A pheromone composition and a method for its preparation

Abstract

The present application belongs to the technical field of veterinary medicine, and relates to a pheromone composition and a preparation method thereof. The pheromone composition can play a soothing role on all mammal species, can be used for relieving stress and anxiety, and can enhance the immune capacity of the mammal by using the component formula of Poria cocos, palmitic acid, linoleic acid, linolenic acid, lauric acid, oleic acid and citronella oil.

Description

Technical Field

[0001] This invention belongs to the field of veterinary drug technology and relates to a pheromone composition and its preparation method. Background Technology

[0002] Pheromones, also known as pheromones, are substances secreted by an individual and detected by other individuals of the same species through their olfactory organs (such as the accessory olfactory bulb and vomeronasal organ), causing the latter to exhibit certain behavioral, emotional, psychological, or physiological changes. They serve a communication function. The presence of pheromones has been proven in almost all animals.

[0003] However, while pheromones currently have a calming effect on mammals (pigs, cattle, cats, dogs, etc.) and birds, no pheromone has a calming effect on all of these animals. Furthermore, current pheromones do not enhance the immunity of mammals and birds, nor do they repel mosquitoes.

[0004] Chinese patent application document (publication number: CN112370528A) discloses a pheromone composition with identification function and its formulation. The pheromone composition includes: component A: androstenone and / or androstenol; component B: one or more of geraniol, cisaldehyde, palmitic acid, oleic acid, linoleic acid, decanoic acid, myristic acid, lauric acid, and 3-methylindole; and component C: a colorant. It uses a combination of multiple pheromones and adds plant-derived synergistic components to improve the estrus rate and reproductive success rate of sows. The addition of a colorant for identification improves the efficiency of estrus detection. However, the addition of geraniol, primarily used to reduce stress in sows, and the presence of ethanol, raises safety concerns. Summary of the Invention

[0005] The purpose of this invention is to address the aforementioned problems in the prior art by proposing a pheromone composition that enhances the immune capacity of mammals and improves the immune effect after vaccination.

[0006] The objective of this invention can be achieved through the following technical solution: the pheromone composition comprises the following raw materials in parts by weight: 1-5 parts Poria cocos, 40-60 parts palmitic acid, 10-30 parts linoleic acid, 10-30 parts linolenic acid, 5-25 parts lauric acid, 1-20 parts oleic acid, and 1-5 parts citronella oil.

[0007] In one of the above-mentioned pheromone compositions, the pheromone composition comprises the following raw materials in parts by weight: 2-3 parts of Poria cocos extract, 45-55 parts of palmitic acid, 15-25 parts of linoleic acid, 15-25 parts of linolenic acid, 10-20 parts of lauric acid, 5-15 parts of oleic acid, and 2-3 parts of lemongrass oil.

[0008] This invention utilizes the synergistic effect of palmitic acid, linoleic acid, linolenic acid, oleic acid, and lauric acid to effectively maintain the sedative activity of mammalian and avian pheromones, while also ensuring good compatibility with Poria cocos components. The polysaccharides in Poria cocos include carboxymethyl polysaccharides, which have sedative and calming effects. Traditional Chinese medicine believes that Poria cocos is effective for relieving chest and rib pain, melancholy, fright, palpitations, epigastric pain, chills and fever, cough, dry mouth and tongue. Poria cocos is diuretic, and long-term use can calm the mind, nourish the spirit, alleviate hunger, and prolong life. Furthermore, by adding a certain amount of citronella oil, this invention provides insect repellent, mosquito repellent, and anti-bite protection, as well as natural antibacterial properties, deodorizing and stimulating effects. This allows the pheromone composition of this invention to purify and uplift mood, thus calming emotions.

[0009] The present invention also provides a method for preparing the above-mentioned pheromone composition, the method comprising the following steps:

[0010] S1. First, soak the Poria cocos, then decoct it with water 1-3 times, and extract the liquid after each decoction.

[0011] S2. Filter the extracted liquid for the first time and let it stand. Take the supernatant and filter it for the second time and let it stand. Then heat and concentrate it.

[0012] S3. After concentration, palmitic acid is added and pre-stirred, followed by the addition of linoleic acid, linolenic acid, lauric acid, oleic acid, and citronella oil, followed by stirring.

[0013] S4. After stirring, the mixture is filtered to obtain the pheromone composition.

[0014] In the above-mentioned method for preparing a pheromone composition, the first filtration and settling screen has a mesh size of 80-120 and a settling time of 6-8 hours.

[0015] In the above-mentioned method for preparing a pheromone composition, the mesh size of the second filtration and settling screen is 180-230, and the settling time is 2-5 hours.

[0016] In the above-mentioned method for preparing a pheromone composition, the heating and concentration temperature is 65-80℃ and the time is 1-3h.

[0017] In the above-mentioned method for preparing a pheromone composition, step S4 involves filtration using a 0.4-0.5 μm filter membrane.

[0018] In the above-mentioned method for preparing a pheromone composition, the solid content of the pheromone composition in step S4 is 20-30%.

[0019] A formulation prepared using the above-mentioned pheromone composition, the formulation comprising at least one of a spray formulation, an aerosol formulation, and a liquid formulation.

[0020] The prepared composition, when used in mammals, can prevent stress and enhance immunity. This invention achieves a calming effect on all mammalian species by controlling the content of each component, and can be used to relieve stress and anxiety, as well as enhance the immune system of mammals.

[0021] This invention applies a pheromone spray composition, in the form of a spray, aerosol, or similar agent, to the area surrounding a stressful event. This leads to a reduction in stress caused by various factors, such as weight gain, social behavior related to other mammals, bodily wounds, particularly ear wounds, salivary cortisol, and heart rate, thereby enhancing the animal's immunity and increasing stress resistance. Therefore, the composition of this invention can be applied to various objects that mammals come into contact with, such as walls, air, and toys.

[0022] Compared with the prior art, the present invention has the following beneficial effects: the present invention achieves a composition that can soothe all mammalian species through the formulation of components such as Poria cocos, palmitic acid, linoleic acid, linolenic acid, lauric acid, oleic acid and citronella oil, and can be used to relieve stress and anxiety, as well as enhance the immune capacity of mammals. Detailed Implementation

[0023] The following are specific embodiments of the present invention, which further describe the technical solution of the present invention, but the present invention is not limited to these embodiments.

[0024] Example 1:

[0025] S1. First, soak Poria cocos in distilled water for 3 hours, with the water level 4 cm above the surface of the herbs. Then, add 10 times the volume of distilled water to the soaked herbs, bring to a boil, and simmer for 2.5 hours. Squeeze out the liquid, then add water again to the surface of the herbs and simmer for another 1.5 hours. Squeeze out the liquid twice, pass it through a 100-mesh sieve, combine the two liquids, refrigerate for 20 minutes, let it stand for 7 hours to settle, take the supernatant, pass it through a 200-mesh sieve, filter, and let it stand for 3 hours. Heat at 70℃ until the water evaporates for 2 hours, stirring continuously. Heat until a thick paste forms, obtaining a medicinal mud. Cool and set aside.

[0026] S2. Take 2 parts of the above-mentioned medicinal mud according to the mass ratio, add 50 parts of palmitic acid and stir for 20 minutes to aid dissolution. Then add 20 parts of linoleic acid, 20 parts of linolenic acid, 15 parts of lauric acid, 10 parts of oleic acid, and 2.5 parts of citronella oil in sequence, and continue stirring at room temperature for 2 hours.

[0027] S4. Finally, filter the solution using a 0.45-micron (Merk-Millipore) filter membrane to obtain a solution with the desired solid content. Before use, dispense the solution into spray nozzles, 100 ml per bottle.

[0028] Example 2:

[0029] S1. First, soak Poria cocos in distilled water for 3 hours, with the water level 4 cm above the surface of the herbs. Then, add 10 times the volume of distilled water to the soaked herbs, bring to a boil, and simmer for 2.5 hours. Squeeze out the liquid, then add water again to the surface of the herbs and simmer for another 1.5 hours. Squeeze out the liquid twice, pass it through an 80-mesh sieve, combine the two liquids, refrigerate for 20 minutes, let it settle for 6 hours, take the supernatant, pass it through an 180-mesh sieve, filter, and let it stand for 2 hours. Heat at 65℃ until the water evaporates for 1 hour, stirring continuously. Heat until a thick paste forms, then cool and set aside.

[0030] S2. Take 2 parts of the above-mentioned medicinal mud according to the mass ratio, add 45 parts of palmitic acid and stir for 20 minutes to aid dissolution. Then add 15 parts of linoleic acid, 15 parts of linolenic acid, 10 parts of lauric acid, 5 parts of oleic acid, and 2 parts of citronella oil in sequence, and continue stirring at room temperature for 2 hours.

[0031] S4. Finally, filter the solution using a 0.40-micron (Merk-Millipore) membrane to obtain a solution with the desired solid content. Before use, dispense the solution into spray nozzles, 100 ml per bottle.

[0032] Example 3:

[0033] S1. First, soak Poria cocos in distilled water for 3 hours, with the water level 4 cm above the surface of the herbs. Then, add 10 times the volume of distilled water to the soaked herbs, bring to a boil, and simmer for 2.5 hours. Squeeze out the liquid, then add water again to the surface of the herbs and simmer for another 1.5 hours. Squeeze out the liquid twice, pass it through a 120-mesh sieve, combine the two liquids, refrigerate for 20 minutes, let it settle for 8 hours, take the supernatant, pass it through a 230-mesh sieve, filter, and let it stand for 5 hours. Heat at 80℃ until the water evaporates for 3 hours, stirring continuously. Heat until a thick paste forms, obtaining a medicinal mud. Cool and set aside.

[0034] S2. Take 3 parts of the above-mentioned medicinal mud according to the mass ratio, add 55 parts of palmitic acid and stir for 20 minutes to aid dissolution. Then add 25 parts of linoleic acid, 25 parts of linolenic acid, 20 parts of lauric acid, 15 parts of oleic acid, and 3 parts of citronella oil in sequence. Continue stirring at room temperature for 2 hours.

[0035] S4. Finally, filter the solution using a 0.50-micron (Merk-Millipore) filter membrane to obtain a solution with the desired solid content. Before use, dispense the solution into spray nozzles, 100 ml per bottle.

[0036] Comparative Example 1:

[0037] The only difference from Example 1 is that palmitic acid was not added in step S2.

[0038] Comparative Example 2:

[0039] The only difference from Example 1 is that linoleic acid was not added in step S2.

[0040] Comparative Example 3:

[0041] The only difference from Example 1 is that linolenic acid was not added in step S2.

[0042] Comparative Example 4:

[0043] The only difference from Example 1 is that lauric acid was not added in step S2.

[0044] Comparative Example 5:

[0045] The only difference from Example 1 is that oleic acid was not added in step S2.

[0046] Comparative Example 6:

[0047] The only difference from Example 1 is that lemongrass oil was not added in step S2.

[0048] Comparative Example 7:

[0049] The only difference from Example 1 is that palmitic acid, linoleic acid, and linolenic acid were not added in step S2.

[0050] Comparative Example 8:

[0051] The only difference from Example 1 is that lauric acid, oleic acid, and citronella oil were not added in step S2.

[0052] Blank control group:

[0053] This refers to any pheromone composition prepared without using Examples 1-3 or Comparative Examples 1-8.

[0054] For each group of experimental animals, 3-5 ml of the above sample was sprayed on the nose or face. Blood was collected 3 days later, the serum was centrifuged, and oxidative stress indicators were detected.

[0055] Establishment and stability testing of the antioxidant model:

[0056] Vitamin C solutions were prepared, and the absorbance of the DPPH solutions was measured at concentrations of 0 μg / mL, 40 μg / mL, 80 μg / mL, 120 μg / mL, 160 μg / mL, and 200 μg / mL. Regression curves and linear relationships were plotted to verify the success and stability of the model.

[0057] The extracted sample aqueous solutions were prepared at concentrations of 1 mg / mL, 10 mg / mL, and 100 mg / mL using a 10-fold dilution method. Color comparisons were performed to determine the linearity interval. Based on the curve, five concentration gradients were established for antioxidant assays.

[0058] Antioxidant screening at a sample concentration of 0.1 g / mL

[0059] (1) Take 0.2 mL of the sample solution of the test substance at the specified concentration and 3.8 mL of 0.2 mol / mL DPPH solution into a test tube, shake well, and react in the dark for 30 min. Use anhydrous ethanol as a blank to measure its absorbance Aa at 517 nm.

[0060] (2) Take 0.2 mL of distilled water and 3.8 mL of 0.2 mol / mL DPPH solution and put them into a test tube. Shake well and react in the dark for 30 min. Use anhydrous ethanol as a blank and measure its absorbance at 517 nm. This is the aqueous solution control. Ab Ra (%) = (1-Aa / Ab)*100% Note: Aa is the absorbance after adding antioxidant; Ab is the control; calculate the IC50 of each sample based on a 50% scavenging rate to evaluate the antioxidant properties of the samples.

[0061] Data Analysis

[0062] The experimental data will be statistically analyzed using the GLM program in SAS 9.4 (SAS Institute, 2008). Each column will serve as the statistical unit for production performance. The Tukey HSD test will be used for multiple comparisons between groups, with a p-value less than or equal to 0.05 indicating a significant difference between groups.

[0063] Table 1: Antioxidant stress effects of pheromone compositions from Examples 1-3 and Comparative Examples 1-8 on pigs

[0064] Example Number of experimental animals unit IC50 value Blank control group 100 μg / ml 1132.85 Example 1 102 μg / ml 159.31 Example 2 110 μg / ml 221.86 Example 3 96 μg / ml 313.61 Comparative Example 1 100 μg / ml 613.19 Comparative Example 2 103 μg / ml 812.62 Comparative Example 3 100 μg / ml 755.97 Comparative Example 4 95 μg / ml 707.2 Comparative Example 5 101 μg / ml 947.78 Comparative Example 6 98 μg / ml 843.09 Comparative Example 7 102 μg / ml 987.65 Comparative Example 8 99 μg / ml 990.48

[0065] Table 2: Antioxidant stress effects of pheromone compositions from Examples 1-3 and Comparative Examples 1-8 on cattle

[0066]

[0067]

[0068] Table 3: Antioxidant stress effects of pheromone compositions from Examples 1-3 and Comparative Examples 1-8 on sheep

[0069] Example Number of experimental animals unit IC50 value Blank control group 48 μg / ml 1278.26 Example 1 52 μg / ml 102.89 Example 2 45 μg / ml 189.93 Example 3 48 μg / ml 195.83 Comparative Example 1 52 μg / ml 308.81 Comparative Example 2 46 μg / ml 355.08 Comparative Example 3 49 μg / ml 534.66 Comparative Example 4 51 μg / ml 603.51 Comparative Example 5 48 μg / ml 648.48 Comparative Example 6 53 μg / ml 663.09 Comparative Example 7 53 μg / ml 850.16 Comparative Example 8 47 μg / ml 939.67

[0070] Table 4: Antioxidant stress effects of pheromone compositions from Examples 1-3 and Comparative Examples 1-8 on chickens

[0071] Example Number of experimental animals unit IC50 value Blank control group 204 μg / ml 1089.51 Example 1 214 μg / ml 178.63 Example 2 192 μg / ml 227.41 Example 3 199 μg / ml 245.23 Comparative Example 1 194 μg / ml 605.56 Comparative Example 2 202 μg / ml 653.77 Comparative Example 3 203 μg / ml 712.07 Comparative Example 4 187 μg / ml 718.41 Comparative Example 5 211 μg / ml 737.31 Comparative Example 6 195 μg / ml 745.66 Comparative Example 7 202 μg / ml 913.02 Comparative Example 8 186 μg / ml 940.01

[0072] Table 5: Antioxidant stress effects of pheromone compositions from Examples 1-3 and Comparative Examples 1-8 on cats

[0073] Example Number of experimental animals unit IC50 value Blank control group 39 μg / ml 1325.48 Example 1 40 μg / ml 141.42 Example 2 36 μg / ml 246.11 Example 3 41 μg / ml 347.62 Comparative Example 1 38 μg / ml 504.69 Comparative Example 2 38 μg / ml 533.54 Comparative Example 3 39 μg / ml 567.37 Comparative Example 4 39 μg / ml 667.37 Comparative Example 5 40 μg / ml 771.62 Comparative Example 6 36 μg / ml 878.66 Comparative Example 7 39 μg / ml 961.36 Comparative Example 8 37 μg / ml 985.43

[0074] Table 6: Antioxidant stress effects of pheromone compositions from Examples 1-3 and Comparative Examples 1-8 on dogs

[0075] Example Number of experimental animals unit IC50 value Blank control group 41 μg / ml 1238.32 Example 1 38 μg / ml 109.59 Example 2 37 μg / ml 199.08 Example 3 40 μg / ml 304.46 Comparative Example 1 38 μg / ml 484.87 Comparative Example 2 41 μg / ml 603.32 Comparative Example 3 40 μg / ml 648.93 Comparative Example 4 39 μg / ml 658.57 Comparative Example 5 38 μg / ml 673.57 Comparative Example 6 35 μg / ml 746.85 Comparative Example 7 38 μg / ml 876.62 Comparative Example 8 42 μg / ml 953.82

[0076] Examples 1-3 and Comparative Examples 1-8: The pheromone compositions were sprayed onto mice daily for one week. After this period, the organ / body weight ratios were measured. After the feeding period, the mice were weighed, euthanized, and the thymus and spleen were removed. The fascia was removed, the blood on the surface of the organs was dried, and their weights were measured. The thymus / body weight ratio and spleen / body weight ratio were calculated. The results are shown in Table 7.

[0077] Table 7: Effects of pheromone compositions from Examples 1-3 and Comparative Examples 1-8 on organ indices of the thymus and spleen in mice.

[0078]

[0079] In summary, the present invention achieves a composition that can soothe all mammalian species through the formulation of Poria cocos, palmitic acid, linoleic acid, linolenic acid, lauric acid, oleic acid, and citronella oil, and can be used to relieve stress and anxiety. Table 7 shows that a higher organ ratio indicates better organ growth and stronger immune performance, indicating that the present invention can also enhance the immune capacity of mammals.

[0080] The embodiments described herein cover any points not exhaustively within the scope of the technical claims of this invention, as well as new technical solutions formed by equivalent substitutions of one or more technical features in the embodiments. These are all within the scope of the claims of this invention. Furthermore, in all listed or unlisted embodiments of this invention, each parameter in the same embodiment merely represents an instance (i.e., a feasible solution) of its technical solution, and there is no strict coordination or limitation relationship between the parameters. The parameters can be substituted for each other without violating axioms and the claims of this invention, unless otherwise stated.

[0081] The technical means disclosed in this invention are not limited to those described above, but also include technical solutions composed of any combination of the above technical features. The above descriptions are specific embodiments of this invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of this invention, and these improvements and modifications are also considered within the scope of protection of this invention.

[0082] The specific embodiments described herein are merely illustrative of the spirit of the invention. Those skilled in the art to which this invention pertains may make various modifications or additions to the described specific embodiments or use similar methods to substitute them, without departing from the spirit of the invention or exceeding the scope defined by the appended claims.

Claims

1. A pheromone composition, characterized in that, The pheromone composition is made from the following raw materials in parts by weight: 2-3 parts Poria cocos extract, 45-55 parts palmitic acid, 15-25 parts linoleic acid, 15-25 parts linolenic acid, 10-20 parts lauric acid, 5-15 parts oleic acid, and 2-3 parts lemongrass oil. The method for preparing the pheromone composition includes the following steps: S1. First, soak the Poria cocos, then decoct it with water 1-3 times, and extract the liquid after each decoction. S2. Filter the extracted liquid for the first time and let it stand. Take the supernatant and filter it for the second time and let it stand. Then heat and concentrate it. S3. After concentration, add palmitic acid and pre-stir, then add linoleic acid, linolenic acid, lauric acid, oleic acid and citronella oil in sequence and stir. S4. After stirring, the mixture is filtered to obtain the pheromone composition.

2. The pheromone composition according to claim 1, characterized in that, The first filtration and settling process involves using a screen with a mesh size of 80-120 and a settling time of 6-8 hours.

3. The pheromone composition according to claim 1, characterized in that, The second filtration and settling process involves using a 180-230 mesh screen and setting the screen for 2-5 hours.

4. The pheromone composition according to claim 1, characterized in that, The heating and concentration temperature is 65-80℃, and the time is 1-3 hours.

5. The pheromone composition according to claim 1, characterized in that, Step S4 involves filtration using a 0.4-0.5 μm filter membrane.

6. The pheromone composition according to claim 1, characterized in that, The solid content of the pheromone composition in step S4 is 20-30%.

7. A formulation prepared using the pheromone composition of claim 1, characterized in that, The formulation is at least one of a spray formulation, an aerosol formulation, and a liquid formulation.

Images