A bile acid synthesis promoter, its preparation method and application

Abstract

The present invention discloses a bile acid synthesis promoter, its preparation method and application, belonging to the field of aquaculture. The above-mentioned fishery bile acid synthesis promoter, by mass, comprises the following components: 10-30 parts of lignans; 2-10 parts of tremella polysaccharide; 460-488 parts of a carrier, and can be used as a feed additive in the aquaculture of aquatic animals. The present invention makes full use of the beneficial effects of lignans, tremella polysaccharide and the carrier, and conducts scientific compatibility thereof. Through in-depth research, the appropriate dosage thereof is mastered, and the regulatory effect and mechanism on bile acid metabolism in fish bodies are clarified, which has strong pertinence. Combined with the growth and physical and chemical indexes of relevant aquaculture experiments, it is proved that the bile acid synthesis promoter prepared by the present invention can significantly promote the biosynthesis of bile acids in fish bodies, improve the bile acid homeostasis in fish bodies, and further maintain the normal metabolic functions of fish bodies.

Description

Technical Field

[0001] The present invention relates to the field of aquaculture, and particularly to a bile acid synthesis promoter, a preparation method thereof, and an application thereof. Background Art

[0002] The protein content in aquaculture feed is relatively high, resulting in high feed costs. Glycolipids have the advantages of wide sources, low prices, etc., and have a protein-saving effect. In production, the protein content in feed is often reduced by increasing the glycolipid content, thereby reducing the feed cost. However, the unreasonable use of the two often leads to fish body metabolic disorders, and further induces nutritional metabolic diseases such as hepatobiliary syndrome, causing huge economic losses to the aquaculture industry. Research shows that bile acids play an important regulatory role in the glycolipid metabolism of fish. Based on this, in production, bile acids are often added to feed to prevent and treat fish liver syndrome. However, bile acids themselves have an unpleasant smell, which can reduce the feed intake of fish to a certain extent, and further reduce their growth performance. In addition, commercial bile acids are mostly extracted from livestock and poultry bile pastes, without considering the differences in bile acid spectra between aquatic animals and livestock and poultry, resulting in weak specificity of the action of bile acids. Based on this, it is of great significance to enhance the bile acid biosynthesis ability and reabsorption function of fish bodies through nutritional regulation means.

[0003] Under normal circumstances, cholesterol is converted into primary bile in the liver under the catalytic action of cholesterol 7α-hydroxylase (CYP7A1). The latter enters the gallbladder for storage through the bile salt export pump and is secreted into the intestine after eating. Under the action of intestinal flora, primary bile acids are converted into secondary bile acids. More than 95% of the secondary bile acids will be reabsorbed in the ileum and returned to the liver through the vein, and then used for the resynthesis of bile acids. The above process is called the enterohepatic circulation of bile acids. In the process of bile acid metabolism, organs such as the liver and intestine play important roles. In view of this, by means of nutritional regulation to improve the bile acid biosynthesis function of fish liver and the bile acid reabsorption function of the intestine, the enterohepatic circulation of bile acids can be significantly enhanced, and then the metabolic function of fish can be improved. Summary of the Invention

[0004] Object of the Invention: The present invention aims to provide a bile acid synthesis promoter that can improve the bile acid biosynthesis ability, improve the metabolic function of animal bodies, and further enhance the utilization rate of glycolipids in feed by animal bodies. Moreover, the present invention also provides a preparation method and an application of the bile acid synthesis promoter.

[0005] Technical Solution: The bile acid synthesis promoter provided by the present invention, by mass, comprises the following components: 10-30 parts of lignans; 2-10 parts of tremella polysaccharide; 460-488 parts of a carrier.

[0006] Preferably, the bile acid synthesis promoter comprises the following components by mass: 15-25 parts of lignan; 2-8 parts of tremella polysaccharide; 467-483 parts of carrier.

[0007] More preferably, the bile acid synthesis promoter comprises the following components by mass: 20 parts of lignan; 5 parts of tremella polysaccharide; 475 parts of carrier.

[0008] Furthermore, the carrier is rice husk powder.

[0009] The preparation method of the bile acid synthesis promoter provided by the present invention comprises the steps of: respectively pulverizing lignan, tremella polysaccharide and carrier, sieving them, and then mixing them to obtain the bile acid synthesis promoter.

[0010] Furthermore, the pulverizing and mixing conditions of the lignan are both: light intensity < 800 lx, temperature < 40 °C.

[0011] Furthermore, the mesh number of the sieving is 100-120.

[0012] The application of the bile acid synthesis promoter provided by the present invention as a feed additive in farmed animals.

[0013] Furthermore, the farmed animals are aquatic animals, preferably cyprinid fish.

[0014] Furthermore, the addition amount of the bile acid synthesis promoter is 1.0-5.0%, which can be appropriately adjusted according to the types, growth stages, specifications, farming methods and feed nutritional levels of aquatic animals, etc.; the feed is high-sugar or high-fat feed.

[0015] Invention principle: In the present invention, lignan, as a natural polyphenol substance, can up-regulate the expression of the bile acid synthesis rate-limiting enzyme CYP7A1, promote the biosynthesis of bile acids in the liver, and thus reduce the cholesterol content in fish. Tremella polysaccharide is a kind of functional polymer sugar, which can promote the proliferation of beneficial bacteria in the intestine and inhibit the growth of harmful bacteria, maintain the intestinal microecological balance of fish, and thus create good conditions for the formation of secondary bile acids and the reabsorption of bile acids. Rice husk powder has good fluidity and adsorption, can carry lignan and tremella polysaccharide, and make the two evenly dispersed in the feed. Based on this, the present invention fully combines the beneficial effects of the two in promoting bile acid synthesis and reabsorption, etc., scientifically formulates them and uses them in aquatic feed, which will help to improve the biosynthesis and enterohepatic circulation of bile acids in fish, and thus improve the glycolipid metabolism function, and reduce the incidence of nutritional metabolic diseases such as hepatobiliary syndrome, and finally promote the healthy and sustainable development of the aquaculture industry.

[0016] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages:

[0017] (1) The bile acid promoter of the present invention uses lignans and tremella polysaccharide as the core components, and rice husk powder as the carrier, fully combining the beneficial effects of lignans and tremella polysaccharide in promoting bile acid biosynthesis and reabsorption. Through in-depth research, the appropriate dosage and compatibility ratio have been mastered, and its regulatory effects and mechanisms on fish bile acid metabolism have been clarified, with strong pertinence.

[0018] (2) Both lignans and tremella polysaccharide have been widely used in human dietary supplements, and their physiological effects, beneficial effects and safety have been confirmed. As a carrier for feed additives, rice husk powder has been widely used in the feed processing industry. All three have no toxic or side effects on aquatic animals, livestock and poultry, and humans.

[0019] (3) Combining the growth and physical-chemical indexes of relevant breeding tests, the present invention proves that the feed additive can significantly promote the biosynthesis of bile acids in the fish liver, enhance the reabsorption of bile acids in the intestine, and thus increase the bile acid secretion of fish; it can significantly reduce the cholesterol level of fish, improve the sugar tolerance of fish, improve the lipid and carbohydrate metabolism of fish, and significantly reduce the incidence of nutritional metabolic diseases such as hepatobiliary syndrome; it can significantly improve the utilization rate of non-protein energy in feed by fish, reduce the protein dosage in feed and the ammonia nitrogen emissions of aquatic animals, and thus reduce the feed production cost and improve water quality, ultimately promoting the healthy and sustainable development of aquaculture. Description of the Drawings

[0020] Figure 1 Shows the effects of different feeds in Application Example 2 on the plasma biochemical indexes of juvenile blunt snout bream;

[0021] Figure 2 Shows the effects of different feeds in Application Example 2 on the liver biochemical indexes of juvenile blunt snout bream;

[0022] Figure 3 Shows the effects of different feeds in Application Example 2 on the posterior intestine biochemical indexes of juvenile blunt snout bream;

[0023] Figure 4 Shows the effects of different feeds in Application Example 2 on the CYP7A1 expression level in the liver of blunt snout bream. Detailed Embodiments

[0024] The technical solutions of the present invention will be further described below in conjunction with the embodiments and the drawings.

[0025] Embodiment 1: The bile acid synthesis promoter provided in this embodiment includes the following components by mass: 10 g of lignans, 10 g of tremella polysaccharide, and 480 g of rice husk powder.

[0026] The preparation method is as follows: Grind the lignan, tremella polysaccharide and rice husk powder respectively and sieve them through a 120-mesh sieve, and set aside; then mix 10 g of the sieved lignan, 10 g of tremella polysaccharide and 480 g of rice husk powder evenly in a step-by-step manner. Ensure the uniformity and stability of the mixture during mixing, and 500 g of the bile acid synthesis promoter is thus obtained.

[0027] Example 2: The bile acid synthesis promoter provided in this example, by mass, comprises the following components: 15 g of lignan, 10 g of tremella polysaccharide and 475 g of rice husk powder.

[0028] The preparation method is as follows: Grind the lignan, tremella polysaccharide and rice husk powder respectively and sieve them through a 120-mesh sieve, and set aside; then mix 15 g of the sieved lignan, 10 g of tremella polysaccharide and 475 g of rice husk powder evenly in a step-by-step manner. Ensure the uniformity and stability of the mixture during mixing, and 500 g of the bile acid synthesis promoter is thus obtained.

[0029] Example 3: The bile acid synthesis promoter provided in this example, by mass, comprises the following components: 20 g of lignan, 5 g of tremella polysaccharide and 475 g of rice husk powder.

[0030] The preparation method is as follows: Grind the lignan, tremella polysaccharide and rice husk powder respectively and sieve them through a 120-mesh sieve, and set aside; after mixing 20 g of the sieved lignan and 5 g of tremella polysaccharide evenly, gradually add them to 475 g of the sieved rice husk powder, and continuously stir and mix during the addition to ensure the uniformity and stability of the mixture, and 500 g of the bile acid synthesis promoter is thus obtained.

[0031] Example 4: The bile acid synthesis promoter provided in this example, by mass, comprises the following components: 25 g of lignan, 5 g of tremella polysaccharide and 470 g of rice husk powder.

[0032] The preparation method is as follows: Grind the lignan, tremella polysaccharide and rice husk powder respectively and sieve them through a 120-mesh sieve, and set aside; then mix 25 g of the sieved lignan, 5 g of tremella polysaccharide and 470 g of rice husk powder evenly in a step-by-step manner. Ensure the uniformity and stability of the mixture during mixing, and 500 g of the bile acid synthesis promoter is thus obtained.

[0033] Example 5: The bile acid synthesis promoter provided in this example, by mass, comprises the following components: 30 g of lignan, 2 g of tremella polysaccharide and 468 g of rice husk powder.

[0034] The preparation method is as follows: Grind the lignan, tremella polysaccharide and rice husk powder respectively and sieve them through a 120-mesh sieve, and set aside; after mixing 30 g of the sieved lignan and 2 g of tremella polysaccharide evenly, gradually add them to 468 g of the sieved rice husk powder, and continuously stir and mix during the addition to ensure the uniformity and stability of the mixture, and 500 g of the bile acid synthesis promoter is thus obtained.

[0035] Comparative Example 1: The bile acid synthesis promoter provided in this comparative example, by mass, includes the following components: 20 g of lignan and 480 g of rice husk powder.

[0036] Preparation method: Crush the lignan and rice husk powder separately and pass through a 120-mesh sieve for standby; gradually add 20 g of the sieved lignan to 480 g of the sieved rice husk powder, and continuously stir and mix during the addition to ensure the uniformity and stability of the mixture, thus obtaining 500 g of bile acid synthesis promoter 1.

[0037] Comparative Example 2: The bile acid synthesis promoter provided in this comparative example, by mass, includes the following components: 5 g of tremella polysaccharide and 495 g of rice husk powder.

[0038] Preparation method: Crush the tremella polysaccharide and rice husk powder separately and pass through a 120-mesh sieve for standby; gradually add 5 g of the sieved tremella polysaccharide to 495 g of the sieved rice husk powder, and continuously stir and mix during the addition to ensure the uniformity and stability of the mixture, thus obtaining 500 g of bile acid synthesis promoter 2.

[0039] Application Example 1: Carp breeding experiment and result analysis

[0040] The breeding experiment was carried out in a pond (specification: 100 m × 50 m) at the Aquaculture Teaching and Research Base of Nanjing Agricultural University (Xingdian Town, Pukou District, Nanjing). 400 experimental fish with uniform specifications (average weight: 28.95 ± 0.29 g) were randomly divided into 20 net cages (specification: 2 m × 1 m × 1 m), and 20 experimental fish were fed in each net cage. Among them, 1) the control group was fed a basal diet with a sugar level of 31.6%; 2) the high-sugar group was fed a high-sugar diet with a sugar level of 45%; 3) the high-sugar + regulator 1 group was fed a high-sugar diet + 1% of the bile acid synthesis promoter 1 prepared in Comparative Example 1 as a feed additive; 4) the high-sugar + regulator 2 group was fed a high-sugar diet + 1% of the bile acid synthesis promoter 2 prepared in Comparative Example 2 as a feed additive; 5) the high-sugar + compound regulator group was fed a high-sugar diet + 1% of the bile acid synthesis promoter prepared in Experimental Example 3 as a feed additive. The feed formula is shown in Table 1. The test period was 12 weeks. During the breeding experiment, 6 test feeds were artificially fed (daily feeding times: 7:10, 12:10, and 16:10) until the fish showed an obvious satiated state, and the growth of blunt snout bream was observed. After the experiment ended, samples were collected for further analysis and determination. The test results are shown in Tables 2 - 4.

[0041] Table 1 Test feed formula

[0042]

[0043] Table 2 Effects of different feeds on the growth and feed utilization rate of juvenile carp

[0044]

[0045] Table 3 Effects of different feeds on plasma biochemical parameters of carp fry

[0046]

[0047] Note: Data in the same industry with the same letters in the superscript do not differ significantly.

[0048] Table 4 Effects of different feeds on biochemical parameters in liver and hindgut of carp fry

[0049]

[0050] As shown in Table 2, different feed treatments had no significant effect on the final weight, weight gain rate, specific growth rate, feed intake, feed-to-weight ratio and protein efficiency ratio of carp fry (P>0.05).

[0051] As shown in Table 3, compared with the control group, the plasma glucose, glycosylated serum protein, triglyceride and total cholesterol content of carp fry in the high sugar group were significantly increased, and the bile acid content was significantly decreased (P<0.05). Compared with the high sugar group, high sugar + regulator 1 and high sugar + regulator 2 both had a significant effect on reducing plasma glucose, and high sugar + compound regulator further improved this effect (P<0.05). In addition, the compound regulator significantly reduced the content of glycosylated serum protein, triglyceride and total cholesterol (P<0.05), while the use of regulator 1 (lignan) or regulator 2 (Sophora japonica polysaccharide) alone did not have a significant effect on these two indicators (P>0.05). Adding regulator 1 to the high sugar feed had a significant effect on increasing the plasma bile acid content (P<0.05), and the plasma bile acid content of the compound regulator group was further increased (P<0.05).

[0052] As shown in Table 4, the liver triglyceride and total cholesterol content of the experimental fish in the high sugar group were significantly higher than those in the control group, while the bile acid content and the activity of the key enzyme CYP7A1 in bile acid synthesis were significantly lower than those in the control group (P<0.05). Adding regulator 1 to the high sugar feed significantly reduced the liver triglyceride content, and regulator 2 significantly reduced the cholesterol content. Both regulator 1 and regulator 2 alone increased the bile acid content and CYP7A1 enzyme activity (P<0.05). Compared with the regulator 1 and regulator 2 groups, the experimental fish in the compound regulator group had lower levels of triglyceride and total cholesterol content, and higher bile acid content and CYP7A1 enzyme activity (P<0.05).

[0053] The above results indicate that the combination of regulator 1 and regulator 2 has a significant effect on improving the biosynthesis of bile acids in fish, thereby exerting a stronger effect on improving glucose and lipid metabolism than using regulator 1 or regulator 2 alone.

[0054] Application Example 2: Culture Experiment and Result Analysis of Juvenile Blunt Snout Bream:

[0055] The culture experiment was carried out in a pond (specification: 100m×50m) at the Aquaculture Teaching and Research Base of Nanjing Agricultural University (Xingdian Town, Pukou District, Nanjing). Juvenile blunt snout bream were purchased from the original fry farm in Ezhou, Hubei, China. After domestication, 240 juvenile fish with uniform specifications (average weight: 35.04±0.14g) were randomly divided into 12 net cages (specification: 2m×1m×1m), and 20 blunt snout bream were fed in each net cage. Among them, 1) the control group was fed with the basal diet with a sugar level of 31.6%; 2) the high-sugar group was fed with the high-sugar diet with a sugar level of 45%; 3) the high-sugar + additive group was fed with the high-sugar diet + 1% of the bile acid synthesis promoter prepared in Example 3 as a feed additive. The feed formula is shown in Table 5.

[0056] The test period was 12 weeks in total. During the culture experiment, three test feeds were artificially fed (daily feeding time: 7:10, 12:10, and 16:10) until the fish showed an obvious satiated state. The growth of blunt snout bream was observed. After the experiment ended, samples were collected for further analysis and determination. The specific results are shown in Table 6 and Figures 1-4 as follows.

[0057] Table 5 Test Feed Formula

[0058]

[0059] Table 6 Effects of Different Feeds on the Growth and Feed Utilization Rate of Juvenile Blunt Snout Bream

[0060]

[0061] Note: Data in the same row with the same superscript letter have no significant difference.

[0062] As can be seen from Table 6, different feed treatments had little effect on the final weight, weight gain rate, specific growth rate (SGR), feed intake, feed conversion ratio, protein efficiency ratio, and viscerosomatic index of blunt snout bream (P>0.05). The hepatosomatic index and abdominal fat rate of blunt snout bream in the high-sugar group were significantly higher than those in the control group (P<0.05). After adding the bile acid synthesis promoter to the high-sugar group, the hepatosomatic index decreased significantly (P<0.05). This indicates that adding the bile acid synthesis promoter to the high-sugar group will not have a negative impact on the growth performance of fish. In addition, after adding the bile acid synthesis promoter, the abdominal fat accumulation and liver enlargement of fish were improved.

[0063] From Figure 1 , Figure 2 and Figure 3It can be seen that there were no significant differences in the blood glucose, triglyceride, and high-density lipoprotein cholesterol levels in the plasma of Megalobrama amblycephala among different feed treatments (P>0.05). The triglyceride and cholesterol levels in the plasma, liver, and hindgut of Megalobrama amblycephala in the high-sugar group were significantly higher than those in the control group (P<0.05), while the addition of a bile acid synthesis promoter improved this phenomenon. The trend of change in the total bile acid content was opposite (P<0.05).

[0064] It can be seen from Figure 4 that, compared with the control group, the expression level of CYP7A1 in the liver of Megalobrama amblycephala in the high-sugar group was significantly decreased (P<0.05), and the transcriptional level of CYP7A1 was significantly increased after the addition of a bile acid synthesis promoter (P<0.05). This indicates that, compared with the control group, the cholesterol accumulation in the liver and hindgut of the high-sugar group was significantly increased, and the bile acid synthesis was reduced. High sugar may cause hypercholesterolemia and hypertriglyceridemia in fish, while the bile acid synthesis promoter can alleviate this phenomenon. In addition, long-term feeding of a high-sugar diet can reduce the bile acid synthesis level of Megalobrama amblycephala, while the addition of a bile acid synthesis promoter will significantly increase the bile acid synthesis level of Megalobrama amblycephala.

[0065] Application Example 3: Juvenile Megalobrama amblycephala culture experiment and result analysis:

[0066] The culture experiment was carried out in a pond (specification: 100m×50m) at the Aquaculture Teaching and Research Base of Nanjing Agricultural University (Xingdian Town, Pukou District, Nanjing City). Juvenile Megalobrama amblycephala were purchased from the original fry farm in Ezhou, Hubei, China. After domestication, 240 juvenile fish with uniform specifications (average weight: 40.12±0.35g) were randomly divided into 12 net cages (specification: 2m×1m×1m), and 20 Megalobrama amblycephala were fed in each net cage. Among them, 1) the control group was fed a basal diet with a fat level of 6%; 2) the high-fat group was fed a high-fat diet with a fat level of 12%; 3) the high-fat + additive group was fed a high-fat diet + 1% of the bile acid synthesis promoter prepared in Example 3 as a feed additive. The feed formula is shown in Table 7.

[0067] The test period was 12 weeks. During the culture experiment, the three test feeds were artificially fed (daily feeding times: 7:10, 12:10, and 16:10) until the fish showed obvious satiety. The growth of Megalobrama amblycephala was observed. After the experiment, samples were collected for further analysis and determination. The results are shown in Tables 8 - 10.

[0068] Table 7 Test feed formula

[0069]

[0070] Table 8 Effects of different feeds on the growth and feed utilization rate of juvenile Megalobrama amblycephala

[0071]

[0072] Note: There is no significant difference in the superscripts of the same-row data with the same letter.

[0073] As can be seen from Table 8, different feed treatments had no significant effect on the final weight, weight gain rate, specific growth rate (SGR), feed intake, feed conversion ratio, and protein efficiency ratio of blunt snout bream (P>0.05). The hepatosomatic index, intraperitoneal fat ratio, and viscerosomatic ratio of blunt snout bream in the high-fat group were significantly higher than those in the control group (P<0.05), while the addition of bile acid synthesis promoter to the high-fat feed significantly reduced the above indexes (P<0.05). This indicates that the addition of bile acid synthesis promoter to high-fat feed will not have a negative impact on the growth performance and feed utilization ability of fish. In addition, bile acid synthesis promoter can significantly improve the excessive deposition of visceral fat in fish caused by high fat.

[0074] Table 9 Effects of different feeds on plasma biochemical indexes of juvenile blunt snout bream

[0075]

[0076] Note: There is no significant difference in the superscripts of the same-row data with the same letter.

[0077] As can be seen from Table 9, compared with the control group, the plasma glucose, triglyceride, and cholesterol contents of blunt snout bream fed with high-fat feed were significantly increased, and the bile acid content was significantly decreased (P<0.05), while the addition of bile acid synthesis promoter significantly reduced the glucose, triglyceride, and total cholesterol contents and increased the bile acid content (P<0.05).

[0078] Table 10 Effects of different feeds on biochemical indexes of liver and hindgut of juvenile blunt snout bream

[0079]

[0080] Note: There is no significant difference in the superscripts of the same-row data with the same letter.

[0081] As can be seen from Table 10, the triglyceride and total cholesterol contents in the plasma and hindgut of blunt snout bream in the high-fat group were significantly higher than those in the control group, while the bile acid content was significantly lower than that in the control group (P<0.05). Bile acid synthesis promoter had a significant improvement effect on these phenomena (P<0.05). Compared with the control group, the activity of the key enzyme CYP7A1 for bile acid synthesis in the liver of blunt snout bream in the high-fat group was significantly decreased, while the addition of bile acid synthesis promoter significantly increased the activity of CYP7A1 enzyme (P<0.05). The above results show that bile acid synthesis promoter can effectively improve the excessive deposition of triglyceride and cholesterol induced by high fat by alleviating bile acid synthesis disorder.

[0082] The above experimental results show that adding an appropriate dose of the bile acid synthesis promoter prepared by the present invention to the feed of cultured fish can significantly reduce lipid accumulation in fish, increase bile acid synthesis in fish, and improve the utilization rate of high-energy feed by fish. Moreover, the bile acid synthesis promoter group prepared by the present invention has no adverse effects on fish as an additive. Lignans, tremella polysaccharide and rice husk powder can all be produced on a large scale, and their prices are also low, so they have broad application prospects in the aquaculture industry.

Claims

1. A bile acid synthesis promoter for fish feed additives, characterized in that: The composition comprises the following components by weight: 10-30 parts of lignans; 2-10 parts of Sophora japonica polysaccharides; 460~488 copies of vector.

2. The bile acid synthesis promoter according to claim 1, characterized in that The composition comprises the following components by weight: 15-25 parts of lignans; 2-8 parts of Sophora japonica polysaccharides; and 467-483 parts of carriers.

3. The bile acid synthesis promoter according to claim 2, characterized in that The invention comprises the following components by weight: 20 parts of lignans; 5 parts of Sophora japonica polysaccharides; and 475 parts of carriers.

4. The bile acid synthesis promoter according to claim 1, characterized in that The carrier is rice husk powder.

5. The bile acid synthesis promoter according to claim 1, characterized in that The bile acid synthesis promoter is added in an amount of 1.0-5.0% in the feed.

6. A method for preparing the bile acid synthesis promoter according to claim 1, characterized in that: The method comprises the following steps: lignan, Sophora japonica polysaccharide and a carrier are crushed and sieved respectively, and then mixed to obtain a bile acid synthesis promoter.

7. The preparation method according to claim 6, characterized in that: The conditions for crushing and mixing the lignans are: light intensity <800lx, temperature <40°C.

8. The preparation method according to claim 6, characterized in that: The mesh number of the sieving is 100-120.

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