Extraction method of perilla leaf oil, perilla leaf oil and application of perilla leaf oil

Abstract

The invention discloses an extraction method of perilla leaf oil, the perilla leaf oil and application of the perilla leaf oil. The extraction method comprises the following steps: chopping fresh perilla leaves to obtain crushed leaves; spraying a composite enzymatic hydrolysate to the crushed leaves, placing the crushed leaves in a heat preservation box, and performing low-temperature enzymolysis; putting the leaves subjected to enzymolysis into a reduced-pressure extraction tank, adding water, and performing negative-pressure extraction; condensing and collecting steam evaporated under negative pressure into an oil-water separator by using a condenser; separating and collecting an oil layer, and adding a drying agent to remove excessive moisture to obtain the perilla leaf oil. The yield of the perilla leaf oil extracted by the method disclosed by the invention is high. Only enzyme and a high-boiling-point solvent are added in the process, and no residual risk exists after distillation. The production process is simple, and the production period is short; the requirement on equipment performance is not high, and the energy consumption is low. The extracted perilla leaf oil can be added with auxiliary materials to be prepared into essence or a cream preparation for use, and the perilla leaf oil can be used in medicines and cosmetics with related effects of resisting oxidation, resisting inflammation and the like.

Description

Technical Field

[0001] This invention relates to the field of plant essential oils and their extraction technology, and in particular to a method for extracting perilla leaf oil, perilla leaf oil and its applications. Background Technology

[0002] Perilla leaf oil and perillaldehyde are important active ingredients in perilla leaves. Perilla leaf oil mainly possesses antioxidant, antibacterial, anti-inflammatory, vasodilatory, antitumor, and antidepressant effects. Its chemical composition is complex, primarily including perillaldehyde, perillone, perillyl alcohol, D-limonene, and β-caryophyllene. These components endow perilla leaf oil with development value in the food, pharmaceutical, and cosmetic fields.

[0003] Perillaldehyde is the main active ingredient in perilla leaf oil, exhibiting significant anti-inflammatory, antibacterial, and antioxidant effects. It also demonstrates certain pharmacological activities in the cardiovascular, nervous, and digestive systems. Studies have shown that perillaldehyde can significantly reduce the expression of pro-inflammatory factors (such as IL-6 and TNF-α) by inhibiting the NF-κB pathway, showing significant efficacy against inflammatory diseases such as ulcerative colitis and dermatitis. Its broad-spectrum antibacterial properties (e.g., a MIC as low as 0.1-5 μL / mL against Staphylococcus aureus) have led to its application in food preservation and antifungal treatment. In terms of antioxidant activity, perilla leaf oil can enhance SOD activity and scavenge free radicals; animal experiments have shown that it can improve cognitive function in aging models.

[0004] In skincare products, perilla leaf oil is more commonly used, as it can exert antioxidant and anti-inflammatory soothing effects, as well as provide moisturizing benefits. Perilla aldehyde, on the other hand, is more irritating and is rarely used alone. It is usually combined with other ingredients to reduce its irritation and enhance its anti-inflammatory effects.

[0005] Currently, research on perilla leaf oil and perillaldehyde mainly focuses on chemical composition analysis, extraction process optimization, and pharmacological effects. Although scholars both domestically and internationally have conducted extensive research on the chemical composition and pharmacological effects of perilla volatile oil, systematic research remains insufficient, especially regarding the extraction efficiency and stability of perillaldehyde, where technical bottlenecks persist. Furthermore, the content of perillaldehyde may vary depending on factors such as perilla germplasm, growth environment, and extraction method, and transformation may occur during extraction, further complicating its research and application.

[0006] Current extraction techniques for perilla leaf oil still primarily rely on steam distillation (yields are only 0.2%–1.5%), which suffers from the drawback of high temperatures damaging heat-sensitive components. While supercritical CO2 extraction and molecular distillation can increase yields to over 5%, the equipment costs are high. Furthermore, the synthesis of perillaldehyde depends on β-pinene oxidation, resulting in significant pollution, and microbial fermentation synthesis is still in the laboratory stage.

[0007] As disclosed in Chinese patent CN201710532622.2, a method for extracting perilla oil from perilla leaves and the perilla oil itself are described, this method requires adjusting the steam pressure twice and involves prolonged steam heating (up to 3 hours), resulting in high energy consumption. High temperatures may also cause oxidation or degradation of some heat-sensitive active ingredients in perilla oil (such as perillaldehyde).

[0008] Chinese patent CN201910866799.5 discloses a method for extracting perillaldehyde from perilla, achieving an extraction rate of 24.76% and a purity of over 99.3%. However, this method requires multiple steps (such as grinding antioxidants, enzyme activation, catalyst preparation, and adsorbent modification), making it cumbersome and time-consuming. Subsequent extraction with ethyl acetate, manganese dioxide oxidant, and macroporous resin adsorption may introduce solvent or chemical residues, affecting the natural purity of perillaldehyde. Furthermore, the preparation of the composite enzyme, catalyst, and modified adsorbent requires additional resources, potentially increasing the cost of industrial production.

[0009] Chinese patent CN201810535059.9 discloses an extraction process for perilla oil. This method uses petroleum ether and ethanol as extractants, posing a risk of flammability and explosion, and solvent residue may affect the safety of the perilla oil (requiring subsequent solvent removal). Furthermore, it requires pulverization at low temperatures (-50 to -90°C) and controlled pressure (0.9-1.1 MPa), placing strict demands on equipment performance and process control, thus increasing investment costs. Although soybean oil is added to improve the extraction rate, the mixed solvents may cause perilla oil to coexist with other lipid components, requiring further separation and purification.

[0010] In summary, there are many existing technologies for extracting perilla leaf oil from perilla leaves, but all of them have shortcomings. It is worthwhile to study an extraction process that integrates high extraction rate, high efficiency, high safety, and low cost. Summary of the Invention

[0011] Purpose of the invention: In order to overcome the problems existing in the prior art, the present invention proposes an extraction method for perilla leaf oil, perilla leaf oil and its application, which has a higher yield, a higher utilization rate of raw materials, and a simple process with low energy consumption, low equipment requirements, and the components are not easily damaged by high temperature.

[0012] Technical solution: To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows:

[0013] A method for extracting perilla leaf oil includes the following steps:

[0014] 1) Chop the fresh perilla leaves into small pieces;

[0015] 2) After spraying the crushed leaves with the compound enzymatic hydrolysate, place them in an incubator for low-temperature enzymatic hydrolysis;

[0016] 3) Place the enzymatically hydrolyzed leaves in a vacuum extraction tank, add water, and extract under negative pressure;

[0017] 4) Use a condenser to condense and collect the steam that evaporates under negative pressure into an oil-water separator;

[0018] 5) Separate and collect the oil layer, add a desiccant to remove excess water, and obtain perilla leaf oil.

[0019] Furthermore, in step 1), the average diameter of the chopped perilla leaves is 1-5 cm.

[0020] Furthermore, in step 2), the compound enzymatic hydrolysate is a mixed aqueous solution containing cellulase, pectinase, triacetin, and Tween 80, and the spraying amount is 0.5 to 3 times the weight of the leaves.

[0021] Furthermore, the content of each component by mass is as follows: cellulase 1%–10%, pectinase 1%–10%, triacetin 2%–8%, and Tween 80 2%–10%.

[0022] Furthermore, in step 2), the enzymatic hydrolysis temperature is 30–50°C, and the enzymatic hydrolysis time is 1–6 hours.

[0023] Furthermore, in step 3), the amount of water added is 8 to 20 times the weight of the perilla leaves, the vacuum gauge reading for the negative pressure extraction is -0.081 to -0.054 MPa, the extraction temperature is 60 to 80°C, and the extraction time is 0.5 to 2 hours.

[0024] Furthermore, in step 5), the desiccant is selected from one or a combination of anhydrous sodium sulfate, anhydrous magnesium sulfate, molecular sieve, and silica gel desiccant, and the amount used is 5% to 20% of the oil layer weight.

[0025] Furthermore, the extraction method includes the following steps:

[0026] Chop fresh perilla leaves into pieces with an average diameter of 2-3 cm;

[0027] Spray the crushed leaves with a compound enzymatic hydrolysate at 1 to 2 times its weight, wherein the compound enzymatic hydrolysate is a mixed aqueous solution containing 5% to 10% cellulase, 1% to 4% pectinase, 2% to 5% triacetin, and 4% to 6% Tween 80.

[0028] Place in an incubator and perform low-temperature enzymatic hydrolysis at 35–45°C for 2–5 hours;

[0029] The enzymatically hydrolyzed leaves were placed in a vacuum extraction vessel, and 10 to 15 times the weight of the perilla leaves in water were added. The extraction was carried out at a vacuum gauge reading of -0.081 to -0.070 MPa and a temperature of 60 to 70°C for 0.5 to 1 hour.

[0030] The steam that evaporates under negative pressure is condensed and collected in the oil-water separator by the condenser;

[0031] The oil layer is separated and collected, and 8% to 15% of anhydrous sodium sulfate or silica gel desiccant is added to remove excess moisture to obtain perilla leaf oil.

[0032] The present invention also discloses perilla leaf oil extracted by the above extraction method, wherein the perillaldehyde content is 33.6%-52.8%.

[0033] The present invention also discloses the application of the above-extracted perilla leaf oil, specifically, it is formulated with excipients into an essence or cream preparation for use in pharmaceuticals and cosmetics.

[0034] Beneficial effects:

[0035] 1) Significantly improved extraction efficiency and higher product purity: This invention achieves a perilla leaf oil yield of up to 1.27% through the synergistic effect of low-temperature enzymatic hydrolysis (30-50℃) and negative pressure distillation (60-80℃), with a perillaldehyde content as high as 52.8%, far exceeding conventional methods (usually <40%), effectively preserving the integrity of the active ingredients; the compound enzymatic hydrolysate (containing triacetylglycerol + Tween 80) has made a breakthrough in solving the technical problems of perillaldehyde volatility and insufficient extraction in traditional processes.

[0036] 2) No organic solvents are involved in the entire process, only water and food-grade enzyme preparations are used, eliminating the risk of solvent residue. The product can be directly used in the fields of medicine and cosmetics. Compared with supercritical CO2 extraction (high pressure equipment) and traditional distillation (above 100℃), this invention completes the extraction at a low temperature of 60-80℃, which greatly saves energy consumption. Moreover, the equipment requirements are simple: only a conventional vacuum distillation device is needed, without the need for expensive supercritical equipment, and small and medium-sized enterprises can also quickly start production.

[0037] 3) Superior Product Performance: The bioactivity of perilla leaf oil is fully preserved in the resulting essence / cream formulations; the perilla leaf oil extracted in this invention can be used in the products without further purification. The efficacy of perilla leaf oil is influenced by other components (such as perillaldehyde and caryophyllene), resulting in a wider range of effects. Its overall efficacy relies more on the synergistic effect of its various chemical components. It has a broader application range than perillaldehyde.

[0038] 4) Significant economic benefits: Improved raw material utilization; reduced overall costs: No need for subsequent processing steps such as solvent recovery and high-temperature sterilization. Attached Figure Description

[0039] Figure 1 This is the liquid chromatogram of perillaldehyde standard reference.

[0040] Figure 2 The liquid chromatogram of perillaldehyde in the perilla leaf oil obtained in Example 1;

[0041] Figure 3 The liquid chromatogram of perillaldehyde in the perilla leaf oil obtained in Example 2;

[0042] Figure 4 The liquid chromatogram of perillaldehyde in the perilla leaf oil obtained in Example 3;

[0043] Figure 5 The liquid chromatogram of perillaldehyde in the perilla leaf oil obtained in Example 4;

[0044] Figure 6 The liquid chromatogram of perillaldehyde in the perilla leaf oil obtained in Example 5;

[0045] Figure 7 This is a schematic diagram comparing the extraction yield differences between Examples 1-5 and Comparative Examples 1-5 using the traditional water steaming method;

[0046] Figure 8 A schematic diagram comparing the differences in perillaldehyde content between Examples 1-5 and Comparative Examples 1-5 using the traditional water steaming method;

[0047] Figure 9 A comparative diagram showing the effect of enzymatic hydrolysate formulation on extraction yield;

[0048] Figure 10 A comparative diagram showing the effect of enzymatic hydrolysate formulation on perillaldehyde content;

[0049] Figure 11 A diagram comparing the antioxidant capacity of perilla leaf oil and vitamin C. Detailed Implementation

[0050] The present invention will be further described in detail below with reference to the embodiments:

[0051] The yield of perilla leaf oil described in this invention is calculated relative to the weight of dried perilla leaves; specifically, it refers to the ratio of the obtained perilla leaf oil to the amount of fresh perilla leaves used.

[0052] The drying method is as follows: Fresh perilla leaves are placed in an oven at 105℃ for 30 minutes to kill the green, and then dried at 60℃ until constant weight. Chromatographic conditions for HPLC determination of perillaldehyde content:

[0053] Shimadzu column: Shim-pack VP-ODS C18 (4.6mm × 250mm, 5μm);

[0054] Mobile phase: methanol-water (68:32); flow rate: 0.8 mL / min; detection wavelength: 230 nm; column temperature: 30 °C; injection volume: 10 μL.

[0055] Example 1:

[0056] The extraction method for perilla leaf oil is as follows:

[0057] (1) Chop 10kg of fresh perilla leaves into pieces with an average diameter of 2-3cm;

[0058] (2) Spray 15 kg of compound enzymatic hydrolysate onto the crushed leaves. The enzymatic hydrolysate is a mixed aqueous solution containing 8% cellulase, 2% pectinase, 3% triacetylglycerol, and 5% Tween 80.

[0059] (3) Place in an incubator and enzymatically hydrolyze at 45℃ for 3 hours;

[0060] (4) Place the enzymatically hydrolyzed leaves in a vacuum extraction tank, add 120 kg of water, and extract for 1 h at a vacuum gauge reading of -0.076 MPa and a temperature of 65 °C.

[0061] (5) The steam evaporated under negative pressure is condensed and collected in the oil-water separator by the condenser;

[0062] (6) Separate and collect the oil layer, add 13.0g of silica gel desiccant to remove excess moisture, and obtain 124g of perilla leaf oil with a yield of 1.24% and a perilla aldehyde content of 52.8%.

[0063] Example 2:

[0064] The extraction method for perilla leaf oil is as follows:

[0065] (1) Chop 10kg of fresh perilla leaves into pieces with an average diameter of 2-3cm;

[0066] (2) Spray 20 kg of compound enzymatic hydrolysate onto the crushed leaves. The enzymatic hydrolysate is a mixed aqueous solution containing 10% cellulase, 1% pectinase, 2% triacetylglycerol and 4% Tween 80.

[0067] (3) Place in an incubator and enzymatically hydrolyze at 45℃ for 2 hours;

[0068] (4) Place the enzymatically hydrolyzed leaves in a vacuum extraction tank, add 100 kg of water, and extract for 0.5 h at a vacuum gauge reading of -0.070 MPa and a temperature of 70 °C.

[0069] (5) The steam evaporated under negative pressure is condensed and collected in the oil-water separator by the condenser;

[0070] (6) Separate and collect the oil layer, add 13.0g of anhydrous sodium sulfate to remove excess water, and obtain 82g of perilla leaf oil with a yield of 0.82% and a perilla aldehyde content of 43.7%.

[0071] Example 3:

[0072] The extraction method for perilla leaf oil is as follows:

[0073] (1) Chop 10kg of fresh perilla leaves into pieces with an average diameter of 2-3cm;

[0074] (2) Spray 10 kg of compound enzymatic hydrolysate onto the crushed leaves. The enzymatic hydrolysate is a mixed aqueous solution containing 5% cellulase, 4% pectinase, 5% triacetylglycerol, and 6% Tween 80.

[0075] (3) Place in an incubator and enzymatically hydrolyze at 35°C for 5 hours;

[0076] (4) Place the enzymatically hydrolyzed leaves in a vacuum extraction tank, add 150 kg of water, and extract for 1 h at a vacuum gauge reading of -0.081 MPa and a temperature of 60 °C.

[0077] (5) The steam evaporated under negative pressure is condensed and collected in the oil-water separator by the condenser;

[0078] (6) Separate and collect the oil layer, add 6.0g of silica gel desiccant to remove excess water, and obtain 74g of perilla leaf oil with a yield of 0.74% and a perilla aldehyde content of 42.5%.

[0079] Embodiment 4:

[0080] The extraction method for perilla leaf oil is as follows:

[0081] (1) Chop 10kg of fresh perilla leaves into pieces with an average diameter of 1-2cm;

[0082] (2) Spray 30 kg of compound enzymatic hydrolysate onto the crushed leaves. The enzymatic hydrolysate is a mixed aqueous solution containing 1% cellulase, 1% pectinase, 2% triacetylglycerol, and 2% Tween 80.

[0083] (3) Place in an incubator and enzymatically hydrolyze at 50°C for 1 hour;

[0084] (4) Place the enzymatically hydrolyzed leaves in a vacuum extraction tank, add 80 kg of water, and extract for 0.5 h at a vacuum gauge reading of -0.054 MPa and a temperature of 80 °C.

[0085] (5) The steam evaporated under negative pressure is condensed and collected in the oil-water separator by the condenser;

[0086] (6) Separate and collect the oil layer, add 3.5g of molecular sieve to remove excess water, and obtain 68g of perilla leaf oil with a yield of 0.68% and a perilla aldehyde content of 38.9%.

[0087] Example 5:

[0088] The extraction method for perilla leaf oil is as follows:

[0089] (1) Chop 10kg of fresh perilla leaves into pieces with an average diameter of 3-5cm;

[0090] (2) Spray 5 kg of compound enzymatic hydrolysate onto the crushed leaves. The enzymatic hydrolysate is a mixed aqueous solution containing 10% cellulase, 10% pectinase, 8% triacetin, and 10% Tween 80.

[0091] (3) Place in an incubator and enzymatically hydrolyze at 30°C for 6 hours;

[0092] (4) Place the enzymatically hydrolyzed leaves in a vacuum extraction tank, add 200 kg of water, and extract for 2 h at a vacuum gauge reading of -0.081 MPa and a temperature of 60 °C.

[0093] (5) The steam evaporated under negative pressure is condensed and collected in the oil-water separator by the condenser;

[0094] (6) Separate and collect the oil layer, add 9.0g of anhydrous sodium magnesium sulfate to remove excess water, and obtain 43g of perilla leaf oil with a yield of 0.43% and a perilla aldehyde content of 33.6%.

[0095] Please refer to Figure 1 The figure shown is a liquid chromatogram of perillaldehyde standard reference. Figure 2-6 The figures are liquid chromatograms of perillaldehyde in the perilla leaf oil obtained in Examples 1-5, respectively.

[0096] Comparative Experiment 1: Comparison of Extraction Methods

[0097] Based on Examples 1-5, the enzymatic hydrolysis process step was removed, and the following steps were taken as the traditional steam distillation method for easy comparison. Please refer to Table 1 for specific parameters.

[0098] (1) Chop fresh perilla leaves into pieces of a certain diameter range;

[0099] (2) Place the crushed leaves in an extraction tank, add water in a certain multiple of the weight of the perilla leaves, and boil to extract for a period of time.

[0100] (3) The steam that is evaporated is condensed and collected by the condenser and then collected in the oil-water separator.

[0101] (4) Separate and collect the oil layer, add a certain proportion of silica gel desiccant by weight of the oil layer to remove excess moisture, obtain perilla leaf oil, calculate the yield, and detect the content of perilla aldehyde.

[0102] Table 1. Process parameters and results of the traditional steaming method

[0103]

[0104] Based on the data in Table 1 and the data from Examples 1-5, a comparative diagram of the extraction yield differences between Examples 1-5 and the traditional water steaming method, as well as a comparative diagram of the perillaldehyde content differences, are drawn as follows: Figure 7 , Figure 8 As shown in the comparison results, it can be seen that the process of the present invention is superior to the traditional water steaming method, which also shows that the enzymatic hydrolysis process of the present invention can significantly improve the yield of perilla leaf oil and the content of perillaldehyde is also increased.

[0105] Comparative Experiment 2: Screening of Enzymatic Hydrolysate Formulation.

[0106] Based on Examples 1-5, without changing other process conditions, the effects of different enzymatic hydrolysate formulations on the yield and perillaldehyde content of perilla leaf oil were compared. Four comparative formulations were set up for each example for comparison.

[0107] Comparison Formula 1: 8% cellulase, 2% pectinase;

[0108] Comparative formulation 2: 8% cellulase, 3% triacetin, 5% Tween 80;

[0109] Comparative formulation 3: 2% pectinase, 3% triacetin, 5% Tween 80;

[0110] Comparative formulation 4: 3% triacetylglycerol, 5% Tween 80.

[0111] Table 2. Effect of enzyme hydrolysate formulation on extraction

[0112]

[0113] As shown in Table 2, the enzymatic hydrolysate formulated with four components in Examples 1-5 is superior to other enzymatic hydrolysate formulations. Based on the data in Table 2 and the data from Examples 1-5, a comparative diagram illustrating the effect of the enzymatic hydrolysate formulation on the extraction yield and the effect of the enzymatic hydrolysate formulation on the perillaldehyde content is presented; specifically as follows... Figure 9 , Figure 10 The comparison results show that the enzymatic hydrolysates formulated with four components in Examples 1 to 5 are all superior to the other comparative enzymatic hydrolysate formulations.

[0114] Comparative Experiment 3: Effect of Extraction Temperature

[0115] Based on Example 1, the effect of extraction temperature was compared without changing other process conditions.

[0116] Table 3 Effect of temperature on extraction

[0117]

[0118]

[0119] As shown in Table 3, temperature has a significant impact on the extraction results. Appropriately lowering the temperature is beneficial to increasing the yield of perilla leaf oil and the content of perillaldehyde.

[0120] Application Example 1:

[0121] Formula: 0.5g perilla leaf oil, 10g dipropylene glycol, 3g PEG-40 hydrogenated castor oil, 0.2g propylene glycol, 0.2g capryloyl hydroxamic acid, 0.2g ethylhexylglycerin, 0.2g 1,2-hexanediol, deionized water to 100g.

[0122] Preparation process: Add the above-mentioned perilla leaf oil and other excipients to deionized water, heat to 70-80℃ to dissolve and mix evenly, and cool to room temperature to obtain the essence preparation.

[0123] Application Example 2:

[0124] Formula: Perilla leaf oil 0.5g, isononyl isononanoate 6g, glycerin 5g, cyclopentamethoxysiloxane 5g, ethylhexyl palmitate 5g, squalane 3g, butylene glycol 3g, polydimethylsiloxane 2g, polysorbate-60 1.5g, propylene glycol 0.2g, ethylhexylglycerin 0.2g, 1,2-hexanediol 0.2g, capryloyl hydroxamic acid 0.2g, deionized water to 100g.

[0125] Preparation process: Perilla leaf oil and other excipients are added to deionized water. The mixture is heated to 85°C to ensure complete dissolution and the absence of undissolved solutes. After cooling to room temperature, a cream formulation is obtained.

[0126] Test Example 1: Antioxidant test of perilla leaf oil.

[0127] DPPH free radical scavenging ability test:

[0128] The perilla leaf oil obtained in Example 1 was prepared into solutions of different concentrations (0.1 mg / mL, 0.2 mg / mL, 0.4 mg / mL, 0.6 mg / mL, 0.8 mg / mL, 1.0 mg / mL, 1.2 mg / mL), and 2.0 mL of each solution was placed in three sets of test tubes.

[0129] The first group was treated with 2.0 mL of 0.1 mmol / L DPPH solution, mixed thoroughly, and reacted in the dark for 30 min. The absorbance A1 was then measured at 517 nm.

[0130] The second group used 2.0 mL of DPPH solution plus 2.0 mL of distilled water as a model control group, and the absorbance measured at 517 nm was recorded as A2.

[0131] The third group used 2.0 mL of sample solution plus 2.0 mL of anhydrous ethanol as a blank control group. The absorbance value measured at 527 nm was recorded as A3.

[0132] The same method was used, with vitamin C solutions of the same concentration gradient as positive controls. The scavenging rate of the samples against DPPH free radicals was calculated using the following formula:

[0133] DPPH free radical scavenging rate (%) = [A2 - (A1 + A3)] / A2 × 100%.

[0134] Experimental results:

[0135] Comparison results of antioxidant capacity are as follows Figure 11 As shown, from Figure 11 It can be seen that the ability of perilla leaf oil to scavenge DPPH free radicals increases linearly with increasing concentration, indicating that it has a certain antioxidant effect.

[0136] Test Example 2: Comparison of Antioxidant Effects of Applied Products

[0137] The testing method is the same as in Test Example 1. This method requires measuring absorbance and has certain requirements on the light transmittance of the sample. Therefore, the sample is based on the essence of Test Example 1, rather than a cream.

[0138] Test group: 0.5% perilla leaf oil (prepared in Example 1);

[0139] Control group: 0.5% tea tree oil;

[0140] Blank group: Water;

[0141] Table 4 Comparison of Antioxidant Effects

[0142] Group DPPH free radical scavenging rate (%) Test Group 36.7 control group 25.3 Blank group 0

[0143] As shown in Table 4, the perilla leaf oil of the present invention still has certain antioxidant properties after being applied to the product, and is superior to tea tree oil.

[0144] Test Example 3: Comparison of Anti-inflammatory Effects of Applied Products

[0145] Test method: A xylene-induced sensitization and inflammation model of mouse auricle was used. Male mice weighing 20-25g were randomly divided into 3 groups of 10 mice each.

[0146] The sample was formulated based on the cream from Application Example 2.

[0147] Test group: 0.5% perilla leaf oil (prepared in Example 1);

[0148] Control group: 0.5% tea tree oil;

[0149] Blank group: Water;

[0150] 0.2g of the corresponding sample was applied to the right ear of each group of mice. 30 minutes later, approximately 0.04mL of xylene was applied to both sides of the right auricle of each mouse to sensitize it; the left auricle served as a normal control. 30 minutes after sensitization, the corresponding sample was applied again to each group. 1 hour later, round ear pieces were punched from the same location in both ears using a 6mm punch, and weighed using a precision balance. The difference in weight (mg) between the two ear pieces was used as the degree of swelling, and the differences between the groups were compared.

[0151] Table 5 Comparison of anti-inflammatory effects

[0152] Swelling degree (x±s, mg) Test Group 2.15±1.84 control group 2.46±2.01 Blank group 5.93±2.68

[0153] As shown in Table 5, the perilla leaf oil of the present invention has certain anti-inflammatory effects when applied to the product, and is superior to tea tree oil.

[0154] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A method for extracting perilla leaf oil, characterized in that... The steps include: 1) Chop the fresh perilla leaves into small pieces; 2) After spraying the crushed leaves with the compound enzymatic hydrolysate, place them in an incubator for low-temperature enzymatic hydrolysis; 3) Place the enzymatically hydrolyzed leaves in a vacuum extraction tank, add water, and extract under negative pressure; 4) Use a condenser to condense and collect the steam that evaporates under negative pressure into an oil-water separator; 5) Separate and collect the oil layer, add a desiccant to remove excess water, and obtain perilla leaf oil.

2. The method for extracting perilla leaf oil according to claim 1, characterized in that: The average diameter of the chopped perilla leaves in step 1) is 1-5cm.

3. The method for extracting perilla leaf oil according to claim 1, characterized in that: In step 2), the compound enzymatic hydrolysate is a mixed aqueous solution containing cellulase, pectinase, triacetin, and Tween 80, and the spraying amount is 0.5 to 3 times the weight of the leaves.

4. The method for extracting perilla leaf oil according to claim 3, characterized in that: The content of each component by mass is as follows: cellulase 1%–10%, pectinase 1%–10%, triacetin 2%–8%, and Tween 80 2%–10%.

5. The method for extracting perilla leaf oil according to claim 1, characterized in that: In step 2), the temperature for enzymatic hydrolysis is 30–50°C, and the hydrolysis time is 1–6 hours.

6. The method for extracting perilla leaf oil according to claim 1, characterized in that: In step 3), the amount of water added is 8 to 20 times the weight of the perilla leaves. The vacuum gauge reading for the negative pressure extraction is -0.081 to -0.054 MPa, the extraction temperature is 60 to 80℃, and the extraction time is 0.5 to 2 hours.

7. The method for extracting perilla leaf oil according to claim 1, characterized in that: In step 5), the desiccant is selected from one or a combination of anhydrous sodium sulfate, anhydrous magnesium sulfate, molecular sieve, and silica gel desiccant, and the amount used is 5% to 20% of the oil layer weight.

8. The method for extracting perilla leaf oil according to claim 1, characterized in that... The steps include: 1) Chop fresh perilla leaves into pieces with an average diameter of 2-3 cm; 2) Spray the crushed leaves with a compound enzymatic hydrolysate at 1 to 2 times its weight, wherein the compound enzymatic hydrolysate is a mixed aqueous solution containing 5% to 10% cellulase, 1% to 4% pectinase, 2% to 5% triacetylglycerol, and 4% to 6% Tween 80. 3) Place in an incubator and perform low-temperature enzymatic hydrolysis at 35–45℃ for 2–5 hours; 4) Place the enzymatically hydrolyzed leaves in a vacuum extraction tank, add 10 to 15 times the weight of the perilla leaves in water, and extract for 0.5 to 1 hour at a vacuum gauge reading of -0.081 to -0.070 MPa and a temperature of 60 to 70°C. 5) The steam evaporated under negative pressure is condensed and collected in the oil-water separator by the condenser; 6) Separate and collect the oil layer, and add 8% to 15% of anhydrous sodium sulfate or silica gel desiccant by weight of the oil layer to remove excess moisture, thus obtaining perilla leaf oil.

9. Perilla leaf oil extracted by the extraction method according to any one of claims 1-8, characterized in that... The content of perillaldehyde is 33.6%-52.8%.

10. An application of the perilla leaf oil according to claim 9, characterized in that... When combined with excipients, it is formulated into an essence or cream for use in pharmaceuticals and cosmetics.

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