Composite fungal lipid extract, preparation method and application thereof in promoting growth of fattening pigs

Abstract

The present invention relates to the technical field of feeds specially suitable for animals, and specifically discloses a composite fungal lipid extract, a preparation method and an application thereof in promoting the growth of fattening pigs. The composite fungal lipid extract is obtained by processing, extracting and purifying ganoderma lucidum spores, shiitake mushrooms and black fungus and then mixing them with sesamol derivatives. Adding the composite fungal lipid extract to feed can not only effectively promote the growth of fattening pigs, but also be safe and have no toxic side effects.

Description

Technical Field

[0001] The invention relates to the technical field of feeding feeds specially suitable for animals, and in particular to a composite fungal lipid extract, a preparation method and application thereof in promoting the growth of fattening pigs. Background Art

[0002] Feed additives are the key material basis for promoting the industrialization of the feed industry and the modernization of the breeding industry. They are added to feed according to specific goals and play a vital role in promoting the development of these two industries. At present, the additives widely used in the feed industry are mainly chemically synthesized, which may have certain side effects, such as leaving residues in pigs or causing certain bacteria in pigs to develop drug resistance. These problems may eventually affect human health through the food chain. In addition, due to the use of these chemical additives, the stress resistance and disease resistance of animals may be affected, resulting in a decline in meat quality. At the same time, the content of drug residues and heavy metals such as copper and zinc is also continuing to increase. Therefore, in order to protect human health and improve the quality of meat products, it is necessary to conduct in-depth research and improvement on existing feed additives and seek safer and more environmentally friendly alternatives. This will not only help to improve the overall level of the breeding industry, but also ensure food safety and promote sustainable development.

[0003] CN202211375749.5 provides a composite feed additive for improving the immune and antioxidant function of fattening pigs, which belongs to the field of aquaculture feed technology. The raw materials used in the composite feed additive are measured by weight percentage, including: composite probiotics 23.5-27%, boric acid 0.5-1%, tea polyphenols 22-28%, ganoderma lucidum polysaccharides 10-14%, oligoxylose 20-28%, and white carbon black 10-15%. During daily feeding, the composite feed additive needs to be used as a composite feed with a basic feed. The application creatively selects composite probiotics, tea polyphenols and ganoderma lucidum polysaccharides from a variety of materials with immune-enhancing and antioxidant effects as the main raw materials of the additive. The resulting composite additive can effectively improve the antibody level in the serum and immune organ tissues of growing and fattening pigs, improve the antioxidant function of immune organs, and help to further improve the production performance and muscle quality of pigs.

[0004] CN202011540638.6 discloses a compound Chinese herbal feed additive for daily health care and growth promotion of poultry and livestock and its preparation method, which is made of 10-20 parts of Radix Isatidis enzymatic polysaccharides, 1-2 parts of Sophora alopecuroides alkaloids, 5-15 parts of Salvia miltiorrhiza dregs fermented polysaccharides, 5-10 parts of grapefruit peel polysaccharides, and 5-15 parts of honeysuckle vine polysaccharides. The invention shows through experiments that the compound Chinese herbal feed additive can enhance animal appetite, promote animal growth, and improve animal immunity. And the invention has the characteristics of environmentally friendly materials, low cost, process innovation, and good practicality. When added to daily feed for breeding, it can improve the animal's own immune function, balance the gastrointestinal microecology, and form an excellent health care effect, thereby reducing the incidence of animals, reducing or even not using medication during the breeding process, improving feed remuneration, reducing feed-to-meat ratio, improving animal growth rate, ensuring product quality, enhancing competitiveness, and improving the economic benefits of farmers (farms).

[0005] Traditional feed additives not only have serious side effects, but also pose a threat to human health. Lipid extracts from natural substances can well replace traditional additives such as antibiotics. However, lipids are easily oxidized, causing deterioration and resulting in ineffectiveness. Summary of the invention

[0006] In view of the above-mentioned defects of the prior art, the technical problem to be solved by the present invention is to provide a composite fungal lipid extract, a preparation method and its application in promoting the growth of fattening pigs.

[0007] Ganoderma lucidum is a fungus of the Polyporaceae family and the genus Ganoderma. It is an important edible and medicinal fungus and traditional Chinese medicine. The traditional medicinal part is the fruiting body. Ganoderma spores are oval reproductive cells ejected from the gills of Ganoderma during the growth and maturity period. The cell wall contains a layer of chitin, which is strong and acid-resistant. The dissolution rate of its active substances can be increased by breaking the cell wall. The lipid extract of Ganoderma contains components such as polysaccharides, fatty acids and sterols, and has multiple effects such as anti-tumor, regulating immunity and protecting the liver. Lentinus edodes is an edible and medicinal fungus. Its extract contains a variety of bioactive ingredients, including polysaccharides, proteins, triterpenes and other lipid components. Among them, polysaccharides can enhance immunity, regulate antioxidant capacity, improve hormone levels, help fattening pigs break down excess fat in the body, avoid fat deposition, and improve the intestinal flora structure of fattening pigs, reduce inflammatory reactions, and reduce diarrhea rates, thereby promoting the growth of fattening pigs. Auricularia auricula is an edible and medicinal fungus, and its lipid composition includes phospholipids, fatty acids, polysaccharides, etc., and phospholipids have a certain effect on activating cells, maintaining metabolism and enhancing immunity, and polysaccharides can have the potential of anti-obesity and regulating intestinal flora. The lipid extracts of these types of fungi are added to the feed of fattening pigs, which can play an important role in its growth regulation, and as a natural ingredient, it will not affect the meat quality. But a large amount of unsaturated fatty acids are included in the lipid extract, which contains double bonds and is easily affected by environmental influences and thus oxidized. Therefore, a natural antioxidant is prepared in the present invention, and the antioxidant is obtained by modifying the natural ingredient sesamol, which not only has excellent antioxidant properties, but also can improve the defect that sesamol itself is easy to form dimers. After the antioxidant is mixed with the fungal lipid extract, it can effectively prevent its effective ingredients from being oxidized and deteriorating, and the mixture is added to the fattening pig feed, which can effectively promote the growth of fattening pigs and improve immunity.

[0008] To achieve the above object, the present invention provides a method for preparing a composite fungal lipid extract, comprising the following steps:

[0009] S1, washing and drying the ganoderma lucidum spores, shiitake mushrooms and black fungus, crushing them and passing them through a 0.1-2 mm sieve to obtain a mixture;

[0010] S2, mixing the mixture obtained in S1 with a chloroform-methanol mixture, performing ultrasonic extraction for 4 to 6 hours, diluting with water and centrifuging, washing the lower precipitate with chloroform, extracting and concentrating to dryness to obtain a residue, and subjecting the residue to solid phase extraction to enrich and purify to obtain a fungal lipid extract;

[0011] S3. Mix the fungal lipid extract and the sesamol derivative in a mass ratio of 100:0.5-2 to obtain a composite fungal lipid extract.

[0012] Furthermore, the mass ratio of the Ganoderma lucidum spores, shiitake mushrooms and black fungus is 1:1:1.

[0013] Furthermore, the volume ratio of chloroform to methanol in the chloroform-methanol mixture is 2:1.

[0014] Furthermore, the solid-liquid ratio of the mixture obtained in S1 to the chloroform-methanol mixed solution is 1:30-40 g / mL.

[0015] Furthermore, the mass ratio of the fungal lipid extract to the sesamol derivative is 100:0.5-2.

[0016] Furthermore, the preparation method of the sesamol derivative comprises the following steps:

[0017] X1. Add sesamol to water, add formaldehyde aqueous solution and calcium oxide after cooling, stir for 1 to 2 hours, add saturated ammonium chloride aqueous solution, mix well, add ether for extraction, separate the liquids, combine the organic phases, dry and concentrate to obtain a residue, and purify it by column chromatography for the next step;

[0018] X2. Mix the product of the previous step with benzaldehyde and dimethylamine aqueous solution, and heat with stirring for 2 to 3 hours. After stirring, cool to room temperature and purify by column chromatography to obtain a sesamol derivative.

[0019] Preferably, the temperature range of cooling in step X1 is 0-5°C.

[0020] Preferably, the heating temperature in step X2 ranges from 50 to 65°C.

[0021] More preferably, the preparation method of the cannabinol derivative comprises the following steps:

[0022] X1. Add 4-10 parts by weight of sesamol to 50-150 parts by weight of water, cool it down, add 4-10 parts by weight of formaldehyde solution and 1-2 parts by weight of calcium oxide, stir for 1-2 hours, add saturated ammonium chloride solution, mix well, add ether for extraction, separate the liquids, combine the organic phases, dry and concentrate to obtain a residue, and purify it by column chromatography for the next step;

[0023] X2. Mix 4-10 parts by weight of the product of the previous step, 4-6 parts by weight of benzaldehyde and 4-8 parts by weight of a dimethylamine aqueous solution, and heat with stirring for 2-3 hours. After stirring, cool to room temperature and purify by column chromatography to obtain a sesamol derivative.

[0024] The present invention also provides a composite fungal lipid extract prepared by the above method, and application of the composite fungal lipid extract in promoting the growth of fattening pigs.

[0025] Beneficial effects of the present invention:

[0026] The present invention prepares a natural antioxidant, which is obtained by modifying the natural ingredient sesamol, and not only has excellent antioxidant properties, but also can improve the defect of sesamol itself that it is easy to form dimers. After the antioxidant is mixed with various fungal lipid extracts, the oxidation and deterioration of the effective ingredients can be effectively prevented, and the mixture is added to fattening pig feed, which can effectively promote the growth of fattening pigs and improve immunity. DETAILED DESCRIPTION

[0027] Formaldehyde aqueous solution, 37wt%.

[0028] Dimethylamine aqueous solution, 40 wt%.

[0029] Comparative Example 1

[0030] A method for preparing a composite fungal lipid extract comprises the following steps:

[0031] S1, washing and drying the ganoderma spores, shiitake mushrooms and black fungus, wherein the mass ratio of the ganoderma spores, shiitake mushrooms and black fungus is 1:1:1, and crushing and passing through a 1.5 mm sieve to obtain a mixture;

[0032] S2, the mixture obtained in S1 and chloroform methanol (V 氯仿 :V 甲醇 =2:1) ​​mixed solution, the solid-liquid ratio is 1:35g / mL, 400W, 45kHz ultrasonic extraction for 6h, diluted with water and centrifuged, the lower precipitate is washed with chloroform, extracted and concentrated to dryness to obtain a residue, the residue is subjected to solid phase extraction to enrich and purify to obtain a fungal lipid extract;

[0033] S3. Mix the fungal lipid extract and sesamol in a mass ratio of 100:1 to obtain a composite fungal lipid extract.

[0034] Comparative Example 2

[0035] A method for preparing a composite fungal lipid extract comprises the following steps:

[0036] S1, washing and drying the ganoderma spores, shiitake mushrooms and black fungus, wherein the mass ratio of the ganoderma spores, shiitake mushrooms and black fungus is 1:1:1, and crushing and passing through a 1.5 mm sieve to obtain a mixture;

[0037] S2, the mixture obtained in S1 and chloroform methanol (V 氯仿 :V 甲醇=2:1) ​​mixed solution, the material-liquid ratio is 1:35g / mL, 400W, 45kHz ultrasonic extraction for 6h, diluted with water and centrifuged, the lower precipitate is washed with chloroform, extracted and concentrated to dryness to obtain the residue, the residue is enriched and purified by solid phase extraction.

[0038] Example 1

[0039] A method for preparing a composite fungal lipid extract comprises the following steps:

[0040] S1, washing and drying the ganoderma spores, shiitake mushrooms and black fungus, wherein the mass ratio of the ganoderma spores, shiitake mushrooms and black fungus is 1:1:1, and crushing and passing through a 1.5 mm sieve to obtain a mixture;

[0041] S2, the mixture obtained in S1 and chloroform methanol (V 氯仿 :V 甲醇 =2:1) ​​mixed solution, the solid-liquid ratio is 1:35g / mL, 400W, 45kHz ultrasonic extraction for 6h, diluted with water and centrifuged, the lower precipitate is washed with chloroform, extracted and concentrated to dryness to obtain a residue, the residue is subjected to solid phase extraction to enrich and purify to obtain a fungal lipid extract;

[0042] S3. Mix the fungal lipid extract and the sesamol derivative in a mass ratio of 100:1 to obtain a composite fungal lipid extract.

[0043] The preparation method of the sesamol derivative comprises the following steps:

[0044] X1. Add 5 g of sesamol to 100 mL of water, cool to 0°C, add 5 mL of formaldehyde solution and 1 g of calcium oxide, stir for 2 h, add saturated ammonium chloride solution, mix well, add ether for extraction, separate the liquids, combine the organic phases, dry and concentrate to obtain a residue, and purify by column chromatography for the next step;

[0045] X2. Mix 6 g of the product from the previous step, 5 g of benzaldehyde and 6 mL of dimethylamine aqueous solution, heat to 60°C and stir for 3 h. After stirring, cool to room temperature and purify by column chromatography to obtain a sesamol derivative.

[0046] Example 2

[0047] A method for preparing a composite fungal lipid extract comprises the following steps:

[0048] S1, washing and drying the ganoderma spores, shiitake mushrooms and black fungus, wherein the mass ratio of the ganoderma spores, shiitake mushrooms and black fungus is 1:1:1, and crushing and passing through a 1.5 mm sieve to obtain a mixture;

[0049] S2, the mixture obtained in S1 and chloroform methanol (V 氯仿 :V 甲醇=2:1) ​​mixed solution, the solid-liquid ratio is 1:35g / mL, 400W, 45kHz ultrasonic extraction for 6h, diluted with water and centrifuged, the lower precipitate is washed with chloroform, extracted and concentrated to dryness to obtain a residue, the residue is subjected to solid phase extraction to enrich and purify to obtain a fungal lipid extract;

[0050] S3. Mix the fungal lipid extract and the sesamol derivative in a mass ratio of 100:1 to obtain a composite fungal lipid extract.

[0051] The preparation method of the sesamol derivative comprises the following steps:

[0052] Add 5 g of sesamol to 100 mL of water, cool to 0°C, add 5 mL of formaldehyde aqueous solution and 1 g of calcium oxide, stir for 2 h, add saturated ammonium chloride aqueous solution, mix well, add ether for extraction, separate the liquids, combine the organic phases, dry and concentrate to obtain the residue, and then purify by column chromatography to obtain the product.

[0053] Example 3

[0054] A method for preparing a composite fungal lipid extract comprises the following steps:

[0055] S1, washing and drying the ganoderma spores, shiitake mushrooms and black fungus, wherein the mass ratio of the ganoderma spores, shiitake mushrooms and black fungus is 1:1:1, and crushing and passing through a 1.5 mm sieve to obtain a mixture;

[0056] S2, the mixture obtained in S1 and chloroform methanol (V 氯仿 :V 甲醇 =2:1) ​​mixed solution, the solid-liquid ratio is 1:35g / mL, 400W, 45kHz ultrasonic extraction for 6h, diluted with water and centrifuged, the lower precipitate is washed with chloroform, extracted and concentrated to dryness to obtain a residue, the residue is subjected to solid phase extraction to enrich and purify to obtain a fungal lipid extract;

[0057] S3. Mix the fungal lipid extract and the sesamol derivative in a mass ratio of 100:1 to obtain a composite fungal lipid extract.

[0058] The preparation method of the sesamol derivative comprises the following steps:

[0059] Mix 5.8 g of sesamol, 5 g of benzaldehyde and 6 mL of dimethylamine aqueous solution, heat to 60°C and stir for 3 h. After stirring, cool to room temperature and purify by column chromatography to obtain a sesamol derivative.

[0060] Example 4

[0061] The same as Example 1, the only difference is that the material-liquid ratio in step S2 is 1:30 g / mL.

[0062] Example 5

[0063] The same as Example 1, the only difference is that the material-liquid ratio in step S2 is 1:40 g / mL.

[0064] Test Example 1

[0065] 120 three-way growing and fattening pigs (half male and half female) with similar health and weight were selected and randomly divided into 6 groups, 5 test groups and 1 blank group, and fed according to the blank control, reference example and embodiment. All 6 groups were fed with a basic diet. Among them, the basic diet used by the 5 test groups was supplemented with the composite fungal lipid extract prepared by the reference example and the embodiment as a feed additive, and the addition ratio of the feed additive was 2% of the basic diet. The feeding test was carried out according to the above requirements, with a pre-feeding period of 7d and a test period of 60d. During the test, feeding management and immunization were carried out as usual, and the feed consumption during the test period was recorded and weighed. The specific data are shown in Table 1.

[0066] Table 1 Growth performance data of fattening pigs

[0067]

[0068]

[0069] The lipid extract of Ganoderma lucidum contains components such as polysaccharides, fatty acids and sterols, and has multiple effects such as anti-tumor, regulating immunity and protecting the liver. Lentinus edodes extract contains polysaccharides, triterpenes and other lipid components, among which polysaccharides can enhance immunity, regulate antioxidant capacity, improve hormone levels, help fattening pigs decompose excess fat in the body, avoid fat precipitation, and improve the intestinal flora structure of fattening pigs, reduce inflammatory reactions, and reduce diarrhea rates, thereby promoting the growth of fattening pigs. The lipid components of fungus include phospholipids, fatty acids, polysaccharides, etc. Phospholipids have a certain effect on activating cells, maintaining metabolism and enhancing immunity, and polysaccharides can have the potential to resist obesity and regulate intestinal flora. It can be seen from the growth indicators of fattening pigs in Table 1 that the lipid extracts of these types of fungi are compounded and added to the feed of fattening pigs, which can play an important role in the growth regulation of fattening pigs. In Example 1, the antioxidant is obtained by modifying the natural ingredient sesamol, which not only has excellent antioxidant properties, but also can improve the defect that sesamol itself is easy to form dimers. After the antioxidant is mixed with the fungal lipid extract, it can effectively prevent the effective ingredients from being oxidized and deteriorated. Adding the mixture to fattening pig feed can effectively promote the growth of fattening pigs, so its weight gain effect on pigs is the most obvious. In Control Example 2, no antioxidant is added. The lipid extract is very easy to be oxidized and deteriorated due to a large amount of unsaturated fatty acids, which not only affects the taste of the feed, but also may cause vomiting and diarrhea in pigs, so the weight gain effect is the worst. In Control Example 1, sesamol is added as an antioxidant, and sesamol is prone to self-polymerization to form dimers, thereby affecting the antioxidant properties. In Examples 2 to 3, hydroxyl groups and alkylamino groups are introduced respectively, and the electron-donating group effect is not as good as in Example 1. Therefore, the growth-promoting performance of the composite fungal lipid extract in Example 1 is the best.

[0070] Test Example 2

[0071] The composite fungal lipid extracts prepared in the control example and the example were placed in a test box and maintained at 35°C. Samples were taken at 0d and 7d for determination of peroxide value. The specific results are shown in Table 2.

[0072] Table 2 Antioxidant test of composite lipid extracts

[0073] Experimental protocol 0d peroxide value / meq / kg 7d peroxide value / meq / kg Comparative Example 1 3.5 12.7 Comparative Example 2 3.6 19.2 Example 1 4.0 6.2 Example 2 3.9 7.5 Example 3 4.5 9.6

[0074] From the antioxidant test, it can be seen that the lipid extract is very easy to be oxidized to produce peroxides due to the presence of many unsaturated fatty acids and other substances, and sesamol, as a natural antioxidant, has good antioxidant properties, but its defect is that it is easy to self-polymerize to form dimers. Therefore, in the antioxidant test, the antioxidant effect of Control Example 1 is not as good as that of Examples 1 to 3. In Example 1, by modifying sesamol, not only the defect of self-polymerization is solved, but also the antioxidant property is enhanced by the introduction of strong electron-donating groups such as hydroxyl and alkylamino groups, so the antioxidant property is stronger than that of Examples 2 to 3.

[0075] The preferred specific embodiments of the present invention are described in detail above. It should be understood that a person skilled in the art can make many modifications and changes based on the concept of the present invention without creative work. Therefore, any technical solution that can be obtained by a person skilled in the art through logical analysis, reasoning or limited experiments based on the concept of the present invention on the basis of the prior art should be within the scope of protection determined by the claims.

Claims

1. A method for preparing a composite fungal lipid extract, characterized in that: The steps include: S1, washing and drying the ganoderma lucidum spores, shiitake mushrooms and black fungus, crushing them and passing them through a 0.1-2 mm sieve to obtain a mixture; S2, mixing the mixture obtained in S1 with a chloroform-methanol mixture, performing ultrasonic extraction for 4-6 hours, diluting with water and centrifuging, washing the lower precipitate with chloroform, extracting and concentrating to dryness to obtain a residue, and subjecting the residue to solid phase extraction to enrich and purify to obtain a fungal lipid extract; S3, mixing the fungal lipid extract with the sesamol derivative to obtain a composite fungal lipid extract; The preparation method of the sesamol derivative comprises the following steps: X1. Add 4-10 parts by weight of sesamol to 50-150 parts by weight of water, cool down, add 4-10 parts by weight of formaldehyde solution and 1-2 parts by weight of calcium oxide, stir for 1-2 hours, add saturated ammonium chloride solution, mix well, add ether for extraction, separate the liquids, combine the organic phases, dry and concentrate to obtain a residue, and purify by column chromatography for the next step; X2. Mix 4-10 parts by weight of the product of the previous step, 4-6 parts by weight of benzaldehyde and 4-8 parts by weight of a dimethylamine aqueous solution, and heat with stirring for 2-3 hours. After stirring, cool to room temperature and purify by column chromatography to obtain a sesamol derivative.

2. The method for preparing the composite fungal lipid extract according to claim 1, characterized in that: The mass ratio of the ganoderma lucidum spores, shiitake mushrooms and black fungus is 1:1:

1.

3. The method for preparing the composite fungal lipid extract according to claim 1, characterized in that: The volume ratio of chloroform to methanol in the chloroform-methanol mixture is 2:

1.

4. The method for preparing the composite fungal lipid extract according to claim 1, wherein The solid-liquid ratio of the mixture obtained in S1 to the chloroform-methanol mixed solution is 1:30-40 g / mL.

5. The method for preparing the composite fungal lipid extract according to claim 1, characterized in that: The mass ratio of the fungal lipid extract to the sesamol derivative is 100:0.5-2.

6. The method for preparing the composite fungal lipid extract according to claim 1, characterized in that: The temperature range of cooling in step X1 is 0-5°C.

7. The method for preparing the composite fungal lipid extract according to claim 1, characterized in that: The heating temperature in step X2 is in the range of 50-65°C.

8. A composite fungal lipid extract, characterized in that Prepared by the method according to any one of claims 1 to 7.

9. Use of the composite fungal lipid extract as claimed in claim 8 in promoting growth of fattening pigs.

Images