CN113498861A - Tea extract and preparation method thereof - Google Patents
Tea extract and preparation method thereof Download PDFInfo
- Publication number
- CN113498861A CN113498861A CN202110814641.0A CN202110814641A CN113498861A CN 113498861 A CN113498861 A CN 113498861A CN 202110814641 A CN202110814641 A CN 202110814641A CN 113498861 A CN113498861 A CN 113498861A
- Authority
- CN
- China
- Prior art keywords
- tea
- eluent
- distilled water
- filtrate
- theabrownin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/16—Tea extraction; Tea extracts; Treating tea extract; Making instant tea
- A23F3/163—Liquid or semi-liquid tea extract preparations, e.g. gels, liquid extracts in solid capsules
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/16—Tea extraction; Tea extracts; Treating tea extract; Making instant tea
- A23F3/18—Extraction of water soluble tea constituents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/82—Theaceae (Tea family), e.g. camellia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Diabetes (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Toxicology (AREA)
- Mycology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Medical Informatics (AREA)
- Obesity (AREA)
- Biochemistry (AREA)
- Alternative & Traditional Medicine (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Endocrinology (AREA)
- Hematology (AREA)
- Microbiology (AREA)
- Emergency Medicine (AREA)
- Epidemiology (AREA)
- Nutrition Science (AREA)
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- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
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Abstract
The invention provides a tea-derived product and a preparation method thereof, which extracts essential components beneficial to the body from natural tea leaves, particularly Ceylon black tea, keeps a proper proportion and exerts the maximum effect. According to the method, the raw material of the Ceylon black tea is subjected to screening pretreatment, is extracted in a high-temperature and high-pressure extraction tank with bubble stirring, is subjected to ultrafiltration membrane and chromatographic column to remove macromolecular impurities and micromolecular theophylline, and is subjected to spray drying to obtain the theanine with the contents of theabrownin, tea polyphenol and caffeine of more than 90%. The invention can prepare theabrownin, tea polyphenol and caffeine products from single tea at one time, greatly improves the purity and activity of the theabrownin, reduces the preparation cost, improves the recovery rate, is suitable for industrial production, does not use toxic and harmful reagents and residual related heavy metal ions in the whole process, and ensures the safety and reliability of the products.
Description
Technical Field
The invention relates to the field of tea processing, in particular to a tea extract product rich in tea pigment and a preparation method thereof.
Background
The tea is one of three traditional beverages in the world, and has multiple functions of nutrition, health care and the like. The tea leaves are classified into non-fermented tea, such as green tea, according to different fermentation degrees in the preparation process; "semi-fermented" teas, such as oolong tea; "Whole fermented" teas, such as black tea. The difference between them is the degree of "enzymatic oxidation" of tea polyphenols, the most major component of tea leaves, in the action of polyphenol oxidase. The green tea has not been subjected to enzymatic oxidation and the polyphenol oxidase is destroyed by heating. The 'semi-fermented' tea is partially oxidized enzymatically, and the oxidation reaction is terminated when the tea is dried. The 'fully fermented' black tea is stopped after the enzymatic oxidation reaction is completed. The enzymatic oxidation process comprises the steps of firstly oxidizing EGCG and the like in the theanine, converting the EGCG and the like into intermediate theaflavin, further oxidizing the EGCG and the like into secondary intermediate thearubigins, and finally converting the secondary intermediate thearubigins into the final product theabrownin with a stable structure. The extract of tea contains tea polyphenols, theanine, alkaloid, etc. Tea polyphenol is a polyphenol compound taking catechins as main components, is an ideal natural food antioxidant, has pharmacological functions of resisting aging and radiation, eliminating free radicals of a human body, reducing blood sugar, inhibiting tumor cells and the like, and has important application in the fields of daily chemical industry, food processing, medicines and the like. At present, the method for extracting tea polyphenol from tea at home and abroad mainly comprises the following steps: organic solvent extraction, metal ion salt precipitation, resin adsorption and supercritical fluid extraction. The organic solvent extraction method has simple and convenient process, maturity and stability, higher product purity and no strict requirement on tea raw materials. The organic solvent extraction method usually uses toxic solvents which are volatile to cause environmental pollution, has low selectivity, and simultaneously adopts a solvent extraction method to have high caffeine content. The ion precipitation extraction method uses heavy metal which is toxic to human bodies as a precipitator, and the product of the method is unacceptable in the food and medicine industries. The resin adsorption method is lack of equipment for large-scale continuous production and is only suitable for small-scale production; the adsorbent with high adsorption capacity and strong tea polyphenol selectivity is needed, the dosage of the adsorbent is large, resin deactivation is caused because protein, polysaccharide and the like in tea easily block resin gaps, resin activation and regeneration are troublesome, the resin price is high, and the investment is large. The supercritical extraction method has extremely low extraction rate, and large-scale industrial production has many difficulties.
Theabrownin is a water-soluble phenolic pigment extracted from tea leaves, and modern researches show that theabrownin has pharmacological effects of reducing blood fat (regulating blood fat in two directions), reducing blood viscosity, reducing blood sugar (regulating blood sugar in two directions), resisting atherosclerosis, resisting blood coagulation, promoting fibrinolysis, resisting lipid peroxidation (resisting free radical), regulating microcirculation, enhancing immunologic function, inhibiting tumors and the like. The theabrownin can also clean garbage substances suspended in blood and deposited on the inner wall of blood vessels, dredge blocked capillary vessels, clean various metabolic wastes and exogenous toxins inside and outside cells of the whole body, and clean various viscous substances attached to the surfaces of red blood cells, and is an internationally recognized blood vessel scavenger. In addition, toxicological studies on theabrownin prove that the results of acute toxicity tests, chronic toxicity tests and special toxicity tests (mutagenic toxicity tests and teratogenic toxicity tests) are negative, and the theaflavin is proved to be safe and edible food and also reflects the medicinal and edible property of the theabrownin.
The alkaloid in tea leaf includes caffeine, theobromine, theophylline, etc., and mainly caffeine. Caffeine has a wide range of medical applications, such as: can be used as heart and respiratory stimulant, and has effects of exciting central nervous system; is also an important antipyretic analgesic, and is one of the main components of compound aspirin, kelike, pain relieving tablet, quick-acting capsule for common cold, etc.; it also has diuretic effect. The method for extracting caffeine from tea mainly comprises solvent extraction, sublimation, ion precipitation, column chromatography, ultrasonic extraction, supercritical extraction, etc.
Although tea polyphenol, theabrownin and caffeine extracted from tea have various effects on human health, how to use the essence components in a matching way and exert the benefits of the tea on the human body to the maximum extent by regulating and controlling a reasonable proportion is urgently pursued by people. In addition, the contents of tea polyphenol, theabrownin and caffeine in various tea leaves are different, and usually, related substances are respectively extracted from different tea leaves rich in the components, or the substances are extracted from one tea leaf step by step and then are proportioned to obtain a final tea product, the process is complex and not purely natural, when brewing is carried out, partial substances from different tea leaf extracts can react to influence the product preparation of the tea, and the large-scale production is easily limited by the production place and the yield of the tea, so that how to extract a tea source product which can exert the maximum benefit on the body from one tea raw leaf at one time is a current urgent need.
Disclosure of Invention
In view of the disadvantages and shortcomings of the prior art, the primary object of the present invention is to provide a tea-derived substance, i.e. a tea product extracted from a single tea leaf and containing a proper proportion of essential components, which can maximize the benefits of the tea leaves on the body. The specific scheme is as follows:
a tea extract is characterized in that: comprises 60-65% of theabrownin, 20-25% of tea polyphenol, 3-5% of caffeine and 3-5% of dextrin by weight percentage.
Preferably, the tea-derived essence comprises 65% of tea pigment, 25% of tea polyphenol and 5% of caffeine by weight percentage.
The application also provides a preparation method of the theanine, which can extract a large amount of theabrownin and tea polyphenol and alkaloid with corresponding proportions from single tea leaves at one time, is natural, does not need to introduce toxic solvents and heavy metal ions, and can be used as an ideal medicinal and edible tea product.
In order to achieve the purpose, the invention adopts the following specific technical scheme:
a preparation method of tea extract comprises the following steps:
(1) taking dry tea leaves as raw materials, sieving to remove tea fine powder to obtain short strip-shaped tea leaves;
(2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, and soaking in a high-temperature and high-pressure environment to obtain tea soup;
(3) filtering the tea soup by a membrane twice, and transferring the obtained filtrate into a feed liquid storage tank;
(4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane;
(5) adsorbing the filtrate after removing impurities with ultrafiltration membrane with chromatographic column, and separating small molecular substances such as theophylline;
(6) concentrating the eluate after chromatography with reverse osmosis membrane, adding dextrin, and spray drying to obtain tea extract granule.
Further, the dried tea leaves in step (1) are preferably black tea, more preferably black tea of Ceylon.
Further, the size of the sieve holes sieved in the step (1) is 2 mm.
Furthermore, the adding amount of the distilled water relative to the tea in the step (2) is 10-30mL:1g, the soaking temperature is 95-105 ℃, the pressure is 0.12-0.18MPa, and the time is 4-5 h.
Further, agitation by bubbles is performed for 4 to 5 minutes, preferably oxygen, every 1 hour interval during the soaking in the step (2).
Further, the aperture of the filter membrane for membrane filtration in the step (3) is 1-5 μm.
Further, the cutting molecular weight of the ultrafiltration membrane in the step (4) is 5-10 ten thousand.
Further, the adsorption process of the chromatographic column in the step (5) comprises the steps of firstly carrying out cross-linked dextran agarose gel column chromatography adsorption, eluting by using 0.7-1BV of distilled water, and collecting eluent A; eluting with 3-4 BV of 60 ℃ distilled water, collecting the eluent, concentrating under reduced pressure, loading to an alumina column for chromatography, eluting with 2.5-3 BV of distilled water, collecting the eluent B, and combining the eluent A and the eluent B for preparing the tea extract.
Further, it is preferable that the filtrate of step (4) is controlled at 20 to 30 ℃ and then applied to a Sephadex column for adsorption, and the eluate is cooled to 5 to 10 ℃ before being subjected to alumina column chromatography.
Further, the flow rate of the eluent in the step (5) is controlled to be 0.5-2 BV/h.
Furthermore, the alumina chromatographic column after being eluted by distilled water can be repeatedly used after being eluted by methanol.
Further, a sampling test is further included between the step (5) and the step (6), and if the content of the substances with the larger molecular weight is too high, the step (4) is performed again; and (5) if the content of the substances with smaller molecular weight is too high, performing the step again until all the components are balanced, and then performing spray drying.
The invention adopts black tea as an extraction raw material, which contains rich theabrownin, in particular to the Ceylon black tea. The invention is different from the traditional process, and the prior art can crush tea leaves and then leach the tea leaves to obtain higher extraction rate of theabrownin and tea polyphenol. The tea raw materials which are not needed in the invention are crushed, and the dried tea leaves are firstly screened before extraction to remove the crushed tea powder, so that the impurities in the tea powder are more, and the tea powder is easy to harden and block in the filtering link, thereby influencing the extraction efficiency.
The method adopts an extraction tank with high temperature and high pressure and bottom blowing for extraction, the pressure is controlled to be between 1 and 2 atmospheric pressures, the temperature is not too high and is controlled to be about 100 ℃, the boiling point is not reached in a corresponding pressure range, the tea cannot roll along with the boiling of distilled water in the extraction tank and is stacked together, so that the method can ensure that the tea rolls and is dispersed properly by blowing oxygen and oxygen flow through the blowing holes arranged at the bottom of the extraction tank, and the pressure in the extraction tank is kept stable through a safety exhaust valve. On one hand, the tea leaves are fully dispersed under the stirring of bubbles, which is beneficial to leaching the essence components in the tea leaves; on the other hand, the oxygen atmosphere can convert the residual catechin in the black tea into tea pigment, particularly greatly improve the content of theabrownin, and the component proportion for preparing the tea source element required by the invention is achieved.
According to the invention, macromolecular impurities such as pectin, protein and polysaccharide can be separated out by ultrafiltration membrane filtration, so that the content of essence components such as tea polyphenol and theabrownin in tea leaves is increased. Because the alkaloid obtained by ultrafiltration also contains a small amount of theophylline, the theophylline has strong effects of strengthening heart, promoting urination, expanding coronary artery, relaxing bronchial smooth muscle and reducing excitation of a human nervous system, and is opposite to the performance of the caffeine, and the theophylline can easily damage gastric mucosa of a human body.
The invention can remove the micromolecule impurities and part of tea polyphenol of tea leaves by two-step chromatographic column adsorption and controlling the temperature, flow rate, elution amount and time of chromatography, so that the rest components mainly comprise theabrownin and reach the optimized proportion of 60-65% of theabrownin, 20-25% of tea polyphenol and 3-5% of caffeine, and then natural products can be obtained by one-step spray drying, and can be directly brewed and drunk to avoid impurity introduction when other single components are added for adapting the proportion.
Compared with the prior art, the invention has the following beneficial effects:
according to the invention, through screening pretreatment of tea raw materials and leaching in a high-temperature and high-pressure extraction tank with bubble stirring, under the condition that the contents of impurities and heavy metals are not increased, the extraction rate of main components of tea is improved, the extraction rate of theabrownin is increased, macromolecular impurities and micromolecular theophylline are removed through an ultrafiltration membrane and a chromatographic column, the contents of theabrownin, tea polyphenol and caffeine in a tea product obtained by spray drying reach more than 90%, and the components are properly matched, so that the traditional Chinese medicine blending requirement of 'monarch, minister and assistant' is met. The method can prepare the theabrownin, the tea polyphenol and the caffeine products from the single tea at one time, greatly improves the yield, the purity and the activity of the theabrownin, reduces the preparation cost, improves the recovery rate, is suitable for industrial production, does not use toxic and harmful reagents and residual related heavy metal ions in the whole process, and ensures the safety and the reliability of the products.
Drawings
FIG. 1 is a process flow diagram of the preparation of tea extract according to the present invention
Detailed Description
In order to better explain the present invention and to facilitate the understanding of the technical solutions of the present invention, the present invention is further described in detail below. However, the following examples are only simple examples of the present invention and do not represent or limit the scope of the present invention, which is defined by the claims.
Example 1
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 10mL:1g, soaking for 4 hours at the high-temperature high-pressure environment, the soaking temperature is 95 ℃, the pressure is 0.12MPa, and introducing oxygen every 1 hour in the soaking process to perform bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 4 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 5 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 20 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 0.7BV of distilled water, collecting eluent A, eluting by 3BV of 60 ℃ distilled water, collecting eluent, concentrating under reduced pressure, cooling to 10 ℃, performing alumina column chromatography, eluting by distilled water, collecting 3BV of eluent B, controlling the flow rate of the eluent in the elution process to be 0.5BV/h, combining the eluent A and the eluent B, sampling and testing, containing no theophylline, concentrating qualified eluent by a reverse osmosis membrane, adding 6g of dextrin, and performing spray drying to obtain 175.7g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 113.7g of theabrownin, 42.5g of tea polyphenol and 7.8g of caffeine.
Example 2
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the addition amount of the distilled water relative to the tea leaves is 20mL:1g, soaking for 5 hours at the high-temperature high-pressure environment, the soaking temperature is 105 ℃, the pressure is 0.15MPa, and introducing oxygen every 1 hour in the soaking process to perform bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes with the filter membrane aperture of 5 μm and 2 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 8 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 25 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 0.9BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating under reduced pressure, cooling to 8 ℃, performing alumina column chromatography, eluting by distilled water, collecting 2.8BV of eluent B, controlling the flow rate of eluent in the elution process to be 2BV/h, combining the eluent A and the eluent B, sampling and testing, containing no theophylline, concentrating qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 188g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 120.2g of theabrownin, 44.4g of tea polyphenol and 8.9g of caffeine.
Example 3
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 30mL:1g, soaking for 5 hours at the high-temperature high-pressure environment, the soaking temperature is 100 ℃, the pressure is 0.18MPa, and introducing oxygen every 1 hour in the soaking process to perform bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 5 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 10 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 30 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 1BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating the eluent under reduced pressure, cooling to 5 ℃, performing alumina column chromatography, eluting by distilled water, collecting 2.5BV of eluent B, controlling the flow rate of eluent in the elution process to be 1BV/h, combining the eluent A and the eluent B, sampling and testing, wherein theophylline is not contained, concentrating qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 199.1g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 128.1g of theabrownin, 48.7g of tea polyphenol and 9.5g of caffeine.
Example 4
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 30mL:1g, soaking for 5 hours at the high-temperature high-pressure environment, the soaking temperature is 100 ℃, the pressure is 0.1MPa, and introducing oxygen every 1 hour in the soaking process to perform bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 5 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 10 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 30 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 1BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating the eluent under reduced pressure, cooling to 5 ℃, performing alumina column chromatography, eluting by distilled water, collecting 2.5BV of eluent B, controlling the flow rate of eluent in the elution process to be 1BV/h, combining the eluent A and the eluent B, sampling and testing, wherein theophylline is not contained, concentrating qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 183.1g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 107.5g of theabrownin, 41.5g of tea polyphenol and 7.9g of caffeine.
Example 5
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 30mL:1g, soaking for 5 hours at the high-temperature high-pressure environment, the soaking temperature is 60 ℃, the pressure is 0.18MPa, and introducing oxygen every 1 hour in the soaking process to perform bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 5 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 10 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 30 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 1BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating the eluent under reduced pressure, cooling to 5 ℃, performing alumina column chromatography, eluting by distilled water, collecting 2.5BV of eluent B, controlling the flow rate of eluent in the elution process to be 1BV/h, combining the eluent A and the eluent B, sampling and testing, wherein theophylline is not contained, concentrating qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 144.9g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 92.6g of theabrownin, 32.7g of tea polyphenol and 6.8g of caffeine.
Example 6
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 30mL:1g, soaking for 5 hours in a high-temperature and high-pressure environment, the soaking temperature is 100 ℃, the pressure is 0.18MPa, and introducing air at intervals of 1 hour during the soaking process to perform bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 5 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 10 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 30 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 1BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating the eluent under reduced pressure, cooling to 5 ℃, performing alumina column chromatography, eluting by distilled water, collecting 2.5BV of eluent B, controlling the flow rate of eluent in the elution process to be 1BV/h, combining the eluent A and the eluent B, sampling and testing, wherein theophylline is not contained, concentrating qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 191.2g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 121.9g of theabrownin, 46.3g of tea polyphenol and 8.4g of caffeine.
Example 7
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 30mL:1g, soaking for 5 hours in a high-temperature and high-pressure environment, the soaking temperature is 100 ℃, the pressure is 0.18MPa, and continuously introducing oxygen to carry out bubble stirring in the soaking process to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 5 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 10 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 30 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 1BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating the eluent under reduced pressure, cooling to 5 ℃, performing alumina column chromatography, eluting by distilled water, collecting 2.5BV of eluent B, controlling the flow rate of eluent in the elution process to be 1BV/h, combining the eluent A and the eluent B, sampling and testing, wherein theophylline is not contained, concentrating qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 202.9g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 124g of theabrownin, 47.4g of tea polyphenol and 8.4g of caffeine.
Example 8
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 30mL:1g, soaking for 5 hours at the high-temperature high-pressure environment, the soaking temperature is 100 ℃, the pressure is 0.18MPa, and introducing oxygen every 1 hour in the soaking process to carry out bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 5 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 10 ten thousand; (5) cooling the filtrate subjected to impurity removal by an ultrafiltration membrane to 10 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 1BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating the eluent under reduced pressure, cooling to 5 ℃, then performing alumina column chromatography, eluting by distilled water, collecting 2.5BV of eluent B, controlling the flow rate of the eluent in the elution process to be 1BV/h, combining the eluent A and the eluent B, sampling, testing, containing no theophylline, concentrating the qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 190.5g of the tea-derived hormone particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 116.5g of theabrownin, 56.1g of tea polyphenol and 5.6g of caffeine.
Example 9
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 30mL:1g, soaking for 5 hours at the high-temperature high-pressure environment, the soaking temperature is 100 ℃, the pressure is 0.18MPa, and introducing oxygen every 1 hour in the soaking process to carry out bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 5 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 10 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 50 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 1BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating the eluent under reduced pressure, cooling to 5 ℃, performing alumina column chromatography, eluting by distilled water, collecting 2.5BV of eluent B, controlling the flow rate of eluent in the elution process to be 1BV/h, combining the eluent A and the eluent B, sampling and testing, wherein theophylline is not contained, concentrating qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 198.5g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 124.7g of theabrownin, 44.5g of tea polyphenol and 8.4g of caffeine.
Example 10
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 30mL:1g, soaking for 5 hours at the high-temperature high-pressure environment, the soaking temperature is 100 ℃, the pressure is 0.18MPa, and introducing oxygen every 1 hour in the soaking process to carry out bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 5 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 10 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 20 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 1BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating under reduced pressure, controlling the temperature to be 20 ℃, then performing alumina column chromatography, eluting by distilled water, collecting 2.5BV of eluent B, controlling the flow rate of eluent in the elution process to be 1BV/h, combining the eluent A and the eluent B, sampling and testing, wherein theophylline is not contained, concentrating qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 195.8g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 123.2g of theabrownin, 42.8g of tea polyphenol and 7.8g of caffeine.
Example 11
A preparation method of tea extract comprises the following steps: (1) taking 1000g of Ceylon black tea as a raw material, and removing tea fine powder by a screen with the size of a sieve hole of 2mm to obtain short strip-shaped tea leaves; (2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, wherein the adding amount of the distilled water relative to the tea leaves is 30mL:1g, soaking for 5 hours at the high-temperature high-pressure environment, the soaking temperature is 100 ℃, the pressure is 0.18MPa, and introducing oxygen every 1 hour in the soaking process to carry out bubble stirring for 4 minutes to obtain tea soup; (3) filtering the tea soup by two membranes, wherein the pore diameters of the filter membranes are 5 μm and 1 μm respectively, and transferring the obtained filtrate into a feed liquid storage tank; (4) separating macromolecular impurities such as tea saponin, protein and polysaccharide from the filtrate by using an ultrafiltration membrane with the cutting molecular weight of 10 ten thousand; (5) controlling the temperature of filtrate subjected to impurity removal by an ultrafiltration membrane to be 30 ℃, then loading the filtrate to a sephadex sepharose column for chromatography and adsorption, eluting by 1.5BV of distilled water, collecting eluent A, eluting by 4BV of 60 ℃ distilled water, collecting eluent, concentrating under reduced pressure, cooling to 5 ℃, performing alumina column chromatography, eluting by distilled water, collecting 1.5BV of eluent B, controlling the flow rate of eluent in the elution process to be 1BV/h, combining the eluent A and the eluent B, sampling and testing, containing no theophylline, concentrating qualified eluent by a reverse osmosis membrane, adding 8g of dextrin, and performing spray drying to obtain 190.4g of tea-derived essence particles. The product is subjected to quantitative analysis and detection of tea polyphenol and caffeine according to methods such as QB 2154-95 food additive tea polyphenol, and the like, and the content and purity of tea pigment are detected by high performance liquid chromatography, so that the product contains 118.4g of theabrownin, 52.8g of tea polyphenol and 6.2g of caffeine.
The tea extract product prepared by the invention is subjected to mouse acute toxicity experiments, and the result shows that the oral LD50 and the 95% confidence limit thereof are as follows: 21.68g/kg (20.81-22.59 g/kg), and the tea source element (LD50 is more than 5g/kg) is practically nontoxic according to the acute toxicity (LD50) dose classification of the GB15193.3-2014 national standard method. The mice receiving samples at 17.7g/kg dose groups and 20.2g/kg day 14 of the mice with tea-derived essence are normal in renal function and liver function indexes. The safety of the tea extract product is very high, even higher than that of common tea, and the existing experiment proves that the LD50 of the Yunnan big leaf green-baked green tea is 7.5 g/kg; the LD50 of the Pu' er tea stored for 1 year is 9.7 g/kg; the LD50 of the Pu' er tea for 5 years is 11.2 g/kg; the LD50 of Pu' er tea for 10 years is 12.2 g/kg. The larger the LD50 value, the higher the security.
Animal experiments on the effect of protecting hyperuricemia prove that the intervention of the tea-derived essence can reduce the blood uric acid, the kidney and the liver have certain protection effect, and compared with the positive medicament allopurinol, the medium dose of 400mg/kg of the holy calyx seu fructus physalis tea-derived essence has better kidney protection effect, which shows that the tea-derived essence can reduce the uric acid, and has better protection effect on the kidney than the common allopurinol for reducing the uric acid, namely, the side effect is smaller.
The related experiments of the tea-derived hormone on the high-fat diet induced obesity show that the tea-derived hormone has a remarkable inhibiting effect on the weight gain of mice caused by the high-fat diet.
Animal research on the tea-derived extract with the effects of relieving hyperlipidemia, hyperglycemia and hyperviscosity shows that the tea-derived extract can improve glucose tolerance of the mice in a high-fat diet group, improve sugar metabolism of an organism, and improve the insulin sensitivity of the mice in the high-fat diet group without influencing the food intake of the mice in the high-fat diet group by the effect of inhibiting blood sugar; slightly reducing the weight of liver tissues of the mice in the tea-derived essence medium/high dose treatment group, and obviously reducing the weight of white adipose tissues of the mice in the high fat diet group; the tea extract can reduce hyperlipemia caused by high fat diet, and effectively improve lipid drop accumulation in liver cells caused by high fat diet, and decrease fat cell number and increase volume. The experiment shows that the tea extract has obvious effects of reducing blood sugar, blood fat and fatty liver.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A tea extract is characterized in that: comprises 60-65% of theabrownin, 20-25% of tea polyphenol, 3-5% of caffeine and 3-5% of dextrin by weight percentage.
2. The theanine of claim 1 comprising, in weight percent, 65% theabrownin, 25% theapolyphenol, and 5% caffeine.
3. A process for the preparation of a theagenin according to claim 1 or claim 2, comprising the steps of:
(1) taking dry tea leaves as raw materials, sieving to remove tea fine powder to obtain short strip-shaped tea leaves;
(2) feeding the short strip-shaped tea leaves into an extraction tank through a feeding port, injecting distilled water, and soaking in a high-temperature and high-pressure environment to obtain tea soup;
(3) filtering the tea soup by a membrane twice, and transferring the obtained filtrate into a feed liquid storage tank;
(4) separating macromolecular impurities such as tea saponin, protein and tea polysaccharide from the filtrate by using an ultrafiltration membrane;
(5) adsorbing the filtrate after impurity removal by a chromatographic column, separating theophylline micromolecule substances, and blending the components;
(6) concentrating the filtrate after chromatography with reverse osmosis membrane, adding dextrin, and spray drying to obtain tea extract granule.
4. The method for preparing theanin according to claim 3, wherein: the dried tea leaves in step (1) are preferably black tea, more preferably black tea of Ceylon.
5. The method for preparing theanin according to claim 3, wherein: the size of the sieve mesh sieved in the step (1) is 2 mm; in the step (2), the adding amount of the distilled water relative to the tea leaves is 10-30mL:1g, the soaking temperature is 95-105 ℃, the pressure is 0.12-0.18MPa, and the time is 4-5 h.
6. The method for preparing theanin according to claim 3, wherein: stirring by bubbles is carried out for 4-5 minutes, preferably oxygen, every 1 hour interval during said soaking in step (2).
7. The method for preparing theanin according to claim 3, wherein: the aperture of the filter membrane for membrane filtration in the step (3) is 1-5 μm, and the cutting molecular weight of the ultrafiltration membrane in the step (4) is 5-10 ten thousand.
8. The method for preparing theanin according to claim 3, wherein: the adsorption process of the chromatographic column in the step (5) comprises the steps of firstly carrying out cross-linked dextran agarose gel column chromatography adsorption, eluting by using 0.7-1BV of distilled water, and collecting eluent A; eluting with 3-4 BV of 60 ℃ distilled water, collecting the eluent, concentrating under reduced pressure, loading to an alumina column for chromatography, eluting with 2.5-3 BV of distilled water, collecting the eluent B, and combining the eluent A and the eluent B for preparing the tea extract.
9. The method for preparing theanin according to claim 3, wherein: preferably, the filtrate of step (4) is controlled at 20-30 deg.C, and loaded on Sephadex column for adsorption, and the eluate is cooled to 5-10 deg.C before being subjected to alumina column chromatography.
10. The method for preparing theanin according to claim 3, wherein: a sampling test is also included between the step (5) and the step (6), and if the content of substances with larger molecular weight is too high, the step (4) is carried out again; and (5) if the content of the substances with smaller molecular weight is too high, performing the step again until all the components are balanced, and then performing spray drying.
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