Highly active and safe substance, its preparation method and its use in medicine

Abstract

A highly active and safe substance, its manufacturing method and its use are disclosed. The substance is obtained from crude rubber seed oil through alkali neutralization, centrifugation, adsorption filtration and deodorization. The substance is not only highly active and has a higher activity for preventing and treating atherosclerosis than crude rubber seed oil, but also has a high safety and can meet the requirements of national food safety standards. The substance can be applied in medicine to prevent and treat atherosclerosis in patients with cardiovascular and cerebrovascular diseases.

Description

[Technical field]

[0001] The present invention relates to the field of biomedicine. [Background technology]

[0002] Drugs, especially those for treating chronic diseases, may have certain side effects, but the smaller the side effects, the better. Patients with chronic diseases need to take drugs for a long time, so small side effects can accumulate over time and cause major problems. Cardiovascular and cerebrovascular diseases caused by atherosclerosis (also known as atherosclerosis) are typical chronic diseases, and the course of the disease is long, requiring patients to take medication for the rest of their lives. If a drug with strong therapeutic effects for atherosclerosis and high safety could be developed, it would be good for patients with cardiovascular and cerebrovascular diseases.

[0003] Cardiocerebrovascular disease caused by atherosclerosis (As) is the number one cause of death in Chinese people and is very harmful, but currently there is no particularly effective drug to prevent and treat atherosclerosis. Currently, drugs for treating atherosclerosis are classified into six categories, namely lipid-lowering drugs, antiplatelet drugs, anticoagulant-thrombotic drugs, antioxidant drugs, and anti-inflammatory drugs, and there are many varieties. However, these drugs can only slow down the formation rate of atherosclerosis and stabilize atherosclerotic plaques, but cannot completely reverse and ablate atherosclerotic plaques, so the therapeutic effect is limited.

[0004] According to medical studies (Non-Patent Documents 1 to 5), rubber seed oil has significant effects in preventing / treating atherosclerosis and has potential for drug development.

[0005] Non-Patent Documents 1-5 speculate that rubber seed oil is rich in unsaturated fatty acids and has a blood lipid lowering effect, which in turn leads to a reversal and regression effect on atherosclerosis. However, this speculation is merely speculation, and it is impossible to confirm or deny the relationship between the unsaturated fatty acids contained in rubber seed oil and the reversal and regression effect on atherosclerosis. To date, there has been no research that has clarified the correlation between specific components contained in rubber seed oil and their biological activity (reversal and regression of atherosclerotic activity).

[0006] Non-Patent Documents 1 to 5 are medical research, but none of them mention the manufacturing method of rubber seed oil. Based on the publication date of the documents, the national situation at the time, and our research, rubber seed oil should be crude oil or crude oil of rubber seeds in food science, that is, a crude extract of fat-soluble components of rubber seeds in phytochemistry.

[0007] Animal experiments have confirmed that rubber seed oil, despite its obvious biological activity of reversing and ablating atherosclerosis, is unhealthy and contains many toxic and harmful components. Both Non-Patent Documents 6 and 12 point out that rubber seed oil has a high acid value, is complex and contains many impurities, and contains special impurities such as rubber, resin, lipoproteins, and cyanosides in addition to the impurities found in general crude oils, such as mechanical impurities, phospholipids, mucilage, and glycolipids, and furthermore, is highly corrosive, resulting in the formation of metal ion chelate compounds.

[0008] In food science, the process of removing components harmful to health from crude vegetable oils is called fat refining. Several food science papers have investigated the various methods of refining crude rubber seed oils. These papers are listed in Non-Patent Documents 6 to 16. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Liu Chaoran, Chen Guozhen, Li Yunshan et al., Study on regression of atherosclerosis in monkeys [J]. Chinese Science (Series B), 1987(02):66-74 [Non-Patent Document 2] Liu Chaoran, Yang Liang, Chen Guozhen, et al., Study on the effect of rubber seed oil on atherosclerosis-I, Effects on the formation and regression of experimental aortic atherosclerosis in rabbits [J], Kunming Journal of Medical Sciences, 1980(3) [Non-Patent Document 3] Liu Chaoran, Li Yunshan, Chen Guozhen, et al., Effects of rubber seed oil on blood lipids and cardiac function during modeling of atherosclerosis in monkeys [J], Journal of Tropical Crops, 1985(2):1 [Non-Patent Document 4] Liu Chaoran, Chen Guozhen, Li Yunshan, Chen Longshun, Tang Chaocai, Zhou Shuyun, Zhang Zhixiong, Study on the preventive effect of rubber seed oil on atherosclerosis in monkeys [J], Kunming Medical Journal, 1985(02):8-22 [Non-Patent Document 5] Liu Hanjun, Zhang Hongfei, Liu Aiwu et al., Ultrastructural Observation of Experimental Atherosclerosis in Monkeys Induced by Rubber Seed Oil [J]. Kunming Medical University Bulletin, 1986(2) [Non-Patent Document 6] Luo Xiaolan, Zhu Wen Xin, He Jian et al., Research and Practice of Refining Rubber Seed Oil [J]. Grain Oil Processing, 2008, 000(011):46-49 [Non-Patent Document 7] Shen Shandeng, Experimental Study on Physical Refining of Rubber Seed Oil[J], China Oil and Fat, 1992(06):18-20 [Non-Patent Document 8] Hu Xiaohong, Liu Dachuan, Zhang Xincai, Study on the production and refining process of rubber seed oil [J]. China Oil and Fats, 2005(11):66-68 [Non-Patent Document 9] Wu Weizhong, Industrial Uses of Rubber Seed Oil[J], China Oil and Fats, 1988(05):62-63 [Non-Patent Document 10] Jia Wei, Technology and Practice of Rubber Seed Oil Refining[J], China Oil and Fat, 2006(02):12-14 [Non-Patent Document 11] Zuting Yue, Study on Extraction of Rubber Seed Oil by Water Enzyme Method and Preparation of Its Microemulsion, Jiangnan University, 2013 [Non-Patent Document 12] Li Linkai, Li Xin, Tao Yin, Research on short-stage / molecular distillation process of rubber seed oil [J]. Grain oil processing (electronic version), 2015(09):26-28 [Non-Patent Document 13] Guo Xiong, Research on the Production of Rubber Seed Oil and Protein [D], Wuhan University of Light Technology, 2018 [Non-Patent Document 14] Lan Qinmu, Storage test report of refined rubber seed oil [J], Oil and Fat Science, 1982(05):12-17 [Non-Patent Document 15] Tian Hua, Huang Tao, Su Minghua, Research on decolorization technology of rubber seed oil[J], Bulletin of Wuhan University of Light Industry, 2007, 26(002):9-11 [Non-Patent Document 16] Wang Xiaoli, Shen Lin, Research on Refining Rubber Seed Oil [J], China Oil and Fats, 2000, 25(004):10-11 Summary of the Invention [Problem to be solved by the invention]

[0010] These refining methods are different, and the resulting refined oils are better or worse from a food science perspective. Food science often considers the following aspects when judging the quality of a refining method: 1. Food safety: whether it meets the requirements of GB 2716-2018 National Food Safety Standard for Vegetable Oil; 2. Yield of the finished product: the weight of the raw material is taken as 100%, and what weight percentage of the finished product can be obtained in the end? Naturally, the higher the yield, the better; 3. Production costs, mainly including the complexity of the process, the high requirements for equipment, the length of the production cycle, the consumption of energy and auxiliary materials, etc., and naturally, the more economical the better.

[0011] Regardless of whether these refining methods are good or bad from a food perspective, the commonality is that none of the rubber seed refined oils produced by these methods are related to their biological activity, i.e., the effect of reversing and ablating atherosclerosis. The present inventors did not know whether the rubber seed refined oils produced by these different refining methods have the effect of reversing and ablating atherosclerosis as described in Non-Patent Documents 1-5.

[0012] In order to investigate the effect of these refining methods on the biological activity (atherosclerosis reversal and ablation effect) of rubber seed oil, the present inventors repeated the technical proposals of Non-Patent Documents 6 to 15. Through numerous animal experiments, the present inventors found that the technical proposals of Non-Patent Documents 6 to 15 have almost no biological activity, regardless of whether the safety standards are met, the yield is high, and the production costs. For details, please refer to Examples 4 to 9.

[0013] Although crude rubber seed oil has biological activity in preventing and treating atherosclerosis, it contains a large amount of unhealthy, toxic and harmful substances; after the unhealthy, toxic and harmful components are removed by oil refining, the refined oil simultaneously loses its biological activity, which is a major problem restricting the development and utilization of rubber seed oil.

[0014] In order to solve the problem of developing and utilizing rubber seed oil, the present invention focuses on the biological activity of rubber seed oil from biological and medical points of view, while also satisfying the safety requirements in food science.

[0015] The objective of the present invention is to provide a novel rubber seed oil, the safety of which meets the GB 2716-2018 National Food Safety Standard for Vegetable Oils, and the biological activity of which meets or exceeds that of the raw crude oil.

[0016] Another object of the present invention is to provide a method for producing such a highly active and safe substance.

[0017] Yet another object of the present invention is to provide a use of such a highly active and safe substance in medicine, which can be applied in medicine for the prevention and treatment of atherosclerosis. [Means for solving the problem]

[0018] In order to achieve the above object, the present invention provides the following technical solutions: It is made from crude rubber seed oil through alkali neutralization, centrifugation, adsorption filtration and deodorization. A substance that is highly active and highly safe, characterized by:

[0019] The present invention also provides a method for producing the highly active and safe substance, characterized in that the substance is obtained by using crude rubber seed oil as a raw material and going through alkali neutralization, centrifugation, adsorptive filtration and deodorization.

[0020] In the alkali neutralization step, an aqueous solution of alkali is added and mixed, the alkali is an edible grade alkali, and the amount of alkali added is equal to the molar mass of the fatty acids contained in the crude oil.

[0021] In the centrifugation step, the temperature of the mixture is 70° C. or higher, and the centrifugal force is 5000 g or higher.

[0022] In order to realize such centrifugation conditions in large-scale industrial production, the present invention further provides a specific realization method: centrifugation is performed in two stages, one before the other, and the other after the other, in the first stage, centrifugation is performed using a tubular centrifuge, and in the second stage, centrifugation is performed using a disk centrifuge or a tubular centrifuge. In the first stage, the conditions for centrifugation using a tubular centrifuge are that the mixture's centrifugation temperature is 70-95°C, the centrifugal force is 5000-30000g, preferably 6000-18000g, to remove fatty acid salts and separate the oil phase; in the second stage, the conditions for centrifugation using a disk centrifuge or a tubular centrifuge are that the oil phase material's centrifugation temperature is 70°C or higher, the centrifugal force is 5000-30000g, preferably 6000-18000g, to remove fatty acid salts and separate the oil phase.

[0023] The oil phase substances are adsorbed, filtered and deodorized, and what remains is the final product.

[0024] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the highly active and safe rubber seed oil and a pharma- ceutical acceptable carrier.

[0025] The present invention also provides the use of said highly active and safe rubber seed oil in the manufacture of a medicament for preventing and treating atherosclerosis, said substance being the only active ingredient.

[0026] Compared with the conventional technical solutions, the rubber seed oil obtained in the present invention has a fundamental difference in activity and also in the method of realization. Effect of the Invention

[0027] From the experimental data, the present invention has the following advantages:

[0028] The present invention provides a novel substance whose safety meets the "GB 2716-2018 National Food Safety Standard for Vegetable Oils" and whose activity in preventing and treating atherosclerosis meets or exceeds that of the raw material, rubber seed oil (Examples 10 to 11).

[0029] Furthermore, the present invention is not limited to laboratory production, but provides a method for industrially and large-scale production of such highly active and highly safe substances (Examples 1 and 2).

[0030] The essential contents of the present invention will be further explained below with reference to the drawings and examples of the present invention, but the present invention is not limited thereto. [Brief description of the drawings]

[0031] [Figure 1] 1 shows the grades of atherosclerotic lesions in different groups of mice in Example 9. [Diagram 2] 1 is a graph showing typical atherosclerotic plaques in rabbit aorta in Example 10. From left to right are taken from normal diet group, high fat diet group, crude rubber seed oil + high fat diet group, present invention + high fat diet group, and simvastatin + high fat diet group, respectively. [Diagram 3] 1 shows the ratio of atherosclerotic plaque to aortic area in different groups of rabbits in Example 10. [Figure 4] The P values ​​of the differences between different groups in Example 10 are shown. When P is less than 0.05, it is a significant difference, and when P is less than 0.01, it is a highly significant difference. [Diagram 5] 1 shows the ratio of atherosclerotic plaque to aortic area in different groups of rabbits in Example 10. [Figure 6] The figure showing typical atherosclerotic plaques in the aorta of rabbits in Example 11. From left to right, the images are taken from the normal diet group, the atherosclerosis modeling group, the modeling + normal feed group, the modeling + crude oil treatment group, the modeling + the present invention treatment group, and the modeling + simvastatin treatment group. [Figure 7] 1 shows the ratio of atherosclerotic plaque to aortic area in different groups of rabbits in Example 11. [Figure 8]The P values ​​of the differences between different groups in Example 11 are shown. When P is less than 0.05, it is a significant difference, and when P is less than 0.01, it is a highly significant difference. [Figure 9] FIG. 9 shows the ratio of atherosclerotic plaque to aortic area in different groups of rabbits in Example 11. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0032] Example 1 According to the method of the present invention, a highly active and safe rubber seed oil has been produced.

[0033] 1000 kg of crude rubber seed oil was used as the raw material (the measured acid value was 30 mg KOH / g, which required 21.4 kg of solid NaOH). The solid alkali was dissolved in 78.6 kg of water to prepare 100 kg of alkali solution with a weight percentage concentration of 21.4%. The crude oil and the alkali solution were mixed in a ratio of 10:1, heated to 80°C, and then injected into a tubular centrifuge for continuous separation at 13,000g, with the flow rate adjusted so that the residence time of the mixture in the centrifuge was 1.5 minutes. The separated oil phase was cached, heated to 90°C, and injected into a disk centrifuge for continued centrifugation. The centrifugal force was 8000g, and the centrifugation residence time was 1 minute. The oil phase after centrifugation was vacuum heated and dehydrated for 60 minutes at a vacuum pressure of 10 kPa and a heating temperature of 100°C, after which 1.2% of the oil weight of adsorbent (the adsorbent is activated clay) was added, stirred for 10 minutes, and filtered to obtain a clear oil. This clear oil was heated to 150°C and deodorized for 30 minutes by introducing steam under a vacuum environment of 1 kPa, after which it was stopped, the temperature was lowered, and the pressure was released to obtain 660 kg of the final product. The yield was 66%. This product is a pale yellow, clear and transparent oily liquid, free of foreign matter, has the smell and taste of rubber seed oil itself, and has no unpleasant odor.

[0034] The final products obtained were tested according to the standard of "GB 2716-2018 Food Safety National Standard Vegetable Oil". All passed the test. Some of the test results are shown below.

[0035] [Table 1]

[0036] The above shows that the safety of the rubber seed oil of the present invention meets the national safety standards for food (edible vegetable oil).

[0037] Example 2 According to the method of the present invention, a highly active and safe rubber seed oil has been produced.

[0038] 1000 kg of crude rubber seed oil was used as the raw material (the measured acid value was 30 mg KOH / g, which required 21.4 kg of solid NaOH). The solid alkali was dissolved in 78.6 kg of water to prepare 100 kg of alkali solution with a weight percentage concentration of 21.4%. The crude oil and the alkali solution were mixed in a ratio of 10:1, heated to 95°C, and then injected into a tubular centrifuge for continuous separation at 18,000g, with the flow rate adjusted so that the residence time of the mixture in the centrifuge was 1.5 minutes. The separated oil phase was cached, heated to 70°C, and injected into a disk centrifuge for continued centrifugation. The centrifugal force was 6000g, and the centrifugation residence time was 1.5 minutes. The oil phase after centrifugation was vacuum heated and dehydrated at a vacuum pressure of 10 kPa and a heating temperature of 100°C for 60 minutes, after which 1.5% of the oil weight of adsorbent (the adsorbent is a mixture of activated clay and diatomaceous earth) was added, stirred for 10 minutes, and filtered to obtain a clear oil. This clear oil was heated to 160°C and deodorized for 30 minutes by introducing steam under a vacuum environment of 0.5 kPa, then stopped, the temperature was lowered, and the pressure was released to obtain 650 kg of the final product. The yield was 65%. The product was a pale yellow, clear and transparent oily liquid, free of foreign matter, had the original smell and taste of rubber seed oil, and had no unpleasant odor.

[0039] The final products obtained were tested according to the standard of "GB 2716-2018 Food Safety National Standard Vegetable Oil". All passed the test. Some of the test results are shown below.

[0040] [Table 2]

[0041] The above shows that the safety of the rubber seed oil of the present invention meets the national safety standards for food (edible vegetable oil).

[0042] Example 3 The conditions and methods for achieving centrifugation in large-scale industrial production are investigated.

[0043] 3.1 Direct centrifugation using a disc centrifuge. 1000 kg of crude rubber seed oil was used as the raw material (the measured acid value was 30 mg KOH / g, which required 21.4 kg of solid NaOH). The solid alkali was dissolved in 78.6 kg of water to prepare 100 kg of alkali solution with a weight percentage concentration of 21.4%. The crude oil and the alkali solution were mixed in a ratio of 10:1, heated to 80°C, and then injected into a disk centrifuge, but separation was not possible. Adjusting the flow rate, changing the ratio of crude oil to alkali solution, adjusting the centrifugal force, adjusting the temperature and other technical parameters all failed.

[0044] 3.2 Direct centrifugation using a tube centrifuge. 1000 kg of crude rubber seed oil was used as the raw material (the measured acid value was 30 mg KOH / g, which required 21.4 kg of solid NaOH). The solid alkali was dissolved in 78.6 kg of water to prepare 100 kg of alkali solution with a weight percentage concentration of 21.4%. The crude oil and the alkali solution were mixed in a ratio of 10:1, heated to 80°C, and then injected into a tubular centrifuge and continuously separated under a condition of 10,000g, with the flow rate adjusted so that the residence time of the mixture in the centrifuge was 1.5 minutes. The separated oil phase was vacuum heated and dehydrated, and then subjected to adsorption filtration, but it was not possible to filter it. No matter how the adsorbent composition, amount used, filtration temperature, pressure, and other conditions were changed, all attempts failed.

[0045] 3.3 Centrifugal force in the first stage. 1000 kg of rubber seed crude oil was used as raw material (the acid value actually measured was 30 mg KOH / g, and 21.4 kg of NaOH solid was required by calculation). Solid alkali was dissolved in 78.6 kg of water to prepare 100 kg of alkali solution with a weight percentage concentration of 21.4%. Crude oil and alkali solution were mixed in a ratio of 10:1, heated to 80°C, and then injected into a tubular centrifuge. From 1000g to 40000g, the centrifugal force was gradually increased by 1000g. It was found that effective separation could not be achieved below 5000g. In the situation above 5000g (including 5000g), effective separation was achieved, and the centrifugal force was increased, improving the processing efficiency of the equipment. Above 30,000g, the material requirements, processing precision requirements, wear rate, and operation difficulty of the equipment are significantly increased, but the effect is limited. When the centrifugal force is between 6000 and 18000g, the overall performance is optimal.

[0046] 3.3. Centrifugal force in the second stage. 1000 kg of rubber seed crude oil was used as the raw material (the measured acid value was 30 mg KOH / g, and 21.4 kg of NaOH solid was required). Solid alkali was dissolved in 78.6 kg of water to prepare 100 kg of alkali solution with a weight percentage concentration of 21.4%. The crude oil and the alkali solution were mixed in a ratio of 10:1, heated to 80°C, and then injected into a tubular centrifuge for continuous separation at 15,000 g. The separated oil phase was cached, heated to 95°C, and injected into a tubular centrifuge or a disk centrifuge for continued centrifugation. From 1000 g to 40,000 g, the centrifugal force was gradually increased by 1000 g. It was found that effective separation could not be achieved below 5000 g. In the situation where the pressure was more than 5000 g (including 5000 g), effective separation was achieved, and the centrifugal force was increased, improving the processing efficiency of the device. When the centrifugal force exceeds 30,000g, the material requirements, processing precision requirements, wear rate and operation difficulty of the equipment increase significantly, but the effect is limited. When the centrifugal force is between 6000 and 18000g, the overall performance is optimal.

[0047] Example 4 According to the method of Non-Patent Document 6, refined rubber seed oil was produced. 1000 kg of rubber seed crude oil was used as a raw material (the acid value actually measured was 30 mgKOH / g). According to the method optimized in Non-Patent Document 6, the oil temperature was adjusted to 40°C, 3 kg of phosphoric acid with a concentration of 85% was added, and after thorough stirring and mixing, the temperature was raised to 80°C, and stirring was carried out quickly, 80 kg of softened water (water from which ions such as calcium and magnesium have been removed) at 80°C was added, water was added, and then the mixture was stirred gently. After the temperature was raised to 85°C, stirring was continued for 30 minutes, stirring was stopped, and the mixture was allowed to stand and settle for 6 hours or more, and the gelatin, wastewater, and waste oil were released, and then vacuum dehydration was carried out. The vacuum dehydration pressure was 0.005 MPa (0.08 MPa to 0.075 MPa) and the temperature was 90°C. After the dehydration was completed, the oil temperature was cooled to 40°C or less in a vacuum state. The pre-bleached oil was heated to 110°C, and 25 kg of activated clay was added under a vacuum of 0.005 MPa (0.08 MPa to 0.075 MPa), and 1.25 kg of activated carbon was added to the clay. When the re-bleaching filtration was half complete, the process was switched to pre-bleaching of the raw oil. When the pressure of the blade filter reached the upper limit of the device, the filtration was stopped, the crepe was blown out, and one filtration cycle was completed. The entire filtration process was difficult, and the filtration speed was very slow. The decolorized oil entered a packed deacidification tower (the pressure was kept below 100 Pa, the temperature of the oil entering the tower was heated to 240 to 250°C, the temperature of the oil discharged from the tower was controlled to ≧230°C, and the acid value of the oil discharged from the tower was controlled to ≦1 mgKOH / g). The temperature of the oil discharged from the tower was cooled to room temperature, and 810 kg of refined oil product produced by the method of Non-Patent Document 6 was obtained. The yield was 81%. The refined oil is red in color, clear and transparent, and has no unpleasant odor.

[0048] The refined oil was measured based on the standard of "GB 2716-2018 National Food Safety Standard for Vegetable Oils" and was determined to meet the standard.

[0049] Example 5 Purified rubber seed oil was produced according to the method of Non-Patent Document 10. 1000 kg of crude rubber seed oil was used as the raw material (the acid value actually measured was 30 mgKOH / g). According to the method of Non-Patent Document 10, the oil temperature was heated to 60°C, 2 kg of phosphoric acid with a concentration of 85% was added, reacted for 18 minutes, allowed to stand and precipitated, the liquid in the lower layer was discharged, water was added and washed once, and a relatively clear emulsification phenomenon was observed. The oil was heated to 65°C, 1 kg of formic acid was added, reacted for 20 to 25 minutes, allowed to stand and precipitated, and washed with water three times, and each time a clear emulsification phenomenon appeared and the oil turned milky white. Under a vacuum pressure of 0.6 kPa, the oil was heated to 112°C and dried and dehydrated. After the dehydration was completed, 40 kg of activated clay and 10 kg of activated carbon were added and stirred for 30 minutes while maintaining the temperature at 100°C. Then, it was filtered while hot, and the clear oil was filtered. Filtration was relatively difficult, and the filtration speed was very slow. The filtered clear oil was extracted for the first time by adding 75% ethanol in a ratio of oil:alcohol = 1:1.5, mixed and stirred, and the temperature was brought to 58 ° C., and then left to stand for layer separation, and the lower oil phase was separated, and then extracted for the second time according to the conditions of the first time. In this way, 15 extractions were repeated, and the extraction was terminated after the acid value was reduced to within 3. The oil phase was heated under reduced pressure to evaporate the ethanol, and 690 kg of refined oil product was obtained according to the method of Non-Patent Document 10. The yield was 69%. The refined oil was red, clear and transparent, and had an ethanol smell when left in the container for a long time.

[0050] The refined oil was tested according to the GB 2716-2018 National Food Safety Standard for Vegetable Oils, and two indicators were found to be unqualified, while the remaining indicators were found to be acceptable. The unqualified indicators were odor, taste, and residual solvent.

[0051] Example 6 Refined rubber seed oil was produced according to the method of Non-Patent Document 12. 1000 kg of crude rubber seed oil was used as the raw material (the acid value actually measured was 30 mgKOH / g). According to the method optimized in Non-Patent Document 12, the oil temperature was adjusted to 40°C, 3 kg of phosphoric acid with a concentration of 85% was added, and after thorough stirring and mixing, the temperature was raised to 80°C, and stirring was continued quickly, 80 kg of softened water (water from which ions such as calcium and magnesium have been removed) at 80°C was added, water was added, and then the mixture was stirred gently. After heating to 85°C, stirring was continued for 30 minutes, stirring was stopped, and the mixture was allowed to stand and settle for 6 hours or more. The upper oil layer and the lower gelatin and wastewater were separated, and then dehydration was performed at normal pressure. The oil temperature was heated to 110°C under normal pressure to prevent air bubbles from appearing. After dehydration was completed, the oil was cooled to 40°C or less. The oil was heated to 110°C, 50g of activated clay was added, and 2.5g of activated carbon was added to the clay at the same time, stirred for 30 minutes, and then filtered through a three-layer filter paper. In order to increase the filtration rate, suction filtration was performed with a Brinell funnel. Nevertheless, the filtration rate was still very slow. The filtered oil was put into a molecular distillation apparatus to carry out molecular distillation. The conditions for molecular distillation were: feed pump rotation speed 20Hz, blade rotor rotation speed 300rpm, cooling liquid temperature 25°C, heat transfer oil temperature 200°C, and vacuum pump vacuum degree 1.33Pa.

[0052] The acid value of one distillation directly according to the method of Non-Patent Document 12 is still high at 12 mg KOH / g, which is higher than the standard of 3 mg KOH / g in "GB 2716-2018 National Food Safety Standard Vegetable Oil". After three molecular distillations, the acid value decreased to below 3 mg KOH / g, and finally 830 g of the product produced by the method of Non-Patent Document 12 was obtained. The yield was 83%.

[0053] This refined oil was measured based on the standards of GB 2716-2018 National Food Safety Standard for Vegetable Oils and was determined to meet the standards.

[0054] Example 7 Refined rubber seed oil was produced according to the method of Non-Patent Document 14. 1000 kg of rubber seed crude oil was used as the raw material (the acid value actually measured was 30 mgKOH / g). The crude oil was heated to 70°C, 3 kg of phosphoric acid with a concentration of 85% was added, 20 kg of saturated NaCl aqueous solution was added, and the mixture was allowed to stand, the hydrate in the lower layer was separated, and the oil phase in the upper layer was retained. 21.4 kg of solid NaOH was added to 78.6 kg of saturated NaCl solution to prepare 100 kg of NaOH-saturated NaCl solution (hereinafter simply referred to as base-sodium chloride solution) with a concentration of 21.4% by weight. 100 kg of base-sodium chloride solution was added to the oil phase, and the entire reaction system was heated to 70°C. At this time, the oil phase and the base-sodium chloride solution were separated into two layers, upper and lower, and no acid-base neutralization reaction occurred. The oil phase was slowly stirred to contact with the alkaline solution and react. The base-sodium chloride solution was gradually brought into contact with the oil phase, causing an acid-base neutralization reaction to produce a large amount of fatty acid sodium, and then an emulsification phenomenon occurred, and the oil phase-fatty acid sodium-base-sodium chloride solution was emulsified together to form a brown, viscous, and uniform emulsion system. After 24 hours of incubation and standing, the upper layer emulsion was collected, added with saturated saline and washed with water at 100°C, but it was still heavily emulsified. Even after 24 hours of incubation and standing, no obvious stratification was observed. The upper layer emulsion was collected and heated to 120°C for drying and dehydration. Dehydration was extremely difficult, and a large amount of foam overflowed. After the dehydration was completed, 120 kg of refined oil produced by the method of Non-Patent Document 14 was obtained. The yield was 12%. The refined oil was viscous jelly-like, red-black, turbid, and had a strong unpleasant odor.

[0055] The refined oil in question was tested in accordance with the standards of GB 2716-2018 National Food Safety Standard for Vegetable Oils and determined to be a product that did not meet the standards.

[0056] Example 8 Purified rubber seed oil was produced according to the method of Non-Patent Document 13. 1000 kg of rubber seed crude oil was used as the raw material (the acid value actually measured was 30 mg KOH / g). According to the method optimized in Non-Patent Document 13, 1000 g of rubber seed crude oil was dissolved in 1703 g of n-hexane to prepare a 37 mass % rubber seed crude oil-n-hexane solution. Using a filtration membrane with a pore size of 10 kDa, the crude oil-n-hexane solution was passed through the filtration membrane at a positive pressure of 0.23 MPa. The filtered crude oil-n-hexane solution was heated and the n-hexane solvent was removed under negative pressure to obtain 950 g of the product produced by the method of Non-Patent Document 13. The yield was 95%. The product was a dark red, opaque oily liquid with no significant difference from the raw crude oil.

[0057] The product was measured according to the standard of "GB 2716-2018 National Food Safety Standard Vegetable Oil", and the results were similar to those of the direct test of crude oil, with many indexes not meeting the standards, especially the acid value, which is high at 27 mg KOH / g, and no matter how many times it is filtered through membrane, the acid value remains above 25 mg KOH / g and cannot fall below 3 mg KOH / g. The material obtained by this method has no significant difference in each physical and chemical index from crude oil.

[0058] Example 9 Animal experiments on the effects of crude rubber seed oil, the rubber seed oil of the present invention, and the rubber seed oils produced in Example 4 (Non-Patent Document 6), Example 5 (Non-Patent Document 10), Example 6 (Non-Patent Document 12), and Example 7 (Non-Patent Document 14) on the formation of atherosclerotic plaque. The physicochemical indexes of the rubber seed oil produced in Example 8 (Non-Patent Document 13) were not significantly different from those of crude rubber seed oil, and therefore it was not selected for the animal experiments.

[0059] Twelve 8-week-old male wide type C57BL / 6J mice were selected; 84 8-week-old male APOE- / - mice were selected; weighing 24±2 g. The APOE- / - mice were randomly divided into 7 groups, each with 12 mice, and 12 wide type C57BL / 6J mice in each group, for a total of 8 groups. The groups were as follows: (1) Normal diet group: 12 male wild-type C57BL / 6J mice aged 8 weeks, fed with basal diet and administered saline intragastrically; (2) High-fat diet group: 12 APOE- / - mice, fed high-fat diet + intragastric administration of peanut oil, intragastric dose 6.2 g / kg / d (based on adult and animal weight, body surface area and Km factor); (3) Rubber seed oil + high-fat diet group: 12 APOE- / - mice, fed with high-fat diet + intragastric administration of rubber seed oil, intragastric dose 6.2 g / kg / d (based on adult and animal body weight, body surface area and Km factor); (4) present invention group + high-fat diet group: 12 APOE- / - mice, fed with high-fat diet + intragastric administration of the product prepared in Example 1-2, intragastric dose 6.2g / kg / d (based on the body weight, body surface area and Km factor of adults and animals); (5) Non-Patent Document 6 Group + High Fat Diet Group: 12 APOE- / - mice, fed with high fat diet + intragastric administration of the product prepared in Example 4, intragastric dose 6.2 g / kg / d (based on the body weight, body surface area and Km factor of adults and animals); (6) Non-Patent Document 10 group + high-fat diet group: 12 APOE- / - mice, fed with high-fat diet + intragastric administration of the product prepared in Example 5, gastric dose 6.2 g / kg / d (based on the body weight, body surface area and Km factor of adults and animals); (7) Non-Patent Document 12 group + high-fat diet group: 12 APOE- / - mice, fed with high-fat diet + intragastric administration of the product prepared in Example 6, gastric dose 6.2 g / kg / d (based on the body weight, body surface area and Km factor of adults and animals); (8) Non-Patent Document 14 group + high-fat diet group: 12 APOE- / - mice, fed with high-fat diet + intragastric administration of the product prepared in Example 7, with a gastric injection dose of 6.2 g / kg / d (based on the body weight, body surface area and Km factor of adults and animals);

[0060] All mice in each group were given intragastric administration of high-fat feed for 8 consecutive weeks. The composition of the high-fat feed was: basal feed: 70%, lard: 20%, sucrose: 5%, milk powder: 4%, bile salts: 1%, cholesterol: 0.15%.

[0061] The mice were sacrificed, the mouse thoracic cavity was cut, the mouse blood vessels were perfused with pre-cooled saline, the aorta was exfoliated, completely cut, washed with PBS, and fixed by immersion in 10% formaldehyde solution, and the degree of atherosclerotic lesions was detected using the classification method:

[0062] Grade 0: smooth intimal surface, no cream-colored changes, i.e., no plaque; Grade 0.5: There are extensive cream-colored or milky changes in the intima, but no plaques protruding to the surface; Grade 1: There is a cream-colored protruding plaque in the intima, and the plaque area is 3mm 2 Grade 2: There is obvious cream-colored protruding plaque in the intima, but there is no phenomenon of fusion into a sheet, and the maximum plaque area is 3 mm 2 Greater than; Grade 3: There are many plaques of different sizes, some of which have merged into sheets, and the area of ​​the largest plaque is 3 mm 2 Grade 4: The intimal surface is mostly covered with consolidated plaque.

[0063] The results of the animal studies are shown in the table below and in FIG. [Table 3]

[0064] Test Results: (1) The rubber seed oil of the present invention has the effect of preventing and inhibiting atherosclerotic plaque. After adding the rubber seed oil of the present invention, the degree of atherosclerosis progression in APOE- / - mice fed a high-fat diet was grade 1; the degree of atherosclerosis in APOE- / - mice fed a high-fat diet without supplementation was grade 3 to 4; after adding the rubber seed oil of the present invention, the degree of atherosclerosis progression in APOE- / - mice was obviously slowed down. This shows that the rubber seed oil of the present invention has the effect of preventing and inhibiting atherosclerotic plaque; (2) The rubber seed oil produced in Examples 4, 5, 6, and 7 did not show any significant effect of preventing and inhibiting atherosclerotic plaques. After adding the rubber seed oil produced in Non-Patent Document 6 (Example 4), Non-Patent Document 10 (Example 5), Non-Patent Document 12 (Example 6), and Non-Patent Document 14 (Example 7), the degree of atherosclerosis in APOE- / - mice on a high-fat diet was 3 to 4 grades; the degree of atherosclerosis in APOE- / - mice on a high-fat diet without supplementation was also 3 to 4 grades; no significant change was observed in the degree of atherosclerosis in APOE- / - mice, regardless of whether the rubber seed oil produced in Non-Patent Document 6 (Example 4), Non-Patent Document 10 (Example 5), Non-Patent Document 12 (Example 6), and Non-Patent Document 14 (Example 7) was added. As can be seen, none of the rubber seed oils produced in Non-Patent Document 6 (Example 4), Non-Patent Document 10 (Example 5), Non-Patent Document 12 (Example 6), and Non-Patent Document 14 (Example 7) showed any significant preventive or inhibitory effect on atherosclerotic plaque.

[0065] Example 10 Animal studies of the effect of crude rubber seed oil, the rubber seed oil of the present invention, and simvastatin on atherosclerotic plaque formation. Rabbits are an ideal animal model for the study of dietary induction of atherosclerosis, and the pathogenesis of this condition is closer to that of humans than that of gene-deficient mice and rats. Forty 4-month-old male Japanese white rabbits weighing 2.0±0.2 kg were selected and randomly divided into five groups as follows: (1) Normal diet group: 8 male Japanese white rabbits fed with basal diet; (2) High-fat diet group: 8 male Japanese white rabbits, fed with peanut oil + high-fat enriched + basal diet, peanut oil feeding dose 1.5g / kg / d (based on adult and animal weight, body surface area and Km factor); (3) Rubber seed oil + high fat diet group: 8 male Japanese white rabbits, fed with rubber seed oil + high fat supplemented + basal diet, with a feeding dose of rubber seed oil of 1.5 g / kg / d (based on the weight, body surface area and Km factor of adults and animals); (4) The present invention + high-fat diet group: 8 male Japanese white rabbits, fed with the product prepared in Example 1-2 + high-fat enriched + basal diet, the feeding dose of the product prepared in Example 1-2 was 1.5g / kg / d (based on the weight, body surface area and Km factor of adults and animals); (5) Simvastatin + high-fat diet group: 8 male Japanese white rabbits, fed with simvastatin + high-fat enriched + basal diet, with a feeding dose of simvastatin of 2.0 mg / kg / d (based on the weight, body surface area and Km factor of adults and animals);

[0066] The high-fat enriched diet contained 0.5 g cholesterol, 2.0 g lard, and 20 g egg yolk per rabbit per day.

[0067] After feeding for 6 weeks according to the above grouping and feeding conditions, the animals were sacrificed. After the animals were sacrificed, the aorta was completely cut from the aortic valve orifice to the beginning of the common iliac artery, the aortic wall was incised longitudinally along the abdominal midline, and fixed with 10% formaldehyde solution, the distribution map of the aortic lesion was recorded by tracing method, the area of ​​atherosclerotic plaque and the total area of ​​the lumen in which the aorta was expanded were measured, and the percentage of the plaque area was calculated.

[0068] Test Results: (1) The rubber seed oil of the present invention has obvious effects of preventing and inhibiting atherosclerotic plaque (the plaque area of ​​the present invention + high-fat diet group is 14.3±5.0%, which is smaller than that of the high-fat diet group, 42.6±9.7%, P<0.01, the difference is very significant); (2) The rubber seed oil of the present invention has a stronger effect of preventing and inhibiting atherosclerotic plaque than crude rubber seed oil (the plaque area of ​​the present invention + high-fat diet group was 14.3±5.0%, which was smaller than that of the crude rubber seed oil + high-fat diet group (19.7±4.7%, P<0.05, the difference was significant); (3) The rubber seed oil of the present invention at a dose of 1.5 g / kg / d has a stronger effect of preventing and inhibiting atherosclerotic plaque than simvastatin at a dose of 2.0 mg / kg / d (the plaque area of ​​the present invention + high-fat diet group was 14.3 ± 5.0%, which was smaller than that of the simvastatin + high-fat diet group at 20.2 ± 4.2%, P < 0.05, the difference was significant).

[0069] Example 11 Animal studies of the effects of crude rubber seed oil, the rubber seed oil of the present invention, and simvastatin on formed atherosclerotic plaques. Rabbits are an ideal animal model for the study of dietary induction of atherosclerosis, and the pathogenesis of this condition is closer to that of humans than that of gene-deficient mice and rats. Forty 4-month-old male Japanese white rabbits weighing 2.0±0.2 kg were selected and randomly divided into two groups as follows: (1) Normal diet group: 8 male Japanese white rabbits fed with basal diet; (2) Atherosclerosis modeling group: 40 male Japanese white rabbits, fed high fat enriched + basal diet, with peanut oil at a feeding dose of 1.5g / kg / d (based on adult and animal body weight, body surface area and Km factor);

[0070] The high fat enriched diet consisted of 0.5g cholesterol, 4.0g lard, and 20g egg yolk per rabbit per day. After six weeks of feeding according to the above grouping and feeding conditions, one rabbit died in the normal diet group.

[0071] After 6 weeks, 8 rabbits in the atherosclerosis modeling group were randomly selected and sacrificed. Whether the establishment of the atherosclerosis model was successful was examined. The examination method was as follows: after the animals were sacrificed, the aorta was completely cut from the aortic valve orifice to the beginning of the common iliac artery, the aortic wall was incised longitudinally along the abdominal midline, and fixed with 10% formaldehyde solution, the aortic lesion distribution map was recorded by tracing method, the area of ​​atherosclerotic plaque and the total area of ​​the lumen in which the aorta was deployed were measured, and the percentage of the plaque area was calculated. In contrast, all 8 rabbits in the normal diet group were also sacrificed, and the percentage of the area of ​​aortic atherosclerotic plaque was measured.

[0072] After the rabbits in the atherosclerosis modeling group were successfully modeled, the remaining 32 rabbits in the remaining modeling group were randomly divided into 4 groups, each with 8 rabbits, as follows:

[0073] (3) Modeling + normal feed group: 8 male Japanese white rabbits successfully modeled on atherosclerosis, fed with peanut oil + basal feed, with a peanut oil feeding dose of 1.5g / kg / d (based on the weight, body surface area and Km factor of adults and animals); (4) Modeling + crude rubber seed oil treatment group: 8 male Japanese white rabbits with successful atherosclerosis modeling, fed with crude rubber seed oil + basal diet, with a feeding dose of 1.5 g / kg / d of crude rubber seed oil (based on the weight, body surface area and Km factor of adults and animals); (5) Modeling + treatment group of the present invention: 8 male Japanese white rabbits successfully modeled on atherosclerosis, fed with the product prepared in Example 1-2 + basic feed, with the feeding dose of the product prepared in Example 1-2 being 1.5g / kg / d (based on the weight, body surface area and Km factor of adults and animals); (6) Modeling + simvastatin treatment group: 8 male Japanese white rabbits successfully modeled on atherosclerosis, fed with simvastatin + basal diet, with a feeding dose of simvastatin of 2.0 mg / kg / d (based on the weight, body surface area and Km factor of adults and animals);

[0074] The animals were fed according to the above grouping and feeding conditions for three months, during which one rabbit died in each of the modeling + normal feed group and the modeling + simvastatin treatment group. After three months of feeding, all the animals were sacrificed.

[0075] After the animals were sacrificed, the aorta was completely excised from the aortic valve orifice to the beginning of the common iliac artery, the aortic wall was incised longitudinally along the abdominal midline, and fixed with 10% formaldehyde solution. The distribution map of aortic lesions was recorded by the tracing method, the area of ​​atherosclerotic plaque and the total area of ​​the lumen in which the aorta was deployed were measured, and the percentage of plaque area was calculated.

[0076] The experimental results are shown in Figures 6-9.

[0077] Test Results: (1) The rubber seed oil of the present invention can not only inhibit the increasing tendency of atherosclerosis (the plaque area of ​​the modeling + present invention group was 28.1±6.4%, which was smaller than that of the modeling + normal feed group at 59.9±8.9%, P<0.01, the difference was significant), but also ablate the already formed atherosclerotic plaque (the plaque area of ​​the modeling + present invention group was 28.1±6.4%, which was smaller than that of the modeling group at 45.8±9.9%, P<0.01, the difference was significant); (2) The rubber seed oil of the present invention has a better effect of inhibiting atherosclerotic plaque than crude rubber seed oil (the plaque area of ​​the modeling + present invention group was 28.1±6.4%, which was smaller than that of the modeling + crude oil group (36.2±7.9%, P<0.05, the difference was significant); (3) The rubber seed oil of the present invention at a dose of 1.5 g / kg / d has a stronger effect of inhibiting atherosclerotic plaque than simvastatin at a dose of 2.0 mg / kg / d (the plaque area of ​​the present invention group was 28.1±6.4%, which was smaller than that of the simvastatin group at 47.1±8.3%, P<0.01, the difference being significant).

[0078] Example 12: According to the methods of Examples 1 and 2, the highly active and safe substance of the present invention was produced. The substance was granulated and compressed into tablets by adding an excipient so that the weight ratio of the substance to the excipient was 1:1 or 1:2. The tablets can be used as a drug.

[0079] Example 13: According to the methods of Examples 1 and 2, the highly active and safe substance of the present invention was produced. The substance was encapsulated in a soft capsule material, and a soft capsule was produced according to a normal capsule preparation method. The soft capsule can be used as a drug.

[0080] Example 14: The highly active and safe substance of the present invention was produced according to the methods of Examples 1 and 2. An emulsifier, an osmotic pressure regulator and water were added to the substance, and the mixture was subjected to high-pressure homogeneous emulsification to obtain an emulsion for injection.

[0081] Example 15: The highly active and safe rubber seed oil of the present invention was produced according to the methods of Examples 1 and 2. The rubber seed oil was made into tablets as follows. Tablet: 100 mg of rubber seed oil of the present invention Starch (appropriate amount) Corn slurry (as needed) Magnesium stearate (appropriate amount) Preparation method: The rubber seed oil of the present invention is mixed with auxiliaries, sieved, and mixed uniformly in a suitable container, and the resulting mixture is made into tablets.

[0082] Example 16: The highly active and safe rubber seed oil of the present invention was produced according to the methods of Examples 1 and 2. The rubber seed oil was made into a soft capsule as follows. Soft capsule: 1000mg of the rubber seed oil of the present invention Gelatin (appropriate amount) Glycerin (appropriate amount) Appropriate amount of water Manufacturing method: Gelatin and glycerin were soaked in distilled water to expand and soften the gelatin, and then the mixture was stirred and mixed uniformly to obtain a rubber liquid for the capsule material. The prepared rubber liquid for the capsule material was taken out and applied to a flat plate surface to make the thickness uniform, and then heated at a temperature of about 90°C to evaporate the moisture on the surface, resulting in a soft rubber sheet with a certain degree of toughness and elasticity. Finally, the rubber seed oil of the present invention (1000 mg) was filled into the soft capsule shell using a rolling mold or an automatic rotating capsule machine to produce a soft capsule.

[0083] Example 17 The highly active and safe rubber seed oil of the present invention was produced according to the methods of Examples 1 and 2. The rubber seed oil was used to prepare an emulsion for injection as follows. Injectable emulsion: 200 g of rubber seed oil according to the invention Lecithin (appropriate amount) Glycerin (appropriate amount) Sodium chloride (appropriate amount) Appropriate amount of water Manufacturing method: According to the prescription, the rubber seed oil, lecithin, and glycerin of the present invention are mixed, water is added to 1000mL, an appropriate amount of sodium chloride is added to adjust the osmotic pressure, high-pressure homogeneous emulsification is performed, the emulsion is sterilized at 121°C, and cooled to room temperature to obtain an emulsion for injection.

[0084] Example 18: Tablet: 10 mg of the rubber seed oil of the present invention produced according to the methods of Examples 1-2, 180 mg of lactose, 55 mg of starch, and 5 mg of magnesium stearate. Preparation method: the rubber seed oil of the present invention prepared according to the method of Examples 1-2, lactose and starch were mixed, uniformly moistened with water, the moistened mixture was sieved, dried, then sieved, magnesium stearate was added, and then the mixture was tableted with a weight of 250 mg per tablet, and the content of the rubber seed oil of the present invention was 10 mg.

[0085] Example 19: Capsules: 2000 g of the highly active and safe rubber seed oil of the present invention produced according to the methods of Examples 1 and 2, 1000 g of gelatin, 500 g of glycerin, and 1000 g of water. Manufacturing method: 1000g of gelatin and 500g of glycerin were put into 1000g of water, and the gelatin was immersed to expand and soften, and then the mixture was stirred and mixed uniformly to obtain a rubber liquid for the capsule material. The prepared rubber liquid for the capsule material was taken out and applied to a flat plate surface to make the thickness uniform, and then heated at a temperature of about 90°C to evaporate the moisture on the surface, resulting in a soft rubber sheet with a certain degree of toughness and elasticity. Finally, 1000mg of the rubber seed oil of the present invention was filled into the soft capsule shell using a rolling mold or an automatic rotating capsule machine to produce a soft capsule. The weight per capsule was 1.4g. The content of the rubber seed oil of the present invention was 1000mg.

[0086] Example 20: Injectable emulsion: 200 g of the highly active and safe rubber seed oil of the present invention prepared according to the methods of Examples 1 and 2, 25 g of glycerin, 12 g of lecithin, 1000 g of water, and 1 g of sodium chloride;

[0087] Manufacturing method: 200g of the rubber seed oil of the present invention manufactured according to the method of Examples 1-2, 25g of glycerin, and 12g of lecithin were precisely weighed, mixed, added to 1000mL of water, and sodium chloride was added to adjust the osmotic pressure to 350mOsm / kg H2O. An emulsion was obtained by high-pressure homogeneous emulsification, sterilized at 121°C, cooled to room temperature, and filled and sealed in a 250mL sterile glass bottle under aseptic conditions. The standard of the emulsion for injection per bottle was 250mL. The content of the rubber seed oil of the present invention was 50g.

Claims

1. It is made from crude rubber seed oil through alkali neutralization, centrifugation, adsorption filtration and deodorization. A substance that is highly active and highly safe, characterized by:

2. The substance is made from crude rubber seed oil and is obtained through alkali neutralization, centrifugation, adsorption filtration and deodorization. The method for producing a highly active and safe substance according to claim 1.

3. In the alkali neutralization step, an aqueous alkali solution is added and mixed, the alkali is an edible-grade alkali, and the amount of the alkali added is equal to the molar mass of the fatty acid contained in the crude oil; The method according to claim 2 .

4. In the centrifugation step, the temperature of the mixture is 70° C. or higher, and the centrifugal force is 5000 g or higher. The method according to claim 2 .

5. In the centrifugation step, centrifugation is performed in two stages, a first stage using a tubular centrifuge, and a second stage using a disk centrifuge or a tubular centrifuge. The method according to claim 4 .

6. In the first stage, the centrifugation is performed using a tubular centrifuge under the conditions of a mixture centrifugation temperature of 70 to 95° C. and a centrifugal force of 5,000 to 30,000 g, preferably 6,000 to 18,000 g, to remove fatty acid salts and separate an oil phase. The method according to claim 5 .

7. In the second stage, the conditions for centrifuging using a disk centrifuge or a tubular centrifuge are that the centrifugation temperature of the oil phase material is 70° C. or higher and the centrifugal force is 5,000 to 30,000 g, preferably 6,000 to 18,000 g, to remove the fatty acid salt and separate the oil phase; The method according to claim 5 .

8. A therapeutically effective amount of the highly active and safe substance according to claim 1; A pharma- ceutically acceptable carrier; 23. A pharmaceutical composition comprising:

9. 13. Use of the highly active and safe substance according to claim 1 in the manufacture of a medicament for preventing and treating atherosclerosis.

10. The highly active and highly safe substance is the only active ingredient in the drug; 10. The use according to claim 9.

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