CN109498702B

CN109498702B - Fermented traditional Chinese medicine preparation and preparation method and application thereof

Info

Publication number: CN109498702B
Application number: CN201811598452.9A
Authority: CN
Inventors: 谢志恒; 李成应; 周淑贞; 付良凯; 陈静虹
Original assignee: Foshan Nanhai Eastern Along Pharmaceutical Co ltd
Current assignee: Foshan Nanhai Eastern Along Pharmaceutical Co ltd
Priority date: 2018-12-26
Filing date: 2018-12-26
Publication date: 2021-07-27
Anticipated expiration: 2038-12-26
Also published as: CN109498702A

Abstract

The invention relates to a fermented traditional Chinese medicine preparation and a preparation method and application thereof. The preparation method of the fermented traditional Chinese medicine preparation comprises the following steps: pulverizing the Chinese medicinal herbs, and sieving to obtain mixed powder; the Chinese herbal medicine comprises radix Codonopsis and radix rehmanniae Preparata; uniformly mixing the mixed powder and a fermentation culture medium, inoculating probiotics, and fermenting at 36-38 ℃ for 24-48 h to obtain a fermentation mixture; filtering the fermented mixture, and respectively collecting filtrate and filter residue; carrying out spray drying treatment on the filtrate at 50-65 ℃ to obtain first powder; drying and crushing the filter residue to obtain second powder; and uniformly mixing the first powder and the second powder to obtain the fermented traditional Chinese medicine preparation. The method uses probiotics to ferment Chinese herbal medicine raw materials, and adopts low temperature drying fermentation mixture, not only can improve the content of effective active ingredients in the Chinese herbal medicine fermentation liquor, but also can maintain the activity of probiotics in the product, and has the effects of enhancing the immunity function of animal organism and promoting the healthy growth of animals.

Description

Fermented traditional Chinese medicine preparation and preparation method and application thereof

Technical Field

The invention relates to the technical field of livestock and poultry additives, in particular to a fermented traditional Chinese medicine preparation and a preparation method and application thereof.

Background

About 10 million tons of antibiotics are used in animal husbandry every year in China, and 90% of the antibiotics are used as feed additives. The antibiotic as the feed additive can promote the growth of animals and improve the immunity of the animals, but the excessive use of the antibiotic can cause the problems of the drug resistance enhancement of pathogenic bacteria, drug residue and the like, and brings great harm to the animal husbandry and human health. The problem of feed safety is increasingly and widely concerned with the rapid development of the feed industry, and the fermented Chinese herbal medicine feed additive which is pure natural, pollution-free, free of antibiotic residue and comprehensive in function becomes a hot point of research at home and abroad.

As a source of traditional Chinese medicine, China has systematic traditional Chinese medicine theory and extremely rich natural medicine resources, and Chinese herbal medicines contain alkaloid, polysaccharide, glycosides, amino acid, volatile oil and other substances and have certain effects of enhancing the immunity of animals and promoting the growth of the animals. However, although facing the favorable situation of the strong increase of the sound of the natural medicine market, the internationalization is slow, except that the traditional Chinese medicine theory and the modern scientific theory can not be well connected, the fundamental reason is that the modernization level of the traditional Chinese medicine is still low, and the processing and application of the Chinese herbal medicine are in urgent need of development.

Chinese fermentation technology has a long history of application and is one of the important methods for processing Chinese herbal medicines, but the traditional Chinese medicine fermentation is performed by utilizing natural strains such as mould, yeast, bacteria and the like, and the medicine effect and the quality stability of the fermented Chinese herbal medicines cannot be ensured due to the factors of weak pertinence, difficult control of fermentation conditions and the like.

Disclosure of Invention

Based on the above, a preparation method of a fermented traditional Chinese medicine preparation capable of enhancing animal immunity and promoting healthy growth of animals is needed.

A preparation method of a fermented traditional Chinese medicine preparation comprises the following steps:

pulverizing the Chinese medicinal herbs, and sieving to obtain mixed powder; wherein said herbal materials comprise radix Codonopsis and radix rehmanniae Preparata;

uniformly mixing the mixed powder and a fermentation culture medium, inoculating probiotics, and fermenting at 36-38 ℃ for 24-48 h to obtain a fermentation mixture;

filtering the fermentation mixture, and respectively collecting filtrate and filter residue; carrying out spray drying treatment on the filtrate at 50-65 ℃ to obtain first powder; drying and crushing the filter residue to obtain second powder;

uniformly mixing the first powder and the second powder to obtain the fermented traditional Chinese medicine preparation;

wherein the formula of the fermentation medium is as follows: 5-10 g of wheat bran, 6-12 g of peptone, 3-10 g of beef powder, 3-7 g of yeast powder, 10-15 g of glucose, 0.5-2 mL of tween, 1-4 g of dipotassium hydrogen phosphate, 5-9 g of sodium acetate, 2-6 g of triammonium citrate, 0.05-0.3 g of magnesium sulfate, 0.01-0.05 g of manganese sulfate and 1000mL of water.

The codonopsis pilosula is sweet and neutral in nature, and has the effects of tonifying middle-jiao and Qi, harmonizing stomach and promoting fluid production.

The prepared rehmannia root is a processed product of rehmannia root, has black and glossy surface, soft and flexible quality, sweet taste and mild nature, has the effects of nourishing yin and supplementing blood, replenishing vital essence and replenishing marrow and the like, and also has the effect of enhancing the immunologic function.

The probiotics are active microorganisms beneficial to a host, and can improve the microecological balance of the gastrointestinal tract of animals and promote the intestinal health.

The preparation method takes pure green Chinese herbal medicines such as codonopsis pilosula, prepared rehmannia root and the like as raw materials, utilizes probiotic fermentation, optimizes a fermentation culture formula, promotes the cell walls of the codonopsis pilosula, the prepared rehmannia root and the like to be broken by controlling the fermentation temperature and time, improves the content of effective active ingredients in the Chinese herbal medicine fermentation liquor, then filters the fermentation mixture, independently dries filter residues and filtrate at low temperature and then mixes the filter residues and the filtrate, and spray-dries the filtrate at 50-65 ℃, so that the activity of the active ingredients can be maintained, the activity of probiotics can be maintained, and the prepared fermented Chinese herbal medicine preparation has high content of the effective ingredients of the Chinese herbal medicines and contains the active probiotics; the prepared fermented traditional Chinese medicine preparation has the effects of tonifying qi and blood, promoting the micro-ecological balance of the gastrointestinal tract of animals, enhancing the immune function of animal organisms and promoting the healthy growth of animals; meanwhile, the active ingredients of the traditional Chinese medicine are protected from being damaged through the fermentation effect of the probiotics, the drug effect is enhanced, and the medicinal material resources can be saved.

In one embodiment, the probiotic bacteria are selected from at least one of lactobacillus acidophilus, lactobacillus plantarum, streptococcus faecalis, lactobacillus bulgaricus, enterococcus faecium.

Preferably, the probiotic is lactobacillus acidophilus.

In one embodiment, the fermentation is carried out in a fermentor under conditions selected from the group consisting of: the pressure is 0.02 kPa-0.1 kPa, the stirring speed is 10 r/min-30 r/min, and the pH value is 6.0-7.0.

Therefore, by controlling the fermentation pressure, the stirring speed and the pH value, proper fermentation conditions such as oxygen concentration, pH value, temperature and the like are provided for the probiotics, so that the Chinese herbal medicine raw materials are promoted to be subjected to microbial metabolic decomposition, the content of effective active ingredients in the fermented mixture is improved, and the concentration of viable bacteria with higher concentration is maintained.

In one embodiment, the mass ratio of the radix codonopsis pilosulae to the prepared rehmannia root is (1-2) to (2-4). Preferably, the mass ratio of the radix codonopsis pilosulae to the prepared rehmannia root is 1: (1.8 to 2.2)

Therefore, the radix codonopsitis and the prepared rehmannia root have good synergistic effect with probiotics and improve the drug effect.

In one embodiment, the step of pulverizing and sieving the Chinese herbal medicines to obtain the mixed powder comprises the following steps:

mixing the Chinese herbal medicines with the soybean meal, crushing, and sieving with a 30-80-mesh sieve to obtain the mixed powder; the weight ratio of the Chinese herbal medicines to the soybean meal is (2-5): 10.

the inventor finds that the prepared rehmannia root is sticky and not easy to crush in the crushing process. Therefore, after a large number of experiments, the soybean meal is screened out as an auxiliary raw material. Compared with other vegetable oil meal such as cottonseed meal, peanut meal, rapeseed meal and the like, the crushing effect is best, the problem that prepared rehmannia roots are crushed and sticky can be well solved, and a nitrogen source can be provided for probiotic fermentation.

Therefore, the radix codonopsitis and the prepared rehmannia root which are used as raw materials of the fermented traditional Chinese medicine preparation are crushed together with the bean pulp, so that the crushing effect and efficiency can be improved, the bean pulp can provide sufficient nitrogen source for subsequent fermentation, the fermentation process is promoted, and the efficacy of the fermented traditional Chinese medicine preparation is improved.

Further, the prepared rehmannia root is dried for 3-4 hours at the temperature of 50-55 ℃, and then is mixed with the codonopsis pilosula and the soybean meal, crushed and sieved.

In one embodiment, the adding ratio of the mixed powder to the fermentation medium is (8-20) g: 1L.

Therefore, when the probiotics are utilized for fermentation, 8g to 20g of mixed powder of the Chinese herbal medicines is added into 1L of fermentation medium, during the fermentation, the fermentation medium can provide enough nutrients such as nitrogen source, carbon source, phosphorus source and the like for the growth and the propagation of the probiotics, and maintain a relatively stable pH environment until the Chinese herbal medicines are fully fermented and decomposed, thereby ensuring the fermentation effect.

In one embodiment, the inoculated probiotic is a secondary seed solution of a probiotic species; the preparation steps of the secondary seed liquid comprise:

activating probiotic strains, inoculating the activated probiotic strains into an expanding culture medium, and statically culturing for 12-24 h at 36-38 ℃ to obtain a primary seed solution;

inoculating the primary seed solution into an expanding culture medium according to the inoculation amount of 2-10 vol%, and performing static culture at 36-38 ℃ for 12-24 h to obtain a secondary seed solution;

and inoculating the secondary seed liquid according to the volume ratio of the secondary seed liquid to the fermentation culture medium of 3-8 vol%.

Further, the formula of the expanding culture medium is as follows: 10g of tryptone, 5g of beef powder, 4g of yeast powder, 20g of glucose, 1mL of tween, 2g of dimethyl hydrogen phosphate heptahydrate, 5g of sodium acetate trihydrate, 2g of triammonium citrate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1000mL of distilled water and pH value of 6.0-6.5.

In one embodiment, the method further comprises the step of activating the probiotic bacterial species:

inoculating probiotic strains to an activation culture medium, and culturing at 36-38 ℃ for 24-48 h for activation;

wherein the formula of the activation medium is as follows: 10g of peptone, 5g of beef powder, 4g of yeast powder, 20g of glucose, 1mL of tween, 2g of dipotassium phosphate heptahydrate, 5g of sodium acetate trihydrate, 2g of triammonium citrate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 15g of agar, 1000mL of water and pH value of 6.0-6.5.

The invention also aims to provide the fermented traditional Chinese medicine preparation prepared by the preparation method.

The fermented traditional Chinese medicine preparation utilizes the fermentation effect of probiotics to decompose macromolecular substances of the Chinese herbal medicines such as the codonopsis pilosula, the prepared rehmannia root and the like, controls the fermentation conditions in the fermentation process, adopts low-temperature drying after fermentation to maintain the activity of the probiotics, not only improves the content of the effective components of the traditional Chinese medicines, but also organically combines the Chinese herbal medicines such as the codonopsis pilosula, the prepared rehmannia root and the probiotics such as lactobacillus acidophilus, lactobacillus plantarum, streptococcus faecalis, lactobacillus bulgaricus, enterococcus faecium and the like, and enhances the drug effect; it is rich in alkaloid, flavonoid, iridoid, monoterpene, active oligosaccharide and polypeptide, and has effects of invigorating qi and blood, enhancing macrophage phagocytic ability, promoting lymphocyte transformation, enhancing antibody cell function, increasing antibody titer and improving resistance of organism to harmful stimulation.

The invention also aims to provide application of the fermented traditional Chinese medicine preparation in preparation of animal feed.

In one embodiment, the fermented traditional Chinese medicine preparation is used in the preparation of sow feed.

Further, the fermented traditional Chinese medicine preparation is added into the feed for the sows in the lactation period.

The fermented traditional Chinese medicine preparation is applied to the feed of the sows in the lactation period and is fed to the lactating sows, so that the immunity and the breast milk quality of the sows can be obviously improved, the content of immunoglobulin in the breast milk is improved, the passive immune function of piglets can be further improved, the health of the piglets is enhanced, and the survival rate of the piglets is improved.

Detailed Description

In order that the invention may be more fully understood, a more particular description of the invention will now be rendered by reference to specific embodiments thereof that are illustrated in the appended drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

The following are specific embodiments.

The raw materials used in the examples of the present invention are commercially available.

Example 1

Fermented traditional Chinese medicine preparation and preparation method thereof

1. Strain: lactobacillus acidophilus.

2. The mixed powder of the compound Chinese herbal medicines comprises the following components: weighing 1 part of radix codonopsitis, 2 parts of prepared rehmannia root and 10 parts of bean pulp, drying the prepared rehmannia root in an oven at 50-55 ℃ for 3-4 hours, mixing with the radix codonopsitis and the bean pulp, crushing, and sieving by a 60-mesh sieve to obtain mixed powder for later use.

3. Culture medium formula

Activating a culture medium: 10g of tryptone, 5g of beef powder, 4g of yeast powder, 20g of glucose, 801 mL of tween, 2g of dimethyl hydrogen phosphate heptahydrate, 5g of sodium acetate trihydrate, 2g of triammonium citrate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1000mL of distilled water, 15g of agar and 6.2 of pH value.

Expanding culture medium: 10g of tryptone, 5g of beef powder, 4g of yeast powder, 20g of glucose, 801 mL of tween, 2g of dimethyl hydrogen phosphate heptahydrate, 5g of sodium acetate trihydrate, 2g of triammonium citrate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1000mL of distilled water and 6.2 of pH value.

Fermentation medium: 5g of wheat bran, 8g of peptone, 4g of beef powder, 5g of yeast powder, 14g of glucose, 801 mL of tween, 2.2g of dipotassium phosphate, 5.1g of sodium acetate, 2g of triammonium citrate, 0.21g of magnesium sulfate, 0.06g of manganese sulfate and 1000mL of pure water. 20g of mixed powder of Chinese herbal medicines is added into 1L of fermentation medium.

4. Strain activation

Cooling the sterilized activation culture medium to 50-60 ℃, and pouring the activated culture medium into a flat plate. After the culture medium is solidified, the lactobacillus acidophilus strain is picked by an inoculating loop and streaked on a plate culture medium, and then the plate is placed upside down at 36-38 ℃ for culture for 36 h.

5. Seed liquid culture

The lactobacillus acidophilus strain activated on a plate culture medium is selected by an inoculating loop, inoculated in an expanding culture medium after sterilization and cooling, and statically cultured in an incubator for 12-24 h at 36-38 ℃ to serve as a primary seed solution.

Absorbing the first-stage seed liquid according to the inoculation amount of 8vol%, and inoculating the first-stage seed liquid into an expanding culture medium. Static culturing for 12-24 h at 36-38 deg.C in incubator as secondary seed liquid.

6. Sterilization

Sterilizing the air filter, introducing air into the air filter after sterilization, and maintaining the pressure to be 0.1-0.15 MPa for quality inspection to prevent bacterial contamination; the tank body is jacketed and preheated to more than 85 ℃, after the temperature reaches, a jacket air inlet switch is closed, hot steam is introduced to heat the tank body, and when the temperature reaches 121 ℃ and the pressure intensity reaches 0.11MPa, the temperature and the pressure are maintained for 20 min. After sterilization, the steam inlet valve is closed, and the air outlet valve is opened to release pressure.

After the empty tank is sterilized and decompressed, adding a fermentation culture medium from a feed inlet, and stirring at 100 r/min-150 r/min; after the feeding is finished, the pH value of the culture medium is adjusted, and the feeding hole is closed. The tank body is jacketed and preheated to more than 85 ℃, after the temperature reaches, a jacket air inlet switch is closed, hot steam is introduced to heat the tank body, and when the temperature reaches 121 ℃ and the pressure intensity reaches 0.11MPa, the temperature and the pressure are maintained for 20 min. After sterilization, closing the steam inlet valve, opening the big air release valve, and opening the vent valve when the pressure is reduced to 0.08MPa, so that the pressure in the tank body is kept at about 0.05 MPa; and opening the cold water valve to cool the tank body.

7. Inoculating and fermenting

When the temperature of the culture medium in the tank is reduced to 37 ℃, the cold water valve is closed. Keeping the pressure of the jar at 0.02MPa, scrubbing the inoculation opening with alcohol, putting the inoculation ring on the inoculation opening, and igniting. Opening the feed port, burning the bottle mouth of the secondary seed liquid on flame according to the inoculation amount of 5 vol%, opening the bottle mouth, rapidly pouring the seed liquid into the fermentation tank under the protection of the flame, closing the feed port, extinguishing the flame and cleaning with alcohol.

Controlling fermentation conditions: the temperature is 37 ℃, the pressure is 0.02kPa, the pH value is 6.2, the stirring speed is 20r/min, and the culture time is 44 h.

8. Filtering, drying, and pulverizing

And after the fermentation is finished, putting the fermentation liquor into a tank to collect the fermentation liquor, adding a flocculating agent into the fermentation liquor, transferring the fermentation liquor to a plate-and-frame filter to be filtered, respectively collecting filtrate and filter residue, drying the filtrate in a low-temperature spray dryer with the nozzle caliber of 0.5-1 mm and the temperature of 50-65 ℃, drying the filter residue in a blast oven for 4-5 h at the temperature of 60-80 ℃, crushing the filter residue, sieving the filter residue with a 30-40-mesh sieve, combining and uniformly mixing the filter residue and the filter residue to obtain the fermented traditional Chinese medicine preparation.

Example 2

The example 2 is basically the same as the example 1, except that the addition amount of the fermentation medium and the mixed powder of the Chinese herbal medicines and the fermentation conditions are different, specifically:

the fermentation medium is as follows: 8g of wheat bran, 10g of peptone, 6g of beef powder, 3g of yeast powder, 12g of glucose, 801.3 mL of tween, 2.5g of dipotassium hydrogen phosphate, 6g of sodium acetate, 3g of triammonium citrate, 0.2g of magnesium sulfate, 0.05g of manganese sulfate and 1000mL of pure water; 12g of mixed powder of Chinese herbal medicines is added into 1L of fermentation medium.

The fermentation conditions were: the temperature is 38 ℃, the pressure is 0.06kPa, the pH value is 6.6, the stirring speed is 30r/min, and the culture time is 38 h.

Example 3

The example 3 is basically the same as the example 1, except that the addition amount of the fermentation medium and the mixed powder of the Chinese herbal medicines and the fermentation conditions are different, specifically:

the fermentation medium is as follows: 10g of wheat bran, 8g of peptone, 6g of beef powder, 6g of yeast powder, 13g of glucose, 801.2 mL of tween, 2g of dipotassium hydrogen phosphate, 5g of sodium acetate, 2.5g of triammonium citrate, 0.18g of magnesium sulfate, 0.08g of manganese sulfate and 1000mL of pure water; 16g of mixed powder of Chinese herbal medicines is added into 1L of fermentation medium.

The fermentation conditions were: the temperature is 36 ℃, the pressure is 0.05kpa, the pH value is 6.4, the stirring speed is 30r/min, and the culture time is 48 h.

Example 4

Example 4 is substantially the same as example 1 except that the fermentation strain is different, specifically, the fermentation strain of example 4 is lactobacillus plantarum.

Example 5

Example 5 is substantially the same as example 1 except that the fermentation strain is different, specifically, the fermentation strain of example 5 is streptococcus faecalis.

Example 6

Example 6 is basically the same as example 1 except that the fermentation type is different, specifically, the fermentation type of example 6 is enterococcus faecium.

Comparative example 1

Comparative example 1 is substantially the same as example 1 except that the amounts of the fermentation medium and the mixed powder of the Chinese herbal medicines were different, specifically:

the fermentation medium is as follows: 3g of wheat bran, 15g of peptone, 10g of beef powder, 12g of yeast powder, 20g of glucose, 803 mL of tween, 6g of dipotassium hydrogen phosphate, 9g of sodium acetate, 7g of triammonium citrate, 0.2g of magnesium sulfate, 0.25g of manganese sulfate and 1000mL of pure water; 5g of mixed powder of Chinese herbal medicines is added into 1L of fermentation medium.

Comparative example 2

Comparative example 2 is substantially the same as example 1 except that the amounts of the fermentation medium and the mixed powder of the Chinese herbal medicines were different, specifically:

the fermentation medium is as follows: 15g of wheat bran, 5g of peptone, 4g of beef powder, 2g of yeast powder, 15g of glucose, 802 mL of tween, 1g of dipotassium hydrogen phosphate, 4g of sodium acetate, 3g of triammonium citrate, 0.04g of magnesium sulfate, 0.07g of manganese sulfate and 1000mL of pure water; 25g of mixed powder of Chinese herbal medicines is added into 1L of fermentation medium.

Comparative example 3

Comparative example 3 is substantially the same as example 1 except that the fermentation conditions are different, and the specific fermentation conditions are as follows: the temperature is 38 ℃, the pressure is 0.1kpa, the pH value is 7.0, the stirring speed is 50r/min, and the culture time is 48 h.

Comparative example 4

Comparative example 4 is substantially the same as example 1 except that the fermentation conditions are different, and the specific fermentation conditions are: the temperature is 35 ℃, the pressure is 0.3kpa, the pH value is 5.5, the stirring speed is 80r/min, and the culture time is 60 hours.

Traits and detection

The fermented traditional Chinese medicine preparation prepared in the embodiments 1-3 of the invention is gray to brown gray powder, has uniform color, no peculiar smell, no foreign substances and fermented smell.

The fermented traditional Chinese medicine preparation samples prepared in the examples and the comparative preparation are subjected to stachyose, total sugar, probiotic viable count and moisture content determination, and the method comprises the following steps:

1) and (3) measuring moisture: the water content of the samples in the examples and the comparative examples is less than or equal to 10.0 percent by measuring the water content in the feed of GB/T6435-.

2) Determination of stachyose content

Preparing a stachyose standard solution: accurately weighing 25mg of stachyose standard substance into a 25ml volumetric flask, adding distilled water to dissolve and fix the volume to obtain standard solution stock solution.

Preparation of a standard curve: taking 0.5ml, 1.0ml, 2.0ml, 4.0ml and 6.0ml of prepared stachyose standard solution in a 10ml volumetric flask respectively, and adding distilled water to constant volume.

Preparing a test solution: accurately weighing 0.025g (accurate to 0.0001g) of sample in a 10ml volumetric flask, adding a proper amount of distilled water for dissolving, placing in an ultrasonic cleaning machine for ultrasonic treatment for 15min, cooling, adding distilled water for constant volume to scale, and taking a part of solution to pass through a membrane to be used as a test solution.

The analysis method comprises the following steps: liquid chromatography

Chromatographic conditions are as follows:

mobile phase: methanol: water (80: 20).

A chromatographic column: amino column or other type of column with equivalent separation effect.

Carrier gas: nitrogen or compressed air.

Flow rate of carrier gas: 4.0 ml/min.

Calculation and results: and (3) drawing by taking the peak area integral value of the standard curve as a vertical coordinate and the concentration integral value of the standard curve as a horizontal coordinate, calculating a regression equation, substituting the peak area integral value of the test sample for calculating the concentration of the test sample, obtaining the actual concentration by inverse logarithm of the concentration of the test sample, and finally calculating the content, wherein the content is expressed in percentage. Taking the arithmetic mean value of the results of the parallel measurement as the measurement result, and taking the relative deviation of the results of the two parallel measurements as no more than 5 percent.

3) Determination of the Total sugar content

Sample treatment: accurately weighing 0.2500g (accurate to 0.0001g) of sample, adding ether 100ml, heating and refluxing at 40 deg.C for 1 hr, standing, cooling, carefully discarding ether solution, and volatilizing ether from residue in water bath. Adding 100ml of 80% ethanol, heating and refluxing for 1 hour in a water bath at 85 ℃, filtering while hot, placing filter residue and filter paper in a flask, adding 120ml of water, and heating and refluxing for 2 hours. Filtering while hot, washing the filter with a small amount of water, combining the filtrate and the washing solution, cooling, transferring into a 250ml measuring flask, diluting with water to a scale, shaking up, adding 10.0ml of the solution into a 50ml volumetric flask, and diluting with water to a scale to obtain the test solution.

Preparation of a standard curve: precisely measuring reference substance solutions 0.2ml, 0.4ml, 0.6ml, 0.8ml and 1.0ml, respectively placing in a test tube with a plug, respectively adding water to 2.0ml, respectively adding 5% phenol solution 1ml, shaking up, rapidly and precisely adding sulfuric acid 5ml, shaking up, standing for 10min, placing in a 40 deg.C water bath, keeping the temperature for 30 min, taking out, rapidly cooling to room temperature, and taking corresponding reagents as blank. And (3) measuring the absorbance at 490nm according to an ultraviolet-visible spectrophotometry, drawing a standard curve by taking the absorbance as a vertical coordinate and the mass of the glucose as a horizontal coordinate, and solving a regression equation.

And (3) sample determination: precisely measuring 1.0mL, placing in a test tube with a plug, adding 1.0mL of water, and preparing according to the method under the preparation item of the standard curve, wherein the method comprises the following steps: after 1.0mL of each 5% phenol solution was added precisely, the absorbance was measured by the method, and the mass (mg) of glucose in the sample solution was read from the standard curve and calculated.

Calculation and results

The result of the polysaccharide content (X) in the sample is expressed in mass fraction (%) and calculated according to formula (1):

in the formula:

c-concentration read on the standard curve in milligrams per milliliter (mg/ml);

v-sample volume to volume in milliliters (ml);

m-sample weight in grams (g).

The final result retains three significant digits; taking the arithmetic mean value of the results of the parallel measurement as the measurement result, and taking the relative deviation of the results of the two parallel measurements as no more than 5 percent.

4) And (3) measuring the viable count: a flat plate viable count method is adopted.

The results of the measurements are shown in Table 1 below.

TABLE 1

As can be seen from the above Table 1, the content of stachyose and the content of viable bacteria in the examples 1-6 of the present invention are significantly higher than those in the comparative examples 1-4, especially, the content of stachyose in the samples of the examples 1-3 is higher than 16.5 wt%, and the content of viable bacteria is greater than 2.2 × 10¹⁰cfu/g, which shows that the content of effective active ingredients in the product can be improved by optimizing the fermentation medium and the fermentation conditions in the embodiment of the invention, and meanwhile, the number of effective viable bacteria in the product can be maintained at a higher level, so that the quality of the product is ensured.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A preparation method of a fermented traditional Chinese medicine preparation is characterized by comprising the following steps:

pulverizing the Chinese medicinal herbs, and sieving to obtain mixed powder; wherein said herbal medicine is prepared from radix Codonopsis and radix rehmanniae Preparata;

uniformly mixing the mixed powder and a fermentation medium, inoculating probiotics, and fermenting at 36-38 ℃ for 24-48 h to obtain a fermentation mixture;

wherein the formula of the fermentation medium is as follows: 5-10 g of wheat bran, 6-12 g of peptone, 3-10 g of beef powder, 3-7 g of yeast powder, 10-15 g of glucose, 0.5-2 mL of Tween, 1-4 g of dipotassium hydrogen phosphate, 5-9 g of sodium acetate, 2-6 g of triammonium citrate, 0.05-0.3 g of magnesium sulfate, 0.01-0.05 g of manganese sulfate and 1000mL of water; the adding ratio of the mixed powder to the fermentation medium is (8-20) g: 1L; the fermentation is carried out in a fermentation tank, and the conditions of the fermentation are as follows: the pressure is 0.02 kPa-0.1 kPa, the stirring speed is 10 r/min-30 r/min, and the pH value is 6.0-7.0.

2. The method of claim 1, wherein the probiotic bacteria are at least one selected from the group consisting of lactobacillus acidophilus, lactobacillus plantarum, streptococcus faecalis, lactobacillus bulgaricus, and enterococcus faecium.

3. The method of claim 2, wherein the probiotic bacteria is Lactobacillus acidophilus.

4. The method according to claim 1, wherein the mass ratio of the radix Codonopsis to the radix rehmanniae Preparata is (1-2) to (2-4).

5. The preparation method according to claim 4, wherein the mass ratio of the radix codonopsis pilosulae to the prepared rehmannia root is 1: (1.8-2.2).

6. The preparation method according to claim 1, wherein the step of pulverizing and sieving the Chinese herbal medicines to obtain a mixed powder comprises:

7. the method according to any one of claims 1 to 6, wherein the inoculated probiotic is a secondary seed solution of a probiotic species; the preparation steps of the secondary seed liquid comprise:

activating probiotic strains, inoculating the activated probiotic strains into an expanding culture medium, and statically culturing for 12-24 hours at 36-38 ℃ to obtain a first-stage seed solution;

inoculating the primary seed liquid into an expanding culture medium according to the inoculation amount of 2-10 vol%, and performing static culture at 36-38 ℃ for 12-24 h to obtain a secondary seed liquid;

and inoculating the secondary seed liquid according to the volume ratio of the secondary seed liquid to the fermentation medium of 3-8 vol%.

8. The method according to claim 7, wherein the formula of the expanding culture medium is as follows: 10g of tryptone, 5g of beef powder, 4g of yeast powder, 20g of glucose, 1mL of tween, 2g of dimethyl hydrogen phosphate heptahydrate, 5g of sodium acetate trihydrate, 2g of triammonium citrate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 1000mL of distilled water and pH value of 6.0-6.5.

9. The fermented traditional Chinese medicine preparation prepared by the preparation method of any one of claims 1 to 8.

10. Use of the fermented Chinese medicinal preparation of claim 9 in the preparation of an animal feed additive.