Fermented bighead atractylodes rhizome black tea, preparation method and application

Abstract

The invention belongs to the technical field of health-care tea processing, and particularly relates to fermented atractylodes macrocephala koidz black tea, a preparation method and application. The invention provides a preparation method of fermented rhizoma atractylodis macrocephalae black tea, which comprises the following steps: mixing fresh rhizoma atractylodis macrocephalae and fresh tea leaves, and sequentially performing three-stage rolling and triple fermentation to obtain the fermented rhizoma atractylodis macrocephalae black tea. A black tea fermentation process is utilized, a flavor regulation and control technology is innovated, the bitter taste of the traditional bighead atractylodes rhizome black tea is improved, and the flavor characteristic of'black tea honey fragrance and bighead atractylodes rhizome medicine fragrance fused herbal sweet aftertaste 'is formed; meanwhile, the prepared fermented atractylodes macrocephala koidz black tea is high in atractylenolide and polysaccharide content, and has the health-care effects of regulating intestinal flora balance and enhancing immunity.

Description

Technical Field

[0001] This invention belongs to the field of health tea processing technology, specifically relating to a fermented Atractylodes macrocephala black tea, its preparation method, and its application. Background Technology

[0002] Atractylodes macrocephala, the dried rhizome of the plant Atractylodes macrocephala Koidz. (family Asteraceae), is a common Chinese medicinal herb with a long history of use. In traditional medicine, Atractylodes macrocephala is used to invigorate the spleen and replenish qi, dry dampness and promote diuresis, stop sweating, and calm the fetus. Modern research shows that Atractylodes macrocephala is rich in volatile oils, atractylodes lactones, polysaccharides, and other active ingredients, exhibiting good pharmacological effects in regulating gastrointestinal function, enhancing immunity, and anti-oxidation. However, its bitter taste and unpleasant palatability when consumed directly limit its widespread use in daily health maintenance.

[0003] Currently, there are some products on the market that combine Chinese medicinal herbs with tea, but most of them are simply mixed and fail to fully utilize the efficacy of the herbs, nor can they guarantee the stability and uniformity of the products. Among products that combine Atractylodes macrocephala with tea, there are issues such as a bitter taste from Atractylodes macrocephala and low levels of its active ingredients, atractylodes lactone and polysaccharides. Summary of the Invention

[0004] The purpose of this invention is to provide a fermented Atractylodes macrocephala black tea, its preparation method, and its application. The fermented Atractylodes macrocephala black tea prepared by the method of this invention has a good taste, reduces the bitterness of Atractylodes macrocephala, and has a high content of Atractylodes macrocephala lactones and polysaccharides.

[0005] To address the aforementioned technical problems, the present invention proposes the following technical solution:

[0006] This invention provides a method for preparing fermented Atractylodes macrocephala black tea, comprising: mixing fresh Atractylodes macrocephala and fresh tea leaves, and then sequentially performing a first stage of kneading, a second stage of kneading and a third stage of kneading, followed by triple fermentation to obtain fermented Atractylodes macrocephala black tea; the triple fermentation includes sequentially performing traditional fermentation, enzymatic hydrolysis and compound fermentation.

[0007] Preferably, the first stage of kneading has a pressure of 0.15-0.2 MPa, a rotation speed of 15-20 rpm, and a time of 10-15 min; the second stage of kneading has a pressure of 0.25-0.3 MPa, a rotation speed of 20-25 rpm, and a time of 10-15 min; and the third stage of kneading has a pressure of 0.15-0.2 MPa, a rotation speed of 10-15 rpm, and a time of 5-10 min.

[0008] Preferably, the compound enzyme used in the enzymatic hydrolysis includes cellulase, pectinase and xylanase; the mass ratio of cellulase, pectinase and xylanase is (2-4):(2-4):(1-3); the amount of the compound enzyme added is 0.05% to 0.15% of the total weight of fresh Atractylodes macrocephala and fresh tea leaves.

[0009] Preferably, the enzymatic hydrolysis temperature is 45–55°C and the time is 70–90 min.

[0010] Preferably, the compound fermentation uses microorganisms and extracts for fermentation together, the extracts including kombucha extract; the microorganisms including Lactobacillus plantarum, and the mass ratio of kombucha extract to Lactobacillus plantarum is 1:(1-5).

[0011] Preferably, the temperature of the compound fermentation is 28-32°C and the time is 16-24 hours.

[0012] Preferably, the compound fermentation process further includes sequentially subjecting the compound fermentation product to hot air drying, far-infrared and microwave synergistic irradiation, and vacuum drying; the hot air drying temperature is 50-60℃, the wind speed is 0.8-3m / s, and the hot air drying is carried out until the moisture content is 15%-18%; the hot air drying includes intermittent drying, and the parameters of the intermittent drying include: drying for 29-31 minutes, followed by a 4-6 minute break;

[0013] The far-infrared irradiation parameters are: wavelength 4–8 μm, intensity 0.7–0.9 W / cm². 2 The duration of combined far-infrared and microwave irradiation is 8–12 minutes; the power of the microwave is 290–310 W.

[0014] The vacuum drying temperature is 45–55°C, the vacuum degree is -0.08–-0.09 MPa, and the vacuum drying is carried out until the moisture content is ≤6%.

[0015] Preferably, the mass ratio of fresh Atractylodes macrocephala to fresh tea leaves during mixing is (5-7):(3-5).

[0016] This invention provides fermented Atractylodes macrocephala black tea prepared by the method described in the above technical solution.

[0017] This invention provides the application of the preparation method described in the above technical solution in at least one of the following:

[0018] 1) Increase the content of active ingredients atractylodes lactone and polysaccharides in fermented Atractylodes macrocephala black tea;

[0019] 2) Reduce the bitterness of fermented Atractylodes macrocephala black tea;

[0020] 3) Enhance the antioxidant effect of fermented Atractylodes macrocephala black tea.

[0021] The beneficial effects of this invention are as follows: This invention provides a method for preparing fermented Atractylodes macrocephala black tea. Fresh Atractylodes macrocephala and fresh tea leaves are mixed and then subjected to a first stage of rolling, a second stage of rolling, and a third stage of rolling, followed by triple fermentation to obtain fermented Atractylodes macrocephala black tea. The triple fermentation includes: traditional fermentation, enzymatic hydrolysis, and compound fermentation. This invention uses three stages of rolling during the fermentation process to break down the cell walls of the fresh Atractylodes macrocephala and tea leaves, allowing tea juice (such as polyphenols and amino acids) to be released, and releasing active ingredients (polysaccharides and atractylodes lactones, etc.) from Atractylodes macrocephala, providing a basis for subsequent fermentation or oxidation reactions. Simultaneously, rolling also activates enzymes in the tea leaves (such as polyphenol oxidase), promoting chemical changes (such as polyphenol oxidation and polymerization), forming the unique color, aroma, and flavor characteristics of the tea. Atractylodes macrocephala contains atractylone, which has drying properties and is easily converted into atractylodes lactone through fermentation. Atractylodes lactone is the main active ingredient in Atractylodes macrocephala for its spleen-strengthening and stomach-invigorating effects.

[0022] Furthermore, enzymatic hydrolysis efficiently breaks down the complex cell walls of Atractylodes macrocephala and black tea, increasing the dissolution rate of polysaccharides and atractylodes lactones in Atractylodes macrocephala. Compound fermentation enhances the flavor of Atractylodes macrocephala black tea, reduces bitterness, and the fermentation metabolites also regulate intestinal flora balance and enhance immunity. This invention, by fermenting Atractylodes macrocephala with fresh tea leaves, helps Atractylodes macrocephala better exert its spleen-strengthening, qi-tonifying, dampness-drying, and diuretic effects, while also improving the richness of the flavor of Atractylodes macrocephala black tea. Detailed Implementation

[0023] This invention provides a method for preparing fermented Atractylodes macrocephala black tea, comprising: mixing fresh Atractylodes macrocephala and fresh tea leaves, and then sequentially performing a first stage of kneading, a second stage of kneading and a third stage of kneading, followed by triple fermentation to obtain fermented Atractylodes macrocephala black tea; the triple fermentation includes sequentially performing traditional fermentation, enzymatic hydrolysis and compound fermentation.

[0024] As an optional implementation, the mass ratio of fresh Atractylodes macrocephala and fresh tea leaves in this invention is (5-7):(3-5), or (5.5-6.5):(3.5-4.5), more preferably 6:4. This mass ratio improves the taste of fermented Atractylodes macrocephala black tea. The fresh Atractylodes macrocephala in this invention refers to Atractylodes macrocephala harvested in winter after all leaves have dried and then refrigerated. The fresh tea leaves in this invention are preferably fresh leaves harvested between Qingming Festival and Guyu Festival. This invention utilizes the processing technology of black tea to process Atractylodes macrocephala, which can convert the drying components of Atractylodes macrocephala into the active ingredient atractylodes lactone. As an optional implementation, before rolling, this invention pre-treats the fresh Atractylodes macrocephala and fresh tea leaves. The pre-treatment includes slicing and withering. This invention does not have specific limitations on the slicing and withering methods; conventional methods are acceptable. This invention only slices the fresh Atractylodes macrocephala, with a slice thickness of 2-4 mm, more preferably 3 mm. The slices meet the requirements of the pharmacopoeia and can promote the dissolution of active ingredients in Atractylodes macrocephala. This invention involves mixing fresh Atractylodes macrocephala slices and fresh tea leaves for compound withering. The pH of the mixture of fresh Atractylodes macrocephala slices and fresh tea leaves is adjusted to 4.5–5.5 before compound withering, more preferably 5. The compound withering temperature is 25–28℃, or 26–27℃; the humidity is 70%–75%, or 72%–74%; and the time is 8–10 hours, or 8.5–9.5 hours. In a specific embodiment of this invention, the moisture content of Atractylodes macrocephala after compound withering is 60%, and the moisture content of the fresh tea leaves is 65%. The purpose of compound withering is to reduce the moisture content of the fresh Atractylodes macrocephala and fresh tea leaves, promote subsequent fermentation, and enhance the aroma of the fermented Atractylodes macrocephala black tea. The fermented Atractylodes macrocephala black tea of ​​this invention is a fermented Atractylodes macrocephala black tea.

[0025] As an optional implementation, the present invention subjectes the composite withering product to a first-stage kneading, a second-stage kneading, and a third-stage kneading to obtain a kneaded product. The first-stage kneading, second-stage kneading, and third-stage kneading of the present invention all employ a dynamic pressure kneading process. The pressure of the first-stage kneading is 0.15–0.2 MPa, or 0.16–0.18 MPa; the rotation speed is 15–20 rpm, or 16–18 rpm; and the time is 10–15 min, or 12–13 min. The pressure of the second-stage kneading is 0.25–0.3 MPa, or 0.26–0.28 MPa; the rotation speed is 20–25 rpm, or 22–23 rpm; and the time is 10–15 min, or 12–13 min. The pressure of the third stage of kneading described in this invention is 0.15–0.2 MPa, or 0.16–0.18 MPa; the rotation speed is 10–15 rpm, or 12–13 rpm; and the time is 5–10 min, or 6–8 min. This invention achieves a light-heavy-light kneading process by sequentially performing three stages of kneading: light kneading, heavy kneading, and then light kneading again. This kneading process can break down the cell walls of Atractylodes macrocephala and black tea, causing polyphenols, amino acids, polysaccharides, and atractylodes lactones in the tea leaves to be released, providing a basis for subsequent fermentation or oxidation reactions. Simultaneously, kneading can activate enzymes in the tea leaves (such as polyphenol oxidase), promoting chemical changes (such as polyphenol oxidative polymerization), forming the unique color, aroma, and flavor characteristics of the tea. It also activates the conversion of atractylone in Atractylodes macrocephala, reducing its drying properties and increasing the activity of atractylodes lactones.

[0026] This invention involves a triple fermentation process on the resulting rolled material. The triple fermentation includes sequential conventional fermentation, enzymatic hydrolysis, and compound fermentation to obtain fermented Atractylodes macrocephala black tea. During this triple fermentation process, the pH is preferably measured every 4 hours to maintain it between 4.8 and 5.2. Fermentation will stop if the pH is too high. Adjusting the pH and controlling the fermentation temperature can optimize the flavor of the resulting fermented product.

[0027] This invention involves traditional fermentation of the obtained kneaded material. The traditional fermentation of this invention involves directly fermenting the obtained kneaded material without adding exogenous microbial agents, utilizing the naturally occurring microorganisms in the raw materials. The traditional fermentation temperature of this invention is 24–26°C, more preferably 25°C; the humidity is 85%–95%, more preferably 90%; the time is 2–3 hours, more preferably 2.5 hours; the dissolved oxygen content during traditional fermentation is controlled at 2–5 mg / L, more preferably 4 mg / L; the function of the traditional fermentation in this invention is to promote the oxidation of polyphenols in black tea and Atractylodes macrocephala.

[0028] After traditional fermentation, the product obtained from traditional fermentation is mixed with a compound enzyme for enzymatic hydrolysis to obtain the hydrolysate. As an optional implementation, the compound enzyme used in the enzymatic hydrolysis of this invention includes cellulase, pectinase, and xylanase. The mass ratio of cellulase, pectinase, and xylanase in the compound enzyme of this invention is (2-4):(2-4):(1-3), more preferably 3:2:1. The cellulase activity before addition is ≥50000 U / g; the pectinase activity before addition is ≥30000 U / g; and the xylanase activity before addition is ≥100000 U / g. This invention does not have specific limitations on the source of purchase for cellulase, pectinase, and xylanase; conventional products are acceptable.

[0029] In a specific embodiment of the present invention, the cellulase activity before addition is 50,000 U / g, brand name Jizhi, product number AC8271-500g; the pectinase activity before addition is 30,000 U / g, brand name Maclean; and the xylanase activity before addition is 100,000 U / g, brand name Maclean. Based on the total mass of fresh Atractylodes macrocephala and fresh tea leaves, the amount of the compound enzymes added in this invention is 0.05% to 0.15% of the total mass of the raw materials, more preferably 0.1%. The Atractylodes macrocephala of the present invention is rich in cellulose. Cellulase can decompose cellulose, hydrolyzing it into small molecule sugars such as glucose and cellobiose. Pectinase can decompose the pectin in Atractylodes macrocephala and black tea to form galacturonic acid, promoting fermentation; xylanase can degrade the xylan in Atractylodes macrocephala and black tea to obtain oligosaccharides and xylose, allowing for better fermentation of Atractylodes macrocephala and black tea. Simultaneously, cellulase and pectinase can digest the cell walls of tea leaves, increasing the content of soluble sugars and water-soluble extracts in the tea. The mass ratio of cellulase, pectinase, and xylanase set in this invention can achieve the synergistic effect of the complex enzymes, further efficiently decomposing the complex structure of plant cell walls.

[0030] As an optional implementation, the enzymatic hydrolysis temperature of the present invention is 45–55°C, or 40–50°C; the time is 70–90 min, more preferably 75–85 min, and even more preferably 80 min. During the enzymatic hydrolysis process of the present invention, an acidic pH adjuster is preferably used to adjust the pH to 4.5–5.5, wherein the acidic pH adjuster comprises a 0.1 mol / L citric acid-sodium citrate buffer system. The purpose of adjusting the pH is to promote the fermentation of tea leaves.

[0031] The synergistic effect of three-stage kneading and triple fermentation enhances the taste of fermented Atractylodes macrocephala black tea, forming the unique color, aroma and flavor characteristics of the tea, and increasing the active ingredient atractylodes lactone.

[0032] After enzymatic hydrolysis, the hydrolysate is mixed with the extract and microorganisms for compound fermentation. As an optional embodiment, the microorganisms used in the compound fermentation include *Lactobacillus plantarum*. As an optional embodiment, the extract includes kombucha extract. Fermentation of the kombucha extract and *Lactobacillus plantarum* can adjust the flavor of fermented Atractylodes macrocephala black tea, reduce bitterness, and simultaneously enable the fermented Atractylodes macrocephala black tea to regulate intestinal flora balance and enhance immunity. This invention does not have specific limitations on the source and preparation method of the kombucha extract and *Lactobacillus plantarum*; conventional products are acceptable. In a specific embodiment of the present invention, the kombucha extract is Kombucha Extract, branded as Selleck, with product number E3279-5mg (see "https: / / www.rjmart.cn / productDetail?productId=200318571054&suppId=80608&source=1&eId=996516217944543241"); the Lactobacillus plantarum agent was purchased from BNCC, product number: BNCC134285.

[0033] As an optional implementation, the present invention mixes the kombucha extract and *Lactobacillus plantarum* inoculum to obtain a mixed inoculum, which is then inoculated into the enzymatic hydrolysate for compound fermentation. The inoculation amount of the kombucha extract in the present invention is 0.3% to 0.8% of the mass of the mixture of fresh *Atractylodes macrocephala* and fresh tea leaves. The mass ratio of kombucha extract to *Lactobacillus plantarum* inoculum in the mixed inoculum is 1:(1-5), or 1:(1-3), more preferably 1:2. This mass ratio setting allows for the synergistic effect of the kombucha extract and *Lactobacillus plantarum* during fermentation, further improving the taste of the fermented *Atractylodes macrocephala* black tea.

[0034] As an optional implementation, the temperature of the compound fermentation in this invention is 28–32°C, or 29–30°C, and the fermentation time is 16–24 hours, or 18–20 hours. During the compound fermentation process, the dissolved oxygen level is controlled at 5–7 mg / L, more preferably 6 mg / L. The compound fermentation of this invention can simultaneously increase the dissolution rate of the active ingredients atractylodes lactone and polysaccharides, improve the taste, resulting in a better taste in fermented Atractylodes macrocephala black tea, reduced bitterness, higher total atractylodes lactone content, and increased reducing sugar content.

[0035] As an optional implementation, the compound fermentation of the present invention further includes sequentially subjecting the compound fermentation product to hot air drying, far-infrared and microwave synergistic treatment, and vacuum drying.

[0036] The hot air drying temperature described in this invention is 50–60°C, or 55–58°C; the air velocity is 0.8–3 m / s, or 1.2–2 m / s; the hot air is dried to a moisture content of 15%–18%, or 16%–17%. Since Atractylodes macrocephala contains volatile oil, the hot air drying temperature cannot exceed 60°C.

[0037] As an optional implementation, the hot air drying of this invention is intermittent drying. Specific parameters for intermittent drying include: drying for 29-31 minutes followed by a 4-6 minute pause. The drying time for intermittent drying in this invention is 29-31 minutes, more preferably 30 minutes; the stop time for intermittent drying in this invention is 4-6 minutes, more preferably 5 minutes. Atractylodes macrocephala is rich in polysaccharides, and continuous high-temperature drying easily leads to surface crusting and difficulty in internal moisture migration. Intermittent drying can avoid surface hardening and internal moisture retention. The function of the hot air drying in this invention is to retain the effective components of fermented Atractylodes macrocephala black tea while achieving preliminary drying. This invention applies far-infrared irradiation and microwave synergistic treatment to the hot air-dried product. The low temperature of far-infrared irradiation can achieve uniform heating and provide gentle and uniform heat conduction, promoting the slow release of aroma precursors (such as glycosides, amino acids, etc.) in tea leaves and Atractylodes macrocephala, avoiding high-temperature damage to volatile components. Combined with the rapid internal heating effect of microwaves, it accelerates moisture evaporation and aroma diffusion, and may also trigger Maillard reactions or caramelization reactions, enhancing the roasted flavor. The synergistic treatment of far-infrared irradiation and microwaves can shorten the aroma enhancement time and improve the aroma of fermented Atractylodes macrocephala black tea. The wavelength of the far-infrared irradiation described in this invention is 4–8 μm, or 6–7 μm; the intensity is 0.7–0.9 W / cm². 2 It can also be 0.8 W / cm 2 The microwave power is 290–310W, more preferably 300W; the synergistic irradiation time of far-infrared and microwave is 8–12 minutes, or 9–11 minutes, more preferably 10 minutes. This invention vacuum-dries the product obtained from far-infrared irradiation, preserving the active ingredients while achieving the shaping of fermented Atractylodes macrocephala black tea.

[0038] As an optional implementation, the vacuum drying temperature of the present invention is 45-55℃, or 48-53℃; the vacuum degree is -0.09--0.08MPa, or -0.087--0.082MPa, and the vacuum drying is carried out until the moisture content is ≤6%. After vacuum drying, the present invention sterilizes and packages the vacuum-dried product, thereby enhancing the quality of the fermented Atractylodes macrocephala black tea and extending its shelf life through sterilization and packaging.

[0039] As an optional implementation method, the sterilization method of the present invention is a combination of UV-C ultraviolet light and ozone, wherein the wavelength of the UV-C ultraviolet light in the present invention is 254nm.

[0040] This invention provides fermented Atractylodes macrocephala black tea prepared by the method described above. The fermented Atractylodes macrocephala black tea of ​​this invention is prepared through a complex enzymatic hydrolysis and multi-stage fermentation process, which improves the release rate of Atractylodes macrocephala's active ingredients and endows the tea with multiple health benefits.

[0041] This invention provides the application of the preparation method described in the above technical solution in at least one of the following:

[0042] 1) Increase the content of active ingredients atractylodes lactone and polysaccharides in fermented Atractylodes macrocephala black tea;

[0043] 2) Reduce the bitterness of fermented Atractylodes macrocephala black tea;

[0044] 3) Enhance the antioxidant effect of fermented Atractylodes macrocephala black tea.

[0045] Atractylodes macrocephala is a commonly used tonic in traditional Chinese medicine, and is known as "Southern Atractylodes and Northern Ginseng". Atractylodes macrocephala is warm in nature, sweet, slightly pungent and bitter in taste, and enters the spleen and stomach meridians. It has the effects of diuresis and swelling reduction, strengthening the exterior and stopping sweating, drying dampness and strengthening the spleen, and anti-oxidation. Traditional Atractylodes macrocephala black tea is usually made from raw Atractylodes macrocephala, stir-fried Atractylodes macrocephala, and charred Atractylodes macrocephala. The taste of Atractylodes macrocephala black tea is obviously bitter and astringent, and consumers have low acceptance. The dissolution rate of the key active ingredient polysaccharide is less than 50% (see "Yang Ying, Wei Mengxin, Wu Yaoye, et al. Research progress on extraction, separation, chemical composition and pharmacological effects of Atractylodes macrocephala polysaccharide [J]. Chinese Traditional and Herbal Drugs, 2021, 52(02):578-584. The polysaccharide yield by water decoction method in the article is 3.13%-35.10%"). The function of single Atractylodes macrocephala raw material is limited and lacks synergistic effect. The fermentation process of black tea only targets the oxidation of tea polyphenols and does not combine it with traditional Chinese medicine. The traditional fermentation process of Atractylodes macrocephala cannot effectively decompose the cell structure of Atractylodes macrocephala, making it difficult to achieve the synergistic transformation of the functional components of black tea and Atractylodes macrocephala. This invention involves a three-stage kneading process between fresh Atractylodes macrocephala and fresh tea leaves, followed by triple fermentation. Enzymatic hydrolysis and oxidation during this triple fermentation decompose the cell structure of Atractylodes macrocephala, promoting the conversion of atractylone to atractylenolide and further decomposing the cell structure. The combined fermentation process, integrating traditional and compound fermentation, offers synergistic advantages in degrading macromolecules. Through the metabolic activities of different microorganisms and the complementarity of enzyme systems, complex organic matter such as cellulose, protein, fat, and polysaccharides can be decomposed more efficiently, reducing the molecular weight of polysaccharides in fermented Atractylodes macrocephala black tea from >50kDa to 10-20kDa, increasing bioavailability by three times. The compound fermentation process modifies the cell structure of Atractylodes macrocephala, increasing the dissolution rate of atractylenolide and polysaccharides to over 80%. The Atractylodes macrocephala black tea obtained by this invention has high contents of atractylenolide I, atractylenolide II, and atractylenolide III, with a total content reaching 2.6mg / g. The combination of black tea and Atractylodes macrocephala polysaccharides in the Atractylodes macrocephala black tea obtained by this invention forms a novel antioxidant complex, enhancing the antioxidant effect of the fermented Atractylodes macrocephala black tea. This invention establishes a scientific ratio system for Atractylodes macrocephala and black tea, achieving multiple benefits including spleen and stomach protection, antioxidant effects, and immune regulation. It also innovates flavor control technology to improve the bitter taste of traditional Atractylodes macrocephala black tea, enabling fermented Atractylodes macrocephala black tea to develop a "herbal sweetness that blends the honey aroma of black tea with the medicinal aroma of Atractylodes macrocephala." This herbal sweetness is attributed to the catechins in the tea leaves, which can mitigate the bitterness of Atractylodes macrocephala and highlight its sweetness; the aroma and polysaccharides of Atractylodes macrocephala may enhance the mellowness of the tea soup and prolong the sweetness.

[0046] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0047] Example 1

[0048] 1. By weight, the raw materials consist of: 6 kg of fresh Atractylodes macrocephala and 4 kg of fresh tea leaves.

[0049] 2. Preparation method:

[0050] (1) Raw material pretreatment:

[0051] Fresh Atractylodes macrocephala is sliced ​​to a thickness of 2-4 mm; then mixed with fresh tea leaves to obtain a mixture.

[0052] (2) Compound withering: The resulting mixture was adjusted to pH 5 and subjected to compound withering at 26°C and 72% humidity for 8 hours until the moisture content of Atractylodes macrocephala was 60% and the moisture content of fresh tea leaves was 65%.

[0053] (3) Synergistic rolling: The withered material obtained in step (2) is subjected to three stages of synergistic rolling using a dynamic pressure rolling process, with the following parameters:

[0054] First stage kneading: light pressure 0.2MPa, speed 20rpm, 15min;

[0055] Second stage kneading: medium pressure 0.3MPa, speed 25rpm, 10min;

[0056] The third stage of kneading: light pressure 0.2MPa, rotation speed 15rpm, 5min.

[0057] (4) The mixture obtained from the synergistic kneading undergoes a triple fermentation, specifically:

[0058] The first stage (the product obtained from kneading black tea and Atractylodes macrocephala is directly fermented without the addition of exogenous microbial agents, i.e., traditional fermentation): the temperature of traditional fermentation is 25℃, the humidity is 90%, and the time is 2.5h; the purpose is to promote the oxidation of polyphenols. After the traditional fermentation is completed, the product of the first stage of fermentation is obtained; the dissolved oxygen content is controlled at 4mg / L.

[0059] The second stage of compound enzymatic hydrolysis: A compound enzyme is added to the fermentation product of the first stage, at a rate of 0.1% of the total mass of fresh Atractylodes macrocephala and fresh tea leaves; during the compound enzymatic hydrolysis process, a 0.1 mol / L citric acid-sodium citrate buffer system is used to control the pH at 4.5-5.5; the dissolved oxygen content is controlled at 6 mg / L.

[0060] The complex enzyme consists of cellulase, pectinase, and xylanase, with a mass ratio of 3:2:1. Enzymatic hydrolysis was performed at 50±1℃ for 80 min to obtain the complex enzymatic hydrolysis product. The enzyme activities of the added cellulase, pectinase, and xylanase were 50000 U / g, 30000 U / g, and 100000 U / g, respectively.

[0061] Third-stage compound fermentation:

[0062] In the second stage of enzymatic hydrolysis, kombucha extract and Lactobacillus plantarum were inoculated into the product for compound fermentation, with the mass ratio of kombucha extract to Lactobacillus plantarum being 1:2.

[0063] The total inoculation amount of kombucha extract and Lactobacillus plantarum was 5% of the total weight of fresh Atractylodes macrocephala and fresh tea leaves;

[0064] The combined fermentation temperature was 28℃, and the fermentation time was 24 hours. The dissolved oxygen level in the third stage was controlled at 6 mg / L.

[0065] During the triple fermentation stage, the pH was measured every 4 hours and maintained at a pH of 4.8–5.2.

[0066] (5) The product obtained from compound fermentation undergoes post-processing, which includes initial drying, aroma enhancement, and shaping. The specific process parameters are as follows:

[0067] Initial drying: After the fermented product is separated into layers, it is dried with hot air at a layer thickness of 3 cm. The product is dried intermittently at 55℃ until the moisture content of the fermented product reaches 15%, and the air velocity is 0.8 m / s. The intermittent hot air drying mode is adopted, with a cycle of 35 minutes. After drying for 30 minutes, there is a 5-minute pause, and the cycle is repeated.

[0068] Aroma enhancement: The product obtained from initial drying is subjected to far-infrared microwave irradiation for 10 minutes. The wavelength of the far-infrared treatment is 4-8 μm, the temperature is 75℃, and the radiation intensity is 0.8 W / cm². 2 The microwave power is 300W.

[0069] Shaping: The aroma-enhancing material is shaped under the following conditions: vacuum drying at 50℃ until the moisture content is ≤6%, the vacuum degree of vacuum drying is -0.085MPa, and the material tray speed is 4rpm, to obtain fermented Atractylodes macrocephala black tea.

[0070] (6) Quality Enhancement Process:

[0071] The resulting Atractylodes macrocephala black tea was sterilized using UV-C ultraviolet light (254nm) combined with ozone and packaged with nitrogen (oxygen residue ≤0.5%).

[0072] Example 2

[0073] 1. By weight, the raw materials consist of: 5.5 kg of fresh Atractylodes macrocephala and 4.5 kg of fresh tea leaves.

[0074] 2. Preparation method: Same as in Example 1.

[0075] Example 3

[0076] 1. By weight, the raw materials consist of: 6.5 kg of fresh Atractylodes macrocephala and 3.5 kg of fresh tea leaves.

[0077] 2. Preparation method: Same as in Example 1.

[0078] Comparative Example 1

[0079] 1. By weight, the raw materials consist of: 6.0 kg of fresh Atractylodes macrocephala and 4.0 kg of fresh tea leaves.

[0080] 2. Preparation method.

[0081] (1) Raw material pretreatment: Same as in Example 1.

[0082] (2) Compound withering: Same as Example 1.

[0083] (3) Synergistic kneading: Same as in Example 1.

[0084] (4) Ferment the product obtained from the combined kneading process, specifically as follows:

[0085] The mixture of black tea and Atractylodes macrocephala is directly fermented without the addition of exogenous microorganisms, i.e., traditional fermentation: the temperature of traditional fermentation is 24-26℃, the humidity is 90%, and the time is 2-3 hours; the purpose is to promote polyphenol oxidation. The pH is measured every 4 hours and maintained at 4.8-5.2.

[0086] (5) The fermented product undergoes post-processing, which includes initial drying, aroma enhancement, and shaping. The specific process parameters are as follows:

[0087] Initial drying: After the fermented product is separated into layers, it is dried with hot air at a layer thickness of 3cm. The product is dried at 55℃ until the moisture content of the fermented product is 15%. The air velocity of the hot air is 0.8m / s. The parameters for intermittent hot air drying are 5 minutes off every 30 minutes.

[0088] Flavoring: Same as in Example 1.

[0089] Finalization: Same as Example 1.

[0090] (6) Quality enhancement process: Same as Example 1.

[0091] Comparative Example 2

[0092] 1. By weight, the raw material composition is: 6.0 kg of fresh Atractylodes macrocephala.

[0093] 2. Preparation method: Same as in Example 1.

[0094] Comparative Example 3: Traditional Fermentation Process

[0095] 1. By weight, the raw materials consist of: 6.0 kg of fresh Atractylodes macrocephala and 4.0 kg of fresh tea leaves.

[0096] 2. The preparation method is as follows:

[0097] The enzyme used in the enzymatic hydrolysis was cellulase, with an addition amount of 0.2%, an enzymatic hydrolysis temperature of 45℃, and an enzymatic hydrolysis time of 6 hours.

[0098] (1) Raw material pretreatment:

[0099] Fresh Atractylodes macrocephala is sliced ​​to a thickness of 2-4 mm; then mixed with fresh tea leaves to obtain a mixture.

[0100] (2) Compound withering: Same as Example 1.

[0101] (3) Synergistic kneading: Same as in Example 1.

[0102] (4) The product obtained from the synergistic kneading undergoes a first-stage and second-stage complex enzymatic hydrolysis transformation, specifically as follows:

[0103] The first stage (the product obtained from kneading black tea and Atractylodes macrocephala is directly fermented without the addition of exogenous microbial agents, i.e., traditional fermentation): the temperature of traditional fermentation is 25℃, the humidity is 90%, and the time is 2.5h; the purpose is to promote the oxidation of polyphenols. After the traditional fermentation is completed, the product of the first stage of fermentation is obtained; the dissolved oxygen content is controlled at 4mg / L.

[0104] The second stage of enzymatic hydrolysis: Cellulase was added to the fermentation product of the first stage at a rate of 0.1% of the total mass of fresh Atractylodes macrocephala and fresh tea leaves. During the enzymatic hydrolysis, a 0.1 mol / L citric acid-sodium citrate buffer system was used to control the pH at 4.5–5.5. The dissolved oxygen content was controlled at 6 mg / L. The enzymatic hydrolysis was carried out at 50 ± 1 °C for 80 min to obtain the enzymatic hydrolysis product. The cellulase source was the same as in Example 1, and the enzyme activity was 50,000 U / g.

[0105] (5) The product obtained from the second stage of compound enzymatic hydrolysis is post-processed. The post-processing process is initial drying, aroma enhancement and shaping. The process parameters are the same as in Example 1.

[0106] (6) Quality enhancement process: Same as Example 1.

[0107] Test results

[0108] The polysaccharide and total phenol contents of the Atractylodes macrocephala black tea obtained in Examples 1-3 and Comparative Examples 1-3 were determined. The determination method was carried out in accordance with the existing technology "Huang Hao, Yu Penghui, Zhao Xi, Zhong Ni, He Yufei, Zheng Hongfa. Analysis of quality components of black tea from different producing areas in Hunan [J]. Tea Communication, 2019, 46(02):208-214."

[0109] The contents of atractylodes I, atractylodes II, and atractylodes III in the Atractylodes macrocephala black tea obtained in Examples 1-3 and Comparative Examples 1-3 were determined by high performance liquid chromatography (HPLC). The determination method is described in "Ding Yixue, Xu Jixiao, Wu Wei, et al. Simultaneous determination of four sesquiterpenes in Atractylodes macrocephala by HPLC [J]. Chinese Traditional and Herbal Drugs, 2020, 42(04):927-931."

[0110] The DPPH scavenging rate, amylase inhibition rate and sensory scores of the Atractylodes macrocephala black tea obtained in Examples 1-3 and Comparative Examples 1-3 were determined. The determination method was carried out in accordance with the existing technology "Zhou Lijing, Hou Caiyun, Qiao Yanhui, Zhao Jing. Study on functional characteristics of Pu'er tea with different storage years [J]. Food Science, 2010, 31(13):19-22."

[0111] The shelf life of the fermented Atractylodes macrocephala black tea obtained in Examples 1-3 and Comparative Examples 1-3 was determined. The determination method was based on the group standard "Shelf Life of Black Tea" (T / CTMA029-2021) and the "Enterprise Consensus on the Implementation of Label Management of Chinese Herbal Medicine Pieces (First Edition)" of Zhejiang Provincial Association of Chinese Medicine Industry.

[0112] The results of the above measurements are shown in Tables 1-3. It can be seen that fermented Atractylodes macrocephala black tea has high contents of atractylodes macrocephala lactone I, II, and III, as well as high polysaccharide content; it also exhibits high DPPH scavenging rate and good antioxidant properties. Furthermore, it has a high amylase inhibition rate, which can assist in blood glucose regulation.

[0113] Table 1 Comparison of the main active ingredient contents of Atractylodes macrocephala black tea obtained in Examples 1-3 and Comparative Examples 1-3

[0114]

[0115] Table 2 Comparison of the functional properties of Atractylodes macrocephala black tea obtained in Examples 1-3 and Comparative Examples 1-3

[0116] Group DPPH removal rate (%) Amylase inhibition rate (%) Sensory rating (1-10 points) Shelf life (months) Example 1 89.7±1.2 65.3±1.5 8.5±0.3 24 Example 2 88.3±1.1 63.8±1.4 8.3±0.3 22 Example 3 92.3±1.3 66.1±1.6 8.6±0.3 25 Comparative Example 1 75.3±1.0 52.4±1.2 6.2±0.4 15 Comparative Example 2 68.3±0.9 58.7±1.3 7.1±0.3 18 Comparative Example 3 60.5±0.8 45.2±1.0 5.8±0.5 12

[0117] Table 3 shows the optimization of compound fermentation parameters for Atractylodes macrocephala black tea obtained in Examples 1-3 and Comparative Examples 1-3.

[0118]

[0119]

[0120] The stability of Atractylodes macrocephala black tea was tested by accelerated testing at 45°C and 75% humidity. The results are shown in Table 4.

[0121] The content of atractylenolide was determined by high performance liquid chromatography. The component content and retention rate were calculated by comparing the peak areas of atractylenolide I, II, and III before and after preparation. The formula was: Retention rate (%) = Atractylenolide content before fermentation / Atractylenolide content after fermentation × 100%.

[0122] Sensory evaluation: The sample was brewed and scored independently by 5 judges, and the average score was taken.

[0123] Microbiological indicators: Refer to GB 4789.2-2016 "National Food Safety Standard - Microbiological Examination of Food - Determination of Total Colony Count".

[0124] Table 4. Stability test results of the Atractylodes macrocephala black tea obtained in Example 1

[0125] Time (month) Atractylodes lactone retention rate (%) Sensory rating Microbiological indicators (CFU / g) 0 100 9.2 <10 3 98.2 9.0 30 6 95.7 8.7 120 8 92.3 7.9 800 12 87.6 7.5 2200

[0126] As shown in Table 4, the Atractylodes macrocephala black tea product of this invention maintains good quality even after 12 months under accelerated conditions, and its shelf life at room temperature is predicted to be more than 48 months.

[0127] The formula for calculating shelf life at room temperature is as follows:

[0128] Shelf life at room temperature = Shelf life at accelerated temperature × Q10 △T / 10

[0129] Where △T is the temperature difference between the acceleration temperature and the room temperature, and Q10 value: the reaction rate doubles for every 10℃ increase in temperature.

[0130] Comparative Example 4

[0131] No complex enzymatic hydrolysis was performed; other parameters were the same as in Example 1.

[0132] Comparative Example 5

[0133] Traditional fermentation was not performed, and other parameters were the same as in Example 1.

[0134] Comparative Example 6

[0135] No compound fermentation was performed; other parameters were the same as in Example 1.

[0136] Comparative Example 7

[0137] The three-stage synergistic kneading was not performed, and other parameters were the same as in Example 1.

[0138] Table 5 shows the atractylodes lactone content, sensory scores, and microbial indicators of the fermented Atractylodes macrocephala black tea obtained from Comparative Examples 4-7 after 24 months of storage.

[0139] Table 5 shows the determination results of fermented Atractylodes macrocephala black tea obtained from Comparative Examples 4-7 and Example 1 after 24 months of storage.

[0140] Storage period (24 months) Atractylodes lactone retention rate (%) Sensory rating (1-10 points) Microbiological indicators (CFU / g) Comparative Example 4 82.2 7.1 870 Comparative Example 5 68.3 6.8 860 Comparative Example 6 67.2 5.8 850 Comparative Example 7 72.5 6.3 910 Example 1 88.6 7.5 850

[0141] In Example 1, the molecular weight of the polysaccharides in the fermented Atractylodes macrocephala black tea was reduced from >50 kDa to 10-20 kDa, and the bioavailability was increased by 3 times. Determination methods: Polysaccharide determination was performed using high-performance gel permeation chromatography (HPGPC); bioavailability was determined using an in vitro simulated digestion model (Caco-2 cell model).

[0142] In Example 1, the viable bacteria count (plate count method) of the fermented Atractylodes macrocephala black tea was determined, showing an 80% increase in the proliferation rate of intestinal probiotics (Lactobacillus plantarum). The viable bacteria count was determined according to GB 4789.35-2016 "Food Microbiology Examination - Lactic Acid Bacteria Examination". The proliferation rate (%) was calculated as: viable bacteria count in the Atractylodes macrocephala black tea group - viable bacteria count in the control group / viable bacteria count in the control group × 100%.

[0143] In summary, this invention establishes a scientific ratio system for Atractylodes macrocephala and black tea. The fermented Atractylodes macrocephala black tea of ​​this invention is a fully fermented tea, made from suitable new tea buds and leaves and fresh Atractylodes macrocephala as raw materials, refined through a series of processes including withering, rolling, fermentation, and drying. During the fermentation process, the tea polyphenols in the fresh tea leaves undergo enzymatic oxidation, producing new components such as thearubigins, which give the black tea its unique color, aroma, and flavor. The fermented black tea has a mellow taste and rich aroma. Fermented Atractylodes macrocephala black tea achieves multiple effects such as strengthening the spleen and stomach, refreshing the mind and relieving fatigue, promoting body fluid production and clearing heat, anti-oxidation, and immune regulation. At the same time, innovative flavor control technology improves the bitter taste of traditional Atractylodes macrocephala black tea, giving fermented Atractylodes macrocephala black tea a flavor characteristic of "a herbal sweetness that blends the honey aroma of black tea and the medicinal aroma of Atractylodes macrocephala".

[0144] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing fermented Atractylodes macrocephala black tea, characterized in that, include: The fresh Atractylodes macrocephala and fresh tea leaves are mixed and then kneaded in three stages: first stage, second stage, and third stage, followed by triple fermentation to obtain fermented Atractylodes macrocephala black tea. The triple fermentation process includes: conventional fermentation, enzymatic hydrolysis, and combined fermentation in sequence.

2. The preparation method according to claim 1, characterized in that, The first stage of kneading has a pressure of 0.15–0.2 MPa, a rotation speed of 15–20 rpm, and a time of 10–15 min; the second stage of kneading has a pressure of 0.25–0.3 MPa, a rotation speed of 20–25 rpm, and a time of 10–15 min; the third stage of kneading has a pressure of 0.15–0.2 MPa, a rotation speed of 10–15 rpm, and a time of 5–10 min.

3. The preparation method according to claim 1, characterized in that, The compound enzyme used in the enzymatic hydrolysis includes cellulase, pectinase, and xylanase; the mass ratio of cellulase, pectinase, and xylanase is (2-4):(2-4):(1-3); the amount of the compound enzyme added is 0.05% to 0.15% of the total weight of fresh Atractylodes macrocephala and fresh tea leaves.

4. The preparation method according to claim 1 or 3, characterized in that, The enzymatic hydrolysis is performed at a temperature of 45–55°C for 70–90 minutes.

5. The preparation method according to claim 1, characterized in that, The compound fermentation process involves fermentation using microorganisms and extracts, the extracts including kombucha extract; the microorganisms including Lactobacillus plantarum, and the mass ratio of kombucha extract to Lactobacillus plantarum is 1:(1-5).

6. The preparation method according to claim 1 or 5, characterized in that, The combined fermentation is carried out at a temperature of 28–32°C for 16–24 hours.

7. The preparation method according to claim 1, characterized in that, The compound fermentation process further includes sequentially subjecting the fermentation product to hot air drying, far-infrared and microwave irradiation, and vacuum drying; the hot air drying temperature is 50-60℃, the wind speed is 0.8-3m / s, and the hot air drying is carried out until the moisture content is 15%-18%; the hot air drying includes intermittent drying, and the parameters of the intermittent drying include: drying for 29-31 minutes, followed by a 4-6 minute break; The far-infrared irradiation parameters are: wavelength 4–8 μm, intensity 0.7–0.9 W / cm². 2 The duration of combined far-infrared and microwave irradiation is 8–12 minutes; the power of the microwave is 290–310 W. The vacuum drying temperature is 45–55°C, the vacuum degree is -0.09–-0.08 MPa, and the vacuum drying is carried out until the moisture content is ≤6%.

8. The preparation method according to claim 1, characterized in that, The mass ratio of fresh Atractylodes macrocephala and fresh tea leaves during the mixing process is (5-7):(3-5).

9. Fermented Atractylodes macrocephala black tea obtained by the preparation method according to any one of claims 1 to 8.

10. The application of the preparation method according to any one of claims 1 to 8 in at least one of the following: 1) increasing the content of atractylodes lactone and polysaccharides, the active ingredients in fermented Atractylodes macrocephala black tea; 2) Reduce the bitterness of fermented Atractylodes macrocephala black tea; 3) Enhance the antioxidant effect of fermented Atractylodes macrocephala black tea.

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