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

Abstract

The present invention relates to the technical field of oil and fat chemistry, and specifically 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 lipase preparation for catalytic reaction to obtain product 1; the raw material oil comprises 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; the lipase preparation is Aspergillus niger lipase and Rhizopus niveus lipase; (2) subjecting product 1 to five-stage molecular distillation to obtain product 2; (3) subjecting product 2 to decolorization and deodorization, that is, obtaining. The diglyceride oil prepared by the present invention has excellent effect in preventing and treating fatty liver.

Description

Technical Field

[0001] The present 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 one of the fatty acids in triacylglycerol (TG) is replaced by a hydroxyl group. DAG is a trace component of natural plant oils and fats and an endogenous intermediate in fat metabolism. In recent years, dietary DAG has been shown to reduce visceral fat, inhibit weight gain, and lower blood lipids, garnering widespread attention.

[0003] Diacerol 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 plant oils and fats, primarily consisting of two isomers: 1,3-DAG and 1,2-DAG, with their content typically less than 5%. In terms of functionality, 1,3-DAG is more effective than 1,2-DAG in inhibiting fat accumulation, regulating blood lipids, preventing arteriosclerosis, preventing obesity and 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 No. CN1544412A discloses a method for producing diglyceride oil, comprising the following steps: (1) adsorbing glycerol onto an adsorption material; (2) mixing the adsorption material obtained in (1) with glycerol and triglycerides, and using immobilized lipase as a catalyst to catalyze the reaction of triglycerides and glycerol at 20-90°C; (3) separating, removing the immobilized lipase and glycerol adsorption material from the product obtained in (2), and then separating and removing monoglycerides and fatty acids to obtain diglyceride oil. This method uses triglycerides and adsorbed glycerol as raw materials, does not add water, and produces diglyceride oil by catalyzing the glycerol decomposition reaction at low temperature by immobilized lipase. The resulting product contains few by-products and is light in color. However, the diglyceride content 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:

[0011] A 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 materials.

[0012] In some embodiments of the present invention, the raw oil comprises, by weight, 0.1-8 parts of sea buckthorn 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.

[0013] In some embodiments of the present invention, the preparation method comprises the following steps:

[0014] (1) mixing raw oil, water, glycerol and a lipase preparation for catalytic reaction to obtain product 1; the lipase preparation is a mixture of Aspergillus niger lipase and Rhizopus niveus lipase;

[0015] (2) molecularly distilling product 1 to obtain product 2;

[0016] (3) Decolorizing product 2 to obtain product 3;

[0017] (4) Deodorize the product 3 to obtain.

[0018] In some embodiments of the present invention, the mass ratio of Aspergillus niger lipase to Rhizopus niveus lipase is 2-4:1.

[0019] In some embodiments of the present invention, the mass ratio of the water to the feedstock oil in step (1) is 1-1.5:1; the mass ratio of the 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.

[0020] In some embodiments of the present invention, the conditions for the catalytic reaction in step (1) are: temperature 50-75° C., time 1-8 h.

[0021] In some embodiments of the present invention, in step (2), product 1 is subjected 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 500 Pa; the temperature of the second stage molecular distillation is 190-230°C and the vacuum degree is less than 10 Pa; the temperature of the third stage molecular distillation is 220-280°C and the vacuum degree is less than 10 Pa; the temperature of the fourth stage molecular distillation is 220-280°C and the vacuum degree is less than 10 Pa; the temperature of the fifth stage molecular distillation is 220-280°C and the vacuum degree is less than 10 Pa; and the heavy phase product is collected. Under these conditions, the target product diglyceride can be separated from the triglyceride.

[0022] In some embodiments of the present invention, the decolorization in step (3) is carried out by using white clay and activated carbon treatment.

[0023] In some preferred embodiments of the present invention, the mass ratio of clay to activated carbon is 0.5-5:1; and the total amount of clay and activated carbon is 0.1-4% of the mass of product 2.

[0024] 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.

[0025] 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 the vacuum degree at 150-240 Pa during the deodorization process.

[0026] The present invention also relates to the use of diglyceride oil in preparing products for preventing and / or treating fatty liver. The diglyceride oil is prepared by the above-mentioned preparation method.

[0027] In some embodiments of the present invention, the product includes food, health care product or medicine.

[0028] The beneficial effects of the present invention are:

[0029] The present invention optimizes the composition of raw oil, the composition of lipase preparation and molecular distillation process to prepare diglyceride oil, which has significant effects in preventing and treating fatty liver. DETAILED DESCRIPTION

[0030] The present invention will be further described below with reference to specific embodiments, and the advantages and features of the present invention will become clearer as the description proceeds. However, these embodiments are merely exemplary 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 solutions of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, and such modifications and replacements fall within the scope of protection of the present invention.

[0031] 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 maple 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., 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.

[0032] Aspergillus niger lipase was purchased from Beijing Ruida Henghui Technology Development Co., Ltd.; Rhizopus niveus lipase was purchased from Hong Kong Gisenberg International Trade Co., Ltd.

[0033] Example 1

[0034] 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.

[0035] The preparation method of diglyceride oil comprises the following steps:

[0036] (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 50°C for 8 h 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;

[0037] (2) Product 1 was subjected to five-stage molecular distillation, with the temperature of the first stage molecular distillation being 100°C and the vacuum being 100 Pa; the temperature of the second stage molecular distillation being 190°C and the vacuum being 1 Pa; the temperature of the third stage molecular distillation being 220°C and the vacuum being 2 Pa; the temperature of the fourth stage molecular distillation being 230°C and the vacuum being 3 Pa; and the temperature of the fifth stage molecular distillation being 240°C and the vacuum being 4 Pa, and the heavy phase product was collected to obtain product 2;

[0038] (3) Product 2 was decolorized using white clay and activated carbon (mass ratio of 0.5:1), with the total amount being 0.1% of the mass of product 2. The decolorization conditions were: decolorization at 80°C and 60 Pa for 50 min to obtain product 3;

[0039] (4) The product 3 was first deodorized at 130°C for 40 min and then at 160°C for 60 min. The vacuum degree was maintained at 150 Pa during the deodorization process.

[0040] Example 2

[0041] 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.

[0042] The preparation method of diglyceride oil comprises the following steps:

[0043] (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 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;

[0044] (2) Product 1 was subjected to five-stage molecular distillation, with the temperature of the first stage molecular distillation being 160°C and the vacuum being 450 Pa; the temperature of the second stage molecular distillation being 230°C and the vacuum being 9 Pa; the temperature of the third stage molecular distillation being 260°C and the vacuum being 7 Pa; the temperature of the fourth stage molecular distillation being 270°C and the vacuum being 8 Pa; and the temperature of the fifth stage molecular distillation being 280°C and the vacuum being 9 Pa, and the heavy phase product was collected to obtain product 2;

[0045] (3) Product 2 was decolorized using white clay and activated carbon (mass ratio of 5:1), with the total amount being 4% of the mass of product 2. The decolorization conditions were: decolorization at 95°C and 150 Pa for 30 min to obtain product 3;

[0046] (4) The product 3 was first deodorized at 145°C for 20 min and then at 180°C for 30 min. The vacuum degree was maintained at 240 Pa during the deodorization process.

[0047] Example 3

[0048] 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.

[0049] The preparation method of diglyceride oil comprises the following steps:

[0050] (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 60°C for 4 h 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;

[0051] (2) Product 1 was subjected to five-stage molecular distillation, with the temperature of the first stage molecular distillation being 130°C and the vacuum being 300 Pa; the temperature of the second stage molecular distillation being 210°C and the vacuum being 5 Pa; the temperature of the third stage molecular distillation being 250°C and the vacuum being 6 Pa; the temperature of the fourth stage molecular distillation being 260°C and the vacuum being 7 Pa; and the temperature of the fifth stage molecular distillation being 270°C and the vacuum being 8 Pa, and the heavy phase product was collected to obtain product 2;

[0052] (3) Product 2 was decolorized using white clay and activated carbon (mass ratio of 3:1), with the total amount being 2% of the mass of product 2. The decolorization conditions were: decolorization at 90°C and 105 Pa for 40 min to obtain product 3;

[0053] (4) The product 3 was first deodorized at 140°C for 30 min and then at 170°C for 45 min. The vacuum degree was maintained at 200 Pa during the deodorization process.

[0054] Comparative Example 1

[0055] The only difference from Example 3 is the composition of the raw oil used to prepare the diglyceride oil. The other conditions are the same, as follows:

[0056] 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.

[0057] Comparative Example 2

[0058] The only difference from Example 3 is the composition of the raw oil used to prepare the diglyceride oil. The other conditions are the same, as follows:

[0059] The raw material oils for preparing the 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.

[0060] Comparative Example 3

[0061] The only difference from Example 3 is the composition of the raw oil used to prepare the diglyceride oil. The other conditions are the same, as follows:

[0062] The raw material oils for preparing the 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.

[0063] Comparative Example 4

[0064] 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. The other conditions are the same.

[0065] Comparative Example 5

[0066] 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. The other conditions are the same.

[0067] Comparative Example 6

[0068] 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:

[0069] Product 1 was subjected to five-stage molecular distillation, with the temperature of the first molecular distillation being 80°C and the vacuum being 300 Pa; the temperature of the second molecular distillation being 160°C and the vacuum being 5 Pa; the temperature of the third molecular distillation being 200°C and the vacuum being 6 Pa; the temperature of the fourth molecular distillation being 210°C and the vacuum being 7 Pa; and the temperature of the fifth molecular distillation being 290°C and the vacuum being 8 Pa. The heavy phase product was collected to obtain product 2.

[0070] Test Example 1

[0071] Fatty Liver Disease Prevention and Treatment Test

[0072] Healthy male Sprague-Dawley rats weighing 200 ± 20 g were selected. A maintenance diet consisted of corn, soybean meal, fish meal, and other ingredients. A high-fat diet was prepared by adding 15% lard to the maintenance diet. A diglyceride diet was prepared by adding 15% diglyceride oil (prepared in the Examples or Comparative Examples) to the maintenance diet. The temperature in the housing was maintained at 22 ± 2°C, the humidity at 55 ± 5%, and the light and dark cycles were adjusted at 12-hour intervals.

[0073] After one week of normal feeding with the maintenance diet, 10 mice were randomly selected as a normal control group and continued to be fed the maintenance diet. The remaining mice were fed a high-fat diet as a high-fat diet group. After 30 days of feeding, they were randomly divided into 10 groups of 10 mice each. One group served as the model group and continued to be fed the high-fat diet, while the remaining 9 groups were fed a diglyceride diet containing the diglyceride oils prepared in Examples 1-3 and Comparative Examples 1-6. Subsequent experiments were performed after 30 days of feeding.

[0074] After the experiment, rats were fasted for 8.5 hours. Each group was anesthetized with 3% sodium pentobarbital (30 mg / kg), weighed, and blood was collected before being sacrificed by cervical dislocation. Blood samples were centrifuged at 4000 rpm, and serum was collected for analysis of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C). Liver tissue was washed with 4°C pre-cooled saline to remove blood, blotted dry with filter paper, and weighed. 10 ml of saline was added per gram of liver tissue, and the mixture was homogenized by electric homogenization. The mixture was centrifuged at 4000 rpm for 10 minutes, repeated three times, and the supernatant was collected for measurement of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the liver tissue.

[0075] Liver index = liver weight / body weight*100%. The results are shown in Table 1.

[0076] 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.

[0077] Table 1 Liver index test results

[0078]

[0079] Note: Compared with the model group, *p<0.05, **p<0.01. Compared with Example 3, each example and comparative example, #p<0.05, ##p<0.01.

[0080] As shown in Table 1, the liver index of the model group was significantly higher than that of the normal control group, indicating that the model group rats had developed fatty liver. However, the liver index of each Example, when treated with diglyceride oil, was significantly lower than that of the model group, indicating that the diglyceride oil prepared in each Example can prevent and treat fatty liver. The liver index of each Example was significantly lower than that of each comparative example, indicating that the diglyceride oil prepared in Examples 1-3 was significantly more effective in preventing fatty liver than the comparative examples.

[0081] Table 2 Test results of various indicators

[0082]

[0083] Note: The same letters between groups in the same column indicate no significant difference between the corresponding groups, and different letters indicate significant difference between the corresponding groups, p < 0.05.

[0084] As shown in Table 2, the model group had significantly higher TC, TG, and LDL-C levels than the normal control group, while HDL-C was significantly lower than the normal control group, indicating that the model group rats had developed fatty liver. AST and ALT, markers of liver damage, also showed significant increases in the model group. In each example, the diglyceride oil intervention significantly altered these indicators, demonstrating that the diglyceride oils prepared in each example can prevent and treat fatty liver. The effects of each indicator in each example were significantly superior to those in the comparative examples, indicating that the diglyceride oils prepared in Examples 1-3 were significantly more effective in preventing fatty liver than those in the comparative examples.

[0085] The above detailed description is a specific description of one feasible embodiment of the present invention. This embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not depart 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 is calculated by weight as follows: 0.1-8 parts of seabuckthorn seed oil, 0.1-8 parts of Acer truncatum seed oil, 60-90 parts of 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; the mass ratio of Aspergillus niger lipase to Rhizopus niveus lipase is 2-4:1; (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 220-260°C, and the vacuum degree is less than 10Pa; the temperature of the fourth-stage molecular distillation is 230-270°C, and the vacuum degree is less than 10Pa; the temperature of the fifth-stage molecular distillation is 240-280°C, and the vacuum degree is less than 10Pa, to obtain product 2; (3) Decolorizing product 2 to obtain product 3; (4) Deodorizing the product 3 to obtain; 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.

2. 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.

3. 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.

4. The preparation method according to claim 3, 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.

5. 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.

6. 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.

7. Use of diglyceride oil in the preparation of a medicine 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 6.

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