CN111602730A - Preparation method of high-purity tea polyphenol compounds in summer and autumn tea leaves - Google Patents
Preparation method of high-purity tea polyphenol compounds in summer and autumn tea leaves Download PDFInfo
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- 241001122767 Theaceae Species 0.000 title claims abstract description 106
- 235000013824 polyphenols Nutrition 0.000 title claims abstract description 46
- -1 polyphenol compounds Chemical class 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 66
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 28
- 238000001035 drying Methods 0.000 claims abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 25
- 238000000605 extraction Methods 0.000 claims abstract description 24
- 238000000746 purification Methods 0.000 claims abstract description 14
- 238000005374 membrane filtration Methods 0.000 claims abstract description 11
- 238000004237 preparative chromatography Methods 0.000 claims abstract description 10
- 239000002904 solvent Substances 0.000 claims abstract description 10
- 239000000243 solution Substances 0.000 claims description 34
- 238000011068 loading method Methods 0.000 claims description 16
- 239000000047 product Substances 0.000 claims description 16
- 238000001914 filtration Methods 0.000 claims description 14
- 238000010828 elution Methods 0.000 claims description 12
- 239000000284 extract Substances 0.000 claims description 10
- 238000010298 pulverizing process Methods 0.000 claims description 8
- 239000000945 filler Substances 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 7
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- 238000007865 diluting Methods 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- 229940092665 tea leaf extract Drugs 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 5
- 239000002031 ethanolic fraction Substances 0.000 claims description 4
- 238000001471 micro-filtration Methods 0.000 claims description 4
- 238000012856 packing Methods 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 4
- 238000000108 ultra-filtration Methods 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 3
- 230000009849 deactivation Effects 0.000 claims description 2
- 239000012153 distilled water Substances 0.000 claims description 2
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 2
- 238000001694 spray drying Methods 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 2
- 239000008215 water for injection Substances 0.000 claims description 2
- 238000011097 chromatography purification Methods 0.000 claims 1
- 150000008442 polyphenolic compounds Chemical class 0.000 abstract description 23
- 238000004519 manufacturing process Methods 0.000 abstract description 10
- 230000008569 process Effects 0.000 abstract description 4
- 231100000252 nontoxic Toxicity 0.000 abstract description 3
- 230000003000 nontoxic effect Effects 0.000 abstract description 3
- 238000012545 processing Methods 0.000 abstract description 2
- 239000012535 impurity Substances 0.000 description 8
- 238000012986 modification Methods 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 239000007864 aqueous solution Substances 0.000 description 4
- 239000000469 ethanolic extract Substances 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 239000012466 permeate Substances 0.000 description 4
- 238000005303 weighing Methods 0.000 description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-catechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- DATAGRPVKZEWHA-YFKPBYRVSA-N N(5)-ethyl-L-glutamine Chemical compound CCNC(=O)CC[C@H]([NH3+])C([O-])=O DATAGRPVKZEWHA-YFKPBYRVSA-N 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- 235000005487 catechin Nutrition 0.000 description 2
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 description 2
- 229950001002 cianidanol Drugs 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
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- 241000894006 Bacteria Species 0.000 description 1
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- CITFYDYEWQIEPX-UHFFFAOYSA-N Flavanol Natural products O1C2=CC(OCC=C(C)C)=CC(O)=C2C(=O)C(O)C1C1=CC=C(O)C=C1 CITFYDYEWQIEPX-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
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- 229930002877 anthocyanin Natural products 0.000 description 1
- 235000010208 anthocyanin Nutrition 0.000 description 1
- 239000004410 anthocyanin Substances 0.000 description 1
- 150000004636 anthocyanins Chemical class 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
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- 239000008280 blood Substances 0.000 description 1
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- 230000036772 blood pressure Effects 0.000 description 1
- 229960001948 caffeine Drugs 0.000 description 1
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
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- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 150000002206 flavan-3-ols Chemical class 0.000 description 1
- 235000011987 flavanols Nutrition 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- HVQAJTFOCKOKIN-UHFFFAOYSA-N flavonol Natural products O1C2=CC=CC=C2C(=O)C(O)=C1C1=CC=CC=C1 HVQAJTFOCKOKIN-UHFFFAOYSA-N 0.000 description 1
- 150000002216 flavonol derivatives Chemical class 0.000 description 1
- 235000011957 flavonols Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000000643 oven drying Methods 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 235000009048 phenolic acids Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000000194 supercritical-fluid extraction Methods 0.000 description 1
- 229940026510 theanine Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
Classifications
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- 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
-
- 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
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Botany (AREA)
- Mycology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of tea processing, and discloses a preparation method of high-purity tea polyphenol compounds in summer and autumn tea, which comprises the following steps: tea pretreatment, extraction and concentration, membrane filtration, high-pressure preparative chromatography purification, concentration and drying, and finally high-purity tea polyphenol is obtained. The preparation method is simple, the production period is short, the cost is low, only safe and nontoxic solvents of water and ethanol are used in the whole process, the production process is green and environment-friendly, the product quality is good, and the method has good commercial application prospect.
Description
Technical Field
The invention belongs to the technical field of tea processing, and particularly relates to a preparation method of high-purity tea polyphenol compounds in summer and autumn tea.
Technical Field
Tea is one of the most important food and health care products in daily life of people and has been widely applied for many years. Researches show that the tea polyphenol has the effects of resisting bacteria, diminishing inflammation, reducing blood fat, reducing blood pressure, preventing cardiovascular and cerebrovascular diseases and the like, and also has stronger cancer prevention, anticancer and anti-aging effects. Has wide application prospect.
Tea polyphenol is a complex of polyphenol compounds with catechin as the main component (wherein catechin accounts for 60-80%, and the content accounts for 18-35% of dry matter), and comprises flavanols, anthocyanins, flavonoids, flavonols, phenolic acids and the like. The scientific and efficient method for preparing high-purity tea polyphenol is an important problem to be solved by scientific researchers, and the research on the extraction and purification method of high-purity tea polyphenol in tea has important significance.
The method for extracting and purifying tea polyphenol mainly comprises organic solvent extraction, column chromatography, supercritical fluid extraction, high-speed countercurrent extraction and the like, wherein the methods are summarized, most of the methods are mainly divided into two steps, and firstly, crude tea polyphenol containing caffeine, theanine and tea polysaccharide is obtained by extracting, concentrating and primarily purifying tea leaves; then the tea polyphenol with higher purity is obtained by solvent extraction, column chromatography for many times and concentration and drying. These prior arts generally use organic solvents as the main solvent, and the solvents usually used include water, methanol, ethanol, acetone, petroleum ether, ethyl acetate, chloroform, etc. These methods have many disadvantages, such as too long production process route, long production period, high production cost, low yield of tea polyphenol product, organic solvent residue, and heavy metal residue. Greatly reduces the quality of tea polyphenol products.
Based on the above, the invention improves the extraction and purification process of tea polyphenol, adopts five steps of tea pretreatment, extraction and concentration, membrane filtration, high-pressure preparative chromatography purification and concentration and drying to prepare the high-purity tea polyphenol product, greatly improves the production speed of tea polyphenol, only uses safe and nontoxic solvent water and ethanol in the whole process, has green and environment-friendly production process and stable product quality, and provides a new idea for the extraction and purification process of tea polyphenol.
Disclosure of Invention
The invention provides a preparation method of high-purity tea polyphenol compounds in tea leaves in summer and autumn, and solves the problems of long process route, low tea polyphenol yield, low product purity, high production cost, residual toxic and harmful solvents, unstable product quality and the like of the tea polyphenol extraction and purification method in the prior art.
The invention provides a preparation method of high-purity tea polyphenol compounds in summer and autumn tea leaves, which is characterized by comprising the following steps:
step 1, tea leaf pretreatment: drying the tea leaves in summer and autumn after enzyme deactivation, and crushing for later use;
step 2, extraction and concentration: extracting the pretreated summer and autumn tea powder with a solvent for multiple times, filtering, concentrating, diluting with water, and stirring to obtain a summer and autumn tea extract water solution;
and 3, membrane filtration: filtering the water solution of the tea leaf extract in summer and autumn by using a membrane to obtain a clarified solution of the tea leaf extract in summer and autumn;
and 4, high-pressure preparative chromatography purification: concentrating the clarified solution of the summer and autumn tea leaf extract under reduced pressure, injecting into a high-pressure preparation chromatograph, performing ethanol-water gradient elution, and collecting 80% ethanol fraction;
and step 5, concentrating and drying: concentrating the 80% ethanol fraction under reduced pressure until no alcohol smell is produced, drying, and pulverizing to obtain high purity tea polyphenols compound product.
Preferably, the summer and autumn tea water-removing mode in the step 1 is manual water-removing, drum type water-removing machine water-removing or steam type water-removing machine water-removing, the water-removing temperature is 100-200 ℃, and the water-removing time is 3-5 min.
Preferably, the summer and autumn tea leaves in the step 1 are dried or dried in the sun, and the drying temperature is 30-70 ℃.
Preferably, in the step 1, the summer and autumn tea leaves are dried and then crushed to 20-60 meshes.
Preferably, the extraction solvent in the step 2 is water or ethanol, the concentration of the ethanol is 70-98%, the ratio of the extraction material to the extraction liquid is 1: 5-10 g/mL, the extraction temperature is 60-90 ℃, the extraction is carried out for 2-4 times, each time lasts for 1-3 hours, the filtration is carried out, the filtrates are combined and concentrated under reduced pressure to 15-20 Baume degrees, and the water addition amount is 4-10 times of the volume of the concentrated solution.
Preferably, the membrane filtration in step 3 is ultrafiltration or microfiltration.
Preferably, the packing material used for the high pressure preparative chromatography purification in the step 4 is C18Or C8The method comprises the following steps of (1) preparing reversed phase spherical filler with the particle size of 10-30 mu m and the column bed packing height of 20-40 cm, wherein the sample loading amount is 2-4 times of the column volume, and the elution gradient procedure is as follows: eluting 1-2 times of column volume by 10% ethanol, eluting 3-5 times of column volume by 80% ethanol, eluting 2-3 times of column volume by 100% ethanol, balancing 2-3 times of column volume by 100% water after elution is finished, loading flow rate is 50-100 cm/h, and elution and balancing linear flow rate is 150-300 cm/h.
Preferably, the concentration in the step 4 is carried out under reduced pressure until the Baume degree is 5-10.
Preferably, the drying mode in the step 5 is microwave drying, vacuum freeze drying or spray drying.
Preferably, the water is deionized water, distilled water or water for injection.
Compared with the prior art, the invention has the beneficial effects that:
the method adopts five steps of tea pretreatment, extraction concentration, membrane filtration, high-pressure preparative chromatography purification and concentration drying to prepare the high-purity tea polyphenol product, greatly improves the production speed of the tea polyphenol, only uses safe and nontoxic solvent water and ethanol in the whole process, has green and environment-friendly production process and stable product quality, and is suitable for the industrial production of the tea polyphenol product with high-end quality requirements.
Detailed Description
In order to make the technical solutions of the present invention better understood and enable those skilled in the art to practice the present invention, the following embodiments are further described, but the present invention is not limited to the following embodiments.
The experimental methods described in the examples of the present invention are all conventional methods unless otherwise specified.
Example 1
A preparation method of high-purity tea polyphenol compounds in summer and autumn tea leaves comprises the following steps:
step 1, tea leaf pretreatment: manually parching summer and autumn tea at 200 deg.C for 3min, oven drying at 60 deg.C, and pulverizing to 20 mesh;
step 2, extraction and concentration: weighing 1kg of pretreated tea powder, extracting with 70% ethanol at 60 deg.C for 2 times (3 hr each time) according to a material-to-liquid ratio of 1:10g/mL (10L), filtering, mixing filtrates, concentrating under reduced pressure to 15 Baume degree, and diluting with 4 times volume of deionized water;
and 3, membrane filtration: filtering the solution with ultrafiltration membrane equipment, and collecting the permeate to obtain a clarified solution of the summer and autumn tea extract;
and 4, high-pressure preparative chromatography purification: concentrating the clarified solution of the summer-autumn tea extract under reduced pressure to 5 Baume degree, and injecting into a high-pressure preparative chromatograph (DAC-50 dynamic axial high-pressure preparative chromatographic column, C)8The grain diameter of the reversed phase spherical filler is 30 mu m, the filling height of a column bed is 20cm), the sample loading is 2 times of the column volume, the sample loading flow rate is 50cm/h, and after the sample loading is finished, 10 percent ethanol solution with 1 time of the column volume is used for eluting and removing impurities; then eluting with 80% ethanol solution with 3 times column volume, and collecting the fraction; eluting with 2 times of 100% ethanol solution to remove weak polar impurities; finally, balancing the chromatographic column by using 2 times of 100 percent aqueous solution of the column volume, wherein the elution and balance linear flow velocity is 150 cm/h;
and step 5, concentrating and drying: vacuum concentrating the 80% ethanol extract until no alcohol smell is produced, microwave drying, and pulverizing to obtain 256.2g tea polyphenols product with purity of 96%.
Example 2
A preparation method of high-purity tea polyphenol compounds in summer and autumn tea leaves comprises the following steps:
step 1, tea leaf pretreatment: deactivating enzyme of the summer and autumn tea by a roller type water-removing machine at 150 ℃ for 4min, drying at 60 ℃, and crushing to 30 meshes for later use;
step 2, extraction and concentration: weighing 2kg of pretreated tea powder, extracting with 80% ethanol at 90 deg.C for 3 times (2 h each time) according to a material-to-liquid ratio of 1:8g/mL (16L), filtering, mixing filtrates, concentrating under reduced pressure to 16 Baume degree, and diluting with 6 times volume of deionized water;
and 3, membrane filtration: filtering the solution with microfiltration membrane equipment, and collecting the permeate to obtain a clarified solution of the summer and autumn tea extract;
and 4, high-pressure preparative chromatography purification: concentrating the clarified solution of the summer-autumn tea extract under reduced pressure to 7 Baume degree, and injecting into a high-pressure preparative chromatograph (DAC-100 dynamic axial high-pressure preparative chromatographic column, C)18The grain diameter of the reversed phase spherical filler is 20 mu m, the filling height of a column bed is 25cm), the sample loading is 3 times of the column volume, the sample loading flow rate is 100cm/h, and after the sample loading is finished, 10 percent ethanol solution 2 times of the column volume is used for eluting and removing impurities; then eluting with 80% ethanol solution with 4 times column volume, and collecting the fraction; eluting with 3 times of 100% ethanol solution to remove weak polar impurities; finally, the chromatographic column is balanced by 100 percent aqueous solution with 3 times of column volume, and the elution and balance linear flow velocity is 200 cm/h;
and step 5, concentrating and drying: vacuum concentrating the 80% ethanol extract until no alcohol smell is present, microwave drying, and pulverizing to obtain 528.5g tea polyphenols product with purity of 95%.
Example 3
A preparation method of high-purity tea polyphenol compounds in summer and autumn tea leaves comprises the following steps:
step 1, tea leaf pretreatment: deactivating enzyme of the summer and autumn tea by a steam type water-removing machine at 100 deg.C for 5min, drying at 60 deg.C, and pulverizing to 40 mesh;
step 2, extraction and concentration: weighing 3kg of pretreated tea powder, extracting with 90% ethanol at 80 deg.C for 4 times (1 h each time) according to a material-to-liquid ratio of 1:6g/mL (18L), filtering, mixing filtrates, concentrating under reduced pressure to 18 Baume degree, and diluting with 8 times volume of deionized water;
and 3, membrane filtration: filtering the solution with microfiltration membrane equipment, and collecting the permeate to obtain the clarified liquid of the summer and autumn tea extract.
And 4, high-pressure preparative chromatography purification: concentrating the clarified solution of the summer-autumn tea extract under reduced pressure to 8 Baume degree, and injecting into a high-pressure preparative chromatograph (DAC-100 dynamic axial high-pressure preparative chromatographic column, C)18Reverse phase spherical filler with the particle diameter of 10 mu m and the column bed packing height of 30cm), loading 4 times of column volume, loading flow rate of 80cm/h, and eluting with 10 percent ethanol solution with 2 times of column volume to remove impurities after loading; then eluting with 5 times of column volume of 80% ethanol solution, and collecting the fraction; eluting with 3 times of 100% ethanol solution to remove weak polar impurities; finally, theEquilibrating the column with 3 column volumes of 100% aqueous solution at an elution and equilibration linear flow rate of 250 cm/h;
and step 5, concentrating and drying: vacuum concentrating the 80% ethanol extract until no alcohol smell is present, microwave drying, and pulverizing to obtain 781.5g tea polyphenols product with purity of 97%.
Example 4
A preparation method of high-purity tea polyphenol compounds in summer and autumn tea leaves comprises the following steps:
step 1, tea leaf pretreatment: deactivating enzyme of the summer and autumn tea by a steam type water-removing machine at 100 ℃ for 5min, drying at 60 ℃, and crushing to 60 meshes for later use;
step 2, extraction and concentration: weighing 5kg of pretreated tea powder, extracting with 98% ethanol at 70 deg.C for 4 times (1 h each time) according to a material-to-liquid ratio of 1:5g/mL (25L), filtering, mixing filtrates, concentrating under reduced pressure to 20 Baume degree, and diluting with 10 times volume of deionized water;
and 3, membrane filtration: filtering the solution with ultrafiltration membrane equipment, and collecting the permeate to obtain a clarified solution of the summer and autumn tea extract;
and 4, high-pressure preparative chromatography purification: concentrating the clarified solution of the summer-autumn tea extract under reduced pressure to 10 Baume degree, and injecting into a high-pressure preparative chromatograph (DAC-100 dynamic axial high-pressure preparative chromatographic column, C)18Reverse phase spherical filler with the particle size of 10 mu m and the column bed filling height of 40cm), loading 4 times of column volume, loading flow rate of 60cm/h, and eluting with 10% ethanol solution with 2 times of column volume to remove impurities after loading; then eluting with 5 times of column volume of 80% ethanol solution, and collecting the fraction; eluting with 3 times of 100% ethanol solution to remove weak polar impurities; finally, the chromatographic column is balanced by 100 percent aqueous solution with 3 times of column volume, and the elution and balance linear flow velocity is 300 cm/h;
and step 5, concentrating and drying: vacuum concentrating the 80% ethanol extract until no alcohol smell is present, microwave drying, and pulverizing to obtain 1235.8g tea polyphenols product with purity of 96%.
It should be noted that when the following claims refer to numerical ranges, it should be understood that both ends of each numerical range and any value between the two ends can be selected, and since the steps and methods used are the same as those in embodiments 1-4, the preferred embodiments of the present invention have been described for the purpose of preventing redundancy, but once the basic inventive concept is known, those skilled in the art can make other changes and modifications to these embodiments. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (10)
1. A preparation method of high-purity tea polyphenol compounds in summer and autumn tea leaves is characterized by comprising the following steps:
step 1, tea leaf pretreatment: drying the tea leaves in summer and autumn after enzyme deactivation, and crushing for later use;
step 2, extraction and concentration: extracting the pretreated summer and autumn tea powder with a solvent for multiple times, filtering, concentrating, diluting with water, and stirring to obtain a summer and autumn tea extract water solution;
and 3, membrane filtration: filtering the water solution of the tea leaf extract in summer and autumn by using a membrane to obtain a clarified solution of the tea leaf extract in summer and autumn;
and 4, high-pressure preparative chromatography purification: concentrating the clarified solution of the summer and autumn tea leaf extract under reduced pressure, injecting into a high-pressure preparation chromatograph, performing ethanol-water gradient elution, and collecting 80% ethanol fraction;
and step 5, concentrating and drying: concentrating the 80% ethanol fraction under reduced pressure until no alcohol smell is produced, drying, and pulverizing to obtain high purity tea polyphenols compound product.
2. The preparation method of the high-purity tea polyphenol compounds in the tea leaves in summer and autumn according to claim 1, wherein the water-removing mode of the tea leaves in summer and autumn in the step 1 is manual water-removing, drum-type water-removing machine water-removing or steam-type water-removing machine water-removing, the water-removing temperature is 100-200 ℃, and the water-removing time is 3-5 min.
3. The method for preparing high-purity tea polyphenol compounds in summer and autumn tea leaves according to claim 1, wherein the summer and autumn tea leaves in the step 1 are dried or dried in the sun, and the drying temperature is 30-70 ℃.
4. The method for preparing high-purity tea polyphenol compounds in summer and autumn tea leaves according to claim 1, wherein the summer and autumn tea leaves in the step 1 are dried and then ground into 20-60 meshes.
5. The preparation method of high-purity tea polyphenol compounds in tea leaves in summer and autumn according to claim 1, wherein the extraction solvent in the step 2 is water or ethanol, the concentration of the ethanol is 70% -98%, the ratio of the extraction material to the extraction liquid is 1: 5-10 g/mL, the extraction temperature is 60-90 ℃, the extraction is carried out for 2-4 times and 1-3 hours each time, the filtration is carried out, the filtrates are combined and concentrated under reduced pressure to 15-20 baume degrees, and the water addition amount is 4-10 times of volume of concentrated solution.
6. The method for preparing high-purity tea polyphenols compounds in tea leaves in summer and autumn as claimed in claim 1, wherein the membrane filtration in step 3 is ultrafiltration or microfiltration.
7. The method for preparing high-purity tea polyphenol compounds in summer and autumn tea leaves as claimed in claim 1, wherein the filler used for the high-pressure preparative chromatographic purification in the step 4 is C18Or C8The method comprises the following steps of (1) preparing reversed phase spherical filler with the particle size of 10-30 mu m and the column bed packing height of 20-40 cm, wherein the sample loading amount is 2-4 times of the column volume, and the elution gradient procedure is as follows: eluting 1-2 times of column volume by 10% ethanol, eluting 3-5 times of column volume by 80% ethanol, eluting 2-3 times of column volume by 100% ethanol, balancing 2-3 times of column volume by 100% water after elution is finished, loading flow rate is 50-100 cm/h, and elution and balancing linear flow rate is 150-300 cm/h.
8. The method for preparing high-purity tea polyphenol compounds in summer and autumn tea leaves as claimed in claim 1, wherein the concentration in step 4 is reduced pressure to 5-10 Baume degree.
9. The method for preparing high-purity tea polyphenol compounds in summer and autumn tea leaves as claimed in claim 1, wherein the drying mode in the step 5 is microwave drying, vacuum freeze drying or spray drying.
10. The method for preparing high-purity tea polyphenol compounds in tea leaves in summer and autumn according to any one of claims 1 to 9, wherein the water is deionized water, distilled water or water for injection.
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