heart infusion broth (BHI), nutrient broth (NB), malt extract, 7.5% sodium chloride broth.

[0034] Main instruments and equipment: biological safety cabinet, vertical pressure steam sterilizer, micropipette, vortex mixer, constant temperature incubator, PB-10 pH meter, Cellstar 96-well cell culture plate, microplate reader, anaerobic / microaerobic culture system.

[0035] Preparation of bacterial suspension: Aerobic bacteria were inoculated on TSA slant medium and cultured at 37℃ for 24 h, and transferred once. A loop of slant colonies was picked up with an inoculation loop and inoculated into 50 mL of tryptone soy broth (TSB), and incubated at 37℃ for 16-18 h; the culture solution after the enrichment was used as the initial culture solution (100).

[0036] Inoculate anaerobic bacteria on BHI plates and culture at 37℃ for 72 hours, transfer once. Pick a loop of colonies on the plate with an inoculation loop and inoculate it into 10 mL of brain heart infusion broth (BHI), incubate at 37℃ for 24-48 hours; the culture solution after the enrichment is used as the initial culture solution (100).

[0037] Preparation of fungal suspension: The fungi were inoculated on SDA plates and cultured at 28°C, yeasts were cultured for 40-48 h, and molds were cultured for 6-10 d, and transferred once. The strains were eluted with 2-3 mL of 0.85% sterile saline, and the spores of molds were eluted with saline containing 0.05% Tween-80, and the spores were placed in 5 mL sterile centrifuge tubes as the initial bacterial solution (100).

[0038] Counting of bacterial suspension: Use a micropipette to pipette 1 mL of the bacterial suspension in steps 1.2.2 and 1.2.3 into 9 mL of sterile saline, and oscillate to mix. This dilution ratio is 1:10. Use a micropipette to pipette 1 mL of the 10-fold diluted bacterial solution into 9 mL of sterile saline, which is a 100-fold dilution. Repeat the same process until the dilution reaches a gradient of 10 to 12. Select the appropriate gradient and count the bacteria respectively.

[0039] Preparation of bacterial culture medium Preparation of TSA medium: Weigh 8g TSA agar, add 200 mL distilled water, heat to dissolve, sterilize at 121℃ for 15 min under high pressure, and cool to about 50℃. Take the above TSA medium out of the autoclave, pour about 20 mL into each sterile culture dish while it is hot, let it stand, and cool to solidify.

[0040] Preparation of BHI medium: weigh 3.7 g BHI and 1.8 g agar, add 150 ml distilled water and heat to dissolve, sterilize at high pressure at 121°C for 15 min and cool to about 50°C.

[0041] Take the above BHI medium out of the sterilizer, pour about 20 mL into each sterile culture dish while it is still hot, let it stand, and cool it to solidify. Pour 40 mL of BHI medium at about 50°C into the bacterial suspension, mix it thoroughly, and use a pipette to transfer about 6 mL (containing 200 μL of Streptococcus mutans liquid) into each culture dish (just cover the surface evenly) to make a bacterial culture medium.

[0042] Table 1: Cola flavor (Formula 1) and control group (Formula 2) were prepared according to the embodiment. Filter paper pieces with a diameter of 4 mm were prepared and sterilized, and then immersed in cola flavor (formula 1) and control group (formula 2) for 2 hours.

[0043] Diameter of the inhibition zone: The diameter of the inhibition zone is determined by the agar plate filter paper method. On the clean bench, use a pipette to draw the test bacteria suspension into the agar plate culture medium. The agar plate must be placed stably and evenly, and the coating rod is used to spread it evenly (note that the coating rod should be sterilized under an alcohol lamp after each use) to ensure that the substance in the agar is evenly diffused. Take the filter paper soaked in cola-flavored toothpaste and stick it on the bacterial plate. Stick 2 to 3 filter paper soaked in cola-flavored toothpaste on each plate. Place the culture plate in a 37℃ constant temperature incubator for 24 hours, and measure the diameter of the inhibition zone. Each group is measured 3 times, and the results are averaged.

[0044] Preparation of MIC plate: Using sterile operation, add solutions of cola flavor essence of different concentrations after multiple dilution into sterilized 96-well polystyrene plates respectively, add sample solution to wells 1 to 8, 100 μL per well, add 100 μL of BHI medium without sample solution to well 9, and add 200 μL of BHI medium without sample solution to well 10.

[0045] Double dilution method: 1 Weigh 3 g and 1.5 g of anhydrous Na2HPO 4 and KH2PO 4, add 1000 mL of distilled water, adjust the pH to 7.2-7.4, sterilize at 121℃ for 20 min, cool and set aside. Weigh 2.83 g and 1.36 g of anhydrous Na2HPO 4 and KH2PO 4, add 1000 mL of distilled water, adjust the pH to 7.2-7.4, sterilize at 121℃ for 20 min, cool and set aside. 2 Take 5 mL of sample with aseptic operation, put it into 45 mL of sterile PBS, shake it thoroughly to dissolve, and prepare a 10% (the initial concentration can be appropriately reduced according to the specific situation) uniform solution. 3 Take 8 sterile 5 mL plastic test tubes. Except for the first tube, add 1.6 mL of BHI broth medium. Add 1 mL of BHI broth medium to each of the remaining tubes. Add 0.5 mL of antibacterial solution to the first tube and mix well. Then pipette 1 mL into the second tube, mix well and then pipette 1 mL into the third tube. Continue to dilute in multiples until the eighth tube, and pipette 1 mL from the eighth tube and discard it.

[0046] Preparation and inoculation of test bacteria: Weigh 0.15 g of malt extract powder into a conical flask, add 10 ml of distilled water to dissolve, and sterilize at high pressure at 121°C for 15 min.

[0047] Weigh 0.9 g of 7.5% sodium chloride broth culture medium into a conical flask, add 10 ml of distilled water, heat to dissolve, and sterilize at high pressure at 121°C for 15 min.

[0048] Weigh 0.36g of NB nutrient broth culture medium into a conical flask, add 20ml of distilled water, heat to dissolve, and sterilize at high pressure at 121℃ for 15 min. The bacterial suspension was diluted with the corresponding culture medium (ten-fold dilution method) until the bacterial content was about 106 CFU / mL.

[0049] 100 μL of bacterial suspension containing approximately 106 CFU / mL was inoculated into a 96-well plate containing a culture medium containing cola-flavored toothpaste flavor (added sequentially from well 9 to well 1), where wells 1 to 8 were experimental groups, well 9 was a positive control, and well 10 was a negative control.

[0050] Culture: Place the sealed 96-well plate in a 37°C incubator for aerobic bacteria for 18 to 24 hours and observe the results; place the anaerobic bacteria in a 37°C anaerobic culture tank for 72 hours and observe the results; place the mold in a 28°C incubator for 72 hours and observe the results. Take 200 μL of Streptococcus mutans ATCC35668 bacterial solution and inoculate it on BHI broth medium, culture it in a constant temperature incubator at 37°C for 24 hours, and observe the growth of colonies. Under a dark background, the lowest drug concentration at which there is no bacterial growth observed by the naked eye, that is, the liquid in the test tube is clear without turbidity or precipitation, is the minimum inhibitory concentration. The determination was repeated 3 times and the results were averaged.

[0051] Evaluation criteria: The results were determined by measuring the absorbance of each well at 600 nm using an ELISA reader with the blank well adjusted to zero. The lowest drug concentration well with an absorbance value that was more than 80% lower than that of the growth control well was taken as the MIC endpoint.

[0052] When there is bacterial growth in the positive control well (turbidity) and no bacterial growth in the negative control well (transparent), the lowest drug concentration that completely inhibits bacterial growth in the well is the MIC.

[0053] Results and Analysis: Table 2: Antibacterial activity of cola-flavored toothpaste flavors. After testing, it was found that without adding white lemon oil, the aroma was defective and unharmonious, the cola flavor was not penetrating, and the antibacterial effect was affected.

[0054] It can be seen that the best concentration of cola flavor formula 1 is 1.2%. Figure 1~Figure 5 As shown, the antibacterial diameter of Escherichia coli is (24.6±1.9) mm ( Figure 1 ), the antibacterial diameter of Staphylococcus aureus was (24.6±0.1) mm ( Figure 2), Aspergillus niger inhibition diameter (19.4±2.2) mm ( Figure 3 ), the inhibitory diameter of Streptococcus mutans was (31.7±5.1) ( Figure 4 ), the inhibition of Pseudomonas aeruginosa was better ( Figure 5 ).

[0055] The minimum inhibitory concentration (MIC) of 4 indicator bacteria and 1 oral anaerobic pathogen was determined by broth microdilution method. The results are shown in the table.

[0056] Table 3: Minimum inhibitory concentration (MIC) of cola flavor (No. 1-4) against 4 indicator bacteria and 1 oral anaerobic pathogen. like Figure 6~Figure 10 As shown, this cola flavor essence has a good inhibitory effect on 4 indicator bacteria and 1 oral anaerobic pathogen. The horizontal axis represents the dilution multiple and the vertical axis represents the absorbance value.

[0057] Stability test of toothpaste with cola flavor.

[0058] Table 4: High temperature stability test results of toothpaste products. The stability of toothpaste with different amounts of added flavors was tested. The results show that the consistency and pH value of groups 1, 2, 3, and 4 are stable, the paste does not produce oil, coarseness, or shelling, and the aroma, color, and stability are normal. All quality indicators meet the requirements of the national standard GB / T 8372-2017.

Claims

1. A cola flavor, characterized in that: The essence is composed of lemon oil, white lime oil, sweet orange oil, cinnamon oil, clove bud oil, pineneol, borneol and menthol.

2. The cola flavor according to claim 1, characterized in that: In terms of mass percentage, lemon oil accounts for 5%-13%, sweet orange oil accounts for 2%-5%, cinnamon oil accounts for 0.2%-0.5%, clove bud oil accounts for 0.2%-0.5%, pineneol accounts for 1.5%-4%, borneol accounts for 0.05%-0.1%, menthol accounts for 6%-20%, and the balance is white lemon oil.

3. The cola flavor according to claim 2, characterized in that: The essence also includes spearmint oil, and the spearmint oil accounts for 2.5%-7% by mass.

4. A toothpaste using the cola flavor according to claim 1, characterized in that: The cola flavor described in claim 1 is applied to toothpaste.

5. An oral cleaning product using the cola flavor according to claim 1, characterized in that: The cola flavor essence described in claim 1 is applied to oral cleaning products.

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