Preparation method and application of diglyceride oil capable of preventing and treating fatty liver

Abstract

The invention relates to the technical field of grease chemistry, in particular to a preparation method and application of diglyceride oil capable of preventing and treating fatty liver. The preparation method comprises the following steps: (1) mixing raw material oil, water, glycerol and a lipase preparation for catalytic reaction to obtain a product 1; the raw material oil comprises 0.1-8 parts of sea buckthorn seed oil, 0.1-8 parts of acer truncatum seed oil, 60-90 parts of low erucic acid rapeseed oil, 15-35 parts of corn oil and 10-25 parts of peanut oil; the lipase preparation is aspergillus niger lipase and rhizopus niger lipase; (2) performing five-stage molecular distillation on the product 1 to obtain a product 2; and (3) decoloring and deodorizing the product 2 to obtain the product. The diglyceride oil prepared by the invention has excellent effects of preventing and treating fatty liver.

Description

Technical Field

[0001] The invention relates to the technical field of oil and fat chemistry, and in particular to a preparation method and application of diglyceride oil capable of preventing and treating fatty liver. Background Art

[0002] Diacylglycerol (DAG) is a structural lipid in which a fatty acid in a triacylglycerol (TG) is replaced by a hydroxyl group. DAG is a trace component of natural plant oils and fats and an endogenous intermediate product of fat metabolism in the body. In recent years, it has been found that dietary DAG has the effects of reducing visceral fat, inhibiting weight gain, and lowering blood lipids, and has therefore received widespread attention.

[0003] Diaglyceride contains dietary DAG. After entering the body, this substance has the effect of reducing visceral fat, can inhibit the excessive accumulation of visceral fat, and also help reduce the incidence of adverse diseases such as fatty liver.

[0004] DAG is a trace component in natural vegetable oils and fats, mainly in the form of 1,3-DAG and 1,2-DAG, with a content usually less than 5%. In terms of functionality, 1,3-DAG has better physiological activities than 1,2-DAG in inhibiting fat accumulation, regulating blood lipids, preventing arteriosclerosis, preventing obesity, fatty liver, and alleviating diabetes.

[0005] In addition, vegetable oils contain other ingredients that can prevent and treat fatty liver, such as unsaturated fatty acids.

[0006] Chinese patent publication number CN1544412A discloses a method for producing diglyceride oil, comprising the following steps: (1) adsorbing glycerol on an adsorption material; (2) mixing the adsorption material obtained in (1) with glycerol and triglyceride, using immobilized lipase as a catalyst, catalyzing the reaction of triglyceride and glycerol at 20-90°C; (3) separation, removing the immobilized lipase and glycerol adsorption material from the product obtained in (2), and then separating and removing monoglyceride and fatty acid to obtain diglyceride oil. This method uses triglyceride and adsorbed glycerol as raw materials, does not add water, and produces diglyceride oil by catalyzing the glycerol hydrolysis reaction at low temperature by immobilized lipase, and the generated product has less by-products and light color. However, the content of diglyceride needs to be further improved.

[0007] Currently, no research has been found on the preparation method of diglyceride oil that can prevent and treat fatty liver.

[0008] Therefore, it is very necessary to develop a preparation method and application of diglyceride oil that can solve the above technical problems and can prevent and treat fatty liver. Summary of the invention

[0009] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a preparation method and application of diglyceride oil with excellent effects in preventing and treating fatty liver.

[0010] The present invention is achieved through the following technical solutions: The first aspect of the present invention provides a method for preparing diglyceride oil capable of preventing and treating fatty liver. The diglyceride oil is prepared using Acer truncatum seed oil, low-erucic acid rapeseed oil, corn oil and peanut oil as raw material oils.

[0011] In some embodiments of the present invention, the raw oil includes, by weight, 0.1-8 parts of seabuckthorn seed oil, 0.1-8 parts of maple seed oil, 60-90 parts of low-erucic acid rapeseed oil, 15-35 parts of corn oil and 10-25 parts of peanut oil.

[0012] In some embodiments of the present invention, the preparation method comprises the following steps: (1) mixing raw oil, water, glycerol and a lipase preparation for catalytic reaction to obtain a product 1; the lipase preparation is a mixture of Aspergillus niger lipase and Rhizopus niveus lipase; (2) subjecting product 1 to molecular distillation to obtain product 2; (3) Decolorizing product 2 to obtain product 3; (4) Deodorize the product 3 to obtain.

[0013] In some embodiments of the present invention, the mass ratio of Aspergillus niger lipase to Rhizopus niveus lipase is 2-4:1.

[0014] In some embodiments of the present invention, in step (1), the mass ratio of water to the feedstock oil is 1-1.5:1; the mass ratio of glycerol to the feedstock oil is 0.3-0.6:1; and the mass of the lipase preparation is 3-8% of the mass of the feedstock oil.

[0015] In some embodiments of the present invention, the conditions of the catalytic reaction in step (1) are: temperature 50-75° C., time 1-8 h.

[0016] In some embodiments of the present invention, in step (2), the product 1 is subjected to five-stage molecular distillation, the temperature of the first-stage molecular distillation is 100-160°C, and the vacuum degree is less than 500Pa; the temperature of the second-stage molecular distillation is 190-230°C, and the vacuum degree is less than 10Pa; the temperature of the third-stage molecular distillation is 220-280°C, and the vacuum degree is less than 10Pa; the temperature of the fourth-stage molecular distillation is 220-280°C, and the vacuum degree is less than 10Pa; the temperature of the fifth-stage molecular distillation is 220-280°C, and the vacuum degree is less than 10Pa; and the heavy phase product is collected. Under these conditions, the target product diglyceride can be separated from triglyceride.

[0017] In some embodiments of the present invention, the decolorization in step (3) is carried out by using white clay and activated carbon treatment.

[0018] In some preferred embodiments of the present invention, the mass ratio of kaolin to activated carbon is 0.5-5:1; the total amount of kaolin and activated carbon is 0.1-4% of the mass of product 2.

[0019] In some embodiments of the present invention, the decolorization temperature in step (3) is 85-95° C.; the decolorization time is 30-50 min; and the vacuum degree is maintained at 60-150 Pa during the decolorization process.

[0020] In some embodiments of the present invention, the specific process of deodorization in step (4) is: first deodorize at 130-145°C for 20-40 min, then deodorize at 160-180°C for 30-60 min, and maintain a vacuum degree of 150-240 Pa during the deodorization process.

[0021] The present invention also relates to the use of diglyceride oil in preparing products for preventing and / or treating fatty liver, wherein the diglyceride oil is prepared by the above-mentioned preparation method.

[0022] In some embodiments of the present invention, the product includes food, health care products or medicines.

[0023] The beneficial effects of the present invention are: The diglyceride oil prepared by the present invention through optimizing the composition of raw oil, the composition of lipase preparation and molecular distillation process has remarkable effect in preventing and treating fatty liver. DETAILED DESCRIPTION

[0024] The present invention will be further described below in conjunction with specific embodiments, and the advantages and features of the present invention will become clearer as the description proceeds. However, these embodiments are exemplary only and do not constitute any limitation to the scope of the present invention. It should be understood by those skilled in the art that the details and forms of the technical solution of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, but these modifications and replacements all fall within the scope of protection of the present invention.

[0025] The sea buckthorn seed oil used in each embodiment of the present invention was purchased from Heze Zhonghe Jianyuan Biotechnology Co., Ltd. with batch number 08T20B48801; the Acer truncatum seed oil was purchased from Heze Zhonghe Jianyuan Biotechnology Co., Ltd. with batch number 22T20B48801; the low-erucic acid rapeseed oil was purchased from Chengdu Xinxing Grain and Oil Co., Ltd., and the corn oil was purchased from Shandong Yuhuang Grain and Oil Food Co., Ltd. with batch number PY240916107; and the peanut oil was purchased from Shandong Jinsheng Grain and Oil Food Co., Ltd. with batch number GY2405.

[0026] Aspergillus niger lipase was purchased from Beijing Ruida Henghui Technology Development Co., Ltd.; Rhizopus niveus lipase was purchased from Hong Kong Gisenberg International Trading Co., Ltd.

[0027] Example 1 The raw material oils for preparing diglyceride oil include, by weight, 0.1 parts of seabuckthorn seed oil, 0.1 parts of maple seed oil, 60 parts of low-erucic acid rapeseed oil, 15 parts of corn oil and 10 parts of peanut oil.

[0028] The preparation method of diglyceride oil comprises the following steps: (1) The raw material oil, water, glycerol and lipase preparation were mixed in a mass ratio of 1:1:0.3:0.03 for catalytic reaction at a temperature of 50°C for 8 hours to obtain product 1; the lipase preparation was a mixture of Aspergillus niger lipase and Rhizopus niveus lipase in a mass ratio of 2:1; (2) Subject product 1 to five-stage molecular distillation, wherein the temperature of the first stage molecular distillation is 100°C and the vacuum degree is 100Pa; the temperature of the second stage molecular distillation is 190°C and the vacuum degree is 1Pa; the temperature of the third stage molecular distillation is 220°C and the vacuum degree is 2Pa; the temperature of the fourth stage molecular distillation is 230°C and the vacuum degree is 3Pa; the temperature of the fifth stage molecular distillation is 240°C and the vacuum degree is 4Pa, and the heavy phase product is collected to obtain product 2; (3) Product 2 was decolorized by using white clay and activated carbon (mass ratio of 0.5:1), the total amount was 0.1% of the mass of product 2, and the decolorization conditions were: decolorization at 80°C and 60Pa for 50min, to obtain product 3; (4) The product 3 was first deodorized at 130°C for 40 min and then deodorized at 160°C for 60 min. During the deodorization process, the vacuum degree was maintained at 150 Pa.

[0029] Example 2 The raw material oils for preparing diglyceride oil include, by weight, 8 parts of seabuckthorn seed oil, 8 parts of maple seed oil, 90 parts of low-erucic acid rapeseed oil, 35 parts of corn oil and 25 parts of peanut oil.

[0030] The preparation method of diglyceride oil comprises the following steps: (1) The raw material oil, water, glycerol and lipase preparation were mixed in a mass ratio of 1:1.5:0.6:0.08 for catalytic reaction at a temperature of 75°C for 1 h to obtain product 1; the lipase preparation was a mixture of Aspergillus niger lipase and Rhizopus niveus lipase in a mass ratio of 4:1; (2) subjecting the product 1 to five-stage molecular distillation, wherein the temperature of the first stage molecular distillation is 160°C and the vacuum degree is 450Pa; the temperature of the second stage molecular distillation is 230°C and the vacuum degree is 9Pa; the temperature of the third stage molecular distillation is 260°C and the vacuum degree is 7Pa; the temperature of the fourth stage molecular distillation is 270°C and the vacuum degree is 8Pa; the temperature of the fifth stage molecular distillation is 280°C and the vacuum degree is 9Pa, and the heavy phase product is collected to obtain the product 2; (3) Product 2 was decolorized by using white clay and activated carbon (mass ratio of 5:1), the total amount was 4% of the mass of product 2, and the decolorization conditions were: decolorization at 95°C and 150Pa for 30min, to obtain product 3; (4) The product 3 was first deodorized at 145°C for 20 min and then deodorized at 180°C for 30 min. During the deodorization process, the vacuum degree was maintained at 240 Pa.

[0031] Example 3 The raw material oils for preparing diglyceride oil include, by weight, 4 parts of seabuckthorn seed oil, 4 parts of maple seed oil, 75 parts of low-erucic acid rapeseed oil, 25 parts of corn oil and 18 parts of peanut oil.

[0032] The preparation method of diglyceride oil comprises the following steps: (1) The raw material oil, water, glycerol and lipase preparation were mixed in a mass ratio of 1:1.2:0.4:0.05 for catalytic reaction at a temperature of 60°C for 4 hours to obtain product 1; the lipase preparation was a mixture of Aspergillus niger lipase and Rhizopus niveus lipase in a mass ratio of 3:1; (2) Subject product 1 to five-stage molecular distillation, wherein the temperature of the first stage molecular distillation is 130°C and the vacuum degree is 300Pa; the temperature of the second stage molecular distillation is 210°C and the vacuum degree is 5Pa; the temperature of the third stage molecular distillation is 250°C and the vacuum degree is 6Pa; the temperature of the fourth stage molecular distillation is 260°C and the vacuum degree is 7Pa; the temperature of the fifth stage molecular distillation is 270°C and the vacuum degree is 8Pa, and the heavy phase product is collected to obtain product 2; (3) Product 2 was decolorized by using white clay and activated carbon (mass ratio of 3:1), the total amount was 2% of the mass of product 2, and the decolorization conditions were: decolorization at 90°C and 105Pa for 40min, to obtain product 3; (4) The product 3 was first deodorized at 140°C for 30 min and then deodorized at 170°C for 45 min. During the deodorization process, the vacuum degree was maintained at 200 Pa.

[0033] Comparative Example 1 The only difference from Example 3 is that the raw material oil composition for preparing diglyceride oil is different, and the other conditions are the same, as follows: The raw material oils for preparing diglyceride oil include, by weight, 4 parts of seabuckthorn seed oil, 78 parts of low-erucic acid rapeseed oil, 26 parts of corn oil and 18 parts of peanut oil.

[0034] Comparative Example 2 The only difference from Example 3 is that the raw material oil composition for preparing diglyceride oil is different, and the other conditions are the same, as follows: The raw material oils for preparing diglyceride oil include, by weight, 4 parts of seabuckthorn seed oil, 14.34 parts of Acer truncatum seed oil, 89.66 parts of corn oil and 18 parts of peanut oil.

[0035] Comparative Example 3 The only difference from Example 3 is that the raw material oil composition for preparing diglyceride oil is different, and the other conditions are the same, as follows: The raw material oils for preparing diglyceride oil include, by weight, 4 parts of seabuckthorn seed oil, 5.266 parts of Acer truncatum seed oil, 98.734 parts of low-erucic acid rapeseed oil and 18 parts of peanut oil.

[0036] Comparative Example 4 The only difference from Example 3 is that the total amount of the lipase preparation remains unchanged, the composition is different, and it is only Aspergillus niger lipase, and the other conditions are the same.

[0037] Comparative Example 5 The only difference from Example 3 is that the total amount of the lipase preparation remains unchanged, the composition is different, and it is only Rhizopus niveus lipase, and the other conditions are the same.

[0038] Comparative Example 6 The only difference from Example 3 is that the molecular distillation process parameters in step (2) are different, and the other conditions are the same, as follows: Product 1 was subjected to five-stage molecular distillation, with the temperature of the first stage molecular distillation being 80°C and the vacuum degree being 300Pa; the temperature of the second stage molecular distillation being 160°C and the vacuum degree being 5Pa; the third stage molecular distillation being 200°C and the vacuum degree being 6Pa; the fourth stage molecular distillation being 210°C and the vacuum degree being 7Pa; the fifth stage molecular distillation being 290°C and the vacuum degree being 8Pa, and the heavy phase product was collected to obtain product 2.

[0039] Test Example 1 Prevention and Treatment of Fatty Liver Disease Test Healthy SD male rats weighing 200±20g were selected. The maintenance feed consisted of raw materials such as corn, soybean cake, and fish meal. The high-fat feed was prepared by adding 15% lard to the maintenance feed. The diglyceride feed was prepared by adding 15% diglyceride oil (the diglyceride oil prepared in each embodiment or comparative example) to the maintenance feed. The temperature of the breeding room was maintained at 22±2°C, the humidity was maintained at 55±5%, and the darkness and light were adjusted at intervals of 12 hours.

[0040] After one week of normal feeding with maintenance feed, 10 mice were randomly selected as the normal control group and continued to be fed with maintenance feed. The remaining mice were fed with high-fat feed as the high-fat diet group. After 30 days of feeding, they were randomly divided into 10 groups, each with 10 mice, of which 1 group was used as the model group and continued to be fed with high-fat feed, and the remaining 9 groups were fed with diglyceride feed containing diglyceride oil prepared in Examples 1-3 and Comparative Examples 1-6. Subsequent experiments were carried out after 30 days of feeding.

[0041] After the experiment, the rats were fasted for 8.5h, and each group was anesthetized with 3% sodium pentobarbital (30mg / kg). The rats were weighed and blood was taken before being killed by cervical dislocation. The blood samples were centrifuged at 4000rpm and serum was taken for the detection of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in blood lipids. The liver tissue was washed with 4℃ pre-cooled saline, and the water was absorbed by filter paper and weighed. 10ml of saline was added to each gram of liver tissue, and the mixture was homogenized by electric stirring, placed in a 4000 r / min centrifuge for 10min, and centrifuged 3 times. The supernatant was taken to measure the content of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the liver tissue.

[0042] Liver index = liver weight / body weight*100%. The results are shown in Table 1.

[0043] The test results of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) are shown in Table 2.

[0044] Table 1 Liver index test results

[0045] Note: Compared with the model group, *p<0.05, **p<0.01. Compared with Example 3, each embodiment and comparative example, #p<0.05, ##p<0.01.

[0046] As shown in Table 1, the liver index of the model group is significantly higher than that of the normal control group, indicating that the rats in the model group have already developed fatty liver. Under the intervention of diglyceride oil, the liver index of each embodiment is significantly lower than that of the model group, indicating that the diglyceride oil prepared in each embodiment can prevent and treat fatty liver. The liver index of each embodiment is significantly lower than that of each comparative example, indicating that the intervention effect of the diglyceride oil prepared in Examples 1 to 3 on fatty liver is significantly better than that of each comparative example.

[0047] Table 2 Test results of various indicators

[0048] Note: The same letters between groups in the same column indicate that there is no significant difference between the corresponding groups, and different letters indicate that there is a significant difference between the corresponding groups, p < 0.05.

[0049] As shown in Table 2, the TC, TG, and LDL-C of the model group were significantly higher than those of the normal control group, and the HDL-C was significantly lower than that of the normal control group, indicating that the rats in the model group had already developed fatty liver, and AST and ALT, which are markers of liver damage, also showed a significant increase in the model group. Under the intervention of diglyceride oil in each embodiment, each index changed significantly, indicating that the diglyceride oil prepared in each embodiment can prevent and treat fatty liver. The effect of each index in each embodiment is significantly better than that of each comparative example, indicating that the intervention effect of the diglyceride oil prepared in Examples 1 to 3 on fatty liver is significantly better than that of each comparative example.

[0050] The above detailed description is a specific description of one feasible embodiment of the present invention. The embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not deviate from the present invention should be included in the scope of the technical solution of the present invention.

Claims

1. A method for preparing diglyceride oil capable of preventing and treating fatty liver, characterized in that: The preparation method comprises the following steps: (1) mixing raw material oil, water, glycerol and lipase preparation for catalytic reaction to obtain product 1; the raw material oil comprises, by weight, 0.1-8 parts of seabuckthorn seed oil, 0.1-8 parts of Acer truncatum seed oil, 60-90 parts of low-erucic acid rapeseed oil, 15-35 parts of corn oil and 10-25 parts of peanut oil; the lipase preparation is a mixture of Aspergillus niger lipase and Rhizopus niveus lipase; (2) subjecting the product 1 to five-stage molecular distillation, wherein the temperature of the first-stage molecular distillation is 100-160°C, and the vacuum degree is less than 500Pa; the temperature of the second-stage molecular distillation is 190-230°C, and the vacuum degree is less than 10Pa; the temperature of the third-stage molecular distillation is 220-280°C, and the vacuum degree is less than 10Pa; the temperature of the fourth-stage molecular distillation is 220-280°C, and the vacuum degree is less than 10Pa; the temperature of the fifth-stage molecular distillation is 220-280°C, and the vacuum degree is less than 10Pa, to obtain product 2; (3) Decolorizing product 2 to obtain product 3; (4) Deodorize the product 3 to obtain.

2. The preparation method according to claim 1, characterized in that: The mass ratio of Aspergillus niger lipase to Rhizopus niveus lipase is 2-4:

1.

3. The preparation method according to claim 1, characterized in that: In step (1), the mass ratio of water to the raw oil is 1-1.5:1; the mass ratio of glycerol to the raw oil is 0.3-0.6:1; and the mass of the lipase preparation is 3-8% of the mass of the raw oil.

4. The preparation method according to claim 1, characterized in that: The conditions of the catalytic reaction in step (1) are: temperature 50-75°C, time 1-8h.

5. The preparation method according to claim 1, characterized in that: The decolorization in step (3) is carried out by using white clay and activated carbon treatment.

6. The preparation method according to claim 5, characterized in that: The mass ratio of kaolin and activated carbon is 0.5-5:1; the total amount of kaolin and activated carbon is 0.1-4% of the mass of product 2.

7. The preparation method according to claim 1, characterized in that: In step (3), the decolorization temperature is 80-95°C; the decolorization time is 30-50 minutes; and the vacuum degree is maintained at 60-150Pa during the decolorization process.

8. The preparation method according to claim 1, characterized in that: The specific process of deodorization in step (4) is: first deodorize at 130-145°C for 20-40 min, then deodorize at 160-180°C for 30-60 min, and maintain the vacuum degree at 150-240 Pa during the deodorization process.

9. Use of diglyceride oil in preparing a product for preventing and / or treating fatty liver, characterized in that: The diglyceride oil is prepared by the preparation method according to any one of claims 1 to 8.

10. The use according to claim 9, characterized in that: The products include food, health products or medicines.

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