CN110963933A - Process for extracting and purifying gamma-aminobutyric acid from tea - Google Patents
Process for extracting and purifying gamma-aminobutyric acid from tea Download PDFInfo
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Abstract
The invention provides a process for extracting and purifying gamma-aminobutyric acid in tea, which comprises the following steps: 1) a raw material pretreatment process; 2) a process of extracting gamma-aminobutyric acid by using ultrasonic-assisted alkali; 3) concentrating the coarse gamma-aminobutyric acid extract; 4) separating and purifying the gamma-aminobutyric acid concentrated solution; 5) drying and storing process of gamma-aminobutyric acid. The invention has the beneficial effects that: the method adopts low-grade tea such as summer and autumn tea and the like as raw materials, and adopts a low-frequency circulating ultrasonic technology to assist in extracting the gamma-aminobutyric acid, the method has high extraction rate and purity of the gamma-aminobutyric acid, the method has low cost, is simple and easy to implement and easy to popularize, the color of the obtained tea protein product is green tea color, and the product quality meets the food sanitation standard. The invention provides a new way for value-added transformation and reutilization of low-grade tea in summer and autumn; develops a novel plant source gamma-aminobutyric acid product, and realizes the comprehensive utilization and the fine and deep processing of tea resources.
Description
Technical Field
The invention relates to the field of development and utilization of plant products, and mainly relates to a process for extracting and purifying gamma-aminobutyric acid from tea.
Background
Tea leaves originate in China, are one of the most popular beverages in the world, and meanwhile, the tea industry is the traditional dominant industry in China. Tea leaves were originally used as sacrificial offerings, and until the west jin became a common beverage popular among people, which has been 6000 years old. The tea contains a plurality of functional components such as tea polyphenol, tea protein, tea polysaccharide, caffeine, gamma-aminobutyric acid and the like, and the functional components have good medicinal value and health-care effect. So far, many studies have been made on various functional components in tea at home and abroad. The tea yield of China is rich, the total tea yield in 2016 reaches about 233.5 ten thousand tons, medium and low-grade tea accounts for a large proportion and is seriously lost in the market, meanwhile, a large amount of summer and autumn tea is not utilized and even is abandoned, and the phenomena greatly influence the development of the tea industry of China and the utilization of tea resources, so that the research and the application of deep processing new process technology on tea are developed, the urgent requirements of improving the utilization rate of the tea resources and keeping the rapid development of the tea industry of China in the international market are met, the tea industry is a necessary way for the traditional tea industry to extend to the industries such as food, medicine and the like, and the tea industry chain is an important means for extending the tea industry chain.
Tea resources in China are rich, tea industry develops rapidly, but spring tea is usually picked in tea gardens, a large amount of summer and autumn tea is not utilized, even is abandoned, and therefore great waste is caused to the tea resources. The tea contains various functional components such as gamma-aminobutyric acid, tea protein and the like, has good health-care effect, and the use of the components in the tea can effectively utilize tea resources.
Gamma-aminobutyric acid is white or yellowish crystalline powder, has strong hygroscopicity, is very soluble in water, is slightly soluble in hot ethanol, is insoluble in organic solvents such as benzene, cold ethanol, diethyl ether and the like, is slightly odorous and has no optical activity, is a polar substance, has a melting point of 203-4H9NO2Dissociation constant of γ -aminobutyric acid: pKCOOH= 4.03，pKNH3= 10.56. Gamma-aminobutyric acid is usually present in zwitterionic form in aqueous solution, and its dissociation depends mainly on the magnitude of pH and its isoelectric point is 7.295. Gamma-aminobutyric acid is a non-protein amino acid, has certain special chemical properties of the amino acid, and can perform a alkylation reaction, an acylation reaction, a ninhydrin color reaction and the like.
In recent years, gamma-aminobutyric acid has been receiving more and more attention as a novel food functional factor. It has been found that gamma-aminobutyric acid can enhance Na pair of nerve cell membrane+Can hyperpolarize the cell membrane and increase the threshold for initiation of action potentials, gamma-aminobutyric acid is therefore a major factor in the central nervous system of humans and mammalsThe inhibitory neurotransmitter can participate in various metabolic activities in organisms, has strong physiological activity, and is a hot research direction concerned in the fields of food, medical animal feed, agriculture and the like in production and application. The physiological functions of gamma-aminobutyric acid are mainly as follows.
a regulation of cardiovascular function
Gamma-aminobutyric acid is used as an important inhibitory transmitter in the central nervous system, has a good regulating effect on cardiovascular activity, and is mainly manifested by regulating heart rate, lowering blood pressure, lowering blood sugar and the like. Gamma-aminobutyric acid has certain pharmacological effect on lowering blood pressure, and the reason may be that specific nerve receptors exist in cerebral blood vessels and can be combined with the gamma-aminobutyric acid, so that the purpose of lowering blood pressure can be achieved when the nerve receptors are combined with the gamma-aminobutyric acid. Experiments of Chinese patent application and the like show that spontaneous hypertension rats can obviously reduce the blood pressure by drinking green tea containing gamma-aminobutyric acid for a long time. Kazami and Inoue K both find that if a patient with hypertension continuously eats a food containing gamma-aminobutyric acid, the blood pressure is obviously reduced; but after the people with normal blood pressure eat the food continuously, the blood pressure can not be changed, other indexes of the body can not be influenced, and no side effect exists. The gamma-aminobutyric acid can also adjust the blood vessel movement center by blocking sympathetic nerves, so that blood vessels are expanded, and the purpose of reducing blood pressure is achieved. Gamma-aminobutyric acid can play a role in reducing blood pressure by inhibiting the activities of angiotensin converting enzyme and vasopressin, an antidiuretic hormone. The gamma-aminobutyric acid can also inhibit the activity of glutamate decarboxylase, and effectively prevent the glutamate decarboxylase from proceeding, so that the blood ammonia content is reduced, and the health of a human body is ensured; meanwhile, the gamma-aminobutyric acid can also promote glutamic acid in blood to be combined with ammonia to generate urea, so that the ammonia toxicity in the body is effectively removed, and the functions of the liver and the kidney are enhanced.
b treating epilepsy
The reported epilepsy patients in China reach about 900 ten thousand, the causes of epilepsy are various, the mechanism is complex, and the epilepsy has close relation with central neurotransmitter. At present, epilepsy becomes the second most common disease in China, and the work and life of people are seriously influenced. And gamma-aminobutyric acid as an important inhibitory neurotransmitter in the human bodyIt can be used for treating epilepsy, insomnia, and Parkinson's disease. There are two types of amino acids, excitatory amino acids and inhibitory amino acids, in the central nervous system, if the content of excitatory amino acids is too high, the neuroinhibitory effect will be reduced, possibly causing epilepsia. Many studies have found that depletion of gamma-aminobutyric acid leads to neuronal hyperexcitation and has specific binding sites in the central nervous system, which increase the Cl-pair of the nerve membrane-Such that the cell membrane potential is at a resting potential level, thereby attenuating synaptic responses to excitatory inputs. Therefore, the gamma-aminobutyric acid can improve the anticonvulsant threshold value of a human body and can effectively treat intractable epilepsy. Okada et al found that gamma-aminobutyric acid can promote sleep, calm nerves, resist anxiety and the like, and contribute to the improvement of symptoms of early-aged mental disorder and female climacteric syndrome by orally administering rice germ food containing gamma-aminobutyric acid, and found that the content of gamma-aminobutyric acid in cerebrospinal fluid is inversely related to the severity of epileptic patients, and the content of gamma-aminobutyric acid is low. Therefore, the epilepsy can be treated by directly increasing the content of the gamma-aminobutyric acid in cerebrospinal fluid. It was found that gamma-aminobutyric acid also reduces the neurogenic excitation caused by glutamate when the brain is severely ischemic, thereby reducing damage to nerve cells. In addition, gamma-aminobutyric acid has certain effect on treating the catalepsy and the Parkinson's disease.
c promoting hormone secretion
The gamma-aminobutyric acid has a regulating effect on the secretion of various hormones such as growth hormone, growth hormone medium, thyroid hormone, sex hormone and the like in an endocrine system, and can effectively improve the metabolic level of a human body. The gamma-aminobutyric acid can directly stimulate the body to secrete the needed hormone, and has higher safety compared with the method of directly taking exogenous hormone. It has been shown that gaba can regulate gonadotropin secretion by inhibiting the excitation of certain nerves in the hypothalamus.
d improving liver and kidney function
The gamma-aminobutyric acid can inhibit decarboxylation of glutamic acid to reduce blood ammonia, and also has the function of diuresis, so that redundant salt in the body is discharged, and the liver and kidney pressure is effectively relieved. It has been shown that gamma-aminobutyric acid can activate kidney function. Meanwhile, gamma-aminobutyric acid can inhibit the activity of alkaline phosphatase, and an important index for evaluating the activity of liver function is the activity of alkaline phosphatase.
e other functions
In addition, the gamma-aminobutyric acid also has the physiological functions of preventing obesity, promoting lipid metabolism, preventing angiosclerosis, repairing skin, delaying aging and the like. Clinically, the gamma-aminobutyric acid can also treat uremia, carbon monoxide poisoning and other diseases. In addition, the gamma-aminobutyric acid also has the effects of killing insects, removing odor and the like.
Therefore, the gamma-aminobutyric acid has strong functions, and must be produced on a large scale in a simple and feasible mode, thereby benefiting mankind.
Disclosure of Invention
The invention aims to provide a process for extracting and purifying gamma-aminobutyric acid from tea aiming at the defects in the existing comprehensive utilization technology of low-grade tea, tea leaves and the like.
The invention also aims to provide the gamma-aminobutyric acid extract prepared by the method, wherein the gamma-aminobutyric acid content of the product is 70-80%, the amino acid content is rich, and the composition is relatively balanced.
The invention adopts the technical scheme for solving the technical problems that: the process for extracting and purifying gamma-aminobutyric acid from tea leaves comprises the following steps:
(1) the raw material pretreatment process comprises the following steps: cleaning tea leaves as a raw material, removing impurities and phenol, deactivating enzymes, drying and crushing for later use;
(2) leaching process of gamma-aminobutyric acid: the method comprises the following steps of (1) carrying out low-frequency ultrasonic assisted alkali extraction to prepare a gamma-aminobutyric acid extracting solution, wherein the specific process comprises the following steps: weighing a certain amount of tea powder, placing the tea powder in a conical flask, adding a certain amount of ultrapure water according to the liquid-material ratio of 20:1 mL/g, shaking uniformly, carrying out ultrasonic leaching for 30min at the temperature of 50 ℃ and the power of 220W, cooling, centrifuging, and carrying out suction filtration to obtain a gamma-aminobutyric acid crude extract;
(3) and (3) a gamma-aminobutyric acid concentration process: and (4) concentrating the gamma-aminobutyric acid crude extract by using a rotary evaporator, and adding water to prepare the required sample solution concentration. (ii) a
(4) The separation and purification process of the gamma-aminobutyric acid concentrated solution comprises the following steps: static adsorption or dynamic elution can be adopted for separation and purification;
(5) drying and storing process of gamma-aminobutyric acid;
preferably: cleaning and airing fresh leaves until the water content is about 80 percent in the raw material pretreatment process in the step (1), respectively treating the fresh leaves for 16 h at the temperature of minus 18 ℃, then carrying out enzyme deactivation for 2-3 min by using a microwave oven (using medium-high grade, uniformly stirring every 30 s), then placing the fresh leaves into a blast drying oven, drying the fresh leaves at the temperature of 80 ℃ until the fresh leaves are completely dried, and crushing the fresh leaves and sieving the fresh leaves by using a 80-mesh sieve.
Preferably: the raw material pretreatment process in the step (1) can also be that fresh leaves are cleaned and aired to the water content of about 80%, the fresh leaves are placed in a vacuum drying oven to be subjected to anaerobic treatment for 2 hours at the vacuum degree of 0.085MPa and the temperature of 30 ℃, then the fresh leaves are spread in the air for 1 hour, the anaerobic treatment is performed for 4 times, the processes are respectively circulated for 4 times, then the green removing is performed for 2-3 min by a microwave oven (the middle-high grade is used, the green removing is performed once every 30 s), then the fresh leaves are placed in a blast drying oven to be dried to the absolute dryness at the temperature of 80.
Preferably: the leaching process of the gamma-aminobutyric acid of the tea in the step (2) is as follows: weighing a certain amount of tea powder, placing the tea powder in a conical flask, adding a certain amount of ultrapure water according to the liquid-material ratio of 20:1 mL/g, shaking up, carrying out ultrasonic leaching for 30min at the temperature of 50 ℃ and the power of 220W, cooling, centrifuging, and carrying out suction filtration to obtain a gamma-aminobutyric acid crude extract.
Preferably: the process of concentrating the gamma-aminobutyric acid extracting solution in the step (3) comprises the following steps: vacuum concentration or rotary evaporation concentration is adopted, the vacuum concentration temperature is controlled at 40-60 ℃, and the concentration is carried out until the volume is 20-50% of the original volume. The rotary evaporation concentration time is 2 h.
Preferably: the static adsorption in the step (4) is specifically as follows: adding 95% ethanol into resin with a certain volume, soaking for 24 h, and washing with water until no floccule and no ethanol smell exist; then soaking the mixture for 5 hours by using 4 percent NaOH, washing the mixture to be neutral by using water, then soaking the mixture for 8 hours by using 5 percent HCl, and washing the mixture to be neutral by using water; the dynamic adsorption elution process comprises the following steps: the flow rate of loading was 3mL/min, the loading concentration was 0.2 mg/mL, the eluent pH =9, and the elution flow rate was 2 mL/min. Under the condition, the retention rate of the gamma-aminobutyric acid is 80.06%, and the purity of the gamma-aminobutyric acid reaches 78.51%.
Preferably: the drying and storing process of the gamma-aminobutyric acid in the step (5) comprises the following steps: adding purified water with the weight being 20-60 times of that of the precipitate into the obtained gamma-aminobutyric acid precipitate, washing for 6-8 times at room temperature, carrying out centrifugal separation for 30min at 11000r/min by 5000-.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples. It should be understood that the examples described herein are for illustrative purposes only and are not intended to limit the present invention.
(1) The raw material pretreatment process comprises the following steps: cleaning tea leaves as a raw material, removing impurities and phenol, deactivating enzymes, drying and crushing for later use; cleaning fresh leaves, air drying to water content of about 80%, treating at-18 deg.C for 16 hr, deactivating enzyme with microwave oven for 2-3 min (using medium-high grade, turning uniformly every 30 s), oven drying at 80 deg.C in blast drying oven, pulverizing, and sieving with 80 mesh sieve. Or cleaning fresh leaves, air drying to water content of about 80%, treating at-18 deg.C for 16 hr, deactivating enzyme with microwave oven for 2-3 min (using medium-high grade, turning uniformly every 30 s), oven drying at 80 deg.C in blast drying oven, pulverizing, and sieving with 80 mesh sieve.
(2) Leaching process of gamma-aminobutyric acid: the method comprises the following steps of (1) carrying out low-frequency ultrasonic assisted alkali extraction to prepare a gamma-aminobutyric acid extracting solution, wherein the specific process comprises the following steps: weighing a certain amount of tea powder, placing the tea powder in a conical flask, adding a certain amount of ultrapure water according to the liquid-material ratio of 20:1 mL/g, shaking uniformly, carrying out ultrasonic leaching for 30min at the temperature of 50 ℃ and the power of 220W, cooling, centrifuging, and carrying out suction filtration to obtain a gamma-aminobutyric acid crude extract;
(3) and (3) a gamma-aminobutyric acid concentration process: and (4) concentrating the gamma-aminobutyric acid crude extract by using a rotary evaporator, and adding water to prepare the required sample solution concentration. Vacuum concentration or rotary evaporation concentration is adopted, the vacuum concentration temperature is controlled at 40-60 ℃, and the concentration is carried out until the volume is 20-50% of the original volume. The rotary evaporation concentration time is 2h
(4) The separation and purification process of the gamma-aminobutyric acid concentrated solution comprises the following steps: static adsorption or dynamic elution can be adopted for separation and purification; the medium static adsorption specifically comprises the following steps: adding 95% ethanol into resin with a certain volume, soaking for 24 h, and washing with water until no floccule and no ethanol smell exist; then soaking the mixture for 5 hours by using 4 percent NaOH, washing the mixture to be neutral by using water, then soaking the mixture for 8 hours by using 5 percent HCl, and washing the mixture to be neutral by using water; the dynamic adsorption elution process comprises the following steps: the flow rate of loading was 3mL/min, the loading concentration was 0.2 mg/mL, the eluent pH =9, and the elution flow rate was 2 mL/min. Under the condition, the retention rate of the gamma-aminobutyric acid is 80.06%, and the purity of the gamma-aminobutyric acid reaches 78.51%.
(5) Drying and storing process of gamma-aminobutyric acid; adding purified water with the weight being 20-60 times of that of the precipitate into the obtained gamma-aminobutyric acid precipitate, washing for 6-8 times at room temperature, carrying out centrifugal separation for 30min at 11000r/min by 5000-.
The gamma-aminobutyric acid obtained by the method has high purity, and original nutrient substances are completely preserved, so that the method is suitable for large-area popularization.
Claims (7)
1. A process for extracting and purifying gamma-aminobutyric acid from tea is characterized by comprising the following steps: the method comprises the following steps:
(1) the raw material pretreatment process comprises the following steps: cleaning tea leaves as a raw material, removing impurities and phenol, deactivating enzymes, drying and crushing for later use;
(2) leaching process of gamma-aminobutyric acid: the method comprises the following steps of (1) carrying out low-frequency ultrasonic assisted alkali extraction to prepare a gamma-aminobutyric acid extracting solution, wherein the specific process comprises the following steps: weighing a certain amount of tea powder, placing the tea powder in a conical flask, adding a certain amount of ultrapure water according to the liquid-material ratio of 20:1 mL/g, shaking uniformly, carrying out ultrasonic leaching for 30min at the temperature of 50 ℃ and the power of 220W, cooling, centrifuging, and carrying out suction filtration to obtain a gamma-aminobutyric acid crude extract;
(3) and (3) a gamma-aminobutyric acid concentration process: concentrating the gamma-aminobutyric acid crude extract by using a rotary evaporator, and adding water to prepare the required sample solution concentration;
(4) the separation and purification process of the gamma-aminobutyric acid concentrated solution comprises the following steps: static adsorption or dynamic elution can be adopted for separation and purification;
(5) drying and storing process of gamma-aminobutyric acid.
2. The process for extracting and purifying gamma-aminobutyric acid from tea leaves as claimed in claim 1, wherein the process comprises the following steps: cleaning and airing fresh leaves until the water content is about 80 percent in the raw material pretreatment process in the step (1), respectively treating the fresh leaves for 16 h at the temperature of minus 18 ℃, then carrying out enzyme deactivation for 2-3 min by using a microwave oven (using medium-high grade, uniformly stirring every 30 s), then placing the fresh leaves into a blast drying oven, drying the fresh leaves at the temperature of 80 ℃ until the fresh leaves are completely dried, and crushing the fresh leaves and sieving the fresh leaves by using a 80-mesh sieve.
3. The process for extracting and purifying gamma-aminobutyric acid from tea leaves as claimed in claim 1, wherein the process comprises the following steps: the raw material pretreatment process in the step (1) can also be that fresh leaves are cleaned and aired to the water content of about 80%, the fresh leaves are placed in a vacuum drying oven to be subjected to anaerobic treatment for 2 hours at the vacuum degree of 0.085MPa and the temperature of 30 ℃, then the fresh leaves are spread in the air for 1 hour, the anaerobic treatment is performed for 4 times, the processes are respectively circulated for 4 times, then the green removing is performed for 2-3 min by a microwave oven (the middle-high grade is used, the green removing is performed once every 30 s), then the fresh leaves are placed in a blast drying oven to be dried to the absolute dryness at the temperature of 80.
4. The process for extracting and purifying gamma-aminobutyric acid from tea leaves as claimed in claim 1, wherein the process comprises the following steps: the leaching process of the gamma-aminobutyric acid of the tea in the step (2) is as follows: weighing a certain amount of tea powder, placing the tea powder in a conical flask, adding a certain amount of ultrapure water according to the liquid-material ratio of 20:1 mL/g, shaking up, carrying out ultrasonic leaching for 30min at the temperature of 50 ℃ and the power of 220W, cooling, centrifuging, and carrying out suction filtration to obtain a gamma-aminobutyric acid crude extract.
5. The process for extracting and purifying gamma-aminobutyric acid from tea leaves as claimed in claim 1, wherein the process comprises the following steps: the process of concentrating the gamma-aminobutyric acid extracting solution in the step (3) comprises the following steps: vacuum concentration or rotary evaporation concentration is adopted, the vacuum concentration temperature is controlled to be 40-60 ℃, and the concentration is carried out until the volume is 20-50% of the original volume; the rotary evaporation concentration time is 2 h.
6. The process for extracting and purifying gamma-aminobutyric acid from tea leaves as claimed in claim 1, wherein the process comprises the following steps: the static adsorption in the step (4) is specifically as follows: adding 95% ethanol into resin with a certain volume, soaking for 24 h, and washing with water until no floccule and no ethanol smell exist; then soaking the mixture for 5 hours by using 4 percent NaOH, washing the mixture to be neutral by using water, then soaking the mixture for 8 hours by using 5 percent HCl, and washing the mixture to be neutral by using water; the dynamic adsorption elution process comprises the following steps: the sample loading flow rate is 3mL/min, the sample loading concentration is 0.2 mg/mL, the eluent pH =9, and the elution flow rate is 2 mL/min;
under the condition, the retention rate of the gamma-aminobutyric acid is 80.06%, and the purity of the gamma-aminobutyric acid reaches 78.51%.
7. The process for extracting and purifying gamma-aminobutyric acid from tea leaves as claimed in claim 1, wherein the process comprises the following steps: the drying and storing process of the gamma-aminobutyric acid in the step (5) comprises the following steps: adding purified water with the weight being 20-60 times of that of the precipitate into the obtained gamma-aminobutyric acid precipitate, washing for 6-8 times at room temperature, carrying out centrifugal separation for 30min at 11000r/min by 5000-.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114343049A (en) * | 2021-12-31 | 2022-04-15 | 淮阴工学院 | Preparation method of water caltrop ice cream |
| CN115477592A (en) * | 2022-09-30 | 2022-12-16 | 西安萃源生物科技有限公司 | Crystallization method of gamma-aminobutyric acid |
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2018
- 2018-09-29 CN CN201811150669.3A patent/CN110963933A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114343049A (en) * | 2021-12-31 | 2022-04-15 | 淮阴工学院 | Preparation method of water caltrop ice cream |
| CN114343049B (en) * | 2021-12-31 | 2023-09-29 | 淮阴工学院 | Preparation method of water chestnut ice cream |
| CN115477592A (en) * | 2022-09-30 | 2022-12-16 | 西安萃源生物科技有限公司 | Crystallization method of gamma-aminobutyric acid |
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