CN107951002B - Polyporus umbellatus fermented extract composition of rosemary and okra and preparation method thereof - Google Patents
Polyporus umbellatus fermented extract composition of rosemary and okra and preparation method thereof Download PDFInfo
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- LCAZOMIGFDQMNC-FORWCCJISA-N rosmanol Chemical compound C1CCC(C)(C)[C@@H]2[C@H]3[C@@H](O)C(C=C(C(=C4O)O)C(C)C)=C4[C@]21C(=O)O3 LCAZOMIGFDQMNC-FORWCCJISA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Virology (AREA)
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- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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- Agricultural Chemicals And Associated Chemicals (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a composition prepared by fermenting and extracting agaric of rosemary and okra and a preparation method thereof. The invention is prepared by reasonably matching rosemary and okra, taking rosemary powder and okra as culture mediums, and refining after biological fermentation and transformation of grifola. The product can be made into refreshing beverage, granule, tablet, capsule, etc. The method solves the problems of low extraction rate of functional components and poor safety in the processing of rosemary and okra products, and simultaneously solves the problems of rosmarinus spicy flavor and okra mucilage taste in the products. The agaric fermented extract composition of rosemary and okra prepared by the method has the functions of improving memory, resisting fatigue and the like.
Description
Technical Field
The invention belongs to the technical field of biological extraction of natural products in bioengineering, and relates to a polyporus umbellatus fermented extract composition of rosemary and okra and a preparation method thereof, in particular to a method for preparing a composition containing active ingredients of rosemary, okra and polyporus umbellatus with health care function by applying polyporus umbellatus deep solid state fermentation and biotransformation technology.
Background
Rosemary (Rosarinus officinalis L.) also known as Oenanthe stolonifera (L.) is a plant of the genus Rosmarinus of the family Labiatae. According to the traditional Chinese medicine, the rosemary is warm in nature and pungent in taste, has the effects of invigorating the stomach, tranquilizing and allaying excitement and the like, and is commonly used for treating various headaches, neurasthenia and the like. Modern pharmacological research proves that rosemary has the effects of enhancing memory, refreshing, resisting oxidation and strengthening liver and heart functions. The main antioxidant components of herba Rosmarini officinalis include rosmanol (1, 8-oleyl phenol), ursolic acid, and rosmanic acid.
The okra belongs to an annual herbaceous plant of okra of Malvaceae, is a vegetable, medicine and flower dual-purpose plant, and has wide application. The main functional components of the health-care food comprise mucopeptidoglycan, flavone, caffeine, selenium, zinc and the like, and the health-care food has the effects of helping digestion, enhancing physical strength, protecting liver, invigorating stomach and regulating intestine and the like. Okra alkaloid (alkaloid) extracted from Abelmoschus esculentus has antifatigue effect.
Polyporus umbellatus, higher medicinal fungus of Grifola of Polyporaceae of Aphyllophorales. The main functional components of the health-care tea are polysaccharide and triterpenes, and the health-care tea has the effects of immunizing, resisting cancer, relieving swelling, benefiting kidney and the like. Abundant fiber degrading enzyme systems can be secreted in a hyphal state, and comprise cellulose degrading enzyme, chitinase, polyphenol oxidase, lipase, protease and the like.
The traditional extraction methods of rosmarinic acid, polysaccharide, alkaloid, flavone and the like comprise a hot water extraction method and an ethanol hot reflux extraction method, and certain physiological active ingredients are often inactivated due to structural change in the heating process. The literature reports that ultrasonic-assisted extraction has some effect, but still does not depart from the conventional hot alcohol process. In other experiments, the cellulase is adopted for assisting in extracting the rosmarinic acid in the perilla, but the release of the rosmarinic acid from plant cells is only solved to a certain extent. Meanwhile, mucopolysaccharide in okra and hard cell wall of rosemary both affect the efficiency of the extraction system.
In addition, from the viewpoint of product development, the pungent taste of rosemary and the mucilage of okra affect the overall acceptance of the product, and a new technology and a new formulation strategy for softening the flavor without chemical addition are needed.
Disclosure of Invention
The invention aims to: overcomes the defects of the prior art, provides a novel agaric fermented extract composition of rosemary and okra without chemical addition (green), which can effectively extract the functional components in the rosemary and the okra and can solve the problem of rosmarinus spicy flavor and okra mucilage taste, and a preparation method thereof.
The invention overcomes the defects in the development of the existing rosemary and okra, and the rosemary and the okra are used as culture mediums, and a agaric solid fermentation biotransformation method is adopted, so that various active components (also called as functional components or functional components) in the rosemary and the okra can be effectively released and enriched on one hand, and a green processing technology for solving the problem of spicy taste of rosewood and the taste of okra mucilage is provided on the other hand. The invention comprehensively solves the technical problems from two aspects of formula and process.
The purpose of the invention is realized by the following technical scheme:
the invention provides a manufacturing method of a agaric fermentation extraction composition of rosemary and okra, which comprises the following steps:
(a) adding rosemary powder, okra powder and glucose powder into water according to a certain proportion, uniformly stirring, heating, and sterilizing at high temperature to obtain a seed culture medium solution;
(b) cooling the seed culture medium solution obtained in the step (a), inoculating grifola, uniformly stirring, and culturing for a plurality of days to obtain a grifola seed culture solution;
(c) mixing and stirring rosemary powder, okra powder and glucose powder uniformly to obtain a solid fermentation material, mixing the solid fermentation material with water uniformly, and sterilizing at high temperature to obtain a solid fermentation culture medium;
(d) and inoculating and fermenting: cooling the solid fermentation culture medium obtained in the step (c) to room temperature, inoculating the polyporus umbellatus strain seed culture solution obtained in the step (b) (namely, uniformly mixing the polyporus umbellatus strain seed culture solution obtained in the step (b) with the solid fermentation culture medium obtained in the step (c) according to a certain proportion), and putting the uniformly mixed solution into a fermentation tank or a fermentation tank for solid gradient temperature-changing multi-stage fermentation to obtain a polyporus umbellatus fermentation composition of rosemary and okra;
(e) adding the agaric fermented composition of rosemary and okra obtained in the step (d) into deionized water, adopting a step-by-step pH value regulation and multi-step crushing homogenization process, regulating the pH value to a certain range step by step, and treating by multi-step physical treatment methods such as crushing, grinding, homogenization and the like (treating by multi-step physical treatment methods such as wet crushing, colloid mill treatment, homogenization and the like) to obtain the agaric fermented extract composition of rosemary and okra (namely the agaric bioconversion and enrichment composition of rosemary and okra);
(f) and (e) analyzing the agaric fermented extract composition of rosemary and okra obtained in the step (e) according to a conventional analysis method to obtain the contents of various functional components (functional components) such as rosmarinic acid, flavone, caffeine, polysaccharide, glycoprotein, triterpenic acid, selenium, zinc and the like.
(g) And (e) preparing the composition obtained in the step (e) into products in various different dosage forms such as granules, capsules, beverages, tablets and the like.
The invention provides a manufacturing method for obtaining bioactive composition by fermenting rosemary and okra with grifola, which optimizes the process routes and process conditions of the steps (a) to (g), such as: the combination (mixing) proportion of rosemary powder and okra powder, solid gradient temperature-changing multi-stage fermentation and enzyme biotransformation of polyporus umbellatus, gradual pH value adjustment, multi-stage crushing and homogenizing processes and the like.
Further, in the step (a), rosemary powder, okra powder and glucose powder are added into water according to a certain proportion, wherein the adding amount of the rosemary powder is 0.1-3%, the adding amount of the okra powder is 0.1-5%, and the adding amount of the glucose powder is 0.5-5%, which are all in mass percentage; the high-temperature sterilization conditions are as follows: sterilizing at 115 ℃ and 125 ℃ for 20-50 minutes.
Further, in the step (b), the method for inoculating grifola in the seed culture solution comprises the following steps: cooling the seed culture medium to 20-40 deg.C, inoculating 0.5-15% Polyporus umbellatus; the stirring conditions were: stirring at the rotating speed of 100-400 rpm; the culture conditions were: culturing at 10-40 deg.C for 3-10 days.
Further, in the step (c), the solid fermentation material comprises the following components in parts by weight: 10-60 parts of rosemary powder, 10-40 parts of okra powder and 1-10 parts of glucose powder; uniformly mixing the solid fermentation material with water at the temperature of 10-60 ℃ according to the mass ratio of 1:1-1: 2; the high-temperature sterilization conditions are as follows: sterilizing at 115 ℃ and 125 ℃ for 20-50 minutes.
Further, the inoculation fermentation in the step (d) is to cool the solid fermentation culture medium obtained in the step (c) to room temperature, inoculate the polyporus umbellatus strain seed culture solution according to the inoculation amount of 1-20% (namely, the polyporus umbellatus strain seed culture solution obtained in the step (b) is mixed with the solid fermentation culture medium obtained in the step (c) according to the mass ratio of 1:99-1:4, namely, in the mixture of the two, the mass ratio of the polyporus umbellatus strain seed culture solution is 1-20%), and the mixture is filled into a fermentation tank or a fermentation tank for solid gradient temperature-changing multi-stage fermentation; the primary fermentation temperature is 20-40 ℃, and the fermentation time is 10-20 days; the secondary fermentation temperature is 5-15 ℃, and the fermentation time is 5-10 days; the third-stage fermentation temperature is 35-55 ℃, and the fermentation time is 2-36 hours; for the fermentation tank, the charging thickness is 0.05 m-0.5 m; for the fermenter, the filling factor is 0.25 to 0.75.
Further, in the step (e), the polyporus umbellatus fermented composition of rosemary and okra obtained in the step (d) is processed by a multi-stage physical method, and the specific steps comprise: step one, adding deionized water into a fermentation composition, adjusting the pH value to 3-4.5 by using alkali, and carrying out wet grinding by using wet grinding equipment; secondly, further adjusting the pH value to 5-6 by alkali, and carrying out levigating treatment by a colloid mill; and thirdly, further regulating the pH value to 7-8 by using alkali, and homogenizing by using a homogenizer. That is: the first stage of physical treatment is crushing with wet crushing equipment, the second stage of physical treatment is colloid mill treatment (further grinding), and the third stage of physical treatment is homogenizing with homogenizer to break cell wall. By adopting the process of gradually adjusting the pH value and multi-stage crushing and homogenizing, after the treatment by the multi-stage physical treatment method, the active substances (functional components) in the composition can be released more easily.
Furthermore, in the step (e), the wet grinding process conditions are that the mass ratio of the fermentation composition to the deionized water is 1:3-1:30, the pH value of the composition is adjusted to 3-8, and the rotation speed of a grinder of a wet grinding device is 1000 revolutions per minute to 5000 revolutions per minute; the action time of the colloid mill is 10 minutes to 2 hours, and the temperature is 10 ℃ to 60 ℃; the homogenizing pressure of the homogenizer is 5MPa-30 MPa.
Further, in the step (f), the measured mass percentage of each functional component is as follows: 1-1.5% of rosmarinic acid, 0.5-2% of okra alkali, 0.5-3% of flavone (okra flavone), 1.5-9% of polysaccharide, 0.5-1.5% of peptidoglycan and 0.1-1.2% of triterpenic acid, wherein the measured contents of trace elements are as follows: selenium 0.5 mug/100 g-5 mug/100 g; 0.3 mu g/100 g-3 mu g/100g of zinc.
In the step (g), the prepared products in various dosage forms such as granules, capsules, beverages, tablets and the like have the common characteristics that: there is no pungent taste of rosemary and the sticky taste of okra.
The invention also provides a composition prepared by fermenting and extracting the agaric of the rosemary and the okra by the preparation method.
It is to be understood that within the scope of the present invention, the above-described technical features of the present invention and the technical features specifically described below (e.g., examples) may be combined with each other to constitute a new or preferred technical solution. Not to be reiterated herein, but to the extent of space.
The invention has the beneficial effects that:
the inventor of the invention has conducted extensive and intensive research, firstly adopts a unique combination of rosemary and okra, utilizes a solid gradient temperature-changing fermentation and biotransformation technology of agaric bacteria, adjusts the pH value step by step, and performs multistage crushing and homogeneous transformation processes, provides a biotransformation and enrichment extraction method of bioactive substances of rosemary and okra, solves the problems of low extraction rate of effective components and poor safety in processing of rosemary and okra products, and simultaneously solves the problems of rosmarinus spicy flavor and okra mucilage taste in the products.
Animal experiments prove that the composition prepared by fermenting and extracting the agaric of the rosemary and the okra by the method has the functions of improving memory and resisting fatigue. In the agaric fermentation extraction composition product of rosemary and okra prepared by the method, the content of two main functional components of rosmarinic acid and okra alkali is high (1-1.5% of rosmarinic acid and 0.5-2% of okra alkali), and animal experiments prove that the two components have high functional display degree on the product, so that the product has strong functions of improving memory and resisting fatigue.
The biological extraction method of biological fermentation conversion of the invention is different from the prior chemical extraction method, does not use chemical solvent, and the prepared product has no chemical substance residue and is harmless to human body.
Detailed Description
The technical solution of the present invention is further illustrated by the following specific examples. The invention will now be further described with reference to specific examples. It should be understood that these specific examples are only for illustrating and explaining the present invention, and are not to be construed as limiting the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Percentages and parts are by weight unless otherwise indicated.
Example 1 seed culture preparation
Adding herba Rosmarini officinalis powder 1%, flos Abelmoschi Manihot powder 2%, and glucose powder 3% into cold water, stirring, heating, sterilizing at 121 deg.C for 30 min, cooling to 30 deg.C, inoculating Polyporus umbellatus 10% under aseptic condition, and culturing at 30 deg.C at rotation speed of 250 rpm for 7 days to obtain Polyporus umbellatus strain seed culture solution. Through inspection, the biomass in the polyporus umbellatus strain seed culture solution is more than 1.8%.
Example 2 seed culture preparation
Adding herba Rosmarini officinalis powder 3%, flos Abelmoschi Manihot powder 5%, and glucose powder 5% into cold water, stirring, heating, sterilizing at 125 deg.C for 50 min, cooling to 20 deg.C, inoculating Polyporus umbellatus 15% under aseptic condition, and culturing at 10 deg.C at 400 rpm for 10 days to obtain Polyporus umbellatus strain seed culture solution. Through inspection, the biomass in the polyporus umbellatus strain seed culture solution is more than 1.8%.
Example 3 seed culture preparation
Adding herba Rosmarini officinalis powder 0.1%, flos Abelmoschi Manihot powder 0.1%, and glucose powder 0.5% into cold water, stirring, heating, sterilizing at 115 deg.C for 20 min, cooling to 40 deg.C, inoculating Polyporus umbellatus 0.5% under aseptic condition, and culturing at 40 deg.C at rotation speed of 100 rpm for 3 days to obtain Polyporus umbellatus strain seed culture solution. Through inspection, the biomass in the polyporus umbellatus strain seed culture solution is more than 1.8%.
Example 4 solid gradient temperature-changing fermentation of Polyporus umbellatus to transform Rosemary and Abelmoschus esculentus and production of granule
Mixing 60 parts of rosemary powder, 35 parts of okra powder and 5 parts of glucose powder in proportion, uniformly stirring to obtain a solid fermentation material, and uniformly mixing the solid fermentation material with water at the temperature of 25 ℃ in a ratio of 1:1 to obtain a solid fermentation culture medium; sterilizing at 121 deg.C for 30 min, cooling to room temperature, inoculating the Polyporus umbellatus seed culture solution obtained in example 1 at an inoculum size of 10%, mixing, and placing into a fermentation tank with a thickness of 0.3 m. Fermenting at 25 deg.C for 20 days, cooling to 10 deg.C, fermenting at constant temperature for 5 days, heating to 45 deg.C, fermenting at constant temperature for 10 hr to obtain fermented composition of Polyporus with herba Rosmarini officinalis and flos Abelmoschi Manihot.
Diluting 1 part of the obtained Polyporus umbellatus fermented composition of herba Rosmarini officinalis and flos Abelmoschi Manihot with 10 parts of deionized water; then adjusting pH to 4.5 with 0.1 mol/L sodium bicarbonate, and pulverizing with 3000 r/min wet pulverizer to obtain pulverized liquid; adjusting pH to 6.0 with 0.1 mol/L sodium bicarbonate, adjusting temperature to 50 deg.C, and treating with colloid mill for 30 min; adjusting pH to 7.0 with 0.1 mol/L sodium bicarbonate, and homogenizing at 20MPa for 5 min to obtain complete Grifola frondosa fermentation conversion composition of herba Rosmarini officinalis and flos Abelmoschi Manihot (i.e. Grifola frondosa fermentation extract composition of herba Rosmarini officinalis and flos Abelmoschi Manihot).
The prepared agaric fermentation extraction composition of rosemary and okra is dried in vacuum at 55 ℃ and crushed to prepare the granular electuary of the agaric fermentation extraction composition of rosemary and okra.
Through conventional analysis, the content of each functional component in the complete agaric fermentation transformed rosemary and okra composition is shown in table 1.
TABLE 1 Polyporus umbellatus fermented extract composition of Rosemary and Abelmoschus esculentus (%)
μ g/100 g; control is conventional heat extraction method
Example 5 solid gradient temperature fermentation of Polyporus umbellatus to transform Rosemary and okra and tablet production
Mixing herba Rosmarini officinalis powder 50 parts, flos Abelmoschi Manihot powder 40 parts, and glucose powder 10 parts, stirring to obtain solid fermentation material, and mixing the solid fermentation material with 60 deg.C water at a ratio of 1:2 to obtain solid fermentation culture medium; sterilizing at 125 deg.C for 20 min, cooling to room temperature, inoculating Polyporus umbellatus strain seed culture solution at an inoculation amount of 1%, mixing, and loading into fermentation tank with a loading coefficient of 0.75. Fermenting at 40 deg.C for 10 days, cooling to 15 deg.C, fermenting at constant temperature for 10 days, heating to 45 deg.C, fermenting at constant temperature for 2 hr to obtain fermented composition of herba Rosmarini officinalis and flos Abelmoschi Manihot.
Diluting 1 part of the obtained Polyporus umbellatus fermented composition of herba Rosmarini officinalis and flos Abelmoschi Manihot with 5 parts of deionized water; then adjusting pH to 4 with 0.1 mol/L sodium bicarbonate, and pulverizing with a wet pulverizer at 1000 rpm to obtain a pulverized liquid; adjusting pH to 5.5 with 0.1 mol/L sodium bicarbonate, adjusting temperature to 50 deg.C, and treating with colloid mill for 120 min; adjusting pH to 7.0 with 0.1 mol/L sodium bicarbonate, and homogenizing under 5MPa for 15 min to obtain complete Grifola frondosa fermentation conversion composition of herba Rosmarini officinalis and flos Abelmoschi Manihot (i.e. Grifola frondosa fermentation extract composition of herba Rosmarini officinalis and flos Abelmoschi Manihot).
The obtained Polyporus umbellatus fermented extract composition of herba Rosmarini officinalis and flos Abelmoschi Manihot is vacuum dried at 55 deg.C and pulverized to obtain granule.
The tablet of the grifola frondosa fermentation extract composition of rosemary and okra is obtained by using 64 percent of grifola frondosa fermentation extract composition granules, adding 18 percent of sucrose powder and 6 percent of dextrin as filling agents, using 10 percent of starch slurry as an adhesive and carrying out conventional processes of crushing, mixing, granulating, drying, tabletting and the like.
Through conventional analysis, the content of each functional component in the complete agaric fermentation transformed rosemary and okra composition is shown in table 2.
TABLE 2 Polyporus umbellatus fermented extract composition of Rosemary and Abelmoschus esculentus (%)
Mu g/100g, control by conventional thermal extraction method
Example 6 solid gradient temperature fermentation of Polyporus umbellatus to transform Rosemary and okra and tablet production
Uniformly stirring 60 parts of rosemary powder, 30 parts of okra powder and 10 parts of glucose powder according to the proportion to obtain a solid fermentation material, and uniformly mixing the solid fermentation material with 10 ℃ water according to the proportion of 1:1.1 to obtain a solid fermentation culture medium; sterilizing at 115 deg.C for 50 min, cooling to room temperature, inoculating Polyporus umbellatus strain seed culture solution at an inoculation amount of 20%, mixing, and loading into fermentation tank with a loading coefficient of 0.5. Fermenting at 10 deg.C for 20 days, cooling to 5 deg.C, fermenting at constant temperature for 10 days, heating to 35 deg.C, fermenting at constant temperature for 36 hr to obtain fermented composition of herba Rosmarini officinalis and flos Abelmoschi Manihot.
Diluting 1 part of the obtained Polyporus umbellatus fermented composition of herba Rosmarini officinalis and flos Abelmoschi Manihot with 15 parts of deionized water; then adjusting pH to 3.0 with 0.1 mol/L hydrochloric acid, and pulverizing with a wet pulverizer at 5000 r/min to obtain a pulverized liquid; adjusting pH to 5.5 with 0.1 mol/L sodium bicarbonate, adjusting temperature to 55 deg.C, and treating with colloid mill for 10 min; adjusting pH to 7.5 with 0.1 mol/L sodium bicarbonate, homogenizing at 30MPa for 10 min with homogenizer to obtain complete Grifola frondosa fermentation conversion composition of herba Rosmarini officinalis and flos Abelmoschi Manihot (i.e. Grifola frondosa fermentation extract composition of herba Rosmarini officinalis and flos Abelmoschi Manihot).
Vacuum drying the obtained Polyporus umbellatus fermented extract composition of herba Rosmarini officinalis and flos Abelmoschi Manihot at 55 deg.C, and pulverizing to obtain Polyporus umbellatus fermented extract composition granule of herba Rosmarini officinalis and flos Abelmoschi Manihot.
The tablet of the grifola frondosa fermentation extract composition of rosemary and okra is obtained by using 64 percent of grifola frondosa fermentation extract composition granules, adding 18 percent of sucrose powder and 6 percent of dextrin as filling agents, using 10 percent of starch slurry as an adhesive and carrying out conventional processes of crushing, mixing, granulating, drying, tabletting and the like.
Through conventional analysis, the content of each functional component in the agaric fermented extract composition of complete rosemary and okra is shown in table 3.
TABLE 3 Polyporus umbellatus fermented extract composition of Rosemary and Abelmoschus esculentus (%)
Mu g/100g, control by conventional thermal extraction method
Example 7 functional test
Memory improvement and fatigue resistance test of agaric fermentation extraction composition of rosemary and okra
1. Sample (I)
The composition granule prepared by fermenting and extracting the agaric of the rosemary and the okra by the method of the embodiment 2 of the invention (hereinafter referred to as the composition granule) comprises 1.17 percent of rosmarinic acid, 0.66 percent of okra alkali, 0.73 percent of okra flavone, 5.87 percent of agaric polysaccharide, 0.91 percent of peptidoglycan, 0.49 percent of triterpenic acid, 3.76 mu g/100g of selenium and 2.07 mu g/100g of zinc.
Control 1-purified Water
Control 2-composition with high content of rosmarinic acid and okra alkali, and other components and content unchanged (rosmarinic acid 1.45%, okra alkali 1.52%, okra flavone 0.73%, Polyporus polysaccharide 5.87%, peptidoglycan 0.91%, triterpenic acid 0.49%, microelement content: selenium 3.76 μ g/100 g; zinc 2.07 μ g/100g)
The reference 3-a composition with unchanged content of rosmarinic acid and okra alkali and increased content of other components (rosmarinic acid 1.17%, okra alkali 0.66%, okra flavone 0.87%, grifola polysaccharide 6.15%, peptidoglycan 1.09%, triterpenic acid 0.58%, trace elements content: selenium 3.81 μ g/100g, zinc 2.23 μ g/100 g).
2. Memory and anti-fatigue test for mice
2.1 test materials and objects
Experimental animals: 160 SPF male Kunming mice with age of 5 weeks and weight of 18.22g are provided by Shanghai Si Laike experimental animal center and are raised in cages according to experimental components; meanwhile, basic feed is purchased, and experiments are carried out after adaptive feeding for one week.
2.2 Experimental methods
2.2.1 animal pretreatment mice were randomly divided into quiet, maze and swim training groups, each group being individually assigned oral composition infusion and control groups. The composition granule group mice were gavaged with 1g/kg body weight of the composition granule daily for 4 weeks, except for feeding with basal diet. Control mice were fed basal diet and gavage with an equal volume of saline to the composition infusion group, and mice were free to ingest water and food. The training group was swimming trained for 4 weeks with water temperature (30 soil 2) C. Swimming for 30 minutes per day on week 1; followed by 10 minutes each week.
2.2.1 hypoxia-tolerant experiments 10 mice per group were placed in 250ml sealed jars after the last gavage, and the survival time of the mice in the quiet group and the trained groups was observed.
2.2.2 swimming experiments 15 mice per group. Mice in the training group and the resting group underwent exhaustive swimming 1 time without load after gastric lavage for 4 weeks, and the swimming time was recorded. The standard of exhaustion judgment is as follows: mice sink in water for more than 10s and are placed on a flat surface and cannot complete a righting reflex.
2.2.3 statistical processing of experimental data analysis of variance for single factor design was performed using SPSS10.0 statistical software.
2.3 Effect of the composition granule on memory, swimming and anoxia tolerance time of mice
TABLE 4 Effect of composition infusion on maze walking of mice (seconds)
| Group of | Time (n 15) |
| Control 1 group | 201±65 |
| Control 2 group | 108±55 |
| Control 3 group | 118±59 |
| Composition granule group | 119±59 |
As can be seen from Table 4, compared with the control group 1, the maze walking time of the composition granule group is averagely shortened by 40.8%, suggesting that the composition granule group has the function of enhancing the memory of the mice. Compared with the composition granule, the maze walking time of the control group 2 is averagely shortened by 9.2%, which indicates that the memory function of the mice can be further enhanced by increasing the rosmarinic acid and the okra alkali in the composition, and indicates that the two have larger influence on enhancing the memory.
TABLE 5 Effect of composition granule on mouse swimming and hypoxia tolerance time (min)
| Group of | Swimming time (n ═ 15) | Hypoxia tolerance time (n ═ 10) |
| Quiet group + control 1 | 96.26±15.59 | 13.12±4.53 |
| Quiet group + control 2 | 146.37±29.82 | 24.05±5.73 |
| Quiet group + control 3 | 140.06±29.85 | 22.01±5.32 |
| Quiet and composition granule group | 139.47±29.71 | 21.17±5.18 |
| Training group + control 1 | 196.29±28.35 | 19.95±5.74 |
| Training group + control 2 | 310.19±40.23 | 29.75±7.94 |
| Training group + control 3 | 299.74±38.25 | 24.09±6.87 |
| Training and composition electuary group | 297.88±37.16 | 23.15±6.67 |
As can be seen from Table 5, the time from swimming to exhaustion and the time to hypoxia tolerance of the mice after administration of the composition granule and training are both significantly prolonged. Compared with a quiet control group, the swimming time and the hypoxia tolerance time of the group added with the composition granules are significantly different (P is less than 0.01, and P is less than 0.05); compared with the group added with the composition electuary, the training group has very significant difference in exhaustive swimming time (P < 0.01); the swimming time and the hypoxia tolerance time of the mice in the training and composition granule group are also significantly different from those in the training control group (P is less than 0.01 and P is less than 0.05), which shows that the composition granule can significantly improve the exercise capacity of the body.
The above experimental results show that: the composition electuary prepared by fermenting and extracting the agaric of the rosemary and the okra can enhance the memory capacity of a mouse, improve the hypoxia tolerance of the mouse and have a certain anti-fatigue effect. Wherein, increasing the content of rosmarinic acid and okra alkali in the composition can obviously improve the hypoxia tolerance of mice.
The above-mentioned% are all mass percentages, i.e. wt%.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent replacements, improvements, etc. within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A manufacturing method of a agaric fermentation extraction composition of rosemary and okra is characterized by comprising the following steps:
(a) adding rosemary powder, okra powder and glucose powder into water according to a certain proportion, uniformly stirring, heating, and sterilizing at high temperature to obtain a seed culture medium solution;
(b) cooling the seed culture medium solution obtained in the step (a), inoculating grifola, uniformly stirring, and culturing for a plurality of days to obtain a grifola seed culture solution;
(c) mixing and stirring rosemary powder, okra powder and glucose powder uniformly to obtain a solid fermentation material, mixing the solid fermentation material with water uniformly, and sterilizing at high temperature to obtain a solid fermentation culture medium;
(d) and inoculating and fermenting: cooling the solid fermentation culture medium obtained in the step (c) to room temperature, inoculating the polyporus umbellatus strain seed culture solution obtained in the step (b) according to the inoculation amount of 1% -20%, uniformly mixing the two, and then putting the mixture into a fermentation tank or a fermentation tank for solid gradient temperature-changing multi-stage fermentation to obtain a polyporus umbellatus fermentation composition of rosemary and okra; the solid-state gradient temperature-changing multi-stage fermentation method comprises the following steps: the primary fermentation temperature is 20-40 ℃, and the fermentation time is 10-20 days; the secondary fermentation temperature is 5-15 ℃, and the fermentation time is 5-10 days; the third-stage fermentation temperature is 35-55 ℃, and the fermentation time is 2-36 hours;
(e) adding the agaric fermented composition of rosemary and okra obtained in the step (d) into deionized water, adopting a step-by-step pH value regulation and multi-step crushing homogenization process, adjusting pH step by step, and carrying out crushing, grinding and homogenization multi-stage physical treatment method for treatment to obtain the agaric fermented extract composition of rosemary and okra; the specific method of the step-by-step pH value adjustment and multi-stage crushing homogenization process comprises the following steps: step one, adding deionized water into a fermentation composition, adjusting the pH value to 3-4.5 by using alkali, and carrying out wet grinding by using wet grinding equipment; secondly, further adjusting the pH value to 5-6 by alkali, and carrying out levigating treatment by a colloid mill; and thirdly, further regulating the pH value to 7-8 by using alkali, and homogenizing by using a homogenizer.
2. The method according to claim 1, wherein in step (a), rosemary powder, okra powder and glucose powder are added into water according to a certain proportion, wherein the addition amount of the rosemary powder is 0.1-3%, the addition amount of the okra powder is 0.1-5%, and the addition amount of the glucose powder is 0.5-5%, which are the mass percentages; the high-temperature sterilization conditions are as follows: sterilizing at 115 ℃ and 125 ℃ for 20-50 minutes.
3. The method of claim 1, wherein the step (b) of inoculating polyporus umbellatus into the seed culture medium comprises: cooling the seed culture medium to 20-40 deg.C, inoculating 0.5-15% Polyporus umbellatus; the stirring conditions were: stirring at the rotating speed of 100-400 rpm; the culture conditions were: culturing at 10-40 deg.C for 3-10 days.
4. The manufacturing method according to claim 1, wherein: in the step (c), the solid fermentation material comprises the following components in parts by weight: 10-60 parts of rosemary powder, 10-40 parts of okra powder and 1-10 parts of glucose powder; uniformly mixing the solid fermentation material with water at the temperature of 10-60 ℃ according to the mass ratio of 1:1-1: 2; the high-temperature sterilization conditions are as follows: sterilizing at 115 ℃ and 125 ℃ for 20-50 minutes.
5. The method of claim 1, wherein the inoculated fermentation in step (d) is carried out in a thickness of 0.05 m to 0.5 m in the fermentation tank; for the fermenter, the filling factor is 0.25 to 0.75.
6. The manufacturing method according to claim 1, wherein in the step (e), the wet pulverization process conditions are: the mass ratio of the fermentation composition to the deionized water is 1:3-1:30, and the rotating speed of a pulverizer of wet pulverizing equipment is 1000 revolutions per minute to 5000 revolutions per minute; the action time of the colloid mill is 10 minutes to 2 hours, and the temperature is 10 ℃ to 60 ℃; the homogenizing pressure of the homogenizer is 5MPa-30 MPa.
7. The method of manufacture of claim 1, 2, 3, 4, 5 or 6, further comprising the steps of: analyzing the agaric fermented extract composition of rosemary and okra obtained in the step (e) according to a conventional analysis method to obtain the following functional components: rosmarinic acid, flavone, caffeine, polysaccharide, glycoprotein, triterpenic acid, selenium and zinc.
8. The method of manufacture of claim 1, 2, 3, 4, 5 or 6, further comprising the steps of: and (e) preparing the agaric fermented extract composition of rosemary and okra obtained in the step (e) into various products in different dosage forms, including granules, capsules, beverages and tablets.
9. The composition prepared by the preparation method of any one of claims 1 to 7 and prepared by fermenting the agaric of rosemary and okra is characterized in that the product comprises the following functional components in percentage by mass: 1 to 1.5 percent of rosmarinic acid, 0.5 to 2 percent of okra alkali, 0.5 to 3 percent of flavone, 1.5 to 9 percent of polysaccharide, 0.5 to 1.5 percent of peptidoglycan and 0.1 to 1.2 percent of triterpenic acid; the product has the following trace element contents: selenium 0.5 mug/100 g-5 mug/100 g; 0.3 mu g/100 g-3 mu g/100g of zinc.
10. The composition prepared by the method of claim 8, wherein the composition is prepared by fermenting the agaric of rosemary and okra, and comprises the following components: the product contains rosmarinic acid, okra alkali, flavone, polysaccharide, peptidoglycan, triterpenic acid, selenium and zinc.
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