A fermentation processing method of baizhuo decoction based on adjusting ammonia nitrogen content in fermentation environment

Abstract

This application relates to a method for fermenting and processing Baiyao Decoction based on adjusting the ammonia nitrogen content of the fermentation environment, and the Baiyao Decoction processed by this fermentation and processing method. The invention provides precise control over the fermentation process of Baiyao Decoction, primarily characterized by using Aspergillus oryzae TJTSW001 instead of yeast for direct fermentation, and using tea juice with added gallnut powder and glutamic acid as the liquid culture medium for Aspergillus oryzae TJTSW001. Compared with traditional fermentation and processing techniques, this method further increases the gallic acid content in the resulting Baiyao Decoction, while also enhancing its antibacterial activity.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine processing technology, specifically relating to a method for processing Baiyao Decoction through fermentation based on adjusting the ammonia nitrogen content in the fermentation environment. Background Technology

[0002] Baiyaojian, also known as Baiyaojian or Baicaojian, is a blocky substance made by fermenting the galls (also known as gallnuts) on the leaves of plants of the Anacardiaceae family, such as Rhus chinensis Mill., Rhus potaninii Maxim., or Rhus punjabensis Stew. var. sinica (Diels) Rehd. Et Wils., together with tea leaves, yeast, etc.

[0003] The properties, flavors, and effects of gallnut change before and after fermentation and processing. Gallnut is sour, astringent, and cold in nature, and enters the lung, large intestine, and kidney meridians. It has the functions of astringing the lungs and reducing fire, astringing the intestines and stopping diarrhea, astringing sweat and stopping bleeding, and astringing dampness and healing sores. It is used to treat chronic cough due to lung deficiency, cough with phlegm due to lung heat, chronic diarrhea and dysentery, night sweats, diabetes, hematochezia, external bleeding, carbuncles and boils, and skin ulcers. On the other hand, Baiyaojian (a traditional Chinese medicine decoction) is sour and sweet, and neutral in nature. In addition to entering the lung meridian, Baiyaojian also enters the heart and stomach meridians. It has the functions of moistening the lungs and resolving phlegm, stopping bleeding and diarrhea, relieving fever and promoting body fluid. It is used for sore throat, chronic cough with excessive phlegm, chronic enteritis, canker sores, gingivitis, bloody dysentery, and thirst due to summer heat.

[0004] Unlike gallnut, fermented gallnut is light and ethereal, with a floating and astringent nature, and a lingering sweet taste. It is particularly suitable for treating coughs, phlegm, thirst, and other ailments of the upper burner (heart and lungs), and is especially beneficial when held in the mouth. Modern research shows that Baiyaojian (a traditional Chinese medicine decoction) has significant therapeutic effects on respiratory and digestive system diseases (such as bronchitis, asthma, pharyngitis, and gastroenteritis). These changes in its properties, flavors, meridian tropism, and efficacy are likely related to its different processing techniques, and the changes in the content and chemical composition of its main active ingredients during processing.

[0005] The main active ingredient in gallnut is tannin, which easily combines with proteins to form precipitates. In the gastrointestinal tract, tannins can irritate the gastrointestinal mucosa, causing adverse reactions such as loss of appetite. Fermentation and processing into a decoction enhances its astringent properties, potentially reducing irritation to the gastrointestinal mucosa and minimizing or eliminating adverse reactions like loss of appetite.

[0006] The processing method of Baiyaojian in Sichuan Province is similar to that in the "Compendium of Materia Medica". In 2021, the Sichuan Provincial Drug Administration issued and implemented a new standard for Baiyaojian, which modified the preparation steps of the test solution in the content determination. The original standard measured the gallic acid content of the finished Baiyaojian after complete hydrolysis of tannins (the content of tannins, calculated as gallic acid, shall not be less than 30.0%). The new standard measures the gallic acid content of the finished Baiyaojian without hydrolysis (the gallic acid content should be 35.0% to 55.0%).

[0007] Baoyaojian has a long history. It was used for leather making in the Song Dynasty and began to be used medicinally in the Ming Dynasty. It was first recorded in "Ben Cao Meng Quan", and Li Shizhen made detailed records on its fermentation and processing methods and indications in "Compendium of Materia Medica". Galla chinensis contains a large amount of tannins. In the gastrointestinal tract, tannins can combine with proteins on the mucosa to form macromolecular precipitates, stimulating the gastrointestinal tract and causing adverse reactions. However, after fermentation and processing, Galla chinensis can reduce irritation. The main active ingredient of Baoyaojian is gallic acid, which is converted from hydrolyzable tannins in Galla chinensis through microbial fermentation and processing. It has activities such as killing trypanosomes, anti-inflammatory, antibacterial, anti-tumor, and antiviral. Therefore, by measuring the content of gallic acid in Baoyaojian, the efficacy can be preliminarily judged.

[0008] In different eras, the processing methods were different. Throughout history, the processing of Baoyaojian mainly focused on brewing, that is, fermentation today, and frying and calcining were less used.

[0009] By consulting previous editions of pharmacopoeias, Hong Kong Chinese medicine material standards, and processing specifications in various regions across the country (Sichuan, Liaoning, Beijing, Fujian, Guangdong, Guangxi Zhuang Autonomous Region, Hunan, Jiangsu, Jiangxi, Ningxia, Shandong, Shanghai, Zhejiang) and national processing specifications, etc., except for the processing methods of Baoyaojian recorded in the Chinese medicine processing specifications of Zhejiang and Jiangxi, Beijing Chinese medicine material standards, and Wuhan Wenbang, previous editions of pharmacopoeias, Hong Kong Chinese medicine material standards and other places only included the cleaning methods of Galla chinensis. The results showed that Baoyaojian is mainly fermented from Galla chinensis, tea leaves, yeast koji, etc. The specific processing techniques and main excipients in different places are different. For example, for koji, baijiu is used in Beijing and rice wine is used in Jiangxi; Wuhan Wenbang uses black tea and smoked plum decoction, and the Beijing school uses licorice, platycodon, green tea, etc. as excipients.

[0010] Currently, most of the literature research on Baoyaojian focuses on its processing technology. The Chinese medicine fermentation method is a method in which the cleaned or processed medicine foams and forms a film under certain temperature and humidity conditions due to the catalytic decomposition of molds and enzymes. Chinese medicine fermentation can change the original properties, produce new therapeutic effects, expand the variety of medicines used (such as Baoyaojian, Jian Shenqu, Baoyaojian, etc.), and can also enhance its efficacy, such as Pinellia ternata fermentata.

[0011] In terms of chemical composition, the main component of Baoyaojian is gallic acid. Baoyaojian is prepared by the fermentation method for gallic acid, which is more than two hundred years earlier than the work of the Swedish pharmacologist Scheele in preparing gallic acid. Modern research on Baoyaojian often uses gallic acid as one of the indicators for further study.

[0012] In terms of efficacy and clinical use, the content of tannins in the raw product of Galla chinensis is relatively high, and its anti-inflammatory and antitussive effects are stronger than those of the fermented product and the hydrolysis product; the content of gallic acid in the fermented product and the hydrolysis product is high, and the antibacterial and expectorant effects are stronger than those of the raw product. Antibacterial experiments show that the fermented product is better than the non-fermented product. Baoyaojian is applied in Chinese patent medicines, such as Jinshuangjian, Jiechang An capsules, and Qingyan Pills composed of Baoyaojian.

[0013] The historical evolution of Baiyaojian (a traditional Chinese medicine decoction) shows that it is mainly prepared using a brewing (fermentation) method, made from gallnuts, tea leaves, and yeast. Different regions vary in their processing techniques, not only in the use of black or green tea and the addition of auxiliary materials such as alum, dried plum, platycodon, and licorice, but also in the proportions of Chinese herbs, yeast, and tea leaves, as well as the specific operational steps. These differences influence the composition and efficacy of Baiyaojian.

[0014] Chinese patent CN103893392A discloses a method for increasing the gallic acid content in Baiyao Decoction. The method includes the following steps: 1) Taking a measured amount of distiller's grains (fermentation starter) and pulverizing it; 2) Boiling tea leaves with water to make tea juice; 3) Mixing the pulverized distiller's grains (fermentation starter) from step 1) and the tea juice obtained from step 2) to form a solution, adding 10%-30% by weight of gallnut powder to this solution, and mixing to obtain a final solution; 4) Cultivating the solution obtained in step 3) at 30-37℃ for 2-3 days to obtain a microbial inoculum; 5) Mixing the microbial inoculum obtained in step 4) with gallnut powder, mixing evenly, and then granulating; 6) Cultivating the granules at 30-37℃ for 3 days; 7) Drying the material cultivated in step 6). This method effectively shortens the production cycle of Baiyao Decoction and significantly increases the gallic acid content in it. Summary of the Invention

[0015] The applicant has conducted in-depth and extensive research on the fermentation process of Baiyao Decoction. The research found that by adjusting the content of ammonia nitrogen in the fermentation environment, the content of gallic acid in the prepared Baiyao Decoction can be further increased compared with that obtained without using this fermentation process, and the antibacterial effect is significantly enhanced.

[0016] This application first discloses a fermentation processing method for Baiyao Decoction based on adjusting the ammonia nitrogen content in the fermentation environment. The fermentation processing method consists of the following steps:

[0017] 1) Boil tea leaves with water to make tea juice, crush gallnuts and remove impurities;

[0018] 2) Inoculate Aspergillus oryzae into slant test tubes and incubate at 35℃ for 5 days. After growth, wash off the surface spores with sterile water and inoculate them into liquid culture medium (tea juice obtained in step 1) with added gallnut powder and glutamic acid. Incubate at 30-37℃ and 180 rpm for 3 days. Use this as the liquid inoculum.

[0019] 3) Mix the liquid seed obtained in step 2) with the gallnut powder, mix evenly, and then granulate;

[0020] 4) Place the particles obtained in step 3) at 30-37℃ and incubate for 3 days;

[0021] 5) Sterilize the material cultured in step 4) and vacuum dry it.

[0022] Preferably, the Aspergillus species used in step 2) is Aspergillus oryzae TJTSW001.

[0023] Preferably, the mass percentage of gallnut powder in step 2) is 10%-30%.

[0024] Preferably, the mass percentage of gallnut powder in step 2) is 10%.

[0025] Preferably, the mass percentage of glutamic acid in step 2) is 1%-10%, preferably 3%.

[0026] As a preferred option, the cultured microbial inoculum is directly added to the gallnut powder as a moisture regulating medium.

[0027] Preferably, the moisture content of the particles obtained in step 3) is adjusted to 50%-100%, with 80% being optimal.

[0028] Preferably, the culture conditions in step 4) are a culture temperature of 30-37℃ and a relative humidity of 60%-90%. More preferably, the culture temperature is 33-37℃ and the relative humidity is 70%-80%, which is conducive to fermentation.

[0029] Preferably, the tea leaves used in step 1) and the gallnut powder added in step 3) are in a mass ratio of 1:20.

[0030] For fermented products, the microbial strain is the primary factor determining the quality of the fermented product. The quantity of microorganisms is the main determinant of the fermentation cycle length. Traditionally, Baiyaojian (a traditional Chinese medicine formula) involves directly mixing gallnuts, tea leaves, and distiller's grains (fermentation starter) for fermentation. In this process, the distiller's grains (fermentation starter) serve as the microbial inoculum, primarily providing the microbial strains required for fermentation. In this application, Aspergillus oryzae TJTSW001 is used instead of the fermentation starter as the microbial inoculum. Compared to using the fermentation starter as the microbial inoculum, the gallic acid content in the resulting Baiyaojian is further increased.

[0031] The advantage of this invention is that it uses Aspergillus oryzae TJTSW001 to replace the yeast for direct fermentation, and adds a certain amount of glutamic acid as ammonia nitrogen to the culture medium. This further increases the gallic acid content in the resulting herbal decoction.

[0032] Tanninase is an inducible enzyme. It catalyzes the conversion of tannins into gallic acid. The amount of gallic acid in the Baiyao decoction is determined by the amount and activity of tanninase produced by the microorganisms in the decoction. Therefore, in this invention, adding a certain amount of gallnut during liquid culture of the strain can induce Aspergillus oryzae to produce a certain amount of tanninase. On the other hand, adding a certain amount of gallnut forms a selective culture medium, inhibiting the growth of other non-active microorganisms. In this invention, the amount of gallnut added to the Aspergillus oryzae liquid culture medium is 10%-30%, preferably 10%. A certain amount of glutamic acid is added to the culture medium as a supplementary nitrogen source to further promote the production of tanninase by Aspergillus oryzae. The amount of glutamic acid added is 1%-10%, preferably 3%.

[0033] In the subsequent fermentation process, to provide oxygen and promote microbial growth, thereby increasing the conversion rate of tannic acid in the herbal decoction, this invention granulates the uniformly mixed material to increase the surface area of ​​the material and the gaps between the particles. For ease of granulation, it is best to adjust the moisture content of the material to 50%-100%, and preferably, to 80%.

[0034] The subsequent culture conditions are a culture temperature of 30-37℃ and a relative humidity of 60%-90%. Preferably, the culture temperature is 33-37℃ and the relative humidity is 70%-80%, which is conducive to fermentation.

[0035] After drying, the Baiyao Decoction was tested for gallic acid according to the 2020 edition of the Pharmacopoeia of the People's Republic of China.

[0036] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0037] This invention features precise control over the fermentation process of Baiyao Decoction. Its main characteristic lies in replacing the traditional yeast starter with Aspergillus oryzae TJTSW001 for direct fermentation, and using tea juice containing gallnut powder and glutamic acid as the liquid culture medium for Aspergillus oryzae TJTSW001. Fermentation with this liquid culture medium containing Aspergillus oryzae TJTSW001 and gallnut powder further increases the gallic acid content in the resulting Baiyao Decoction compared to traditional fermentation processes. Simultaneously, it enhances its antibacterial activity.

[0038] Finally, this application provides a Baiyao Decoction, which is prepared by the aforementioned Baiyao Decoction fermentation and processing process. Detailed Implementation

[0039] The present invention will be further described in detail below through embodiments. These embodiments are illustrative of the invention, but do not limit the invention in any way. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in this technical field.

[0040] Example 1: Preparation of Baiyao Decoction

[0041] Step 1: Wash the gallnuts and tea leaves separately and remove non-medicinal parts, moldy herbs, and impurities;

[0042] Step 2: Crush the cleaned gallnuts using a crusher. The crushed gallnuts will have no intact shells.

[0043] Step 3: Place the crushed gallnuts into a drying oven and dry them at 50-60℃ for 1-2 hours, turning them over during the drying process;

[0044] Step 4: Pass the crushed and dried gallnuts through a 24-mesh sieve to remove the dark brown aphids and gray powdery excrement;

[0045] Step 5: Pass the crushed gallnuts through a 100-mesh sieve. Continue crushing any gallnuts that do not pass through the 100-mesh sieve until all of them pass through it.

[0046] Step 6: Put the tea leaves into the jacketed kettle, add water at a ratio of 15 times the weight of the tea leaves, heat the jacketed kettle, and simmer for 20 minutes after the water boils. Then filter the liquid through an 80-mesh sieve.

[0047] Step 7: Repeat step 6 twice, and combine the three decoctions to obtain tea juice;

[0048] Step 8) Inoculate Aspergillus oryzae TJTSW001 into a slant test tube and incubate at 35℃ for 5 days. After growth, wash off the surface spores with sterile water and inoculate the tube into a liquid culture medium (the tea juice obtained in step 7) containing 10% gallnut powder and 3% glutamic acid. Incubate at 30-37℃ and 180 rpm for 3 days. Use this as the liquid inoculum.

[0049] Step 9) Mix the microbial inoculum obtained in step 8) with the gallnut powder, mix evenly, granulate, dry, and control the moisture content of the granules to 80%.

[0050] Step 10) Place the granules obtained in Step 9) at 33-37℃ and 70%-80% relative humidity. During the fermentation process, after mold grows, continue fermentation until the surface is covered with mold and then remove the granules.

[0051] Step 11) Sterilize the material cultured in Step 10) and vacuum dry it.

[0052] The tea leaves used in step 6 and the gallnut powder used in step 9 are in a mass ratio of 1:20.

[0053] Comparative Example 1: Sample 1 was prepared according to Example 2 of Chinese Patent CN103893392B.

[0054] Example 2: Samples 1-4 were prepared using the same process parameters as in Example 1. The difference between Example 2 and Example 1 lies in the composition of the culture medium in step 8), as shown in the table below.

[0055] sample Culture medium composition Sample 1 20% gallnut powder and 1% glutamic acid Sample 2 30% gallnut powder and 5% glutamic acid Sample 3 10% gallnut powder and 10% glutamic acid Sample 4 10% Gallnut Powder .

[0056] Example 3: Determination of gallic acid content in the Baiyao Decoction samples obtained from Examples 1, 2 and Comparative Example 1

[0057] Determined by high performance liquid chromatography (Appendix 0512, Part IV, Chinese Pharmacopoeia 2020 Edition).

[0058] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase, methanol-0.05% phosphoric acid solution (7:93) was used as the mobile phase, and the detection wavelength was 271 nm. The theoretical plate number, calculated based on the gallic acid peak, should not be less than 4000.

[0059] Preparation of reference solution: Take an appropriate amount of gallic acid reference standard, accurately weigh it, and add 50% methanol to prepare a solution containing 40 μg per ml.

[0060] Preparation and determination of the test solution: Accurately weigh approximately 0.1 g of the powder (passed through a No. 4 sieve), place it in a stoppered conical flask, accurately add 25 ml of mobile phase, seal tightly, shake well, weigh, and sonicate for 20 minutes (power 280 W, frequency 53 kHz, the same below). Cool and weigh again. Make up the weight loss with mobile phase, shake well, filter, accurately measure 1 ml of the filtrate into a 50 ml volumetric flask, dissolve and dilute to the mark with mobile phase, shake well, filter through a microporous membrane, and collect the filtrate to obtain the test solution.

[0061] The assay involves precisely pipetting 20 μl of both the reference solution and the test solution into a high-performance liquid chromatograph and measuring the results.

[0062] This product contains not less than 30.0% gallic acid (C7H6O5) on a dried basis.

[0063] The results of gallic acid content determination in the Baiyao Decoction samples obtained in Examples 1 and 2 and Comparative Example 1 are as follows:

[0064] sample Gallic acid content (%) sample Gallic acid content (%) Example 1 Sample 43.9％ Comparative Example 1 Sample 38.4％ Example 2 Sample 1 41.6％ Example 2 Sample 2 45.4％ Example 2 Sample 3 45.5％ Example 2 Sample 4 39.2％ .

[0065] As shown in the table above, compared with Comparative Example 1 (fermented with yeast), the gallic acid content was higher in all samples of Examples 1 and 2 where Aspergillus oryzae TJTSW001 was used to replace yeast in the preparation of Baiyao Decoction. In all samples of Examples 1 and 2, the gallic acid content increased with increasing glutamic acid content in the culture medium. Specifically, sample 4 of Example 2, which did not contain glutamic acid, had a significantly lower gallic acid content than the other groups. For the samples with glutamic acid added as a component of the culture medium, the gallic acid content increased with increasing glutamic acid concentration. Specifically, the gallic acid content of samples 1-3 of Example 2 increased sequentially.

[0066] Example 4: Determination of the antibacterial properties of the Baiyao Decoction samples obtained from Examples 1, 2 and Comparative Example 1

[0067] Baiyao Decoction is commonly used for pharyngitis and sore throat, chronic cough with excessive phlegm, chronic enteritis, canker sores, gingivitis, bloody dysentery, and thirst due to summer heat. Staphylococcus aureus is the most common pathogen in human purulent infections, easily causing pneumonia and enteritis, and is also a major pathogen in these diseases. Therefore, this experiment used Staphylococcus aureus to conduct an antibacterial test.

[0068] Preliminary experiments determined that the optimal concentration of the drug solution was 0.25 g / L. The concentration should not be too high, as this can cause the inhibition zones to stick together. The optimal dosage was 200 μL; too much or too little solution would negatively impact the experimental results.

[0069] The bacterial solution concentration was uniformly equivalent to a McFarland turbidity of 0.5, and the amount of bacterial solution added was 100 μL.

[0070] 4.1 Sample solution preparation

[0071] The samples of Baiyao Decoction obtained in Examples 1, 2 and Comparative Example 1 were pulverized and passed through an 80-mesh sieve. 25g of each sample was weighed out and decocted twice with 100mL of purified water for 30min each time. The filtrates from the two decoctions were combined and concentrated to a concentration of 0.25g / L for later use.

[0072] 4.2 Preparation of bacterial culture

[0073] Staphylococcus aureus (colony) was prepared into a bacterial solution with a concentration equivalent to 0.5 McFarland oil content.

[0074] 4.3 Oxford Cup Method and Experimental Results

[0075] First, add 20 mL of melted MH agar medium to a sterile Petri dish as the bottom layer. After solidification, add 100 μL of the prepared bacterial solution and spread it evenly with a spreader. Then, evenly place sterilized Oxford cups (stainless steel, 6 mm inner diameter, 7.8 mm outer diameter, and 10 mm height) into the Petri dish. Add 200 μL of the same solution to 6 of the Petri dishes, and add the corresponding solvent (sterile water) to 1 of the Petri dishes as a negative control, and label them accordingly. Place all Petri dishes directly in a 37°C incubator for 24 hours. Observe the colony production, measure the inhibition zone (including the 7.8 mm outer diameter of the Oxford cup) with calipers, calculate the average value, and conduct a comparative study.

[0076] sample Diameter of the inhibition zone (mm) sample Gallic acid content (%) Example 1 Sample 8.7 Comparative Example 1 Sample 8.3 Example 2 Sample 1 8.6 Example 2 Sample 2 9.2 Example 2 Sample 3 9.3 Example 2 Sample 4 8.5 Blank control 7.8 .

[0077] As shown in the table above, compared with the blank control group, all the Baiyao Decoction products exhibited certain antibacterial effects. Compared with Comparative Example 1 (fermentation with yeast), the samples in Examples 1 and 2, which used Aspergillus oryzae TJTSW001 to replace yeast in the preparation of Baiyao Decoction, showed larger inhibition zone diameters, indicating better antibacterial effects. In the samples of Examples 1 and 2, the diameter of the inhibition zone increased with the increase of glutamic acid content in the culture medium. Specifically, the inhibition zone diameter of Sample 4 in Example 2, which did not contain glutamic acid, was significantly lower than that of the other groups. For the samples in each group that added glutamic acid as a component of the culture medium, the diameter of the inhibition zone increased with the increase of glutamic acid concentration. Specifically, the inhibition zone diameters of Samples 1-3 in Example 2 increased sequentially.

[0078] The above experiments demonstrate that when Aspergillus oryzae TJTSW001 is used as the fermentation vitamin for processing Baiyao Decoction, and glutamic acid is added to the culture medium as a nitrogen source, the gallic acid content in the resulting Baiyao Decoction is significantly increased compared with that of Comparative Example 1 without added glutamic acid. At the same time, its antibacterial effect against Staphylococcus aureus is also significantly increased.

[0079] The above description of the invention and embodiments illustrates the basic principles, main features, and advantages of this patent application. Those skilled in the art should understand that this patent application is not limited to the above embodiments. The embodiments and descriptions in the specification are merely the optimal technical solutions of this patent application. Various changes and improvements can be made to this patent application without departing from the spirit and scope of this patent application. That is, the process of fermenting and processing Baiyao Decoction by controlling the ammonia nitrogen content in the fermentation environment through appropriate methods all fall within the scope of this patent application. The scope of protection of this patent application is defined by the appended claims and their equivalents.

Claims

1. A fermentation processing method for Baiyao Decoction based on adjusting the ammonia nitrogen content in the fermentation environment, the fermentation processing method comprising the following steps: 1) Boil tea leaves with water to make tea juice, crush gallnuts and remove impurities; 2) Take Aspergillus oryzae and inoculate it into a slant test tube. Incubate it at 35℃ for 5 days. After it has grown well, wash off the surface spores with sterile water and inoculate it into a liquid culture medium. The liquid culture medium is the tea juice obtained in step 1) with added gallnut powder and glutamic acid. Incubate it at 30-37℃ and 180 rpm for 3 days and use it as a liquid inoculum. 3) Mix the liquid seed obtained in step 2) with the gallnut powder, mix evenly, and then granulate; 4) Place the particles obtained in step 3) at 30-37℃ and incubate for 3 days; 5) Sterilize and vacuum dry the cultured material from step 4); The characteristic feature is that the Aspergillus species in step 2) is Aspergillus oryzae TJTSW001; The mass percentage of gallnut powder in step 2) is 10%-30%; The mass percentage of glutamic acid in step 2) is 1%-10%.

2. The fermentation and processing method as described in claim 1, characterized in that, In step 2), the mass percentage of gallnut powder is 10%.

3. The fermentation and processing method as described in claim 1, characterized in that, The mass percentage of glutamic acid in step 2) is 3%.

4. The fermentation and processing method as described in claim 1, characterized in that, The moisture content of the particles obtained in step 3) is adjusted to 50%-100%.

5. The fermentation and processing method as described in claim 1, characterized in that, The moisture content of the particles obtained in step 3) is adjusted to 80%.

6. The fermentation and processing method as described in claim 1, characterized in that, The tea leaves used in step 1) and the gallnut powder added in step 3) have a mass ratio of 1:

20.

7. The fermentation and processing method as described in claim 1, characterized in that, The cultivation conditions for step 4) are a cultivation temperature of 33-37℃ and a relative humidity of 70%-80%.

8. A decoction of a hundred herbs, characterized in that, Prepared by fermentation using any one of the fermentation methods described in claims 1-7.