Traditional Chinese medicine essential oil and tea oil composition with cough relieving effect and application of traditional Chinese medicine essential oil and tea oil composition

Abstract

The invention discloses a traditional Chinese medicine essential oil and tea oil composition with a cough relieving effect and application thereof, and belongs to the field of combination of food processing and traditional Chinese medicine. According to the product, on the basis of fresh eggs, ginger-processed pericarpium citri reticulatae or orange peel essential oil is prepared through a specific method, and the ginger-processed pericarpium citri reticulatae or orange peel essential oil and tea oil are mixed according to the mass ratio of (0.5-2): (99.5-98) to be used for decocting the eggs. The preparation method of the ginger-processed pericarpium citri reticulatae or orange peel essential oil comprises the steps of ginger juice preparation, processing and essential oil extraction. According to the cough-relieving tea oil egg added with the ginger-processed dried orange peel or the orange peel essential oil, the nutrition of the egg and the nutrition of the tea oil are fused, the ginger-processed dried orange peel or the orange peel essential oil has the cough-relieving and health-care effects, and effective components of the ginger-processed dried orange peel or the orange peel essential oil can reduce respiratory tract inflammation and smooth the respiratory tract. The product is simple in preparation method, can be directly eaten or used in the catering industry, and provides the choice of dietary therapy food-nourishing products with natural nutrition for cough people. Experiments prove that the cough frequency of cough mice fed with the product is obviously reduced, and the product has a good cough relieving effect.

Description

Technical Field

[0001] The present invention belongs to the technical fields of food processing and traditional Chinese medicine, and particularly relates to a traditional Chinese medicine essential oil tea oil composition with a cough-relieving effect and its application. Background Art

[0002] Traditional Chinese medicine essential oils have diverse biological activities and have significant effects such as relieving cough, asthma, and expectoration. For example, eucalyptus essential oil relieves cough symptoms through antibacterial and anti-inflammatory effects, and eucalyptus globulus essential oil has expectorant and antitussive effects. In addition, peppermint essential oil and rosemary essential oil are also widely used to relieve throat discomfort and cough. Traditional Chinese medicine essential oils are usually used by inhalation, steam inhalation, diffuser, or massage. For example, eucalyptus globulus essential oil and peppermint essential oil can relieve cough through steam inhalation, while tea tree essential oil can be used for chest massage to soothe respiratory discomfort. Traditional Chinese medicine essential oils are often used in combination with other therapies, such as steam therapy, honey lemon tea, etc., to enhance the cough-relieving effect. These therapies not only utilize the natural properties of essential oils but also combine the conditioning effects of traditional Chinese medicine. The application of traditional Chinese medicine essential oils in the field of cough relief has made certain progress, and their natural and safe characteristics make them a popular adjuvant treatment method. However, further research is still needed to optimize their formulations and usage methods.

[0003] Tangerine peel essential oil is a complex mixture derived from the pericarp of Rutaceae. Components such as d-limonene, α-pinene, β-pinene, β-myrcene, and γ-terpinene in the essential oil have significant antioxidant activity, inhibiting the generation and release of inflammatory mediators, thereby reducing inflammatory responses and also counteracting allergic reactions. Network pharmacology proves that this combination can effectively act on 310 cough-related targets, and the pathways involved in biological processes are mainly enriched in inflammatory responses, responses to exogenous stimuli, and responses to lipopolysaccharides, etc. Animal experiments also prove that this combination can reduce inflammatory responses and significantly reduce the manifestation of cough, thereby achieving a cough-relieving effect. Population observations have found that it can significantly reduce the frequency of cough.

[0004] Orange peel and tangerine peel have similar chemical compositions, both rich in volatile oils and flavonoids. Research shows that the volatile oil in orange peel can reduce airway inflammation by inhibiting the activation of inflammatory cells and reducing the release of inflammatory factors; its flavonoids have significant antioxidant effects, which can scavenge free radicals and protect cells from oxidative damage.

[0005] Processing is a method in traditional Chinese medicine to enhance the medicinal efficacy. By processing, essential oils are increased to relieve cough, asthma, and phlegm. Ginger is a commonly used auxiliary material in the processing of traditional Chinese medicine. Ginger itself has a pungent taste and a warm nature, and has the effects of relieving exterior syndrome and dispelling cold, warming the middle-jiao to arrest vomiting, resolving phlegm and relieving cough, etc. Using ginger to process aromatic herbs can make them have a better cough-relieving effect. Camellia oil contains rich unsaturated fatty acids, vitamin E, tea polyphenols and other components, and has antioxidant, anti-inflammatory and antibacterial effects. These components can reduce respiratory tract inflammation, relieve cough symptoms, and have a protective effect on the mucous membrane of the throat, with anti-inflammatory and lung-moistening effects; Eggs are rich in protein, minerals (such as calcium, phosphorus, iron), vitamins (such as vitamin A, E) and other nutrients. These components help to enhance immunity, maintain the integrity of the respiratory mucosa, thus relieving cough and enhancing the body's resistance. The protein and lecithin in eggs can repair the respiratory mucosa.

[0006] Therefore, how to use ginger to process tangerine peel or orange peel essential oil, form a composition with camellia oil, and combine the efficacy of eggs to prepare a cough-relieving product is worthy of research. Summary of the Invention

[0007] The technical problem to be solved by the present invention is: aiming at the defects and deficiencies such as less research and application of the therapeutic diet and health preservation functions in the existing poultry egg processing technology, to provide a traditional Chinese medicine essential oil and camellia oil composition with a cough-relieving effect and its application in the preparation of cough-relieving therapeutic diet and health preservation products. The traditional Chinese medicine essential oil composition mainly contains ginger-processed tangerine peel or orange peel essential oil. It is proved by network pharmacology that this combination can effectively act on cough-related targets, and it is also proved by in vitro and in vivo animal experiments that this combination can reduce the inflammatory response and significantly reduce the manifestation of cough, so as to achieve the cough-relieving effect.

[0008] The present invention adopts the following technical solutions to achieve the invention purpose.

[0009] First of all, the present invention provides a traditional Chinese medicine essential oil and camellia oil composition with a cough-relieving effect, including ginger-processed traditional Chinese medicine essential oil and camellia oil, wherein the preparation method of the ginger-processed traditional Chinese medicine essential oil includes the following steps: S1. Preparation of ginger juice: Take fresh ginger, wash it, crush it, add distilled water and decoct it, combine the filtrates and concentrate it under reduced pressure, and take the concentrated solution to obtain ginger juice; S2. Processing tangerine peel or orange peel with ginger juice: Cut tangerine peel or orange peel into granules and dry them in the shade to obtain tangerine peel or orange peel granules, add ginger juice to soak them, and obtain tangerine peel or orange peel granules soaked with ginger juice; S3. Extraction of ginger-processed tangerine peel or orange peel essential oil: Take the tangerine peel or orange peel particles soaked in ginger juice and place them in a distillation container. Add distilled water containing 1-ethyl-3-methylimidazolium tetrafluoroborate ionic liquid, heat to boiling and boil gently under normal pressure. Collect the mixed steam of volatile oil and water, condense and separate to obtain the oil-phase fraction. Add a desiccant for drying, filter to remove the desiccant, and obtain the ginger-processed tangerine peel or orange peel essential oil, that is, the ginger-processed traditional Chinese medicine essential oil;

[0010] The mass ratio of the ginger-processed traditional Chinese medicine essential oil to tea oil is 0.5 - 2:99.5 - 98.

[0011] Preferably, in step S1, the mass-volume ratio of ginger to distilled water is 1:4 - 6, and each decoction is for 15 - 25 min; decoct for 1 - 3 times.

[0012] Preferably, in step S1, concentrate to a crude drug mass concentration of 0.9 - 1.1 g / mL.

[0013] Preferably, in step S2, the dosage of ginger juice is 0.5 - 1.2 times the mass of the tangerine peel or orange peel particles, and soak for 12 - 18 h.

[0014] Preferably, in step S3, the addition amount of the distilled water containing 1-ethyl-3-methylimidazolium tetrafluoroborate ionic liquid is 3 - 8 times the mass of the tangerine peel or orange peel particles, and based on distilled water, 1-ethyl-3-methylimidazolium tetrafluoroborate ionic liquid is 0.5 - 1.5% of the mass of distilled water.

[0015] The present invention also discloses the application of the above composition in the preparation of cough-relieving products.

[0016] Furthermore, the addition amount of the composition accounts for 20 - 40% of the total mass of the cough-relieving product.

[0017] Furthermore, the preparation method of the cough-relieving product is as follows: Select the egg liquid of fresh and intact poultry eggs; Pour the composition into a flat pan, heat to 80 - 100 °C, break in the egg liquid, and decoct until cooked through.

[0018] Beneficial effects: (1) The composition of the ginger-processed tangerine peel or orange peel essential oil, tea oil and eggs of the present invention can significantly play a role in relieving cough.

[0019] (2)The composition of ginger-processed tangerine peel or orange peel essential oil, tea oil and eggs of the present invention can have a more effective and stronger antioxidant capacity and the effect of scavenging hydroxyl oxygen free radicals. Cell experiments have proved that this essential oil combination does not affect cell viability, can more significantly reduce the expression of inflammatory cells IL-6 and IL-12, and more significantly increase the inflammatory levels of IL-4 and IL-10; Animal experiments have also proved that this combination can significantly reduce the inflammatory response, reduce the number of coughs and prolong the cough latency period.

[0020] (3)The traditional Chinese medicine essential oil, tea oil and egg composition can prove through network pharmacology that this combination can effectively act on cough-related targets. Animal experiments have also proved that this combination can reduce the inflammatory response and significantly reduce the manifestation of cough, thereby achieving the effect of relieving cough. Description of the Drawings

[0021] Figure 1 Effect of the essential oil-tea oil composition of each example on the expression of pro-inflammatory factors IL-6 and IL-1β in LPS-induced RAW264.7 cells; Compared with the LPS-induced group, Indicates P < 0.05, Indicates P < 0.01.

[0022] Figure 2 Effect of the essential oil-tea oil composition of each example on the expression of anti-inflammatory factors IL-4 and IL-10 in LPS-induced RAW264.7 cells; Compared with the LPS-induced group, Indicates P < 0.05, Indicates P < 0.01; Compared with the cell model group, ## indicates P < 0.01.

[0023] Figure 3 Effect of the essential oil-tea oil composition of each example on the DPPH· scavenging rate; Figure 4 Effect of the essential oil-tea oil composition of each example on the ABTS + · scavenging rate; Figure 5 Effect of the essential oil-tea oil composition of each example on the O2 - · scavenging rate; Figure 6 TIC diagrams of Example 8 and Example 9, wherein, in the positive ion mode, A: Example 8; B: Example 9; in the negative ion mode, C: Example 8; D: Example 9; Figure 7 Biological process enrichment analysis classification bubble diagram of the intersection target genes of Example 8 and chronic cough; Figure 8 Cell component enrichment analysis classification bubble diagram of the intersection target genes of Example 8 and chronic cough; Figure 9 It is the classification bubble chart of the molecular function enrichment analysis of the intersection target genes of Example 8 and chronic cough; Figure 10 It is the classification bubble chart of the biological process enrichment analysis of the intersection target genes of Example 9 and chronic cough; Figure 11 It is the classification bubble chart of the cellular component enrichment analysis of the intersection target genes of Example 9 and chronic cough; Figure 12 It is the classification bubble chart of the molecular function enrichment analysis of the intersection target genes of Example 9 and chronic cough; Figure 13 It is the KEGG pathway map of the intersection target genes of Example 8 and Example 9 and chronic cough; among them, A: the pathway enrichment map of the intersection target genes of Example 8 and chronic cough; B: the pathway enrichment map of the intersection target genes of Example 9 and chronic cough. Detailed implementation manners

[0024] The technical solution of the present invention will be further described below with specific examples.

[0025] In the following examples, tangerine peel is the dried and aged peel of the fruit of Citrus reticulata Blanco of Rutaceae, and orange peel can also be the peel of citrus fruits.

[0026] Example 1: Preparation of tangerine peel essential oil Cut the tangerine peel into particles with a size of about 0.8 mm in length and width, dry it in the shade, take 400 g and put it into a 5000 mL distillation flask, and add 2400 mL of distilled water; add 1 wt% of 1-ethyl-3-methylimidazolium tetrafluoroborate ionic liquid to the distilled water; heat it to boiling with an electric heating mantle and keep it slightly boiling under normal pressure to generate a mixed vapor of volatile oil and water. Stop distillation when the amount of essential oil in the extraction device hardly increases anymore (the distillation time is about 4 h). After condensation, open the piston at the lower end of the oil-water separator, slowly release the water until the oil layer drops to the scale and then let it stand for a while, and collect the upper light yellow essential oil liquid. Dry it with anhydrous sodium sulfate to obtain 4.2 mL of water-free tangerine peel essential oil. The yield of this tangerine peel essential oil is 1.05%.

[0027] Example 2: Preparation of ginger-processed tangerine peel essential oil Preparation of ginger juice: Take fresh ginger, wash the sand on the outside of the ginger with clean water, wash and slice it, dry it until the surface of the ginger slices is dry. Take 500 g of the dried ginger slices, add 2000 mL of distilled water and heat and decoct it twice, each time for 20 min; combine the filtrates and concentrate them under reduced pressure until the mass concentration of the crude drug contained is 1 g / mL, that is, ginger juice is obtained.

[0028] Processing tangerine peel with ginger juice: Cut tangerine peel into particles about 0.8 mm in length and width and dry them in the shade. Put 400 g of the dried tangerine peel particles into a self-sealing bag, add 400 mL of ginger juice, and place them in a cool place to moisten for 12 h to obtain tangerine peel processed with ginger juice.

[0029] Extraction of ginger-processed tangerine peel essential oil: Put the tangerine peel processed with ginger juice as described above into a 5000 mL distillation flask, and add 2400 mL of distilled water; add 1 wt% of 1-ethyl-3-methylimidazolium tetrafluoroborate ionic liquid to the distilled water; heat with an electric heating mantle until boiling, and keep slightly boiling under atmospheric pressure to generate a mixed vapor of volatile oil and water. Stop distillation when the amount of essential oil in the extraction device hardly increases anymore. After condensation, open the lower piston of the oil-water separator, slowly release the water until the oil layer drops to the scale and then let it stand for a while, and collect the upper light yellow essential oil liquid. Dry and separate with anhydrous sodium sulfate to obtain 6.6 mL of water-free ginger-processed tangerine peel essential oil, and the yield of this ginger-processed tangerine peel essential oil is 1.65%.

[0030] Example 3: Orange peel essential oil Cut the orange peel into particles about 0.8 mm in length and width, dry them in the shade, then take 400 g and put them into a 5000 mL distillation flask, and add 2400 mL of distilled water; add 1 wt% of 1-ethyl-3-methylimidazolium tetrafluoroborate ionic liquid to the distilled water; heat with an electric heating mantle until boiling, and keep slightly boiling under atmospheric pressure to generate a mixed vapor of volatile oil and water. Stop distillation when the amount of essential oil in the extraction device hardly increases anymore (the distillation time is about 4 h). After condensation, open the lower piston of the oil-water separator, slowly release the water until the oil layer drops to the scale and then let it stand for a while, and collect the upper light yellow essential oil liquid. Dry and separate with anhydrous sodium sulfate to obtain 3.92 mL of water-free orange peel essential oil, and the yield of this orange peel essential oil is 0.98%.

[0031] Example 4: Ginger-processed orange peel essential oil Prepare 6.2 mL of ginger-processed orange peel essential oil in the same way as in Example 2, and the yield of this ginger-processed orange peel essential oil is 1.55%.

[0032] Example 5: Extraction and preparation of camellia oil Select mature camellia seeds, remove impurities, and conduct drying treatment to reduce the moisture content to 3 - 5%. Feed the dried camellia seeds into a shelling machine for shelling, and remove metal impurities through magnetic separation. Microwave dry or heat-treat the shelled camellia kernels, and then crush them into powder. Feed the heat-conditioned camellia seed powder into a dedicated low-temperature cold press for pressing, with the pressing temperature controlled between 35°C and 60°C. The crude oil extracted is subjected to rough filtration and fine filtration to remove impurities and insoluble substances. Reduce the moisture content in the oil through a dehydration device under vacuum conditions, then add a small amount of decolorizing agent (diatomaceous earth) for decolorization treatment, followed by deodorization treatment, and then crystallize at low temperature to separate the solid fat crystals from the liquid, and finally obtain the low-temperature cold-pressed finished camellia oil.

[0033] Example 6: Preparation of Ginger-processed Tangerine Peel Essential Oil + Camellia Oil Mix the ginger-processed tangerine peel essential oil obtained in Example 2 and the camellia oil obtained in Example 5 in a mass ratio of 1:99 to form a traditional Chinese medicine essential oil composition of ginger-processed tangerine peel essential oil + camellia oil.

[0034] Example 7: Preparation of Ginger-processed Orange Peel Essential Oil + Camellia Oil Mix the ginger-processed orange peel essential oil obtained in Example 4 and the camellia oil obtained in Example 5 in a mass ratio of 1:99 to form a traditional Chinese medicine essential oil composition of ginger-processed orange peel essential oil + camellia oil.

[0035] Example 8: Preparation of a small sample of fried eggs containing ginger-processed tangerine peel essential oil + camellia oil Ratio: By mass percentage: 1% of ginger-processed tangerine peel essential oil (prepared in Example 2), 99% of camellia oil.

[0036] Preparation method: According to the above formula ratio, first mix the essential oil and camellia oil. Pour an appropriate amount of camellia oil containing ginger-processed tangerine peel essential oil into a frying pan, heat it to 80 - 100°C, pour in the pre-treated egg liquid, and fry until half-cooked.

[0037] Example 9: Preparation of a small sample of fried eggs containing ginger-processed orange peel essential oil + camellia oil Ratio: By mass percentage: 1% of ginger-processed orange peel essential oil (prepared in Example 4), 99% of camellia oil.

[0038] Preparation method: According to the above formula ratio, first mix the essential oil and camellia oil. Pour an appropriate amount of camellia oil containing ginger-processed orange peel essential oil into a frying pan, heat it to 80 - 100°C, pour in the pre-treated egg liquid, and fry until half-cooked.

[0039] Control Example 1: Xingbei Zhike Granules, produced by Jiangsu Kanion Pharmaceutical Co., Ltd., with a specification of 4 g / bag and a product batch number of 240925.

[0040] Experimental Example 1: In vitro antibacterial experiment Measuring the diameter of the antibacterial circle: Pipette 0.5 mL of the bacterial suspension at 1×10 7 CFU / mL, spread it on the agar plate, place the Oxford cup, add 0.75 g / mL (calculated based on crude drug) of the medicinal liquid of each example, use sodium penicillin as the positive control (mass concentration is 10 U / mL), and sterile distilled water as the blank control. After culturing at 37 °C for 24 h, measure the diameter of the antibacterial circle. The results are shown in Table 1. As can be seen from Table 1, the essential oil of tangerine peel or orange peel after being processed with ginger has good antibacterial effects, and the examples combined with tea oil and eggs also show good inhibitory ability against harmful bacteria.

[0041] Table 1 Comparison of in vitro antibacterial effects of each example

[0042] Note: The unit of the diameter of the antibacterial circle: mm

[0043] Experimental Example 2: Effects on inflammatory factors and anti-inflammatory factors in in vitro inflammatory cell experiments (1) Effects on the activity of RAW264.7 cells:

[0044] Culture RAW264.7 cells in high-glucose medium containing 10% fetal bovine serum. When the cells grow to the logarithmic phase, adjust the cell concentration to 2×10 5 cells / mL, inoculate them into 96-well plates, 200 μL per well, and culture them in an incubator at 37 °C with a CO2 concentration of 5% for 24 h. Then replace the medium with different concentrations of Examples 1-9 and Comparative Example 1 to make the final concentrations 10, 20, 40, 80, 100 μg / mL. Set six replicates at each concentration as the drug group; at the same time, culture with serum-free starvation medium without drugs as the control group; culture with serum-free starvation medium without cells as the blank group. After culturing for 24 hours, discard the culture medium, add medium containing 10% MTT to each well, continue to culture for 4 hours, then terminate the culture. Carefully aspirate the medium in the wells, add 150 μL of DMSO to each well, place it on a shaker and shake gently for 10 min, and detect the absorbance of each well at 490 nm on an enzyme-linked immunosorbent assay (ELISA) reader. The results show that compared with the control group, there is no difference in the cell survival rate among each example and Comparative Example 1, and the survival rate is greater than 90%, indicating that Examples 1-9 have no toxic effects on cells at a concentration of 100 μg / mL.

[0045] (2) Detection of inflammatory factors in LPS-induced RAW264.7 cells: Cultivate cells according to the above method, inoculate them in a 96-well plate, and after culturing for 24 h, carefully aspirate the culture medium and wash twice with PBS. Administer the solutions of Examples 1-9 and Comparative Example 1 at a concentration of 80 μg / mL, and after 1 h of administration, administer 1 μg / mL of LPS. At the same time, set up an LPS induction group (administer 1 μg / mL of LPS) and a normal control group (do not administer any drugs), and set 3 replicate wells for each group. After culturing for 24 h, collect the cell supernatant, and detect the inflammatory factors IL-6, IL-1β, IL-4, and IL-10 according to the ELISA kit. Figure 1 Effects of each example on the expression of pro-inflammatory factors IL-6 and IL-1β in LPS-induced RAW264.7 cells; Figure 2 Effects of each example on the expression of anti-inflammatory factors IL-4 and IL-10 in LPS-induced RAW264.7 cells.

[0046] For Figure 1 、 Figure 2 Analysis shows that: compared with the normal control group (cell model group), the contents of IL-6 and IL-1β in the model control group (LPS induction group) increased significantly (P < 0.01), and the contents of IL-4 and IL-10 decreased significantly (P < 0.01). Compared with the LPS induction group, the contents of pro-inflammatory factors IL-6 and IL-1β in each example decreased significantly (P < 0.05 or P < 0.01). Among them, the anti-inflammatory effects of ginger-processed tangerine peel (Example 2) or ginger-processed orange peel (Example 4) were better than those before ginger processing (Examples 1 and 3). The anti-inflammatory effects of Examples 6 and 7 combined with tea oil of Examples 2 and 4, and Examples 8 and 9 formed by the combination of tea oil and eggs were also very significantly different from those of the LPS induction group (P < 0.01), indicating that this combination has a significant anti-inflammatory effect; compared with the LPS induction group, the contents of anti-inflammatory factors IL-4 and IL-10 in each example increased significantly (P < 0.05 or P < 0.01). Among them, the anti-inflammatory effects of ginger-processed tangerine peel (Example 2) or ginger-processed orange peel (Example 4) were better than those before ginger processing (Examples 1 and 3). The anti-inflammatory effects of Examples 6 and 7 combined with tea oil of Examples 2 and 4, and Examples 8 and 9 formed by the combination of tea oil and eggs were also very significantly different from those of the LPS induction group (P < 0.01), further indicating that this combination has a significant anti-inflammatory effect.

[0047] Experimental Example 3 Scavenging ability of free radicals (1) Determination of DPPH· scavenging rate

[0048] Prepare a DPPH free radical ethanol solution (0.1 mmol / L), fill it into a brown bottle and store it in a cool place for later use. The reagent needs to be prepared and used immediately. Measure 0.195 mL of the DPPH ethanol solution and mix it thoroughly with 0.65 mL of the solution samples of each example and Comparative Example 1. Place it in the dark and react at room temperature for 30 min, then measure the absorbance at 517 nm and record it as A (sample). Use 0.65 mL of ethanol solution as the control tube. At the same time, use 0.65 mL of the original Ve solution as the positive control group. After the same treatment, measure its absorbance at 517 nm and record it as A (control). Each sample is measured in parallel 3 times and the average value is taken for calculation.

[0049] Calculate the scavenging rate through the following formula: ; In the formula: A (sample) is the absorbance of the sample to be measured, and A (control) is the absorbance of the standard product.

[0050] The results are shown in Figure 3 , from Figure 3 it can be seen that the processed tangerine peel and orange peel essential oils, as well as Examples 6 and 7 and 8 and 9 added with the above essential oils, all improved the scavenging rate of DPPH·.

[0051] (2) Determination of the scavenging rate of ABTS + · Using an ABTS aqueous solution (7 mmol / L) as the solvent, prepare a potassium persulfate solution (2.45 mmol / L). Place the prepared potassium persulfate solution in the dark for 12 - 16 h to generate the ABTS + · ion. Before the experiment, use ethanol as the dilution medium to dilute the solution containing ABTS + · ion so that its absorbance at a wavelength of 734 nm is 0.700 ± 0.025. Take 0.195 mL of the above solution and mix it with 0.65 mL of each example, Comparative Example 1 and ethanol sample. After reacting thoroughly in the dark for 30 min, measure the absorbance at 734 nm, and the result is recorded as A (sample). Use 0.65 mL of ethanol solution to replace the test examples and the ethanol samples of Comparative Example as the control tube. At the same time, use 0.65 mL of the original Ve solution as the positive control group. Conduct the same treatment and measure its absorbance at a wavelength of 734 nm, and the result is recorded as A (control). Each sample is repeated 3 times and the average value is taken. Calculate the scavenging rate through the following formula: ; In the formula: A (sample) is the absorbance of the sample to be measured, and A (control) is the absorbance of the standard product.

[0052] The results are shown in Figure 4 , and it can be seen from Figure 4 that the processed tangerine peel and orange peel essential oils, as well as Examples 6 and 7 and Examples 8 and 9 added with the above essential oils, all improved the scavenging rate of ABTS + ·.

[0053] (3) Determination of the scavenging rate of O2 - ·

[0054] Using sodium phosphate buffer (0.1 mol / L, pH = 7.4) as the solvent, prepare nitroblue tetrazolium chloride solution (150 μmol / L), nicotinamide adenine dinucleotide solution (468 μmol / L) and phenazine methosulfate solution (60 μmol / L) respectively. Take 0.5 mL of the ethanol solutions of different concentrations of each example and Comparative Example 1 and mix them with 0.5 mL of nitroblue tetrazolium chloride solution and 0.5 mL of nicotinamide adenine dinucleotide solution respectively. After adding 0.5 mL of phenazine methosulfate solution, mix well immediately. After reacting fully for 5 min at room temperature, measure the absorbance at 560 nm, which is A (sample). Replace the sample with 0.5 mL of sodium phosphate buffer, and at the same time use 0.5 mL of the Ve stock solution as the positive control group. After the same treatment, measure the absorbance value at 560 nm as A (control). Each sample is measured 3 times and the average value is taken.

[0055] Calculate the scavenging rate through the following formula: ; In the formula: A (sample) is the absorbance of the sample to be measured, and A (control) is the absorbance of the standard product.

[0056] The results are shown in Figure 5 , and it can be seen from Figure 5 that the essential oil of processed tangerine peel, the essential oil of orange peel, and Examples 6 - 9 added with the above essential oils all improved the scavenging rate of O2 - ·.

[0057] Experimental Example 4 Comparison of the components of Example 8 and Example 9 by LC-MS The components of Example 8 and Example 9 were analyzed using an optimized LC-MS method. After extraction with 80% methanol, the samples were separated on a Waters BEH C18 column (1.7 µm, 2.1×100 mm), with the mobile phase consisting of 0.1% formic acid aqueous solution and acetonitrile / methanol (4:6, containing 0.1% formic acid), and gradient elution was carried out for 13.5 min. The QE-HF-X mass spectrometer collected data in positive and negative ion switching mode (resolution 60,000, m / z 60 - 900). The original data (.raw format) was processed by PeakView 1.2 software to extract the total ion intensity signal, and preprocessing such as baseline correction and Savitzky-Golay smoothing was performed; finally, a total ion chromatogram (TIC) was plotted with retention time as the abscissa and ion intensity as the ordinate. After analysis, the difference in components between the two was not significant, as shown in Figure 6 .

[0058] Experimental Example 5 Analysis of Cough Suppressant Targets and Action Pathways of Example 8 and Example 9 The structural formulas of each component were obtained through the PubChem database, and their corresponding target genes were predicted using the Swiss Target Prediction database. Combining the DisGeNET gene-disease association database and the MalaCards disease database, disease targets related to chronic cough were screened. The target genes of the fried eggs in Examples 8 and 9 were de-duplicated. The Venny online tool was used to generate the intersections of Examples 8 and 9 with disease-related targets respectively, and the drug-related target and disease target datasets were uploaded to construct a Venn diagram. The drug-disease intersection genes were mapped to each node of the Gene Ontology database, and functional annotation was performed using the GO database (http: / / www.geneontology.org / ) and pathway enrichment analysis was performed using the KEGG database (https: / / www.kegg.jp / ). According to the P-value ranking, the top 20 GO terms and the top 20 KEGG pathways were selected for enrichment analysis, and the corresponding bubble charts and bar charts were drawn. The target proteins obtained from the GO function analysis were classified and displayed in three independent ways: biological process (BP), cellular component (CC), and molecular function (MF). The drug-disease intersection targets were imported into the STRING database (https: / / cn.string-db.org / ), the species was limited to "Homo sapiens", and the minimum interaction score threshold was set to 0.400 to obtain protein-protein interaction relationship data. The data was imported into the Cytoscape 3.10.2 software to construct a target protein interaction network. The core targets were screened by analyzing the node degree values. Further, the related components of dried tangerine peel and the associated targets of 8 pathways were imported into the Cytoscape 3.10.2 software to construct a component-target-disease-pathway network diagram.

[0059] GO functional enrichment analysis was performed on the intersection target genes of Example 8 and chronic cough, and a total of 1,280 significantly enriched terms were obtained. The top 20 terms in each classification were selected for visual analysis and a bubble chart was drawn. The results showed that: BP was mainly enriched in inflammatory response, response to external stimuli, and response to lipopolysaccharide, etc.; CC was mainly enriched in plasma membrane, membrane raft, and cell surface, etc.; MF was mainly enriched in protein tyrosine kinase activity, homologue binding, and enzyme binding, etc. See Figures 7 - 9 。

[0060] GO functional enrichment analysis was performed on the intersection target genes of Example 9 and chronic cough, and a total of 1280 significant GO terms were obtained. The top 20 terms with the highest enrichment in the three categories of BP, CC, and MF were selected for visualization analysis, and a bubble plot was constructed. The results showed that: BP was mainly enriched in inflammatory response, response to external stimuli, and response to lipopolysaccharide, etc.; CC was mainly enriched in plasma membrane, membrane raft, and cell surface, etc.; MF was mainly enriched in protein tyrosine kinase activity, homologue binding, and enzyme binding, etc. For details, see Figures 10 - 12 。

[0061] The results of KEGG pathway analysis showed that a total of 228 significantly enriched terms were obtained for the intersection target genes of Example 8 and Example 9 and chronic cough. The top 20 pathways with the highest enrichment were selected for visualization analysis. The results showed that: the intersection target genes of Example 8 and chronic cough were significantly enriched in biological pathways such as cancer pathway, Kaposi sarcoma-associated herpesvirus infection, and HIF-1 signaling pathway; the intersection target genes of Example 9 and chronic cough were mainly involved in signal transduction pathways such as cancer pathway, lipid and atherosclerosis, and HIF-1 signaling pathway. It is worth noting that 15 out of the top 20 significantly enriched pathways of the two cough-relieving eggs were exactly the same, indicating a high similarity in the signal pathways they act on. For details, see Figure 13 。

[0062] According to the test results of the ingredient-target-disease-pathway network, the ingredients of Example 8 mainly included compounds such as ethanolamine, glycine, pyridine, 4-aminobutyric acid, piperidine, and 2-methylpyrrolidine. Its action targets mainly included key genes such as GRIN2A, NOS1, NOS2, NOS3, EGFR, CHRM3, and CYP2D6, and mainly exerted pharmacological effects through MAPK signaling pathway, chemical carcinogenesis-receptor activation pathway, HIF-1 signaling pathway, PD-L1 expression and PD-1 checkpoint pathway in cancer, cancer-related microRNA pathway, and acute myeloid leukemia pathway, etc. The ingredients of Example 9 mainly included ethanolamine, glycine, pyridine, 4-aminobutyric acid, piperidine, 2-methylpyrrolidine, putrescine, etc. Its action targets mainly included ABL1, AGTR1, AKT1, AKT2, ALK, BIRC3, XIAP, etc., and mainly exerted pharmacological effects through EGFR tyrosine kinase inhibitor resistance pathway, MAPK signaling pathway, chemical carcinogenesis-receptor activation pathway, HIF-1 signaling pathway, influenza A-related pathway, and PD-L1 expression and PD-1 cancer checkpoint pathway, etc.

[0063] Experimental Example 6: Cough-relieving experiment on ammonia water-induced coughing mice The adult dosage was set at 2.5 times and 10 times as the low dose and high dose respectively, and the gavage dose was 0.1 mL / kg. Sixty mice were randomly divided into six groups: a model group, a positive control group (pentoxyverine citrate solution group), a low-dose group of Example 8, a high-dose group of Example 8, a low-dose group of Example 9, and a high-dose group of Example 9, with 10 mice in each group. The mice in the groups of Example 7 and Example 8 were continuously gavaged at the designed doses for 14 d, and the model group and the positive control group were gavaged with equal amounts of normal saline and pentoxyverine citrate solution respectively during the same period. One hour after administration on the 15th day, the mice in each group were placed in a 1000 mL container, 0.3 mL of concentrated ammonia water was added, and video recording was immediately started. After continuous exposure for 45 s, the mice were taken out, and the cough behavior and frequency of the mice within 6 min were recorded through video playback, and the latency of the first cough after taking out the mice from the container was statistically analyzed. The judgment criteria for cough behavior were as follows: the mouse opened its mouth accompanied by a cough sound, and at the same time, there were contractions of the chest and abdominal muscles and twitching of the front part of the body.

[0064] Compared with the model group, the cough latency of the mice in the low-dose group and high-dose group of Example 8, the low-dose group and high-dose group of Example 9, and the positive control group was significantly increased (P<0.05, P<0.01), and the increase was most obvious in the high-dose group of Example 8 and the high-dose group of Example 9 (P<0.01). Compared with the positive control group, the latency extension of the low-dose group of Example 8 and the low-dose group of Example 9 was less than that of the positive control group, and the difference was significant (P<0.05), but the increase in the high-dose group of Example 8 and the high-dose group of Example 9 was similar to that of the positive control group, and the difference was not significant (P>0.05); while the cough frequency of the mice in the low-dose group and high-dose group of Example 8, the low-dose group and high-dose group of Example 9, and the positive drug group was reduced, and the difference was statistically significant (P<0.05, P<0.01), among which the cough frequency of the mice in the high-dose group of Example 8 and the high-dose group of Example 9 was reduced most significantly (P<0.01); compared with the positive control group, the reduction of the cough frequency of the mice in the high-dose group of Example 8 and the high-dose group of Example 9 was less than that of the positive control group, and the difference was significant (P<0.01), but the reduction of the cough frequency of the mice in the high-dose group of Example 8 and the high-dose group of Example 9 was not significantly different from that of the positive control group (P>0.05). See Table 2 for details.

[0065] Table 2 Effects of Examples 8 and 9 on cough latency and cough frequency of mice

[0066]

[0067] Note: Compared with the model group, , 。Compared with the positive group, # P< 0.05,## P< 0.01

[0068] In summary, the following conclusions can be drawn: (1) The traditional Chinese medicine essential oil tea oil composition obtained by using ginger-processed tangerine peel essential oil and tea oil as the main components has a significant cough-relieving effect and can be applied to the preparation of cough-relieving dietetic and nutritional products (such as cough-relieving poached eggs or tea eggs).

[0069] (2) Through the in vitro antibacterial test in Experimental Example 1, it is proved that the various essential oils (Examples 1-4) and various essential oil tea oil products (Examples 6-9) prepared and provided by the present invention have the inhibitory ability against harmful bacteria, and there is no significant difference among several groups. Among them, the groups containing ginger-processed tangerine peel and ginger-processed orange peel essential oils (Examples 2, 4), the experimental groups of ginger-processed tangerine peel essential oil tea oil composition and the experimental groups of ginger-processed tangerine peel essential oil tea oil composition (Examples 6, 7) and the related experimental samples (Examples 8, 9) can all significantly inhibit the bacterial level, and the treatment effect is close to that of Comparative Example 1. Especially for the groups of Examples 8-9, because the formulated composition is a compatibility composition of ginger-processed tangerine peel and ginger-processed orange peel essential oils and tea oil, whether compared with the blank control group and the tangerine peel essential oil group or with the orange peel essential oil group, the inhibitory effect of this composition sample on harmful bacteria is more significant (Experimental Example 1), and the antioxidant ability and the effect of scavenging hydroxyl oxygen free radicals in the in vitro experiment (Experimental Example 3) are also stronger, indicating that the compatibility combination of ginger-processed tangerine peel essential oil and ginger-processed orange peel essential oil and tea oil has a synergistic effect.

[0070] (3) The present invention proves through in vitro cell experiments that tangerine peel essential oil and orange peel essential oil can significantly reduce the expression of pro-inflammatory factors IL-6 and IL-12 (P<0.05) and increase the expression of anti-inflammatory factors IL-4 and IL-10 (P<0.05); while ginger-processed tangerine peel essential oil and ginger-processed orange peel essential oil can more significantly reduce the expression of IL-6, IL-12 and increase the expression of IL-4 and IL-10 (P<0.01), which indicates that the tangerine peel essential oil and orange peel essential oil after being processed with ginger juice have a stronger anti-inflammatory effect. Animal experiments prove that the tea oil compositions containing ginger-processed tangerine peel essential oil and ginger-processed orange peel essential oil (Examples 8, 9) can all significantly reduce the inflammatory reaction.

[0071] (4) The present invention proves through animal experiments that the tea oil compositions containing ginger-processed tangerine peel essential oil and ginger-processed orange peel essential oil (Examples 8, 9) can significantly increase the cough induction latency of ammonia water and reduce the number of coughs, indicating that the tea oil composition containing ginger-processed tangerine peel essential oil and ginger-processed orange peel essential oil has a better effect of increasing the cough induction latency of ammonia water and reducing the number of coughs.

[0072] (5) The present invention proves through experiments that the tea oil compositions containing ginger-processed tangerine peel essential oil and ginger-processed orange peel essential oil (Examples 8, 9) can significantly increase the cough latency and reduce the number of coughs, indicating that they have a good cough-relieving effect.

[0073] (6)The preparation method of the ginger-processed tangerine peel, orange peel essential oil and tea oil according to the present invention involves conventional equipment, and has the advantages of unique method, simple process, convenient operation, low cost, high essential oil yield, etc.

[0074] The present invention has conducted experimental exploration and research and development on the cough-relieving effect of the tangerine peel essential oil and tea oil composition before and after processing, providing a theoretical basis for the development and utilization of cough-relieving food therapy and health care products.

[0075] The present invention has a cough-relieving effect by extracting essential oil from ginger-processed tangerine peel and formulating it with tea oil to make egg products. Conducting research and development on it has great practical significance and also provides a theoretical basis for the development and utilization of egg food therapy products.

[0076] The specific embodiments of the present invention have been described in detail above, but they are only examples, and the present invention is not limited to the above-described specific embodiments. For those skilled in the art, any equivalent modifications and substitutions made to the present invention are also within the scope of the present invention. Therefore, all equivalent transformations and modifications made without departing from the spirit and scope of the present invention are covered by the scope of the present invention.

Claims

1. A traditional Chinese medicine essential oil tea oil composition with a cough-relieving effect, characterized in that, It includes ginger-processed traditional Chinese medicine essential oil and tea oil. The preparation method of the ginger-processed traditional Chinese medicine essential oil includes the following steps: S1. Preparation of ginger juice: Take fresh ginger, wash it, crush it, add distilled water and decoct it. Combine the filtrates and concentrate them under reduced pressure. Take the concentrated solution to obtain ginger juice; S2. Process tangerine peel or orange peel with ginger juice: Cut tangerine peel or orange peel into particles and dry them in the shade to obtain tangerine peel or orange peel particles. Add ginger juice to soak them to obtain tangerine peel or orange peel particles soaked with ginger juice; S3. Extraction of ginger-processed tangerine peel or orange peel essential oil: Take the tangerine peel or orange peel particles soaked with ginger juice in a distillation container, add distilled water containing 1-ethyl-3-methylimidazolium tetrafluoroborate ionic liquid, heat to boiling and slightly boil under normal pressure, collect the mixed steam of volatile oil and water, obtain the oil-phase fraction through condensation and liquid separation, add a desiccant to dry it, filter to remove the desiccant to obtain ginger-processed tangerine peel or orange peel essential oil, that is, ginger-processed traditional Chinese medicine essential oil; the mass ratio of the ginger-processed traditional Chinese medicine essential oil to tea oil is 0.5-2:99.5-98.

2. The composition according to claim 1, characterized in that, In step S1, the mass-volume ratio of ginger to distilled water is 1:4-6, and each decoction is for 15-25 min; decoct for 1-3 times.

3. The composition according to claim 1, characterized in that, In step S1, concentrate until the mass concentration of the crude drug contained is 0.9-1.1 g / mL.

4. The composition according to claim 1, characterized in that, In step S2, the dosage of ginger juice is 0.5-1.2 times the mass of tangerine peel or orange peel particles, and soak for 12-18 h.

5. The composition according to claim 1, characterized in that, In step S3, the addition amount of the distilled water containing 1-ethyl-3-methylimidazolium tetrafluoroborate ionic liquid is 3-8 times the mass of tangerine peel or orange peel particles, and calculated by distilled water, 1-ethyl-3-methylimidazolium tetrafluoroborate ionic liquid is 0.5-1.5% of the mass of distilled water.

6. Use of the composition according to any one of claims 1-5 in the preparation of a cough-relieving product.

7. The use according to claim 6, characterized in that, The addition amount of the composition accounts for 20-40% of the total mass of the cough-relieving product.

8. The use according to claim 6, characterized in that, The preparation method of the cough-relieving product is as follows: Select the egg liquid of fresh and intact poultry eggs; Pour the composition into a frying pan, heat to 80-100 °C, break in the egg liquid, and decoct until cooked through.

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