Preparation method of herba aloe arborescens seed oil and application thereof

Abstract

The application discloses a preparation method and application of a herba brevicollis seed oil and relates to the technical field of vegetable oil processing. The preparation method comprises the following steps: (1) variable-temperature cycle treatment: the herba brevicollis seed raw material is subjected to high-temperature treatment at 200-250 DEG C, low-temperature treatment at 8-15 DEG C, and normal-temperature treatment at 22-26 DEG C; (2) wet grinding treatment: the herba brevicollis seed raw material subjected to the variable-temperature treatment is soaked in a eutectic solvent and is crushed and finely ground; (3) primary enzymolysis treatment; and (4) secondary enzymolysis treatment. The herba brevicollis seed oil prepared through the multiple steps can retain the active bacteriostatic components to the maximum extent, so that the herba brevicollis seed oil has a significant antiperspirant effect.

Description

Technical Field

[0001] This invention relates to the field of vegetable oil processing technology, and in particular to a method for preparing and applying tamanu seed oil. Background Technology

[0002] Malus halliana ( Calophyllum inophyllum Linn. ), belonging to the Clusiaceae family and the genus *Linn.* Calophyllum An evergreen tree, scientifically known as *Malus tamarindus*. (Tamanu seed oil) Calophyllum inophyllum Tamanu seed oil, also known as tamanu seed oil, is a traditional Chinese medicine. Studies show that Tamanu seed oil contains 40-73% oil, rich in active substances such as xanthones, coumarins, terpenes, and tamanu fruit acids. It possesses activities promoting wound healing, relieving neuralgia, and exhibiting antibacterial, anti-inflammatory, and anti-aging properties. It has a long history of medicinal use both domestically and internationally and has broad development prospects. In recent years, scholars both at home and abroad have conducted systematic research on Tamanu seed oil, achieving many research results. Currently, the main extraction processes for Tamanu seed oil include mechanical pressing, ultrasonic-assisted extraction, organic solvent extraction, and Soxhlet extraction. However, these methods suffer from problems such as small processing capacity, low oil yield, low levels of active ingredients, and high cost.

[0003] Hyperhidrosis is a medical condition characterized by unpredictable excessive sweating. When excessive sweating affects the hands, feet, and armpits, it is known as primary or focal hyperhidrosis. Primary hyperhidrosis affects approximately 2-3% of the population, but less than 40% of those with this condition seek medical attention. In most cases of primary hyperhidrosis, no underlying cause is identified. The most common form of hyperhidrosis is palmoplantar hyperhidrosis, characterized by excessive sweating of the palms, soles, and armpits. Therefore, it is crucial to develop a Tamanu seed oil that not only improves oil yield but also maximizes the retention of active antibacterial components while possessing antiperspirant properties. Summary of the Invention

[0004] In view of the above-mentioned deficiencies in the prior art, the present invention proposes a method for preparing Tamanuum tamarind seed oil and its application, so as to solve the problems mentioned in the background art.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] A method for preparing Tamanuum tamarind seed oil includes the following steps:

[0007] (1) Variable temperature cycle treatment: The raw material of Malus spectabilis seeds is first treated at a high temperature of 200~250℃, then treated at a low temperature of 8~15℃, and finally treated at a normal temperature of 22~26℃. High temperature-low temperature-normal temperature is one cycle, and the number of cycles is 5~8 times to obtain Malus spectabilis seeds after variable temperature treatment.

[0008] (2) wet grinding treatment: the temperature change treated Euphorbiae humifusae seed material obtained in step (1) is soaked in a eutectic solvent, and is crushed and finely ground to obtain a wet grinding treated Euphorbiae humifusae seed material; the eutectic solvent comprises 10-20 parts of citric acid, 10-15 parts of glucose, 5-8 parts of glycerol, and 50-80 parts of water;

[0009] (3) first enzymolysis treatment: a mixed enzyme is added to the wet grinding treated Euphorbiae humifusae seed material obtained in step (2) for first enzymolysis, and then the enzyme is inactivated and centrifuged to reserve the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of (1-3):(2-5):(1-4); the α-amylase has an enzyme activity of 50000 U / g, the xylanase has an enzyme activity of 42000 IU / g, and the cellulase has an enzyme activity of 50000 U / g;

[0010] (4) second enzymolysis treatment: a compound enzyme is added to the supernatant obtained in step (3) for second enzymolysis, and then the enzyme is inactivated and centrifuged to obtain Euphorbiae humifusae seed oil; the compound enzyme comprises protease and β-glucanase in a mass ratio of (0.5-1.0):(1.5-3.5); the protease has an enzyme activity of 50000 U / g, and the β-glucanase has an enzyme activity of 200000 U / g.

[0011] Preferably, the high-temperature treatment time in step (1) is 60-90 s, the low-temperature treatment time is 30-45 s, and the normal-temperature treatment time is 150-200 s.

[0012] Preferably, the weight ratio of the temperature change treated Euphorbiae humifusae seed material to the eutectic solvent in step (2) is 1:(5-10).

[0013] Preferably, the soaking time in step (2) is 120-300 min.

[0014] Preferably, the eutectic solvent in step (2) comprises 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol, and 60 parts of water.

[0015] Preferably, the addition amount of the mixed enzyme in step (3) is 3-7% of the mass of the wet grinding treated Euphorbiae humifusae seed material.

[0016] Preferably, the first enzymolysis time in step (3) is 240-300 min, and the first enzymolysis temperature is 30-50℃.

[0017] Preferably, the addition amount of the compound enzyme in step (4) is 9-15% of the mass of the supernatant.

[0018] Preferably, the second enzymolysis time in step (4) is 100-170 min, and the second enzymolysis temperature is 40-45℃.

[0019] Preferably, the Euphorbiae Pekinensis Rupr seed oil involved in the present application can be used for preparing antiperspirant products.

[0020] Compared with the prior art, the present application has the following advantages:

[0021] (1) By short-time temperature cycle treatment, the bacteriostatic active ingredients such as oleic acid, linolenic acid and unsaturated fatty acid in the Euphorbiae Pekinensis Rupr seed oil can be retained, the toxicity and irritation of fatty companions such as xanthones, coumarins, terpenes and flavonoids can be reduced, the generation of benzopyrene can be controlled, and the economic value of the Euphorbiae Pekinensis Rupr seed oil can be improved.

[0022] (2) The eutectic solvent has the characteristics of easy preparation, low cost and low toxicity, and operation at room temperature can not only avoid the damage to the heat-sensitive components in the oil, but also improve the quality of the Euphorbiae Pekinensis Rupr seed oil, reduce the impurity content, and improve the oil dissolution rate and the extraction speed of the oil.

[0023] (3) The Euphorbiae Pekinensis Rupr seed oil has the effect of converging the skin, can shrink the pores and sweat glands, thereby reducing the secretion of sweat glands, and the active ingredients of the Euphorbiae Pekinensis Rupr seed oil have strong antibacterial ability, which can inhibit the reproduction of bacteria and reduce the peculiar smell caused by the decomposition of sweat by bacteria, thereby solving the problem of sweat odor. The present application first proposes that the Euphorbiae Pekinensis Rupr seed oil is applied to antiperspirant products, which provides a new direction for the application of the Euphorbiae Pekinensis Rupr seed oil, and has the characteristics of safety and high efficiency, so it has broad market development prospects.

[0024] (4) By multiple enzymatic hydrolysis and stage extraction of the Euphorbiae Pekinensis Rupr seed oil, the efficient catalytic effect of the enzyme can effectively improve the extraction rate of the oil, the mild enzymatic hydrolysis reaction condition can maintain the original structure and activity of each effective component, and meanwhile, by matching the mixed enzyme and the composite enzyme, the proportion of the bacteriostatic components in the prepared Euphorbiae Pekinensis Rupr seed oil can be effectively adjusted, and the antiperspirant effect can be improved. DETAILED DESCRIPTION

[0025] The technical solutions of the present application will be further described in detail in combination with specific embodiments.

[0026] Example 1

[0027] A preparation method of Euphorbiae Pekinensis Rupr seed oil comprises the following steps:

[0028] (1) Temperature cycle treatment: the Euphorbiae Pekinensis Rupr seed raw material is first treated at a high temperature of 200℃ for 60s, then treated at a low temperature of 8℃ for 30s, and finally treated at a normal temperature of 22℃ for 150s, and the high temperature-low temperature-normal temperature is one cycle, and the cycle number is 5 times, to obtain the Euphorbiae Pekinensis Rupr seed raw material after temperature treatment;

[0029] (2) wet grinding treatment: the temperature change treated Euphorbiae humifusae seed material obtained in step (1) is soaked in a deep eutectic solvent, the soaking time is 120 min, and the temperature change treated Euphorbiae humifusae seed material is crushed and finely ground to obtain a wet grinding treated Euphorbiae humifusae seed material; wherein the deep eutectic solvent comprises 10 parts of citric acid, 10 parts of glucose, 5 parts of glycerol, and 50 parts of water; wherein the weight ratio of the temperature change treated Euphorbiae humifusae seed material to the deep eutectic solvent is 1:5;

[0030] (3) first enzymolysis treatment: mixed enzymes are added to the wet grinding treated Euphorbiae humifusae seed material obtained in step (2), and the first enzymolysis is carried out at a temperature of 30℃ for 240 min, and then the enzymes are inactivated and centrifuged to reserve the supernatant; the mixed enzymes comprise α-amylase, xylanase and cellulase in a mass ratio of 1:2:1; wherein the addition amount of the mixed enzymes is 3% of the mass of the wet grinding treated Euphorbiae humifusae seed material;

[0031] (4) second enzymolysis treatment: a compound enzyme with a mass of 9% of the supernatant is added to the supernatant obtained in step (3), and the second enzymolysis is carried out at a temperature of 40℃ for 100 min, and then the enzymes are inactivated and centrifuged to obtain Euphorbiae humifusae seed oil; the compound enzyme comprises protease and β-glucanase in a mass ratio of 0.5:1.5.

[0032] The oil yield of example 1 is 60.08%.

[0033] Example 2

[0034] A preparation method of Euphorbiae humifusae seed oil comprises the following steps:

[0035] (1) temperature change cycle treatment: the Euphorbiae humifusae seed material is first treated at a high temperature of 250℃ for 90s, then treated at a low temperature of 15℃ for 45s, and finally treated at a normal temperature of 26℃ for 200s, which is one cycle of high temperature-low temperature-normal temperature, and the cycle number is 8 times, to obtain a temperature change treated Euphorbiae humifusae seed material;

[0036] (2) wet grinding treatment: the temperature change treated Euphorbiae humifusae seed material obtained in step (1) is soaked in a deep eutectic solvent, the soaking time is 300 min, and the temperature change treated Euphorbiae humifusae seed material is crushed and finely ground to obtain a wet grinding treated Euphorbiae humifusae seed material; wherein the deep eutectic solvent comprises 20 parts of citric acid, 15 parts of glucose, 8 parts of glycerol, and 80 parts of water; wherein the weight ratio of the temperature change treated Euphorbiae humifusae seed material to the deep eutectic solvent is 1:10;

[0037] (3) first enzymolysis treatment: adding mixed enzymes into the wet-milling treated Euphorbiae humifusae seed material obtained in step (2), and performing first enzymolysis for 300 min at a temperature of 50℃, then centrifuging after enzyme inactivation, and reserving the supernatant; the mixed enzymes include α-amylase, xylanase and cellulase with a mass ratio of 3:5:4; wherein the addition amount of the mixed enzymes is 7% of the mass of the wet-milling treated Euphorbiae humifusae seed material;

[0038] (4) second enzymolysis treatment: adding 15% of the supernatant mass of the compound enzymes into the supernatant obtained in step (3), and performing second enzymolysis for 170 min at a temperature of 45℃, then centrifuging after enzyme inactivation, thereby obtaining the Euphorbiae humifusae seed oil; the compound enzymes include protease and β-glucanase with a mass ratio of 1.0:3.5.

[0039] The oil yield of Example 2 is 60.10%.

[0040] Example 3

[0041] A preparation method of Euphorbiae humifusae seed oil includes the following steps:

[0042] (1) temperature cycle treatment: first performing high-temperature treatment of 220℃ for 80s on Euphorbiae humifusae seed material, then performing low-temperature treatment of 10℃ for 40s, and finally performing normal-temperature treatment of 25℃ for 180s, high-temperature-low-temperature-normal temperature being one cycle, and the cycle number being 6, thereby obtaining temperature-treated Euphorbiae humifusae seed material;

[0043] (2) wet-milling treatment: soaking the temperature-treated Euphorbiae humifusae seed material obtained in step (1) in a eutectic solvent, and performing crushing and fine grinding after soaking for 250 min, thereby obtaining wet-milling treated Euphorbiae humifusae seed material; wherein the eutectic solvent includes 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol and 60 parts of water; wherein the weight ratio of the temperature-treated Euphorbiae humifusae seed material to the eutectic solvent is 1:9;

[0044] (3) first enzymolysis treatment: adding mixed enzymes into the wet-milling treated Euphorbiae humifusae seed material obtained in step (2), and performing first enzymolysis for 270 min at a temperature of 40℃, then centrifuging after enzyme inactivation, and reserving the supernatant; the mixed enzymes include α-amylase, xylanase and cellulase with a mass ratio of 2:4:3; wherein the addition amount of the mixed enzymes is 6% of the mass of the wet-milling treated Euphorbiae humifusae seed material;

[0045] (4) second enzymolysis treatment: adding 12% of the supernatant mass of the compound enzymes into the supernatant obtained in step (3), and performing second enzymolysis for 150 min at a temperature of 42℃, then centrifuging after enzyme inactivation, thereby obtaining the Euphorbiae humifusae seed oil; the compound enzymes include protease and β-glucanase with a mass ratio of 0.7:2.0.

[0046] The oil yield of Example 3 is 61.23%.

[0047] Comparative Example 1

[0048] The difference between this comparative example and Example 3 is that step (1) is high-temperature treatment, and the other components and steps remain unchanged.

[0049] A preparation method of Euphorbiae humifusae seed oil, comprising the following steps:

[0050] (1) High-temperature cycle treatment: the Euphorbiae humifusae seed raw material is first treated at 220℃ for 300s to obtain high-temperature treated Euphorbiae humifusae seed raw material;

[0051] (2) Wet grinding treatment: the high-temperature treated Euphorbiae humifusae seed raw material obtained in step (1) is soaked in a eutectic solvent, and after soaking for 250min, it is crushed and finely ground to obtain wet ground Euphorbiae humifusae seed raw material; wherein the eutectic solvent comprises 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol, and 60 parts of water; wherein the weight ratio of the high-temperature treated Euphorbiae humifusae seed raw material to the eutectic solvent is 1:9;

[0052] (3) First enzyme treatment: mixed enzymes are added to the wet ground Euphorbiae humifusae seed raw material obtained in step (2), and first enzyme treatment is carried out at a temperature of 40℃ for 270min, and then the enzymes are inactivated and centrifuged to retain the supernatant; the mixed enzymes comprise α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the amount of mixed enzymes added is 6% of the mass of the wet ground Euphorbiae humifusae seed raw material;

[0053] (4) Second enzyme treatment: 12% of the supernatant mass of the supernatant obtained in step (3) is added to the supernatant, and second enzyme treatment is carried out at a temperature of 42℃ for 150min, and then the enzymes are inactivated and centrifuged to obtain Euphorbiae humifusae seed oil; the complex enzyme comprises protease and β-glucanase in a mass ratio of 0.7:2.0.

[0054] The oil yield of Comparative Example 1 is 38.19%.

[0055] Comparative Example 2

[0056] The difference between this comparative example and Example 3 is that step (1) is low-temperature treatment, and the other components and steps remain unchanged.

[0057] A preparation method of Euphorbiae humifusae seed oil, comprising the following steps:

[0058] (1) Variable-temperature cycle treatment: the Euphorbiae humifusae seed raw material is treated at 10℃ for 300s to obtain low-temperature treated Euphorbiae humifusae seed raw material;

[0059] (2) Wet grinding treatment: the low eutectic solvent is used to soak the low temperature treated Euphorbiae humifusae seed material obtained in step (1), and after soaking for 250 min, the soaking material is crushed and finely ground to obtain the wet grinding treated Euphorbiae humifusae seed material; the low eutectic solvent comprises 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol and 60 parts of water; the weight ratio of the low temperature treated Euphorbiae humifusae seed material to the low eutectic solvent is 1:9;

[0060] (3) First enzymolysis treatment: the mixed enzyme is added to the wet grinding treated Euphorbiae humifusae seed material obtained in step (2), and the first enzymolysis is carried out at 40℃ for 270 min, and then the enzyme is inactivated and centrifuged to reserve the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; the addition amount of the mixed enzyme is 6% of the mass of the wet grinding treated Euphorbiae humifusae seed material;

[0061] (4) Second enzymolysis treatment: the compound enzyme is added to the supernatant obtained in step (3) in an amount of 12% of the mass of the supernatant, and the second enzymolysis is carried out at 42℃ for 150 min, and then the enzyme is inactivated and centrifuged to obtain the Euphorbiae humifusae seed oil; the compound enzyme comprises protease and β-glucanase in a mass ratio of 0.7:2.0.

[0062] The oil yield of the comparative example 2 is 37.25%.

[0063] Comparative example 3

[0064] The difference between the present comparative example and the example 3 is that the ethanol is used instead of the low eutectic solvent in step (2), and the other components and steps are unchanged.

[0065] A preparation method of Euphorbiae humifusae seed oil comprises the following steps:

[0066] (1) Temperature cycle treatment: the Euphorbiae humifusae seed material is first treated at high temperature of 220℃ for 80 s, then treated at low temperature of 10℃ for 40 s, and finally treated at room temperature of 25℃ for 180 s, and the high temperature-low temperature-room temperature is one cycle, and the cycle number is 6 times, to obtain the low temperature treated Euphorbiae humifusae seed material;

[0067] (2) Wet grinding treatment: the low eutectic solvent is used to soak the low eutectic solvent is used to soak the low temperature treated Euphorbiae humifusae seed material obtained in step (1), and after soaking for 250 min, the soaking material is crushed and finely ground to obtain the wet grinding treated Euphorbiae humifusae seed material; the low eutectic solvent comprises 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol and 60 parts of water; the weight ratio of the low temperature treated Euphorbiae humifusae seed material to the low eutectic solvent is 1:9;

[0068] (3) first enzymolysis treatment: adding mixed enzymes into the wet-milling treated Euphorbiae humifusae seed material obtained in step (2), and performing first enzymolysis for 270 min at a temperature of 40℃, then centrifuging after enzyme inactivation, and retaining the supernatant; the mixed enzymes include α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the addition amount of the mixed enzymes is 6% of the mass of the wet-milling treated Euphorbiae humifusae seed material;

[0069] (4) second enzymolysis treatment: adding 12% of the mass of the supernatant of step (3) of compound enzymes into the supernatant, and performing second enzymolysis for 150 min at a temperature of 42℃, then centrifuging after enzyme inactivation, thereby obtaining Euphorbiae humifusae seed oil; the compound enzymes include protease and β-glucanase in a mass ratio of 0.7:2.0.

[0070] The oil yield of Comparative Example 3 is 55.64%.

[0071] Comparative Example 4

[0072] This comparative example is different from Example 3 in that dry-milling treatment is used instead of wet-milling treatment in step (2), and the other components and steps remain unchanged.

[0073] A preparation method of Euphorbiae humifusae seed oil includes the following steps:

[0074] (1) temperature cycle treatment: first performing high-temperature treatment of 220℃ for 80 s on Euphorbiae humifusae seed material, then performing low-temperature treatment of 10℃ for 40 s, and finally performing normal-temperature treatment of 25℃ for 180 s, high-temperature-low-temperature-normal temperature being one cycle, and the cycle number being 6 times, thereby obtaining temperature-treated Euphorbiae humifusae seed material;

[0075] (2) dry-milling treatment: performing crushing and fine grinding on the temperature-treated Euphorbiae humifusae seed material obtained in step (1), thereby obtaining dry-milling treated Euphorbiae humifusae seed material;

[0076] (3) first enzymolysis treatment: adding mixed enzymes into the dry-milling treated Euphorbiae humifusae seed material obtained in step (2), and performing first enzymolysis for 270 min at a temperature of 40℃, then centrifuging after enzyme inactivation, and retaining the supernatant; the mixed enzymes include α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the addition amount of the mixed enzymes is 6% of the mass of the wet-milling treated Euphorbiae humifusae seed material;

[0077] (4) second enzymolysis treatment: adding 12% of the mass of the supernatant of step (3) of compound enzymes into the supernatant, and performing second enzymolysis for 150 min at a temperature of 42℃, then centrifuging after enzyme inactivation, thereby obtaining Euphorbiae humifusae seed oil; the compound enzymes include protease and β-glucanase in a mass ratio of 0.7:2.0.

[0078] The oil yield of Comparative Example 4 was 50.11%.

[0079] Comparative Example 5

[0080] The difference between this comparative example and Example 3 is that the alpha-amylase in step (3) is replaced by pectinase, and the other ingredients and steps remain unchanged.

[0081] A preparation method of Euphorbiae humifusae seed oil, comprising the following steps:

[0082] (1) Variable temperature cycle treatment: the Euphorbiae humifusae seed raw material is first treated at a high temperature of 220℃ for 80s, then treated at a low temperature of 10℃ for 40s, and finally treated at a normal temperature of 25℃ for 180s, high temperature-low temperature-normal temperature being a cycle, and the cycle number being 6 times, to obtain a variable temperature treated Euphorbiae humifusae seed raw material;

[0083] (2) Wet grinding treatment: the variable temperature treated Euphorbiae humifusae seed raw material obtained in step (1) is soaked in a eutectic solvent, and after soaking for 250 min, it is crushed and finely ground to obtain a wet ground Euphorbiae humifusae seed raw material; wherein the eutectic solvent comprises 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol, and 60 parts of water; and the weight ratio of the variable temperature treated Euphorbiae humifusae seed raw material to the eutectic solvent is 1:9;

[0084] (3) First enzyme treatment: mixed enzymes are added to the wet ground Euphorbiae humifusae seed raw material obtained in step (2), and first enzyme treatment is carried out at a temperature of 40℃ for 270 min, and then the enzymes are inactivated and centrifuged to reserve the supernatant; the mixed enzymes comprise pectinase, xylanase and cellulase in a mass ratio of 2:4:3; and the amount of the mixed enzymes added is 6% of the mass of the wet ground Euphorbiae humifusae seed raw material;

[0085] (4) Second enzyme treatment: 12% of the supernatant of the supernatant obtained in step (3) is added to the supernatant, and second enzyme treatment is carried out at a temperature of 42℃ for 150 min, and then the enzymes are inactivated and centrifuged to obtain Euphorbiae humifusae seed oil; the composite enzyme comprises protease and beta-glucanase in a mass ratio of 0.7:2.0.

[0086] The oil yield of Comparative Example 5 was 53.50%.

[0087] Comparative Example 6

[0088] The difference between this comparative example and Example 3 is that the preparation method does not have a second enzyme treatment, and the other ingredients and steps remain unchanged.

[0089] A preparation method of Euphorbiae humifusae seed oil, comprising the following steps:

[0090] (1) variable temperature cycle treatment: the euphorbia antiquorum seed raw material is first treated at a high temperature of 220°C for 80s, then treated at a low temperature of 10°C for 40s, and finally treated at a normal temperature of 25°C for 180s, high temperature-low temperature-normal temperature being one cycle, and the cycle number being 6 times, to obtain a variable temperature treated euphorbia antiquorum seed raw material;

[0091] (2) wet grinding treatment: the variable temperature treated euphorbia antiquorum seed raw material obtained in step (1) is soaked in a eutectic solvent, and after soaking for 250 min, it is crushed and finely ground to obtain a wet ground euphorbia antiquorum seed raw material; wherein the eutectic solvent comprises 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol, and 60 parts of water; wherein the weight ratio of the variable temperature treated euphorbia antiquorum seed raw material to the eutectic solvent is 1:9;

[0092] (3) first enzyme hydrolysis treatment: mixed enzymes are added to the wet ground euphorbia antiquorum seed raw material obtained in step (2), and first enzyme hydrolysis is carried out at a temperature of 40°C for 270 min, and then the enzymes are inactivated and centrifuged to obtain euphorbia antiquorum seed oil; the mixed enzymes comprise α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the amount of mixed enzymes added is 6% of the mass of the wet ground euphorbia antiquorum seed raw material.

[0093] The oil yield of Comparative Example 6 is 42.67%.

[0094] Comparative Example 7

[0095] This comparative example is different from Example 3 in that there is no first enzyme hydrolysis treatment in the preparation method, and the other components and steps remain unchanged.

[0096] A preparation method of euphorbia antiquorum seed oil, comprising the following steps:

[0097] (1) variable temperature cycle treatment: the euphorbia antiquorum seed raw material is first treated at a high temperature of 220°C for 80s, then treated at a low temperature of 10°C for 40s, and finally treated at a normal temperature of 25°C for 180s, high temperature-low temperature-normal temperature being one cycle, and the cycle number being 6 times, to obtain a variable temperature treated euphorbia antiquorum seed raw material;

[0098] (2) wet grinding treatment: the variable temperature treated euphorbia antiquorum seed raw material obtained in step (1) is soaked in a eutectic solvent, and after soaking for 250 min, it is crushed and finely ground to obtain a wet ground euphorbia antiquorum seed raw material; wherein the eutectic solvent comprises 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol, and 60 parts of water; wherein the weight ratio of the variable temperature treated euphorbia antiquorum seed raw material to the eutectic solvent is 1:9;

[0099] (3) enzymatic hydrolysis treatment: 12% of the compound enzyme by mass of the raw material was added to the wet-milling treated Euphorbiae humifusae seed raw material obtained in step (2), and enzymatic hydrolysis was carried out at 42°C for 150 min, and then the enzyme was inactivated and centrifuged, to obtain the Euphorbiae humifusae seed oil; the compound enzyme comprises protease and β-glucanase at a mass ratio of 0.7:2.0.

[0100] The oil yield of Comparative Example 7 was 40.12%.

[0101] Comparative Example 8

[0102] The Euphorbiae humifusae seed oil was prepared by using the patent CN200910166874.3, entitled "Cold-pressing preparation process of Euphorbiae humifusae seed oil", and the oil yield was 39.97%.

[0103] Experimental Example 1 Animal experiment

[0104] 1.1 Animal selection

[0105] Healthy SD rats weighing 200-230 g were selected, with 65 males and 65 females, a total of 130 rats;

[0106] 1.2 Experimental process

[0107] The SD rats were divided into 13 groups, 10 rats in each group (5 males and 5 females), namely, the experimental groups (Examples 1-3 and Comparative Examples 1-8), the control group (a commercially available product, mainly containing 20%-25% aluminum chloride solution, 0.5% aluminum acetate solution, 3%-25% formaldehyde solution, 5% alum solution, and 5% sulfuric acid solution), and the blank group (without any treatment). After the rats were adapted to the environment, the rats were cleaned with alcohol on the toes (modeling), and then the Euphorbiae humifusae seed oil prepared by Examples 1-3 and Comparative Examples 1-8 and the commercially available product were used to treat the experimental groups and the control group, with a coating amount of 1 mL. After spraying, the rats were allowed to move freely within the range, and the rats were sacrificed after 3 h. The skin of the toe part sprayed with the agent was removed, and after slicing, staining, and fixing and mounting, the sweat gland cells were observed under a microscope, and the number of cells and vacuoles was counted. The cell vacuole occurrence rate (%) was calculated by the following formula:

[0108] Cell vacuole occurrence rate (%) = (cell vacuole number / total number of sweat gland cells) × 100%;

[0109] 1.3 Experimental results

[0110] As shown in Table 1:

[0111]

[0112] As shown in Table 1, the cell vacuole occurrence rate of the Euphorbiae humifusae seed oil prepared in Examples 1-3 is lower than that of the control group (commercial product), indicating that the antiperspirant effect thereof is better than that of the control group. Meanwhile, the control group contains various chemical components, and the components thereof are complex, and frequent use thereof can cause skin irritation, while the Euphorbiae humifusae seed oil is a natural plant component, and is safe to use. The antiperspirant effect of Example 3 is the best. As compared with Comparative Examples 1 and 2, it can be seen that all high temperature or low temperature can destroy the active bacteriostatic components of the Euphorbiae humifusae seed, resulting in volatilization of the heat-sensitive components and reduction of the antiperspirant effect thereof. As compared with Comparative Examples 3 and 4, it can be seen that, in the preparation method of the present application, the Euphorbiae humifusae seed oil obtained by using the low eutectic solvent is higher in extraction rate than that obtained by using ethanol extraction, and wet grinding can make the Euphorbiae humifusae seed fully contact with the low eutectic solvent, so as to facilitate subsequent enzymolysis. As compared with Comparative Examples 4-7, it can be seen that the enzymolysis process affects the antiperspirant effect of the Euphorbiae humifusae seed oil, and the selection of the type of enzyme is very important, and replacement of the type of enzyme can reduce the antiperspirant effect. As compared with Comparative Example 8, it can be seen that the Euphorbiae humifusae seed oil prepared in Example 3 has a better antiperspirant effect than the Euphorbiae humifusae seed oil prepared by using the existing cold pressing preparation process.

[0113] The above description is merely preferred embodiments of the present application, but not to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing Averrhoa carambola seed oil, characterized by, The method comprises the following steps: (1) temperature cycle treatment: the Euphorbiae humifusae seed raw material is first treated at a high temperature of 200-250℃, then treated at a low temperature of 8-15℃, and finally treated at a normal temperature of 22-26℃, and the high temperature-low temperature-normal temperature is one cycle, and the cycle number is 5-8 times, to obtain the Euphorbiae humifusae seed raw material treated at a variable temperature; (2) wet grinding treatment: the Euphorbiae humifusae seed raw material treated at a variable temperature obtained in step (1) is soaked in a eutectic solvent, and is crushed and finely ground, to obtain the Euphorbiae humifusae seed raw material treated by wet grinding; wherein the eutectic solvent comprises 10-20 parts of citric acid, 10-15 parts of glucose, 5-8 parts of glycerol, and 50-80 parts of water; (3) first enzyme hydrolysis treatment: mixed enzymes are added to the Euphorbiae humifusae seed raw material treated by wet grinding obtained in step (2) for first enzyme hydrolysis, and then centrifuged after enzyme inactivation, and the supernatant is reserved; the mixed enzymes comprise α-amylase, xylanase and cellulase in a mass ratio of (1-3):(2-5):(1-4); (4) second enzyme hydrolysis treatment: composite enzymes are added to the supernatant obtained in step (3) for second enzyme hydrolysis, and then centrifuged after enzyme inactivation, to obtain the Euphorbiae humifusae seed oil; the composite enzymes comprise protease and β-glucanase in a mass ratio of (0.5-1.0):(1.5-3.5).

2. The method of claim 1, wherein the A. integrifolia seed oil is prepared by cold pressing the seeds of A. integrifolia. In step (1), the high-temperature treatment time is 60-90s, the low-temperature treatment time is 30-45s, and the normal-temperature treatment time is 150-200s.

3. The method of claim 1, wherein the A. integrifolia seed oil is prepared by cold pressing the seeds of A. integrifolia. In step (2), the weight ratio of the Euphorbiae humifusae seed raw material treated at a variable temperature to the eutectic solvent is 1:(5-10).

4. The method of claim 1, wherein the A. integrifolia seed oil is prepared by cold pressing the seeds of A. integrifolia. In step (2), the soaking time is 120-300min.

5. The method of claim 1, wherein the A. integrifolia seed oil is prepared by cold pressing the seeds of A. integrifolia. In step (2), the eutectic solvent comprises 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol, and 60 parts of water.

6. The method of claim 1, wherein the A. integrifolia seed oil is prepared by cold pressing the seeds of A. integrifolia. In step (3), the addition amount of the mixed enzymes is 3-7% of the mass of the Euphorbiae humifusae seed raw material treated by wet grinding.

7. The method of claim 1, wherein the A. integrifolia seed oil is prepared by cold pressing the seeds of A. integrifolia. In step (3), the first enzyme hydrolysis time is 240-300min, and the first enzyme hydrolysis temperature is 30-50℃.

8. The method of claim 1, wherein the A. integrifolia seed oil is prepared by cold pressing the seeds of A. integrifolia. In step (4), the addition amount of the composite enzymes is 9-15% of the mass of the supernatant.

9. The method of claim 1, wherein the A. integrifolia seed oil is prepared by cold pressing the seeds of A. integrifolia. In step (4), the second enzyme hydrolysis time is 100-170min, and the second enzyme hydrolysis temperature is 40-45℃.

10. The application of the Euphorbiae humifusae seed oil prepared by the preparation method in any one of claims 1-9 in the preparation of an antiperspirant product.

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