Preparation method and application of calophyllum inophyllum seed oil

Abstract

The invention discloses a preparation method and application of calophyllum inophyllum seed oil, and relates to the technical field of vegetable fat processing. The preparation method comprises the following steps: (1) variable-temperature circulating treatment: performing high-temperature treatment on a begonia seed raw material at 200-250 DEG C, then performing low-temperature treatment at 8-15 DEG C, and finally performing normal-temperature treatment at 22-26 DEG C; (2) wet grinding treatment: soaking the tamanu seed raw material subjected to variable-temperature treatment in a deep-eutectic solvent, and crushing and levigating the tamanu seed raw material; (3) primary enzymolysis treatment; and (4) secondary enzymolysis treatment. The active antibacterial components of the calophyllum inophyllum seed oil prepared through multiple steps can be reserved to the maximum extent, so that the calophyllum inophyllum seed oil has a remarkable antiperspirant effect.

Description

Technical Field

[0001] The invention relates to the technical field of vegetable oil processing, and in particular to a preparation method of tamanu seed oil and application thereof. Background Art

[0002] Calophylluminophyllum Linn. is an evergreen tree belonging to the genus Calophyllum of the family Clusiaceae. Its scientific name is Calophyllum. At present, more than 200 species of Calophyllum have been found in the world, mainly distributed in China, Malaysia, India, Indonesia, the Philippines, Australia and other countries and tropical regions such as Central America. In my country, it is mainly distributed in Hainan, Guangdong, Guangxi, Taiwan and other places. Calophylluminophyllum seed oil is also known as Tamanu oil. Studies have shown that the oil content of Calophyllum seeds is 40-73%. Calophyllum seed oil is rich in active substances such as xanthones, coumarins, terpenes and malus acid. It has the activities of promoting wound healing, anti-neuralgia, antibacterial, anti-inflammatory and anti-aging. It has a long history of medicinal use among the people at home and abroad, and has broad development prospects. In recent years, scholars at home and abroad have carried out systematic research on Calophyllum seed oil and achieved many research results. At present, the extraction processes of malus taenia seed oil mainly include mechanical pressing, ultrasound-assisted extraction, organic solvent extraction, Soxhlet extraction, etc., but there are problems such as small processing volume, low oil yield, low active ingredients of malus taenia seed oil and high cost.

[0003] Hyperhidrosis is a medical condition in which the body sweats excessively and unpredictably. When excessive sweating affects the hands, feet and underarms, this is called primary or focal hyperhidrosis. Primary hyperhidrosis affects approximately 2-3% of the population, but less than 40% of patients with this condition seek medical attention. In most cases of primary hyperhidrosis, no cause is found. The most common form of hyperhidrosis is palmoplantar hyperhidrosis characterized by excessive sweating on the palms, soles and underarms. Therefore, it is important to develop a tamanu seed oil that not only improves the oil yield, but also maximizes the retention of the active antibacterial components in the oil and also has antiperspirant effects. Summary of the invention

[0004] In view of the above-mentioned defects of the prior art, the present invention provides a preparation method of tamanu seed oil and application thereof to solve the problems raised by the above-mentioned background technology.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] A method for preparing tamanu seed oil comprises the following steps:

[0007] (1) variable temperature cycle treatment: the raw material of the tamarisk seed is first subjected to a high temperature treatment at 200 to 250° C., then subjected to a low temperature treatment at 8 to 15° C., and finally subjected to a room temperature treatment at 22 to 26° C., high temperature-low temperature-room temperature being one cycle, and the number of cycles being 5 to 8 times, to obtain the raw material of the tamarisk seed after variable temperature treatment;

[0008] (2) Wet grinding: soaking the temperature-varied Begonia seed raw material obtained in step (1) in a low eutectic solvent, crushing and grinding, and obtaining a wet-milled Begonia seed raw material; wherein the low eutectic solvent comprises 10 to 20 parts of citric acid, 10 to 15 parts of glucose, 5 to 8 parts of glycerol, and 50 to 80 parts of water;

[0009] (3) primary enzymatic hydrolysis: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2) for primary enzymatic hydrolysis, then inactivating the enzymes and centrifuging to retain 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 activity is 50,000 U / g, the xylanase activity is 42,000 IU / g and the cellulase activity is 50,000 U / g;

[0010] (4) Secondary enzymatic hydrolysis: adding a composite enzyme to the supernatant obtained in step (3) for secondary enzymatic hydrolysis, and then inactivating the enzyme and centrifuging to obtain the malus seed oil; the composite enzyme comprises a protease and a β-glucanase in a mass ratio of (0.5-1.0):(1.5-3.5); the protease activity is 50,000 U / g and the β-glucanase activity is 200,000 U / g.

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

[0012] Preferably, the weight ratio of the Malus officinalis seed raw material after the temperature change treatment in step (2) to the low eutectic solvent is 1:(5-10).

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

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

[0015] Preferably, the amount of the mixed enzyme added in step (3) is 3-7% of the mass of the wet-milled malus seeds.

[0016] Preferably, in step (3), the enzymatic hydrolysis time is 240 to 300 min, and the enzymatic hydrolysis temperature is 30 to 50°C.

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

[0018] Preferably, in step (4), the secondary enzymolysis time is 100 to 170 min, and the secondary enzymolysis temperature is 40 to 45°C.

[0019] Preferably, the tamanu seed oil involved in the present application can be used to prepare antiperspirant products.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] (1) Through short-term temperature-changing cycle treatment, the antibacterial active ingredients such as oleic acid, linolenic acid and unsaturated fatty acids in the seed oil of Malus tamanufacturer can be retained, and the toxicity and irritation of fat accompaniments such as xanthones, coumarins, terpenes and flavonoids can be reduced, the production of benzopyrene can be controlled, and the economic value of the seed oil of Malus tamanufacturer can be improved.

[0022] (2) Low eutectic solvents are easy to prepare, low cost, and low toxicity. Operating at room temperature can not only avoid destroying the heat-sensitive components in the oil, but also improve the quality of Malus taenia seed oil, reduce the impurity content, increase the oil solubility rate, and accelerate the oil extraction rate.

[0023] (3) The tamanu seed oil has the effect of astringing the skin, shrinking pores and sweat glands, thereby reducing sweat gland secretion. The active ingredients of the tamanu seed oil have strong antibacterial ability, which can inhibit bacterial reproduction and reduce the odor caused by bacterial decomposition of sweat, thereby solving the problem of sweat odor. This application first proposes that the tamanu seed oil be used in antiperspirant products, providing a new direction for the application of the tamanu seed oil, which has the characteristics of safety and high efficiency, and therefore has broad market development prospects.

[0024] (4) The tamanu seed oil is extracted in stages through multiple enzymatic hydrolysis. The efficient catalytic effect of the enzyme can effectively improve the oil extraction rate. The enzymatic hydrolysis reaction conditions are mild and can maintain the original structure and activity of each effective ingredient. At the same time, through the combination of mixed enzymes and composite enzymes, the proportion of antibacterial components in the obtained tamanu seed oil can be effectively adjusted, thereby improving its antiperspirant effect. DETAILED DESCRIPTION

[0025] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods.

[0026] Example 1

[0027] A method for preparing tamanu seed oil comprises the following steps:

[0028] (1) variable temperature cycle treatment: the raw material of the malus tamanu seed is first subjected to a high temperature treatment of 200°C for 60 seconds, then subjected to a low temperature treatment of 8°C for 30 seconds, and finally subjected to a room temperature treatment of 22°C for 150 seconds. High temperature-low temperature-room temperature is one cycle, and the number of cycles is 5, thereby obtaining the raw material of the malus tamanu seed after variable temperature treatment;

[0029] (2) Wet grinding: soaking the temperature-variable treated tamanu seed raw material obtained in step (1) in a low eutectic solvent for 120 min, crushing and grinding to obtain a wet-milled tamanu seed raw material; wherein the low 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-variable treated tamanu seed raw material to the low eutectic solvent is 1:5;

[0030] (3) a primary enzymatic hydrolysis treatment: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2), performing a primary enzymatic hydrolysis at a temperature of 30° C. for 240 min, then inactivating the enzyme and centrifuging to retain the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of 1:2:1; wherein the amount of the mixed enzyme added is 3% of the mass of the wet-milled malus seed raw material;

[0031] (4) Secondary enzymatic hydrolysis: adding a composite enzyme of 9% by mass of the supernatant to the supernatant obtained in step (3), performing secondary enzymatic hydrolysis at 40° C. for 100 min, and then inactivating the enzyme and centrifuging to obtain malus seed oil; the composite enzyme comprises a protease and a 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 method for preparing tamanu seed oil comprises the following steps:

[0035] (1) variable temperature cycle treatment: the raw material of the tamarisk seed is first subjected to a high temperature treatment at 250°C for 90 seconds, then subjected to a low temperature treatment at 15°C for 45 seconds, and finally subjected to a room temperature treatment at 26°C for 200 seconds. High temperature-low temperature-room temperature is one cycle, and the number of cycles is 8 times, thereby obtaining the raw material of the tamarisk seed after variable temperature treatment;

[0036] (2) Wet grinding: soaking the temperature-variable treated tamanu seed raw material obtained in step (1) in a low eutectic solvent for 300 minutes, and then crushing and grinding to obtain a wet-milled tamanu seed raw material; wherein the low 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-variable treated tamanu seed raw material to the low eutectic solvent is 1:10;

[0037] (3) a primary enzymatic hydrolysis treatment: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2), performing a primary enzymatic hydrolysis at a temperature of 50° C. for 300 min, then inactivating the enzyme and centrifuging to retain the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of 3:5:4; wherein the amount of the mixed enzyme added is 7% of the mass of the wet-milled malus seed raw material;

[0038] (4) Secondary enzymatic hydrolysis: adding a composite enzyme of 15% of the mass of the supernatant to the supernatant obtained in step (3), performing secondary enzymatic hydrolysis at a temperature of 45° C. for 170 min, and then inactivating the enzyme and centrifuging to obtain the malus seed oil; the composite enzyme comprises a protease and a β-glucanase in a mass ratio of 1.0:3.5.

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

[0040] Example 3

[0041] A method for preparing tamanu seed oil comprises the following steps:

[0042] (1) variable temperature cycle treatment: the raw material of the malus tamanu seed is first subjected to a high temperature treatment of 220° C. for 80 seconds, then subjected to a low temperature treatment of 10° C. for 40 seconds, and finally subjected to a room temperature treatment of 25° C. for 180 seconds. High temperature-low temperature-room temperature is one cycle, and the number of cycles is 6 times, thereby obtaining the raw material of the malus tamanu seed after variable temperature treatment;

[0043] (2) Wet grinding: soaking the temperature-variable treated tamanu seed raw material obtained in step (1) in a low eutectic solvent, soaking for 250 minutes, and then crushing and grinding to obtain a wet-milled tamanu seed raw material; wherein the low 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 temperature-variable treated tamanu seed raw material to the low eutectic solvent is 1:9;

[0044] (3) a primary enzymatic hydrolysis treatment: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2), performing a primary enzymatic hydrolysis at a temperature of 40° C. for 270 min, then inactivating the enzyme and centrifuging to retain the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the amount of the mixed enzyme added is 6% of the mass of the wet-milled malus seed raw material;

[0045] (4) Secondary enzymatic hydrolysis: adding a composite enzyme of 12% of the mass of the supernatant to the supernatant obtained in step (3), performing secondary enzymatic hydrolysis at a temperature of 42° C. for 150 min, and then inactivating the enzyme and centrifuging to obtain the malus seed oil; the composite enzyme comprises a protease and a β-glucanase in 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 a high-temperature treatment, and the other components and steps remain unchanged.

[0049] A method for preparing tamanu seed oil comprises the following steps:

[0050] (1) high temperature cycle treatment: the raw material of the malus tamanu seed is first subjected to a high temperature treatment at 220° C. for 300 seconds to obtain a high temperature treated raw material of the malus tamanu seed;

[0051] (2) Wet grinding: soaking the high-temperature treated malus seed raw material obtained in step (1) in a low eutectic solvent, soaking for 250 minutes, and then crushing and grinding to obtain a wet-milled malus seed raw material; wherein the low 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 temperature-variable treated malus seed raw material to the low eutectic solvent is 1:9;

[0052] (3) a primary enzymatic hydrolysis treatment: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2), performing a primary enzymatic hydrolysis at a temperature of 40° C. for 270 min, then inactivating the enzyme and centrifuging to retain the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the amount of the mixed enzyme added is 6% of the mass of the wet-milled malus seed raw material;

[0053] (4) Secondary enzymatic hydrolysis: adding a composite enzyme of 12% of the mass of the supernatant to the supernatant obtained in step (3), performing secondary enzymatic hydrolysis at a temperature of 42° C. for 150 min, and then inactivating the enzyme and centrifuging to obtain the malus seed oil; the composite enzyme comprises a protease and a β-glucanase in a mass ratio of 0.7:2.0.

[0054] The oil yield of Comparative Example 1 was 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 method for preparing tamanu seed oil comprises the following steps:

[0058] (1) variable temperature cycle treatment: subjecting the raw material of Malus tamanufolia seeds to a low temperature treatment at 10° C. for 300 seconds to obtain a low temperature treated raw material of Malus tamanufolia seeds;

[0059] (2) Wet grinding: soaking the low-temperature treated tamanu seed raw material obtained in step (1) in a low eutectic solvent, soaking for 250 minutes, and then crushing and grinding to obtain a wet-milled tamanu seed raw material; wherein the low 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 temperature-variable treated tamanu seed raw material to the low eutectic solvent is 1:9;

[0060] (3) a primary enzymatic hydrolysis treatment: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2), performing a primary enzymatic hydrolysis at a temperature of 40° C. for 270 min, then inactivating the enzyme and centrifuging to retain the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the amount of the mixed enzyme added is 6% of the mass of the wet-milled malus seed raw material;

[0061] (4) Secondary enzymatic hydrolysis: adding a composite enzyme of 12% of the mass of the supernatant to the supernatant obtained in step (3), performing secondary enzymatic hydrolysis at a temperature of 42° C. for 150 min, and then inactivating the enzyme and centrifuging to obtain the malus seed oil; the composite enzyme comprises a protease and a β-glucanase in a mass ratio of 0.7:2.0.

[0062] The oil yield of Comparative Example 2 was 37.25%.

[0063] Comparative Example 3

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

[0065] A method for preparing tamanu seed oil comprises the following steps:

[0066] (1) variable temperature cycle treatment: the raw material of the malus tamanu seed is first subjected to a high temperature treatment of 220° C. for 80 seconds, then subjected to a low temperature treatment of 10° C. for 40 seconds, and finally subjected to a room temperature treatment of 25° C. for 180 seconds. High temperature-low temperature-room temperature is one cycle, and the number of cycles is 6 times, thereby obtaining the raw material of the malus tamanu seed after variable temperature treatment;

[0067] (2) wet grinding treatment: soaking the temperature-variable treated tamanu seed raw material obtained in step (1) in 98% ethanol by volume for 250 minutes, then crushing and grinding to obtain a wet-milled tamanu seed raw material; wherein the weight ratio of the temperature-variable treated tamanu seed raw material to ethanol is 1:9;

[0068] (3) a primary enzymatic hydrolysis treatment: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2), performing a primary enzymatic hydrolysis at a temperature of 40° C. for 270 min, then inactivating the enzyme and centrifuging to retain the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the amount of the mixed enzyme added is 6% of the mass of the wet-milled malus seed raw material;

[0069] (4) Secondary enzymatic hydrolysis: adding a composite enzyme of 12% of the mass of the supernatant to the supernatant obtained in step (3), performing secondary enzymatic hydrolysis at a temperature of 42° C. for 150 min, and then inactivating the enzyme and centrifuging to obtain the malus seed oil; the composite enzyme comprises a protease and a 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] The difference between this comparative example and Example 3 is that dry grinding is used in step (2) instead of wet grinding, and the other ingredients and steps remain unchanged.

[0073] A method for preparing tamanu seed oil comprises the following steps:

[0074] (1) variable temperature cycle treatment: the raw material of the malus tamanu seed is first subjected to a high temperature treatment of 220° C. for 80 seconds, then subjected to a low temperature treatment of 10° C. for 40 seconds, and finally subjected to a room temperature treatment of 25° C. for 180 seconds. High temperature-low temperature-room temperature is one cycle, and the number of cycles is 6 times, thereby obtaining the raw material of the malus tamanu seed after variable temperature treatment;

[0075] (2) dry grinding treatment: crushing and grinding the raw material of the malus truncatum seeds after the temperature change treatment obtained in step (1) to obtain a raw material of the malus truncatum seeds after dry grinding treatment;

[0076] (3) a primary enzymatic hydrolysis treatment: adding a mixed enzyme to the dry-milled malus seed raw material obtained in step (2), performing a primary enzymatic hydrolysis at a temperature of 40° C. for 270 min, then inactivating the enzyme and centrifuging to retain the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the amount of the mixed enzyme added is 6% of the mass of the wet-milled malus seed raw material;

[0077] (4) Secondary enzymatic hydrolysis: adding a composite enzyme of 12% of the mass of the supernatant to the supernatant obtained in step (3), performing secondary enzymatic hydrolysis at a temperature of 42° C. for 150 min, and then inactivating the enzyme and centrifuging to obtain the malus seed oil; the composite enzyme comprises a protease and a 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 α-amylase in step (3) is replaced with pectinase, and the other components and steps remain unchanged.

[0081] A method for preparing tamanu seed oil comprises the following steps:

[0082] (1) variable temperature cycle treatment: the raw material of the malus tamanu seed is first subjected to a high temperature treatment of 220° C. for 80 seconds, then subjected to a low temperature treatment of 10° C. for 40 seconds, and finally subjected to a room temperature treatment of 25° C. for 180 seconds. High temperature-low temperature-room temperature is one cycle, and the number of cycles is 6 times, thereby obtaining the raw material of the malus tamanu seed after variable temperature treatment;

[0083] (2) Wet grinding: soaking the temperature-variable treated tamanu seed raw material obtained in step (1) in a low eutectic solvent, soaking for 250 minutes, and then crushing and grinding to obtain a wet-milled tamanu seed raw material; wherein the low 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 temperature-variable treated tamanu seed raw material to the low eutectic solvent is 1:9;

[0084] (3) a primary enzymatic hydrolysis treatment: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2), performing a primary enzymatic hydrolysis at a temperature of 40° C. for 270 min, then inactivating the enzyme and centrifuging to retain the supernatant; the mixed enzyme comprises pectinase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the amount of the mixed enzyme added is 6% of the mass of the wet-milled malus seed raw material;

[0085] (4) Secondary enzymatic hydrolysis: adding a composite enzyme of 12% of the mass of the supernatant to the supernatant obtained in step (3), performing secondary enzymatic hydrolysis at a temperature of 42° C. for 150 min, and then inactivating the enzyme and centrifuging to obtain the malus seed oil; the composite enzyme comprises a protease and a β-glucanase in a mass ratio of 0.7:2.0.

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

[0087] Comparative Example 6

[0088] The difference between this comparative example and Example 3 is that the preparation method does not include secondary enzymatic hydrolysis treatment, and the other components and steps remain unchanged.

[0089] A method for preparing tamanu seed oil comprises the following steps:

[0090] (1) variable temperature cycle treatment: the raw material of the malus tamanu seed is first subjected to a high temperature treatment of 220° C. for 80 seconds, then subjected to a low temperature treatment of 10° C. for 40 seconds, and finally subjected to a room temperature treatment of 25° C. for 180 seconds. High temperature-low temperature-room temperature is one cycle, and the number of cycles is 6 times, thereby obtaining the raw material of the malus tamanu seed after variable temperature treatment;

[0091] (2) Wet grinding: soaking the temperature-variable treated tamanu seed raw material obtained in step (1) in a low eutectic solvent, soaking for 250 minutes, and then crushing and grinding to obtain a wet-milled tamanu seed raw material; wherein the low 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 temperature-variable treated tamanu seed raw material to the low eutectic solvent is 1:9;

[0092] (3) primary enzymatic hydrolysis: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2), performing a primary enzymatic hydrolysis at 40° C. for 270 min, and then inactivating the enzyme and centrifuging to obtain malus seed oil; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of 2:4:3; wherein the amount of the mixed enzyme added is 6% of the mass of the wet-milled malus seed raw material.

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

[0094] Comparative Example 7

[0095] The difference between this comparative example and Example 3 is that the preparation method does not include a primary enzymatic hydrolysis treatment, and the other components and steps remain unchanged.

[0096] A method for preparing tamanu seed oil comprises the following steps:

[0097] (1) variable temperature cycle treatment: the raw material of the malus tamanu seed is first subjected to a high temperature treatment of 220° C. for 80 seconds, then subjected to a low temperature treatment of 10° C. for 40 seconds, and finally subjected to a room temperature treatment of 25° C. for 180 seconds. High temperature-low temperature-room temperature is one cycle, and the number of cycles is 6 times, thereby obtaining the raw material of the malus tamanu seed after variable temperature treatment;

[0098] (2) Wet grinding: soaking the temperature-variable treated tamanu seed raw material obtained in step (1) in a low eutectic solvent, soaking for 250 minutes, and then crushing and grinding to obtain a wet-milled tamanu seed raw material; wherein the low 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 temperature-variable treated tamanu seed raw material to the low eutectic solvent is 1:9;

[0099] (3) Enzymatic hydrolysis: adding a composite enzyme of 12% by weight of the raw material to the wet-milled malus seed raw material obtained in step (2), performing enzymatic hydrolysis at a temperature of 42° C. for 150 min, and then inactivating the enzyme and centrifuging to obtain malus seed oil; the composite enzyme comprises a protease and a β-glucanase in a mass ratio of 0.7:2.0.

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

[0101] Comparative Example 8

[0102] The application number CN200910166874.3, patent name: a cold pressing preparation process of tamanu seed oil, is used to prepare tamanu seed oil, with an oil yield of 39.97%.

[0103] Experimental Example 1 Animal Experiment

[0104] 1.1 Animal selection

[0105] Select healthy SD rats weighing 200-230 g, 65 males and 65 females, for a total of 130 rats;

[0106] 1.2 Experimental process

[0107] SD rats were divided into 13 groups, 10 in each group (5 males and 5 females), respectively, experimental group (Examples 1-3 and Comparative Examples 1-8), control group (commercially available product, the main components are 20%-25% aluminum chloride solution, 0.5% aluminum acetate solution, 3%-25% formaldehyde solution, 5% alum solution, 5% sulfuric acid solution) and blank group (no treatment). After the rats are adapted to the environment, the rats' toes are cleaned with alcohol skin (modeling), and then the tamanu seed oil prepared by Examples 1-3 and Comparative Examples 1-8 and commercially available products are used to treat the experimental group and the control group respectively, and the application amount is 1mL. After spraying, the rats are allowed to move freely within the range, and the rats are killed after 3h. The skin of the rats' toes sprayed with the agent is excised, and after sectioning, staining and fixing, the sweat gland cells are observed under a microscope, and the number of cells and the number of vacuoles are counted, and the cell vacuoles incidence (%) is calculated by the following formula:

[0108] Cell vacuolation incidence (%) = (number of cell vacuoles / total number of sweat gland cells) × 100%;

[0109] 1.3 Experimental Results

[0110] As shown in Table 1:

[0111] Table 1 Antiperspirant effect

[0112]

[0113] As shown in Table 1, the cell vacuolation rate of the tamanu seed oil prepared in Examples 1 to 3 is lower than that of the control group (commercially available product), indicating that the antiperspirant effect is better than that of the control group. At the same time, the control group contains multiple chemical components, which are complex and can cause skin irritation when used frequently, while the tamanu seed oil is a natural plant component and is safe to use. Example 3 has the best antiperspirant effect. Comparison between Example 3 and Comparative Examples 1 and 2 shows that all high or low temperatures will destroy the active antibacterial components of the Malus cliffe seeds, causing the volatilization of heat-sensitive components and reducing their antiperspirant effect; comparison between Example 3 and Comparative Examples 3 and 4 shows that, in the preparation method of the present application, the extraction rate of the Malus cliffe seed oil obtained by the low eutectic solvent is higher than that of ethanol extraction, and wet grinding can make the Malus cliffe seeds fully contact with the low eutectic solvent for subsequent enzymatic hydrolysis; comparison between Example 3 and Comparative Examples 4 to 7 shows that the enzymatic hydrolysis process affects the antiperspirant effect of the Malus cliffe seed oil, and the selection of the enzyme type is very important. If the enzyme type is replaced, the antiperspirant effect will be reduced; comparison between Example 3 and Comparative Example 8 shows that the Malus cliffe seed oil prepared in Example 3 has a better antiperspirant effect than the Malus cliffe seed oil prepared by the existing cold pressing preparation process.

[0114] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing tamanu seed oil, characterized in that: The following steps are involved: (1) variable temperature cycle treatment: the raw material of the tamarisk seed is first subjected to a high temperature treatment at 200 to 250° C., then subjected to a low temperature treatment at 8 to 15° C., and finally subjected to a room temperature treatment at 22 to 26° C., high temperature-low temperature-room temperature being one cycle, and the number of cycles being 5 to 8 times, to obtain the raw material of the tamarisk seed after variable temperature treatment; (2) Wet grinding: soaking the temperature-varied Begonia seed raw material obtained in step (1) in a low eutectic solvent, crushing and grinding, and obtaining a wet-milled Begonia seed raw material; wherein the low eutectic solvent comprises 10 to 20 parts of citric acid, 10 to 15 parts of glucose, 5 to 8 parts of glycerol, and 50 to 80 parts of water; (3) primary enzymatic hydrolysis: adding a mixed enzyme to the wet-milled malus seed raw material obtained in step (2) for primary enzymatic hydrolysis, then inactivating the enzyme and centrifuging to retain the supernatant; the mixed enzyme comprises α-amylase, xylanase and cellulase in a mass ratio of (1-3):(2-5):(1-4); (4) Secondary enzymatic hydrolysis: adding a composite enzyme to the supernatant obtained in step (3) for secondary enzymatic hydrolysis, and then inactivating the enzyme and centrifuging to obtain the malus seed oil; the composite enzyme comprises a protease and a β-glucanase in a mass ratio of (0.5-1.0):(1.5-3.5).

2. The method for preparing tamanu seed oil according to claim 1, wherein In step (1), the high temperature treatment time is 60 to 90 seconds, the low temperature treatment time is 30 to 45 seconds, and the room temperature treatment time is 150 to 200 seconds.

3. The method for preparing tamanu seed oil according to claim 1, wherein The weight ratio of the Malus tamanu seed raw material and the low eutectic solvent after the temperature change treatment in step (2) is 1:(5-10).

4. The method for preparing tamanu seed oil according to claim 1, wherein The soaking time in step (2) is 120 to 300 minutes.

5. The method for preparing tamanu seed oil according to claim 1, wherein In step (2), the low eutectic solvent includes 15 parts of citric acid, 12 parts of glucose, 7 parts of glycerol, and 60 parts of water.

6. The method for preparing tamanu seed oil according to claim 1, characterized in that: The amount of the mixed enzyme added in step (3) is 3-7% of the mass of the wet-milled malus seeds.

7. The method for preparing tamanu seed oil according to claim 1, characterized in that: In step (3), the enzymolysis time is 240 to 300 minutes, and the enzymolysis temperature is 30 to 50°C.

8. The method for preparing tamanu seed oil according to claim 1, characterized in that: The amount of the mixed complex enzyme added in step (4) is 9-15% of the mass of the supernatant.

9. The method for preparing tamanu seed oil according to claim 1, characterized in that: In step (4), the secondary enzymolysis time is 100 to 170 minutes, and the secondary enzymolysis temperature is 40 to 45°C.

10. Use of the tamanu seed oil prepared by the preparation method according to any one of claims 1 to 9 in the preparation of antiperspirant products.

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