CN101497906A - Separation and purification preparation as well as hypoglycemic activity of tea polysaccharide - Google Patents
Separation and purification preparation as well as hypoglycemic activity of tea polysaccharide Download PDFInfo
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Abstract
The invention relates to a method for separating, purifying and preparing tea polysaccharide and the hypoglycemic activity of the tea polysaccharide. The method comprises the following processing steps of raw materials preparation, high-pressure draft breakage, screening, extracting microwave secondary solvent, filtering, vacuum enzyme extraction, filtering, Combining the filtrate through repeatedly extracting filter residues, centrifugation, radial highly-efficient chromatographic fractionation, ultrafiltration, vacuum concentration-, freeze drying and obtaining tea polysaccharide. The invention has the technical effect that the content of the tea polysaccharide and the hypoglycemic activity is improved, the extraction time is shortened, the energy consumption is reduced, and the biological activity of the tea polysaccharide is preserved effectively.
Description
Technical field
The present invention relates to a kind of preparation method of tea polysaccharide, relate in particular to a kind of preparation method for separating and purifying and hypoglycemic activity of tea polysaccharide
Background technology
Tealeaves is large agricultural byproducts of China, about 1,000,000 hectares of existing tea place area is produced about about 600,000 tons of tealeaves per year, wherein has coarse tea leaf about 100,000 tons to stagnate approximately to disappear or overstocks, if do not consider the comprehensive utilization of these tea making byproducts, the very big wasting of resources and financial loss will be caused.Begin from the eighties in 20th century, Chinese scholars has been carried out a large amount of research to polysaccharide compound contained in the tealeaves, finds that tea polysaccharide has multiple biological activitys such as the body immunity of increasing, hypoglycemic, reducing blood-fat and minimizing atherosclerosis, antitumor, radioprotective and anti-inflammatory.
The patent report that extracts about tea polysaccharide mainly contains in recent years: Zhang Xiaolin " a kind of method of extracting tea-polyphenol by-product trimethyl-xanthine and tea polysaccharide from tealeaves " (application number 200510042629.3), Li Yongquan " extracts the method for tea polysaccharide " (application number 03114898.0) from the tankage that extracted tea-polyphenol, Wang Shuxiong " extracts the method for tea-polyphenol and byproduct thereof " (application number 96113134.9) in tealeaves, Yan Liangrong " thick; that old tea leaves extracts polysaccharide; polyphenol mixed crystal processing method " (application number 97109115.3), Huang Baosheng " extracting method of tea polysaccharide " (application number 01134065.7), Hui Yongzheng " purifying tea polysaccharide and deduction method " (application number 02110999.0), river and source " the comprehensive tea polysaccharide of extracting from tealeaves; tea-polyphenol; theanine; the method for trimethyl-xanthine " (application number 200410015905.2), Wang Dongfeng " effective constituents in tea preparation technology " (application number 200310114694.3), Zhang Shouzheng " extracts tea-polyphenol; theanine; tea polysaccharide; the method for tea pigment " (application number 200610055145.7) from tealeaves, Yang Hongwu " a kind of tea polysaccharide and its production and use " (application number 200610125174.6), these patent reports have enriched tea polysaccharide greatly and have extracted means of purification, but are not purified into a kind of one-component from tea leaf extract.
The characteristic of present method is to improve in the tea polysaccharide extraction process extraction yield from tealeaves, shorten extraction time, reduce energy consumption, effectively preserve purpose such as tealeaves polysaccharide biological activity, and purifying obtains a polysaccharide and proteic conjugate component (Tea Glycocongujate, and conformation in physico-chemical property, molecular weight, amino acid composition, monose composition, primary structure and the solution of this component etc. has been carried out the Analysis and Identification of system TGC).
α-Dian Fenmei participates in the carbohydrate metabolism process of human body, in small intestine, is amylolysis glucose and oligosaccharides by the α-Dian Fenmei of pancreas.It is generally acknowledged that in the past amylase inhibitor there is no direct effect to the utilization of Regular Insulin and blood sugar, only has the assistant hypoglycemic effect, and does not have direct therapeutic action.Amylase inhibitor is hypoglycemic to be the hypoglycemic new approaches of rising after the seventies.Research is in recent years also found, especially postprandial blood sugar is too high, blood sugar descended is the main reason that causes the many organ injuries of patient's multisystem slowly for the long hyperglycemia of diabetics, and to be that monose reduces level of postprandial blood sugar just apparent to diabetic subject's benefit thereby therefore reduce amylolysis.Amylase inhibitor can suppress digestive tube, especially duodenal amylase activity, reduce or delay digestive tube starch and be hydrolyzed to monose, make the absorption minimizing of monose and lowering blood glucose, so the fat increase that diabetics's postprandial hyperglycemia and obese person's overfeeding carbohydrate are caused has antagonistic action.Late 1980s, China also began the screening and the development of this type of medicine, but product-free appearance still at present.The inhibition effect that this research is lived to the α-Dian Fenmei enzyme by the mensuration product is further estimated the effect of product, instructs and produces.
Summary of the invention
The object of the present invention is to provide a kind of preparation method for separating and purifying and hypoglycemic activity of tea polysaccharide, this method shortens extraction time, reduces energy consumption, effectively preserves tealeaves polysaccharide biological activity.
The present invention is achieved in that its processing step is:
1, raw material: with the tea waste residue behind coarse tea leaf, low-grade tea or the extraction tea-polyphenol is raw material;
2, high pressure draft fragmentation: with raw material after pre-treatment such as 50-60 ℃ of dryings ,-40 ℃ are freezing, the air-flowing type micronizing is handled raw material, it mainly is that impact, shearing, collision and friction by air-flow pulverized material, the requirement bleed pressure is 5MPa, pulverizing pressure is 3-5MPa, thereby the stripping quantity that makes raw material isoreactivity composition improves, powder granularity reduces, to improve in the raw material problems such as yield is low in the tea polysaccharide leaching process.
3, sieve: the raw material after the high pressure draft fragmentation carries out classification 40-80 orders that sieve;
4, microwave-assisted solvent extraction: adopt microwave extraction that the interior temperature of raw cell is risen rapidly, the pressure that the vaporization of intracellular fluid moisture content produces makes cytolemma and cell walls sharply broken, forms small hole, makes the quick stripping of soluble substance.Microwave-assisted solvent extraction optimum process condition is 75 seconds microwave time, microwave intensity 100%, solid-to-liquid ratio 1:15;
5, the vacuum enzyme is carried: adopt Cellubrix
TML cellulase (cellulase 1500NCU/mL, cellobiase 25CbU/mL), Termamyl
The heatproof α-Dian Fenmei, for keeping the activity of tea polysaccharide, employing vacuum and low temperature water is carried, enzyme is put forward the secondary bond method and extracted tea polysaccharide.At 50 ℃, solid-to-liquid ratio 1:15, water extraction 30min, polysaccharide extract rate is 2.33% for the first time, and the extraction yield of Crude polysaccharides (dry weight) is 6.82%.After the filtration, filter residue adds cellulase with citric acid-sodium citrate buffer solution of pH4.6 and extracts, and it is 55 ℃, solid-to-liquid ratio 1:14, enzyme dosage 2.2 μ L/g tealeaves that enzyme is carried optimal processing parameter, 120min;
6, radially high efficiency chromatography separates: the gel column purifying of polysaccharide: the preliminary purification tea polysaccharide of preparation is dissolved in (or directly using without the exsiccant extracting solution) in the distilled water.Cross SephadexG100 gel column (2.6 * 70cm).Dropwise sample liquid is added in the post with the 10ml sampler, with 0.1mol/L NaCl wash-out, flow velocity 1.5mL/min, the every 6min of fraction collection instrument collects a pipe automatically.Detect A by pipe
280nm(protein absorption peak), A
620(absorption peak of the anthrone sulfuric acid derivative of polysaccharide).Collection contains sugar moieties, concentrates, and is standby.Sephadex G 100 gel wet methods dress post: get a certain amount of Sephadex G100 gel, place beaker, add the distilled water immersion of capacity, swelling is 5 to 6 hours in 100 ℃ of water-baths, after the taking-up cooling, continues swelling in 24 hours in the room temperature placement.After swelling finishes, the suspended substance on the surface of inclining, ultrasonic degas can be adorned post.What this patent adopted is the special-purpose chromatography column of tetrafluoroethylene, adding distil water is to certain altitude in post, by the top of funnel the swollen gel is added the post from post, the bottom of post discharges water simultaneously, the limit edged beats shaft, finish until filling out post, attention will keep gel interface to be in below the water surface all the time.After the dress post is finished, use distilled water balance one day earlier, use the elutriant balance at last, flow velocity is less than elution flow rate during balance.HPLC identifies when groping of purity and chromatographic condition thereof: HPLC analyzes, the employing hand sampling, and each sample size is 20 μ L, elution flow rate is 0.5mL/min.Sample solution and elutriant are used the membrane filtration of 0.80 μ m and 0.45 μ m respectively.
7, ultrafiltration: select the tubular fibre material for use, operating pressure is 28psi, and feed temperature is 29 ℃, and material liquid pH is 6.7, and it is 9.98LHM that uf processing 40min, the permeation flux of film reach optimum value;
8, lyophilize: lyophilize condition: behind-80 ℃ of pre-freeze 8hr ,-80 ℃ of vacuum lyophilizations 6 hours.
9, alpha-amylase activity suppresses experiment
The method of calculation that suppress alpha-amylase activity are the spectrophotometer method (method that provides according to Novozymes Company, with " absorbance A-enzyme concn c " appendix table): the phosphoric acid buffer 5.0mL that draws Zulkovsky starch solution 20.0mL and pH6.0 is in test tube, pre-thermal equilibrium is 5 minutes in 70 ℃ of waters bath with thermostatic control, join then in 20 minutes mixing solutions of 55 ℃ of preheatings (contain polysaccharide solution and the 1mL enzyme liquid to be measured of 1mL, enzyme liquid has been diluted to suitable concn).Shake up accurate response 5 minutes.Draw reaction solution 1.00mL immediately, be added in the test tube that fills 0.5mL hydrochloric acid (0.1mol/L) and the rare iodine liquid of 5.00mL in advance, shake up.Replace starch solution to do the reagent blank zeroing with distilled water.660nm place 10mm cuvette its absorbancy of rapid test (A).
In the method for the present invention, the tea polysaccharide solution of extract of preliminary purification is as follows to α-Dian Fenmei enzyme activity system effect:
The different tea polysaccharide solution of extract of table 1 suppress the contrast of effect to the α-Dian Fenmei enzyme activity
The result shows, the second time, the polysaccharide with the inhibiting rate ratio water extraction first time of the tea polysaccharide of cellulase extraction hanged down 21.2%, this may be because variation has to a certain degree taken place for the molecular weight or the structure of polysaccharide, perhaps change has taken place in the molecular weight distribution of this mixing polysaccharide, the polysaccharide of some other molecular weight has mixed, but main kind does not become, and still has the activity of stronger inhibition α-Dian Fenmei.Water is carried the polysaccharide low 76.2% of the inhibiting rate of tea polysaccharide than the water extraction first time for the second time, this may be since under cold condition for the second time the composition of the tea polysaccharide that obtains of water extraction differ greatly with the polysaccharide of water extraction for the first time, wherein may be mainly the bigger polysaccharose substances such as soluble cellulose of molecular weight.
In the method for the present invention, it is as follows to α-Dian Fenmei enzyme activity inhibition effect that differing temps is extracted the tea polysaccharide powder:
Table 2 polysaccharide extracts temperature to suppressing the influence of α-Dian Fenmei effect
Result of study shows that it is high more to extract temperature, though the yield raising may make tea polysaccharide degraded or sex change, makes thereby influence is active and suppresses the effect reduction.It is too low as the normal temperature extraction to extract temperature, may be low owing to active tea polysaccharide content in the extract, and it is also bad to suppress effect.
In the method for the present invention, tea polysaccharide powder consumption is as follows to the influence that it suppresses α-Dian Fenmei enzyme activity effect:
The results are shown in Table 3.Result of study shows, the tea polysaccharide consumption increases, and can improve the inhibiting rate of α-Dian Fenmei, and consumption is increased to a certain degree, and the inhibiting rate increasing degree can reduce.
Table 3 tea polysaccharide consumption is to suppressing the influence of α-Dian Fenmei effect
In the method for the present invention, the protein chain of tea polysaccharide is as follows to the influence that suppresses α-Dian Fenmei enzyme activity effect:
Table 4 protein removal method is to suppressing the influence of effect
Tea polysaccharide is an acid glycoprotein, through different removal albumen tests, find owing to the Degradation of proteolytic enzyme the protein chain of glycoprotein, the tealeaves polysaccharide is reduced greatly to the restraining effect of α-Dian Fenmei, and the Sevag method removes albumen because mild condition, can the protein chain of glycoprotein not exerted an influence,, illustrate that the protein chain of tealeaves glycoprotein cuts much ice to its inhibition activity to α-Dian Fenmei so inhibiting rate is high a lot of than the former.Assorted floating preteins makes the purity drop of polysaccharide, thereby makes that the tea polysaccharide of specific activity through removing the albumen preliminary purification of thick tea polysaccharide is low.
In the method for the present invention, acidic polysaccharose is as follows to the inhibition effect of α-Dian Fenmei:
Be 50 ℃ of water extraction equally, the tea polysaccharide extract that obtains with ethanol sedimentation and the acid tea polysaccharide that obtains with the CTAB precipitation see Table 5 to the inhibition effect of α-Dian Fenmei.
Table 5 acidic polysaccharose and mixing tea polysaccharide are to the comparison of the inhibition effect of α-Dian Fenmei
With the sedimentary acid tea polysaccharide of CTAB to the inhibition effect of α-Dian Fenmei significantly better than the ethanol sedimentation sample, but yield reduces.This may mean that the material that α-Dian Fenmei is played inhibition is certain acidic polysaccharose.
In the method for the present invention, the existence of tea polysaccharide and conformation are as follows to the influence that it suppresses the alpha-amylase activity effect:
The tea polysaccharide powder and the aqueous solution see Table 6 to the comparative result that the α-Dian Fenmei enzyme activity suppresses effect.
The existence of table 6 tea polysaccharide and conformation are to suppressing the influence of effect
The result shows, because the natural radioactivity conformation of tea polysaccharide has obtained maximum preservation in the solution, thereby its inhibition effect to α-Dian Fenmei is better than the Powdered tea polysaccharide of exsiccant far away, illustrate that tea polysaccharide is active closely related with its conformation to the inhibition of α-Dian Fenmei, drying makes tea polysaccharide dewater, native conformation is destroyed, thereby greatly influences its activity.
In the method for the present invention, it is as follows that the different purity tea polysaccharide suppresses α-amylase activity experimental result:
α-Dian Fenmei participates in the carbohydrate metabolism process of human body, and tea polysaccharide suppresses the vigor of α-Dian Fenmeishuixie starch, and the release of delay glucose may be one of its hypoglycemic mechanism.The result shows, raising along with the purity of polysaccharide, the inhibition activity of tea polysaccharide also increases, the inhibition α of tea polysaccharide TGC-diastatic activity is the highest, activity than the tea polysaccharide of preliminary purification exceeds 367.86%, from α-diastatic inhibiting rate (PER), the inhibiting rate of 500mgTGC is up to 97.2%.
Technique effect of the present invention is: improved tea polysaccharide content and hypoglycemic activity, improved the tea polysaccharide extraction yield, shorten extraction time, reduce energy consumption, effectively preserve tealeaves polysaccharide biological activity.
Embodiment
Embodiment one:
With the coarse tea leaf after pre-treatment such as 50 ℃ of dryings ,-40 ℃ are freezing, the air-flowing type micronizing is handled tealeaves, take by weighing 40 order tea powder 100g, the extraction of microwave-assisted solvent, 75 seconds microwave time, microwave intensity 100%, solid-to-liquid ratio 1: 15, double-layer filter cloth filters, and 1. filter residue washs with hot distilled water, merging filtrate and washings obtain extracting solution I; 2. filter residue washs with hot distilled water, and filter residue adds the citric acid-sodium citrate damping fluid of pH4.6, adds a certain amount of enzyme, the heating in water bath hydrolysis, and adjust pH is to the neutral enzyme that presses down, and double-layer filter cloth filters, and filter residue washs with hot distilled water, and merging filtrate and washings get extracting solution II.
Extracting solution I and II is centrifugal respectively, and vacuum concentration adds dehydrated alcohol to certain volume, the low temperature alcohol precipitation that spends the night, with the centrifugation of ethanol sedimentation liquid, precipitation is used dehydrated alcohol, acetone, ether washed twice successively, gets tealeaves Crude polysaccharides I (water is carried) and II (enzyme is carried).
The preliminary purification tea polysaccharide of preparation is dissolved in (or directly using without the exsiccant extracting solution) in the distilled water.Cross Sephadex G100 gel column (2.6 * 70cm).Dropwise sample liquid is added in the post with the 10ml sampler, with 0.1mol/L NaCl wash-out, flow velocity 1.5mL/min, the every 6min of fraction collection instrument collects a pipe automatically.Detect A by pipe
280nm(protein absorption peak), A
620(absorption peak of the anthrone sulfuric acid derivative of polysaccharide).Collection contains sugar moieties, concentrates, and is standby.
Get a certain amount of Sephadex G100 gel, place beaker, add the distilled water immersion of capacity, swelling is 5 hours in 100 ℃ of water-baths, after the taking-up cooling, places 24 hours continuation swellings in room temperature.After swelling finishes, the suspended substance on the surface of inclining, ultrasonic degas can be adorned post.What experiment was adopted is the special-purpose chromatography column of tetrafluoroethylene, adding distil water is to certain altitude in post, by the top of funnel the swollen gel is added the post from post, the bottom of post discharges water simultaneously, the limit edged beats shaft, finish until filling out post, attention will keep gel interface to be in below the water surface all the time.After the dress post is finished, use distilled water balance one day earlier, use the elutriant balance at last, flow velocity is less than elution flow rate during balance.
Embodiment two:
With the coarse tea leaf after pre-treatment such as 60 ℃ of dryings ,-40 ℃ are freezing, the air-flowing type micronizing is handled tealeaves, take by weighing 40 order tea powder 50g, the extraction of microwave-assisted solvent, 75 seconds microwave time, microwave intensity 100%, solid-to-liquid ratio 1:15, double-layer filter cloth filters, and 1. filter residue washs with hot distilled water, merging filtrate and washings obtain extracting solution I; 2. filter residue washs with hot distilled water, and filter residue adds the citric acid-sodium citrate damping fluid of pH4.6, adds a certain amount of enzyme, the heating in water bath hydrolysis, and adjust pH is to the neutral enzyme that presses down, and double-layer filter cloth filters, and filter residue washs with hot distilled water, and merging filtrate and washings get extracting solution II.
Extracting solution I and II is centrifugal respectively, and vacuum concentration adds dehydrated alcohol to certain volume, the low temperature alcohol precipitation that spends the night, with the centrifugation of ethanol sedimentation liquid, precipitation is used dehydrated alcohol, acetone, ether washed twice successively, gets tealeaves Crude polysaccharides I (water is carried) and II (enzyme is carried).
The preliminary purification tea polysaccharide of preparation is dissolved in (or directly using without the exsiccant extracting solution) in the distilled water.Cross Sephadex G100 gel column (2.6 * 70cm).Dropwise sample liquid is added in the post with the 10ml sampler, with 0.1mol/L NaCl wash-out, flow velocity 1.5mL/min, the every 6min of fraction collection instrument collects a pipe automatically.Detect A by pipe
280nm(protein absorption peak), A
620(absorption peak of the anthrone sulfuric acid derivative of polysaccharide).Collection contains sugar moieties, concentrates, and is standby.
Get a certain amount of Sephadex G100 gel, place beaker, add the distilled water immersion of capacity, swelling is 6 hours in 100 ℃ of water-baths, after the taking-up cooling, places 24 hours continuation swellings in room temperature.After swelling finishes, the suspended substance on the surface of inclining, ultrasonic degas can be adorned post.What experiment was adopted is the special-purpose chromatography column of tetrafluoroethylene, adding distil water is to certain altitude in post, by the top of funnel the swollen gel is added the post from post, the bottom of post discharges water simultaneously, the limit edged beats shaft, finish until filling out post, attention will keep gel interface to be in below the water surface all the time.After the dress post is finished, use distilled water balance one day earlier, use the elutriant balance at last, flow velocity is less than elution flow rate during balance.
Claims (2)
1, a kind of preparation method for separating and purifying of tea polysaccharide is characterized in that preparing as follows:
(1) raw material: with the tea waste residue behind coarse tea leaf, low-grade tea or the extraction tea-polyphenol is raw material;
(2) high pressure draft fragmentation: with raw material after pre-treatment such as 50-60 ℃ of dryings ,-40 ℃ are freezing, the air-flowing type micronizing is handled raw material, it mainly is that impact, shearing, collision and friction by air-flow pulverized material, the requirement bleed pressure is 5MPa, pulverizing pressure is 3-5MPa, thereby the stripping quantity that makes raw material isoreactivity composition improves, powder granularity reduces, to improve in the raw material problems such as yield is low in the tea polysaccharide leaching process;
(3) sieve: the raw material after the high pressure draft fragmentation carries out classification 40-80 orders that sieve;
(4) microwave-assisted solvent extraction: adopt microwave extraction that the interior temperature of raw cell is risen rapidly, the pressure that the vaporization of intracellular fluid moisture content produces makes cytolemma and cell walls sharply broken, form small hole, make the quick stripping of soluble substance, microwave-assisted solvent extraction optimum process condition is 75 seconds microwave time, microwave intensity 100%, solid-to-liquid ratio 1:15;
(5) the vacuum enzyme is carried: adopt cellulase, the heatproof α-Dian Fenmei, for keeping the activity of tea polysaccharide, employing vacuum and low temperature water is carried, enzyme is put forward the secondary bond method and extracted tea polysaccharide, for the first time at 50 ℃, solid-to-liquid ratio 1:15, water extraction 30min, the tea polysaccharide extraction yield is 2.33%, the extraction yield of thick tea polysaccharide is 6.82%, with dry weight basis, after the filtration, filter residue adds cellulase with citric acid-sodium citrate buffer solution of pH4.6 and extracts, and it is 55 ℃, solid-to-liquid ratio 1:14, enzyme dosage 2.2 μ L/g tealeaves that enzyme is carried optimal processing parameter, 120min;
(6) radially high efficiency chromatography separates: the gel column purifying of tea polysaccharide: the preliminary purification tea polysaccharide of preparation is dissolved in the distilled water, cross Sephadex G100 gel column, dropwise sample liquid is added in the post with the 10ml sampler, with 0.1mol/L NaCl wash-out, flow velocity 1.5mL/min, automatically the every 6min of fraction collection instrument collects a pipe, detects by pipe
, A
620, collection contains sugar moieties, concentrates, standby, Sephadex G 100 gel wet methods dress post: get a certain amount of SephadexG100 gel, place beaker, add the distilled water immersion of capacity, swelling is 5 to 6 hours in 100 ℃ of water-baths, after taking out cooling, place 24 hours continuation swellings in room temperature, after swelling finishes, surperficial suspended substance inclines, ultrasonic degas can be adorned post, and what this patent adopted is the special-purpose chromatography column of tetrafluoroethylene, adding distil water is to certain altitude in post, by the top of funnel from post the swollen gel is added the post, the bottom of post discharges water simultaneously, and the limit edged beats shaft, finish until filling out post, attention will keep gel interface to be in all the time below the water surface, after the dress post is finished, uses distilled water balance one day earlier, use the elutriant balance at last, flow velocity is less than elution flow rate during balance.HPLC identifies when groping of purity and chromatographic condition thereof: HPLC analyzes, the employing hand sampling, and each sample size is 20 μ L, elution flow rate is 0.5mL/min.Sample solution and elutriant are used the membrane filtration of 0.80 μ m and 0.45 μ m respectively;
(7) ultrafiltration: select the tubular fibre material for use, operating pressure is 28psi, and feed temperature is 29 ℃, and material liquid pH is 6.7, and it is 9.98LHM that uf processing 40min, the permeation flux of film reach optimum value;
(8) lyophilize: the lyophilize condition, behind-80 ℃ of pre-freeze 8hr ,-80 ℃ of vacuum lyophilizations 6 hours.
2, the preparation method for separating and purifying of tea polysaccharide as claimed in claim 1, it is characterized in that resulting tea polysaccharide, the activity that adopts spectrophotometer method to calculate the tea polysaccharide of tea polysaccharide inhibition α-diastatic specific activity preliminary purification exceeds 367.86%, from α-diastatic inhibiting rate, the inhibiting rate of 500mgTGC is up to 97.2%.
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| CN108623705A (en) * | 2018-06-06 | 2018-10-09 | 完美（中国）有限公司 | A kind of preparation method of dendrobium candidum polysaccharide extractive |
| CN109021142A (en) * | 2018-09-27 | 2018-12-18 | 咸宁市农业科学院 | One grows tea the extracting method that milli extracts tea polyphenols |
| CN112844170A (en) * | 2020-12-22 | 2021-05-28 | 福建安溪铁观音集团股份有限公司 | Tea leaf and tea stalk multi-component modularized synchronous extraction production line |
| CN112844170B (en) * | 2020-12-22 | 2022-06-28 | 福建安溪铁观音集团股份有限公司 | Tea leaf and tea stalk multi-component modularized synchronous extraction production line |
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