Extraction of theabrownin in Ya'an Tibetan tea processing by-product and development of theabrownin lozenge

Abstract

The invention belongs to the technical field of extraction of bioactive substances, and particularly relates to extraction of theabrownin in Ya'an Tibetan tea processing by-products and development of theabrownin lozenges, which comprises the following steps: extracting tea by-products to obtain a theabrownin extract; wherein an extraction reagent for the extraction treatment is water, the extraction temperature of the extraction treatment is 60-100 DEG C, the extraction time of the extraction treatment is 60-120 minutes, the material-liquid ratio of the tea by-products to the extraction reagent is (1: 20)-(1: 60), and the unit is g / mL. The method for extracting the theabrownin extract from the tea by-product has the advantages of wide raw material source, low cost, high theabrownin extraction efficiency, short extraction time, small influence on the environment, high biological safety, high application value and the like, and is wide in application range.

Description

Technical Field

[0001] The invention belongs to the technical field of extraction of bioactive substances, and in particular relates to the extraction of theabrownin from Ya'an Tibetan tea processing by-products and the development of theabrownin lozenges. Background Art

[0002] The by-products obtained after refining and removing the stems of tea leaves after fermentation mainly include tea powder, tea flakes, tea stems, etc., which are rich in biologically active ingredients such as theabrownins, tea polyphenols, and flavonoids. If they are directly discarded or incinerated, it will not only cause waste of resources but also pollute the environment. If they are effectively utilized, it will not only reduce production costs, but also transform tea by-products into high-value-added products, thereby improving the economic benefits of the entire tea industry.

[0003] At present, the utilization of tea by-products mainly involves mixing tea stems, tea powder, etc. with refined tea of ​​different grades in proportion and pressing them during the blending process. However, the presence of tea powder will affect the clarity and taste of the tea. As for the extraction of bioactive ingredients in tea by-products, the main method is to use a variety of organic solvents for extraction and purification, which has problems such as high cost, environmental pollution, low biosafety and low extraction efficiency.

[0004] Therefore, there is an urgent need to develop a method for extracting theabrownin from tea by-products with high extraction efficiency, low cost, biosafety and less environmental pollution, so as to achieve high-value utilization of tea by-products. Summary of the Invention

[0005] The present invention aims to, at least to some extent, address at least one of the technical problems existing in the prior art. To this end, the present invention provides methods for extracting theabrownin from a byproduct of Ya'an Tibetan tea processing and the development of theabrownin lozenges. The present method for extracting theabrownin from tea byproducts offers advantages such as a broad source of raw materials, low cost, high theabrownin extraction efficiency, short extraction time, minimal environmental impact, high biosafety, and high application value, thus providing a wide range of applications.

[0006] In the first aspect of the present invention, the present invention proposes a method for extracting theabrownin extract from tea by-products. According to an embodiment of the present invention, the method comprises: subjecting tea by-products to an extraction treatment to obtain a theabrownin extract; wherein, the extraction reagent of the extraction treatment is water, the extraction temperature of the extraction treatment is 60-100°C, the extraction time of the extraction treatment is 60-120min, and the material-liquid ratio of the tea by-products to the extraction reagent is 1:20-1:60, and the unit is g:mL. The method for extracting theabrownin extract from tea by-products according to an embodiment of the present invention has the advantages of a wide source of raw materials, low cost, high theabrownin extraction efficiency, short extraction time, small impact on the environment, high biosafety and high application value, and has a wide range of applications.

[0007] According to an embodiment of the present invention, the above-mentioned extraction method may also have the following additional technical features:

[0008] According to an embodiment of the present invention, the tea by-product is derived from the by-product obtained from the refining and stem removal step after the pile fermentation of Ya'an Tibetan tea.

[0009] According to an embodiment of the present invention, the tea leaves are black tea.

[0010] According to an embodiment of the present invention, the tea is Ya'an Tibetan tea.

[0011] In a second aspect, the present invention provides a theabrownin extract. According to an embodiment of the present invention, the theabrownin extract is produced by the method described in the first aspect. The theabrownin extract according to the embodiment of the present invention has advantages such as high theabrownin content, high biosafety, and high application value, and has a wide range of applications.

[0012] In the third aspect of the present invention, the present invention proposes a method for preparing theabrownin freeze-dried powder. According to an embodiment of the present invention, the method comprises: S1: extracting tea by-products to obtain theabrownin extract; S2: refining the theabrownin extract to obtain theabrownin freeze-dried powder; wherein, the extraction reagent of the extraction treatment is water, the extraction temperature of the extraction treatment is 60-100°C, the extraction time of the extraction treatment is 60-120min, and the material-liquid ratio of the tea by-products to the extraction reagent is 1:20-1:60, and the unit is g / mL. The method for preparing theabrownin freeze-dried powder according to an embodiment of the present invention has the advantages of convenient storage and transportation, maintaining active ingredients and high application value, and has a wide range of applications.

[0013] According to an embodiment of the present invention, the above-mentioned preparation method may also have the following additional technical features:

[0014] According to an embodiment of the present invention, the tea by-product is a by-product obtained by refining and removing the stems of tea leaves after fermentation.

[0015] According to an embodiment of the present invention, the tea leaves are black tea.

[0016] According to an embodiment of the present invention, the tea is Ya'an Tibetan tea.

[0017] In a fourth aspect, the present invention provides a freeze-dried theabrownin powder. According to an embodiment of the present invention, the freeze-dried theabrownin powder is produced by the method described in the third aspect. The freeze-dried theabrownin powder according to the embodiment of the present invention has advantages such as ease of use, good stability, low storage cost, and high application value, and has a wide range of applications.

[0018] In its fifth aspect, the present invention provides a method for preparing theabrownin buccal tablets. According to an embodiment of the present invention, the method comprises: A: mixing the theabrownin extract described in the second aspect and / or the theabrownin lyophilized powder described in the fourth aspect with excipients to obtain a theabrownin mixture; and B: tableting the theabrownin mixture to obtain theabrownin buccal tablets. The method for preparing theabrownin buccal tablets according to an embodiment of the present invention has the advantages of simple preparation, low cost, high biosafety, and high application value. The theabrownin buccal tablets prepared by this method are convenient to carry and have a wide range of applications.

[0019] In a sixth aspect, the present invention provides a theabrownin lozenge. According to an embodiment of the present invention, the theabrownin lozenge is prepared by the method described in the fifth aspect. The theabrownin lozenge according to the embodiment of the present invention has the advantages of high theabrownin content, high product convenience, precise control of theabrownin dosage per intake, high application value, and a wide range of applications.

[0020] In a seventh aspect, the present invention provides a food. According to an embodiment of the present invention, the food comprises the theabrownin extract of the second aspect and / or the theabrownin freeze-dried powder of the fourth aspect.

[0021] It should be noted that the characteristics and advantages described above for theabrownin extract and theabrownin freeze-dried powder are also applicable to this application and will not be repeated here.

[0022] In the eighth aspect of the present invention, the present invention proposes the use of the theabrownin extract described in the second aspect and the theabrownin freeze-dried powder described in the fourth aspect in the preparation of theabrownin lozenges.

[0023] It should be noted that the characteristics and advantages described above for theabrownin extract and theabrownin freeze-dried powder are also applicable to this application and will not be repeated here.

[0024] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the following description of the embodiments with reference to the accompanying drawings, in which:

[0026] Figure 1The response surface diagrams of the other two variables and theabrownin extraction rate when one variable is fixed at the "0" level in Example 16 of the present invention are as follows: A (solid-liquid ratio), B (extraction time), C (extraction temperature), D (theabrownin extraction rate), AB (interaction between solid-liquid ratio and time), BC (interaction between extraction time and temperature), AC (interaction between solid-liquid ratio and temperature). From left to right, they are the response surface diagrams of solid-liquid ratio, extraction time and theabrownin extraction rate when the extraction temperature is fixed at the "0" level, the response surface diagrams of extraction temperature, extraction time and theabrownin extraction rate when the solid-liquid ratio is fixed at the "0" level, and the response surface diagrams of solid-liquid ratio, extraction temperature and theabrownin extraction rate when the extraction time is fixed at the "0" level;

[0027] Figure 2 This is the preparation process of theabrownin freeze-dried powder in Example 17 of the present invention, wherein Figure A is a diagram of a sample of a Ya'an Tibetan tea by-product, Figure B is a diagram of the condition after adding 4 volumes of 95% edible alcohol to the extract XV, Figure C is a diagram of the collected precipitate, and Figure D is a diagram of the prepared theabrownin freeze-dried powder sample I;

[0028] Figure 3 This is a diagram showing the preparation results of theabrownin lozenges in Example 17 of the present invention. DETAILED DESCRIPTION

[0029] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0030] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of the technical features indicated. Therefore, features defined as "first" or "second" may explicitly or implicitly include one or more of such features. Furthermore, in the description of the present invention, unless otherwise specified, "plurality" means two or more.

[0031] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0032] In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present invention, but not excluding other contents.

[0033] As used herein, the terms "optionally," "optional," or "optionally" generally mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0034] Terms and Definitions

[0035] In this article, "dark tea" refers to a type of tea that has undergone a post-fermentation process. Its characteristic is that it undergoes microbial fermentation during the production process, which gives the tea a unique color, aroma and flavor. It includes many different types of tea, and Ya'an Tibetan tea is one of them.

[0036] In this article, "Ya'an Tibetan tea by-product samples" refer to the products produced during the refining and stem removal process after the fermentation of Ya'an Tibetan tea, mainly including tea stems, tea powder, tea leaves, tea residues, etc.

[0037] Extraction method

[0038] The present invention proposes a method for extracting theabrownin extract from tea by-products. According to an embodiment of the present invention, the method comprises: subjecting tea by-products to an extraction treatment to obtain a theabrownin extract; wherein, the extraction reagent of the extraction treatment is water, the extraction temperature of the extraction treatment is 60 to 100°C, the extraction time of the extraction treatment is 60 to 120 minutes, and the material-liquid ratio of the tea by-products to the extraction reagent is 1:20 to 1:60, and the unit is g:mL. The method for extracting theabrownin extract from tea by-products according to an embodiment of the present invention has the advantages of a wide source of raw materials, low cost, high theabrownin extraction efficiency, short extraction time, small impact on the environment, high biosafety and high application value, and has a wide range of applications.

[0039] Exemplarily, the extraction temperature of the extraction process is 60°C, 70°C, 80°C, 90°C, 100°C, preferably 80-100°C, more preferably 90°C; the extraction time of the extraction process is 60min, 70min, 80min, 90min, 100min, 110min, 120min, preferably 100-120min, more preferably 110min; the solid-liquid ratio of the tea by-product to the extraction reagent (in g / mL) is 1:20, 1:30, 1:40, 1:50, 1:60, preferably 1:30-1:50, more preferably 1:40. Thus, by optimizing the extraction temperature, extraction time, and solid-liquid ratio of tea by-products to extraction reagents during the extraction process, the extraction efficiency of theabrownin in tea by-products can be further improved.

[0040] It should be noted that according to the method for extracting theabrownin extract from tea by-products in an embodiment of the present invention, by optimizing the extraction conditions, the obtained theabrownin extract has a high theabrownin content and its protein, tea polyphenols, tea polysaccharides and flavonoids contents are simultaneously greatly improved compared with other extraction conditions.

[0041] According to an embodiment of the present invention, the tea by-products are obtained by-products obtained by refining and removing the stems of tea leaves after fermentation. Thus, by extracting, separating, and purifying the active ingredients in the tea by-products, the added value and utilization rate of tea processing by-products are further increased, resource waste is reduced, and high-value utilization of tea by-products is achieved.

[0042] According to an embodiment of the present invention, the tea is dark tea. Thus, by limiting the source of the tea, it is determined that the obtained tea by-product contains relatively rich theabrownin, ensuring the normal extraction of theabrownin in the subsequent extraction.

[0043] According to an embodiment of the present invention, the tea is Ya'an Tibetan tea. Therefore, as a type of dark tea, the obtained tea by-product is determined to contain rich theabrownin, ensuring the normal extraction of theabrownin in the subsequent process.

[0044] Theabrownin extract

[0045] The present invention provides a theabrownin extract. According to an embodiment of the present invention, the theabrownin extract is prepared by the aforementioned method. The theabrownin extract according to the embodiment of the present invention has advantages such as high theabrownin content, high biosafety, and high application value, and has a wide range of applications.

[0046] Method for preparing freeze-dried theabrownin powder

[0047] The present invention proposes a method for preparing freeze-dried theabrownin powder. According to an embodiment of the present invention, the method comprises: S1: extracting tea by-products to obtain a theabrownin extract; S2: refining the theabrownin extract to obtain freeze-dried theabrownin powder; wherein the extraction reagent of the extraction treatment is water, the extraction temperature of the extraction treatment is 60-100°C, the extraction time of the extraction treatment is 60-120min, and the material-liquid ratio of the tea by-products to the extraction reagent is 1:20-1:60, and the unit is g / mL. The method for preparing freeze-dried theabrownin powder according to an embodiment of the present invention has the advantages of convenient storage and transportation, maintaining active ingredients and high application value, and has a wide range of applications.

[0048] According to an embodiment of the present invention, the tea by-products are obtained by-products obtained by refining and removing the stems of tea leaves after fermentation. Thus, by extracting, separating, and purifying the active ingredients in the tea by-products, the added value and utilization rate of tea processing by-products are further increased, resource waste is reduced, and high-value utilization of tea by-products is achieved.

[0049] According to an embodiment of the present invention, the tea is dark tea. Therefore, by limiting the source of the tea, it is determined that the tea by-products obtained contain relatively rich theabrownins, ensuring the normal subsequent extraction of theabrownins and further improving the quality of the prepared theabrownin freeze-dried powder.

[0050] According to an embodiment of the present invention, the tea is Ya'an Tibetan tea. Therefore, as a type of dark tea, the obtained tea by-product is determined to contain rich theabrownins, ensuring the normal subsequent extraction of theabrownins and further improving the quality of the prepared theabrownin freeze-dried powder.

[0051] Theabrownin freeze-dried powder

[0052] The present invention provides a freeze-dried theabrownin powder. According to an embodiment of the present invention, the freeze-dried theabrownin powder is produced by the aforementioned method. The freeze-dried theabrownin powder according to the embodiment of the present invention has advantages such as ease of use, good stability, low storage cost, and high application value, and has a wide range of applications.

[0053] Method for preparing theabrownin buccal tablets

[0054] The present invention provides a method for preparing theabrownin buccal tablets. According to an embodiment of the present invention, the method comprises: A: mixing the aforementioned theabrownin extract and / or the aforementioned theabrownin lyophilized powder with excipients to obtain a theabrownin mixture; and B: tableting the theabrownin mixture to obtain theabrownin buccal tablets. The method for preparing theabrownin buccal tablets according to an embodiment of the present invention has the advantages of simple preparation, low cost, high biosafety, and high application value. The theabrownin buccal tablets prepared by this method are convenient to carry and have a wide range of applications.

[0055] It should be noted that the theabrownin lozenges of the present invention also contain a variety of excipients, including, for example, compounding agents, maltodextrin, mannitol, microcrystalline cellulose, xylitol, citric acid, magnesium stearate, pigments, flavors, preservatives, etc., but are not limited to those listed. The selection and use of all excipients comply with national food safety standards.

[0056] Theabrownin lozenges

[0057] The present invention provides a theabrownin lozenge. According to an embodiment of the present invention, the theabrownin lozenge is prepared by the aforementioned method. The theabrownin lozenge according to the embodiment of the present invention has advantages such as high theabrownin content, high product convenience, precise control of theabrownin dosage per intake, and high application value, thus having a wide range of applications.

[0058] food

[0059] The present invention provides a food. According to an embodiment of the present invention, the food comprises the aforementioned theabrownin extract and / or the aforementioned theabrownin freeze-dried powder.

[0060] It should be noted that the characteristics and advantages described above for theabrownin extract and theabrownin freeze-dried powder are also applicable to this application and will not be repeated here.

[0061] use

[0062] The present invention proposes the use of the theabrownin extract and the theabrownin freeze-dried powder in preparing theabrownin lozenges.

[0063] It should be noted that the characteristics and advantages described above for theabrownin extract and theabrownin freeze-dried powder are also applicable to this application and will not be repeated here.

[0064] Below, the scheme of the present invention will be explained in conjunction with embodiment.It will be understood by those skilled in the art that the following examples are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention.In the embodiment, if specific technology or conditions are not indicated, the technology or conditions described in the literature in this area or the product instructions are used.The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained by commercial purchase.

[0065] Example 1: Extraction of theabrownin by leaching method

[0066] The theabrownins in Ya'an Tibetan tea by-products were extracted by water extraction. 1 g of Ya'an Tibetan tea by-products (derived from the tea stems and tea foam removed from the Ya'an Tibetan tea refining process) was accurately weighed into a clean conical flask. Drinking water preheated at the extraction temperature was added at a material-liquid ratio of 1:20 (g:mL). The conical flask was placed in a water bath at 80°C for extraction for 80 min, and the extract I was obtained by filtering with gauze.

[0067] The moisture content in Ya'an Tibetan tea by-products was determined according to the first method (direct drying method) of GB5009.3-2016 "Determination of Water in Foods", and the moisture content in Ya'an Tibetan tea by-products was recorded as w in %.

[0068] Pipette 20% of the total volume of the extract I into a 20 mL volumetric flask (for example, when the material-liquid ratio is 1:40, take 8 mL), dilute to volume to obtain a sample solution, and determine the theabrownin content in the extract I according to NY / T 3675-2020 "Determination of thearubigins and theabrownins in black tea - Spectrophotometric method", specifically:

[0069] Take 10mL of sample solution in a 50mL centrifuge tube, add 10mL of n-butanol, vortex continuously at 2500r / min for 10min, and centrifuge at 4500r / min for 10min; transfer to a 30mL separatory funnel, let it stand and separate into layers, then place the lower layer (aqueous layer) into a small beaker, accurately draw 2mL, place it in a 25mL volumetric flask, add 2mL of saturated oxalic acid solution and 6mL of water, and make up to volume with 95% ethanol, which is recorded as the colorimetric solution; using 95% ethanol as a reference, use a 1cm cuvette to measure the absorbance E of the colorimetric solution at 380nm.

[0070]

[0071] Where:

[0072] X TB :Theabrownin content (theabrownin extraction rate), %

[0073] E: Absorbance of colorimetric solution

[0074] m: sample mass, in g

[0075] w: dry matter content (mass fraction), i.e. the moisture content in Ya'an Tibetan tea by-products, unit is %

[0076] 16.944: Roberts empirical coefficient

[0077] Example 2: Optimization of extraction conditions for theabrownin extraction

[0078] The inventors used single-factor experiments to further investigate the effects of material-liquid ratio, extraction temperature, extraction time, etc. on the content level of theabrownin extracted by the extraction method.

[0079] The differences between the parameter settings of Examples 1 to 15 and Example 1 are shown in Table 1, where:

[0080] In Examples 1 to 5, the extraction temperature was fixed at 80° C. and the extraction time was 80 min, and the material-liquid ratio was changed by a single factor to obtain extracts I to V;

[0081] In Examples 6 to 10, the solid-liquid ratio was fixed at 1:40, the extraction time was 80 min, and the extraction temperature was changed by a single factor to obtain extracts VI to X;

[0082] In Examples 11 to 15, the solid-liquid ratio was fixed at 1:40 and the extraction temperature was 80° C., and the extraction time was changed as a single factor to obtain extracts XI to XV.

[0083] Table 1 Extraction conditions for theabrownin extraction by leaching method

[0084]

[0085]

[0086] In Examples 1 to 15, the extraction results of theabrownin using the leaching method are shown in Table 2.

[0087] Table 2 Extraction results of theabrownin by leaching method

[0088]

[0089]

[0090] Among them, theabrownin extraction rates ①, ②, and ③ were repeated three times, and the average of theabrownin extraction rates was the average of three repetitions.

[0091] The results show:

[0092] (1) When the extraction temperature and extraction time were fixed and the solid-liquid ratio was changed as a single factor, the extraction rate of theabrownin gradually increased when the solid-liquid ratio increased from 1:20 to 1:40, while the extraction rate of theabrownin showed a decreasing trend when the solid-liquid ratio increased from 1:40 to 1:60. That is, when the solid-liquid ratio reached a certain threshold, the dissolution of theabrownin reached its upper limit.

[0093] (2) When the material-liquid ratio and extraction time were fixed and the extraction temperature was changed as a single factor, the extraction rate of theabrownin gradually increased when the extraction temperature increased from 60℃ to 90℃, and the extraction rate of theabrownin reached the highest when the extraction temperature was 90℃. The increase in temperature will accelerate the dissolution of theabrownin, and the simple polyphenols in Tibetan tea may also be oxidized to theabrownin in the form of polymerized polyphenols under high temperature conditions; however, when the extraction temperature was increased to 100℃, the extraction rate of theabrownin decreased relatively, which indicates that the boiling water bath may lead to a decrease in the biological activity of theabrownin;

[0094] (3) When the material-liquid ratio and extraction temperature were fixed and the extraction time was changed as a single factor, the extraction rate of theabrownin gradually increased when the extraction time increased from 40 min to 100 min, indicating that the dissolution of theabrownin increased with the increase of extraction time. However, when the extraction time was extended to 120 min, the extraction rate of theabrownin decreased, suggesting that as the extraction time continued to increase, components other than theabrownin would also dissolve, thereby affecting the dissolution of theabrownin. Moreover, as the extraction time continued to increase, the extraction efficiency of theabrownin further decreased.

[0095] The above results show that when theabrownin is extracted by the leaching method, the preferred solid-liquid ratio is set in the range of 1:30-1:50, the preferred extraction temperature is set in the range of 80-100°C, and the preferred extraction time is set in the range of 80-120min. At this time, the extraction of theabrownin is higher.

[0096] Example 16: Calculation and verification of the optimal extraction conditions using response surface optimization

[0097] 1. Calculation of response surface optimization of optimal extraction conditions

[0098] On the basis of single factor experiment, the extraction ratio of material to liquid, extraction temperature and extraction time were selected as three factors, and the content of theabrownin in the extraction solution was used as the dependent variable. The three-factor and three-level Box-Behnken response surface method was designed, and the experimental design and data processing were performed in the software Design-Expert 8.0.6. Finally, the optimal process for extracting theabrownin from the by-products of Ya'an Tibetan tea was determined, in which the factors and levels of the Box-Behnken design are shown in Table 3.

[0099] Table 3 Factors and levels of Box-Behnken design

[0100]

[0101] On the basis of single factor experiment, the extraction ratio of material to liquid, extraction temperature and extraction time were selected as three factors, and the content of theabrownin in the extraction solution was used as the dependent variable. The three-factor and three-level Box-Behnken response surface method was designed, and the experimental design and data processing were performed in the software Design-Expert 8.0.6. Finally, the optimal process for extracting theabrownin from the by-products of Ya'an Tibetan tea was determined, in which the factors and levels of the Box-Behnken design are shown in Table 3.

[0102] Table 4 Box-Behnken experimental design and results

[0103] Serial number Material-liquid ratio (g:mL) Time (min) Temperature (℃) Theabrownin extraction rate (%) 1 1:30 80 90 8.277 2 1:50 100 80 8.287 3 1:40 120 100 8.908 4 1:50 120 90 8.996 5 1:30 100 100 8.326 6 1:40 100 90 9.359 7 1:50 80 90 8.387 8 1:40 100 90 9.306 9 1:30 100 80 7.768 10 1:40 80 80 8.194 11 1:40 80 100 8.513 12 1:30 120 90 8.362 13 1:40 100 90 9.495 14 1:40 100 90 9.422 15 1:40 100 90 9.545 16 1:50 100 100 8.536 17 1:40 120 80 8.526

[0104] The experimental results were analyzed by regression equation to obtain the quadratic multinomial regression equation of the extraction ratio of material to liquid, extraction temperature and extraction time of theabrownin from the by-products of Ya'an Tibetan tea: Y = 9.4254 + 0.184125*A + 0.177625*B + 0.1885*C + 0.131*AB - 0.07725*AC + 0.01575*BC - 0.61295*A 2 -0.30695*B 2 -0.5832*C 2 .

[0105] The variance analysis of the regression equation is shown in Table 5. The P value of this model is less than 0.0001, indicating that the model is very significant as a whole. The R 2 = 0.9916, indicating that the model has good fitting degree. The P value of Lack of Fit is 0.9806, indicating that the model has no significant lack of fit phenomenon, and the model used can well describe the experimental data. The P values of A, B and C are all less than 0.05, indicating that these three factors have significant effects on the response value. 2 , B 2 , C 2The P values of the quadratic terms were all less than 0.0001, indicating that the effects of the quadratic terms on the response values were very significant, and that there was a nonlinear relationship between the response values and the factors; the P value of the AB interaction term was 0.0101, indicating that there was a significant interaction between the liquid-solid ratio and the time; the P value of the AC interaction term was 0.0784, which was close to the significant level (0.05), indicating that the interaction between the liquid-solid ratio and the temperature was weak; and the P value of the BC interaction term was 0.6871, indicating that the interaction between the time and the temperature was not significant.

[0106] Table 5 Analysis of variance of the regression equation

[0107]

[0108]

[0109] The response surface results of the other two variables and the theabrownine extraction rate with one variable fixed at the "0" level are shown in Table 6. Figure 1 .

[0110] The results show that the response surface graph of the liquid-solid ratio and the extraction time is the steepest, and the interaction between the two factors has the most significant effect on the theabrownine extraction rate; the response surface graph of the liquid-solid ratio and the extraction temperature is relatively steep, and the two factors have a relatively significant effect on the theabrownine extraction rate; and the response surface graph of the extraction time and the extraction temperature is relatively smooth, and the two factors have the least effect on the theabrownine extraction rate, which is consistent with the results of the analysis of variance of the regression equation.

[0111] According to the analysis by the Design-Expert 8.0.6 experimental software, the optimal extraction process of theabrownine in the by-products of Ya'an Tibetan tea is as follows: the liquid-solid ratio is 1:41.7623 (g:mL), the extraction time is 106.615 min, and the extraction temperature is 91.539 ℃, and the predicted theoretical value of the theabrownine extraction rate is 9.486%. In the subsequent verification operation extraction process, the above conditions are adjusted as follows: the liquid-solid ratio is 1:42 (g:mL), the extraction time is 107 min, and the extraction temperature is 92 ℃.

[0112] 2. Verification of the optimal extraction conditions optimized by the response surface

[0113] Accurately weigh 30 g of the by-products of Ya'an Tibetan tea, and put them into a conical flask. According to the liquid-solid ratio of 1:42, add Wahaha purified water preheated to 92 ℃ for extraction. The water bath temperature is set to 92 ℃, and the extraction time is set to 107 min. Shake the flask 3 times during the extraction process. After the extraction is completed, filter the extraction liquid XIV while it is hot with a filter bag. Repeat the above extraction method 3 times, and determine the content of theabrownine (theabrownine extraction rate) in the extraction liquid XIV according to NY / T 3675-2020 "Determination of theabrownine and thearubigins in black tea - spectrophotometric method".

[0114] The results showed that the extraction rate of theabrownin was 9.518%±0.208%, which was basically consistent with the theoretical value (9.486%) with a relative error of 0.032%, indicating that the regression equation fitted the actual situation well.

[0115] The above results show that the extraction rate of theabrownin is the highest when the solid-liquid ratio is 1:42 (g:mL), the extraction time is 107min, and the extraction temperature is 92℃.

[0116] Example 17: Preparation of theabrownin freeze-dried powder and theabrownin lozenges

[0117] 1. Preparation of theabrownin freeze-dried powder

[0118] Accurately weigh 750g of Ya'an Tibetan tea by-product sample, put it into a conical flask, and add Wahahaha purified water preheated to 92°C in advance according to the material-liquid ratio of tea powder: water = 1:42 for extraction. The water bath temperature is set to 92°C and the extraction time is set to 107min. During the extraction process, shake the flask 3 times, filter through gauze to obtain an extract, add 95% edible alcohol 4 times the volume of the extract, let it settle for 24h, gently aspirate most of the clear upper liquid, transfer the lower liquid to a centrifuge tube and centrifuge at 4500rpm for 10min, discard the supernatant, collect the precipitate, add Wahahaha drinking water to dissolve, put it in a -80°C refrigerator for pre-freezing for 24h, then freeze-dry the theabrownin crude extract for 24h, weigh and record the obtained theabrownin freeze-dried powder sample I, and perform component analysis on the theabrownin freeze-dried powder sample I, including theabrownin content, tea polysaccharide content, tea polyphenol content, flavonoid content, protein content, etc. The specific method is as follows:

[0119] The theabrownin content in the extract was determined according to NY / T 3675-2020, "Determination of thearubigins and theabrownins in black tea - Spectrophotometric method." Protein content was determined using the Coomassie Brilliant Blue method with bovine serum albumin as the standard. Tea polysaccharides were determined using the phenol-sulfuric acid method with glucose as the standard. Tea polyphenols were determined using the folin-phenol method with gallic acid as the standard. Total flavonoids were determined using the sodium nitrite-aluminum nitrate colorimetric method with rutin as the standard.

[0120] The preparation process of theabrownin freeze-dried powder is shown in Figure 2 .

[0121] The component analysis results of theabrownin freeze-dried powder sample I are shown in Table 6

[0122] Table 6 Component analysis results of theabrownin freeze-dried powder sample 1

[0123] Element Content (w / w, %) Theabrownin 42.59±0.83 Protein (calculated as bovine serum albumin)% 15.56±0.56 Tea polyphenols (calculated as gallic acid)% 20.30±0.35 Tea polysaccharides (calculated as glucose)% 25.62±0.69 Flavonoids (calculated as rutin)% 1.04±0.02

[0124] The results showed that the theabrownin content in theabrownin freeze-dried powder sample I was the highest, accounting for 42.59±0.83%; in addition, the protein content in theabrownin freeze-dried powder sample I was 15.56±0.56%, the tea polyphenol content was 20.30±0.35%, the tea polysaccharide content was 25.62±0.69%, and the flavonoid content was 1.04±0.02%; theabrownin is a macromolecular polymer complexed by proteins, tea polysaccharides, tea polyphenols, etc., and the theabrownin in the theabrownin freeze-dried powder sample I is formed by the complexation of part of the tea polysaccharides, tea polyphenols and proteins.

[0125] 2. Preparation of theabrownin lozenges

[0126] Raw material pretreatment: The main auxiliary materials such as theabrownin, whole milk powder, acai berry powder, broccoli powder, cocoa powder, turmeric powder, maltodextrin, microcrystalline cellulose, sodium carboxymethyl cellulose (CMC), mannitol, xylitol, anhydrous citric acid, etc. are sieved through 24 mesh respectively, sealed and stored away from light to ensure uniform powder fluidity.

[0127] Mixing of excipients: According to the formula of theabrownin lozenges, in each portion (20 tablets) of theabrownin lozenge mixed powder formula, first fix the addition amount of excipients such as maltodextrin, microcrystalline cellulose, sodium carboxymethyl cellulose (CMC), mannitol, xylitol, citric acid, etc., and then add five compounding agents such as whole milk powder (A), acai berry powder (B), broccoli powder (C), cocoa powder (D) and turmeric powder (E) respectively, and mix them. Change the amount of theabrownin added to obtain the theabrownin lozenges in the order of 1, 2, and 3. Finally, add magnesium stearate and gently stir manually to mix.

[0128] Tablet pressing: The mixed powder is compressed using a small manual tablet press to produce tablets of appropriate size.

[0129] Naming of theabrownin lozenges: A total of 15 types of theabrownin lozenges were obtained, named: TB-A1, TB-A2, TB-A3; TB-B1, TB-B2, TB-B3; TB-C1, TB-C2, TB-C3; TB-D1, TB-D2, TB-D3; TB-E1, TB-E2, TB-E3. TB-A is a theabrownin-milk powder lozenge, TB-B is a theabrownin-brazilian powder lozenge, TB-C is a theabrownin-broccoli powder lozenge, TB-D is a theabrownin-cocoa powder lozenge, and TB-E is a theabrownin-turmeric powder lozenge. Serial numbers 1, 2, and 3 indicate theabrownin addition levels of 1g, 1.5g, and 2g per serving, respectively.

[0130] The sources of the raw materials of theabrownin lozenges are shown in Table 7.

[0131] Table 7 Sources of raw materials for theabrownin lozenges

[0132] name Manufacturer whole milk powder New Zealand Fonterra Group (Anchor) maltodextrin Suqian Dongyuda Trading Co., Ltd. Citric acid Suqian Dongyuda Trading Co., Ltd. Sodium carboxymethyl cellulose (CMC) Suqian Dongyuda Trading Co., Ltd. Xylitol Guangzhou Fuzheng Donghai Food Co., Ltd. microcrystalline cellulose Suqian Dongyuda Trading Co., Ltd. magnesium stearate Suqian Dongyuda Trading Co., Ltd. Mannitol Suqian Dongyuda Trading Co., Ltd.

[0133] The formula of theabrownin lozenges is shown in Table 8.

[0134] Table 8 The formula of theabrownin lozenges

[0135]

[0136]

[0137] Among them, based on the existing sensory scoring standards and the characteristic intensity of sensory perception, the inventors designed a sensory scoring standard applicable to the theabrownin probiotic lozenges under the basic formula of the present invention. After controlling the relevant variables, a review panel composed of 18 volunteers conducted corresponding evaluations. The experimental results were taken from the average value of the evaluation results. The basic formula of the theabrownin lozenges was determined based on the sensory evaluation experimental results.

[0138] The sensory evaluation standards of theabrownin lozenges are shown in Table 9.

[0139] Table 9 Sensory evaluation standards for theabrownin lozenges

[0140]

[0141] The sensory test results of theabrownin lozenges are shown in Table 10

[0142] Table 10 Sensory test results of theabrownin probiotic lozenges

[0143]

[0144]

[0145] The results of the preparation of theabrownin tablets are shown in Figure 3 .

[0146] The results showed that theabrownin is dark brown, and its powder is often accompanied by salty, astringent and bitter tastes, resulting in theabrownin lozenges with problems such as insufficient color appeal and bland taste. By adding five compound agents, including whole milk powder (A), acai berry powder (B), broccoli powder (C), cocoa powder (D) and turmeric powder (E), the flavor characteristics of the lozenges were systematically improved, the bitterness and astringency were neutralized, and the functional activity was enhanced.

[0147] In addition, there were differences in the total sensory scores among different formulas and theabrownin addition levels: TB-A (theabrownin-milk powder lozenges) had the highest total score (69.76-61.77), significantly better than the other groups (p<0.05), especially when theabrownin addition was 1g. TB-E (theabrownin-turmeric powder lozenges) had the lowest total score (48.98-45.56), which was approximately 30% lower than TB-A. Furthermore, as theabrownin addition level increased, the total sensory evaluation score showed a downward trend (e.g., TB-A dropped from 69.76 to 61.77), indicating that high concentrations of theabrownin may intensify the release of bitterness and deepen the color.

[0148] TB-A ranked first in both taste (16.71-15.98) and flavor (18.94-14.33). The milk fat in whole milk powder encapsulates theabrownin polyphenols through hydrophobic interactions, significantly reducing bitter receptor activation. Meanwhile, its lactose component provides natural sweetness, balancing the taste experience. TB-E, on the other hand, received the lowest taste (9.67-8.86) and flavor (9.88-8.07) scores due to a synergistic negative effect between the spicy flavor of turmeric powder and the bitterness of theabrownins. Curcumin and theabrownins may undergo a complexation reaction, resulting in a darker color and an enhanced odor. Anthocyanins in acai berry powder can improve color, but their sourness does not effectively mask the bitterness of theabrownins, leading to fluctuations in the flavor score of TB-B (theabrownin-acai berry powder lozenge).

[0149] TB-C (theabrownin-broccoli powder lozenge) had the highest histomorphology score (17.55-17.24). The fibrous structure of broccoli powder enhanced the tablet integrity through physical cross-linking and delayed the structural collapse during disintegration.

[0150] In terms of color, TB-A (17.50-16.68) and TB-C (18.34-19.04) scored higher, as the milky white of whole milk powder and the chlorophyll green of broccoli powder synergistically neutralized the dark brown of theabrownin. TB-D (theabrownin-cocoa powder lozenges) scored lower (16.00-14.96) because its color is similar to that of theabrownin.

[0151] These results indicate that TB-A had the highest total score, while TB-E had the lowest. Furthermore, as theabrownin addition increased, the total sensory score decreased, suggesting that high theabrownin concentrations may exacerbate bitterness and deepen color. TB-A ranked first in both taste and flavor, while TB-E had the lowest taste and flavor scores. TB-C and TB-A had high color scores, while TB-D had the lowest color score.

[0152] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" mean that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification and features of different embodiments or examples without contradiction.

[0153] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A method for extracting theabrownin extract from tea by-products, characterized in that: include: Extracting tea by-products to obtain theabrownin extract; in, The leaching agent of the leaching process is water, The extraction temperature of the leaching process is 60-100°C. The extraction time of the leaching process is 60 to 120 minutes. The material-liquid ratio of the tea by-product to the extraction reagent is 1:20 to 1:60, with the unit being g:mL.

2. The method according to claim 1, characterized in that The tea by-product is a by-product obtained by refining and removing the stems of tea leaves after fermentation; Optionally, the tea leaves are black tea; Optionally, the tea leaves are Ya'an Tibetan tea.

3. A theabrownin extract, characterized in that The theabrownin extract is prepared by the method according to claim 1 or 2.

4. A method for preparing freeze-dried theabrownin powder, characterized in that: The method comprises: S1: extracting tea by-products to obtain theabrownin extract; S2: refining the theabrownin extract to obtain theabrownin freeze-dried powder; Wherein, the leaching agent of the leaching treatment is water, The extraction temperature of the leaching process is 60-100°C. The extraction time of the leaching process is 60 to 120 minutes. The material-liquid ratio of the tea by-product to the extraction reagent is 1:20 to 1:60, with the unit being g:mL.

5. The method according to claim 4, characterized in that The tea by-product is a by-product obtained by refining and removing the stems of tea leaves after fermentation; Optionally, the tea leaves are black tea; Optionally, the tea leaves are Ya'an Tibetan tea.

6. A theabrownin freeze-dried powder, characterized in that: The theabrownin freeze-dried powder is prepared by the method according to claim 4 or 5.

7. A method for preparing theabrownin buccal tablets, characterized in that: The method comprises: A: mixing the theabrownin extract according to claim 3 and / or the theabrownin freeze-dried powder according to claim 6 with excipients to obtain a theabrownin mixture; B: The theabrownin mixture is tableted to obtain theabrownin buccal tablets.

8. A theabrownin buccal tablet, characterized in that: The theabrownin lozenge is prepared by the method according to claim 7.

9. A food, characterized in that The food comprises the theabrownin extract according to claim 3 and / or the theabrownin freeze-dried powder according to claim 6.

10. Use of the theabrownin extract according to claim 3 and the theabrownin freeze-dried powder according to claim 6 in the preparation of theabrownin lozenges.

Images