CN101961426A - Puerh tea extract and extraction method - Google Patents
Puerh tea extract and extraction method Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicinal extract and a preparation method thereof, in particular to a puerh tea extract and preparation thereof. The preparation method comprises the technical means of water extraction, membrane filtration, centrifugation, condensation, filtration, and the like.
Description
Technical field
The present invention relates to a kind of Chinese medicine extract and preparation thereof, particularly the extract of Folium camelliae assamicae and preparation thereof.
Background technology
Folium camelliae assamicae is the distinctive local well-known tea in Yunnan.Be to shine Folium Camelliae sinensis and reprocessing forms two series with the large leaf in original producton location, Yunnan: directly reprocessing be that the life of finished product is general and reprocess form ripe general through artificial rapid-result fermentation back, divides bulk tea and compressed tea two classes on the type system again; All also continue behind the finished product carrying out natural ageing process, have unique quality of Chen Yue perfume (or spice) more.Folium camelliae assamicae is to make with the bright leaf of improved seeds big-leaf species in yunnan, also is called Puer Bulk Tea.The sturdy hypertrophy of its profile bar rope, color and luster Wu Run or brown red is commonly called as and resembles liver-coloured.Mellow time of flavour is sweet, has unique CHENXIANG taste, and the reputation of " beauty treatment tea " is arranged.
Folium camelliae assamicae is the tea of only after fermentation type, harmful material such as its theophylline, tea polyphenols is divided to have melted in secular sweat, so moral character gentleness, human body is not stimulated, can also enhance metabolism, quicken clearing up and transforming of fat, toxin in the health, perplex urbanite's problems such as obesity, three fat height now, Folium camelliae assamicae can both play good mitigation, as toxin expelling, nourishing the stomach, antiinflammatory, cholesterol reducing, the fat that disappears go to be bored with, cosmetic slimming.
Pu'er tea belongs to tea product.Be chocolate, water-soluble, be insoluble to ethanol, methanol, reach chloroform; The main chemical constituent of Pu'er tea, count with butt or dry rate: tea polysaccharide, the abrownin class, protein, all the other are material such as thearubigins, theaflavin and the water soluble pectin etc. of the Folium Camelliae sinensis self of minute quantity.Pu'er tea is of high nutritive value, and biological active substances is abundant.
Chinese patent 200510010871 discloses a kind of Pu'er tea
Preparation process is as follows
A. take by weighing a certain amount of Folium camelliae assamicae, be crushed to 20 orders, the temperature that adds 5-10 times of volume is 80～90 ℃ a distilled water, adds a cover insulation and soaks 40 minutes after-filtration;
B. filtering residue is 30 minutes after-filtration of distilled water immersion of 80～90 ℃ with the temperature of 3-5 times of volume;
C. the temperature of 3 times of volumes of filtering residue reuse is 30 minutes after-filtration of distilled water immersion of 80～90 ℃;
D. merge filtering filtrate three times, adding dehydrated alcohol to concentration of alcohol and be 50%～90% scope precipitates, leave standstill after 24 hours and filter, the collecting precipitation thing, carry out vacuum dehydrating at lower temperature, vacuum and low temperature is 50～60 ℃, is dried to moisture and is about 5～11% and promptly gets Pu'er tea.Contain tea polysaccharide 15-45% in the Folium camelliae assamicae, abrownin 50-70%, protein 4.7-14.0% etc.
Above Pu'er tea preparation method consumption alcohol amount is big, and hold facility is big, and the power consumption of extracting solution Direct Filtration is consuming time again, the inventor is carrying out in the research process Pu'er tea, adopts new isolation technics, tests out one group of new Pu'er tea and preparation method thereof, these Pu'er teas have the curative effect height, good absorbing, steady quality, the preparation method separating effect is good simultaneously, technology is simple, easy to operate, with low cost, be fit to suitability for industrialized production.
Summary of the invention
The invention provides a kind of Pu'er tea.
The main active that described extract contains is a tea polyphenols, tea polysaccharide, abrownin, caffeine.
The percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols 19-41%, tea polysaccharide 15-45%, abrownin 16-25%, caffeine 4-13%, wherein four percentage by weight sums are less than 100%.
Pu'er tea of the present invention, preparation method is as follows:
Get Folium camelliae assamicae superfine powder flour, add water, add water multiple 8-30 doubly, extraction time 1-3 hour in 75-98 ℃ of lixiviate 1-4 time; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with centrifuge, merges centrifugal liquid, and centrifugal liquid nature or heat exchange are cooled to 30-55 ℃, and 10-30 ten thousand molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; The film trapped fluid is concentrated into the concentrated solution that solid content is 5-30%, and concentrated solution is once centrifugal, a centrifugal liquid secondary centrifuging, with membrane filtration liquid and secondary centrifuging liquid separately or merge and be concentrated into proportion 1.0-1.4/55-65 ℃, the condensed cream drying.
Preferred manufacturing procedure is as follows:
Get Folium camelliae assamicae superfine powder flour, add water, add water multiple 12-18 doubly, extraction time 1-3 hour in 75-98 ℃ of lixiviate 2-3 time; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and centrifugal liquid nature or heat exchange are cooled to 30-55 ℃, and 10-30 ten thousand molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, the film trapped fluid is evaporated to the concentrated solution that solid content is 5-30%, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, with membrane filtration liquid and secondary centrifuging liquid separately or merge and be concentrated into proportion 1.13-1.15/55-65 ℃, condensed cream microwave drying, vacuum belt type drying, or spray drying gets final product.
The most preferred preparation method of the present invention in embodiments of the present invention.
The present invention comprises that also described pharmaceutical composition is the pharmaceutical preparation that is prepared into as active constituents of medicine with above-mentioned Folium camelliae assamicae with the pharmaceutical composition of Pu'er tea preparation of the present invention.
Pharmaceutical composition of the present invention can contain the medicine acceptable carrier as required, and wherein Folium camelliae assamicae is as active constituents of medicine, and its shared percentage by weight in preparation can be 0.1-99.9%, and all the other are the medicine acceptable carrier.Pharmaceutical preparations composition of the present invention exists with unit dosage form, and described unit dosage form is meant the unit of preparation, as every of tablet, and capsular every capsules, every bottle of oral liquid, every bag of granule, every of injection etc.
Pharmaceutical composition of the present invention can be any pharmaceutically useful dosage form, and these dosage forms comprise: tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, oral liquid, suck agent, granule, electuary, pill, powder, unguentum, sublimed preparation, suspensoid, powder, solution, injection, suppository, ointment, plaster, cream, spray, drop, patch.
Pharmaceutical composition of the present invention, the preparation of its oral administration can contain excipient commonly used, such as binding agent, filler, diluent, tablet agent, lubricant, disintegrating agent, coloring agent, flavoring agent and wetting agent, can carry out coating to tablet in case of necessity.
The filler that is suitable for comprises cellulose, mannitol, lactose and other similar filler.Suitable disintegrating agent comprises starch, polyvinylpyrrolidone and starch derivatives, for example sodium starch glycollate.Suitable lubricant comprises, for example magnesium stearate.The acceptable wetting agent of appropriate drug comprises sodium lauryl sulphate.
Can fill by mixing, the method that tabletting etc. are commonly used prepares solid oral composition.Mix repeatedly active substance is distributed in those compositionss of a large amount of filleies of whole use.
The form of oral liquid for example can be aqueous or oily suspensions, solution, Emulsion, syrup or elixir, perhaps can be a kind of available water before use or other suitable composite dry products of carrier.This liquid preparation can contain conventional additive, such as suspending agent, for example sorbitol, syrup, methylcellulose, gelatin, hydroxyethyl-cellulose, carboxymethyl cellulose, aluminium stearate gel or hydrogenation edible fat, emulsifying agent, for example lecithin, anhydro sorbitol monooleate or arabic gum; Non-aqueous carrier (they can comprise edible oil), for example almond oil, fractionated coconut oil, such as oily ester, propylene glycol or the ethanol of the ester of glycerol; Antiseptic, for example para hydroxybenzene methyl ester or propyl p-hydroxybenzoate or sorbic acid, and if desired, can contain conventional flavouring agent or coloring agent.
For injection, the liquid unit dosage forms of preparation contains active substance of the present invention and sterile carrier.According to carrier and concentration, this chemical compound can be suspended or dissolving.The preparation of solution is normally by being dissolved in active substance in a kind of carrier filter-sterilized before it is packed into a kind of suitable bottle or ampoule, sealing then.For example a kind of local anesthetic of adjuvant, antiseptic and buffer agent also can be dissolved in this carrier.In order to improve its stability, can be after the bottle of packing into that this compositions is freezing, and under vacuum, water is removed.
Pharmaceutical composition of the present invention, when being prepared into medicament, optionally add suitable medicine acceptable carrier, described medicine acceptable carrier is selected from: mannitol, sorbitol, sodium pyrosulfite, sodium sulfite, sodium thiosulfate, cysteine hydrochloride, TGA, methionine, vitamin C, the EDTA disodium, EDTA calcium sodium, the alkali-metal carbonate of monovalence, acetate, phosphate or its aqueous solution, hydrochloric acid, acetic acid, sulphuric acid, phosphoric acid, aminoacid, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivative, cellulose and derivant thereof, alginate, gelatin, polyvinylpyrrolidone, glycerol, soil temperature 80, agar, calcium carbonate, calcium bicarbonate, surfactant, Polyethylene Glycol, cyclodextrin, beta-schardinger dextrin-, the phospholipid material, Kaolin, Pulvis Talci, calcium stearate, magnesium stearate etc.
Pharmaceutical composition of the present invention is determined usage and dosage according to patient's situation in use, but obeys every day three times, each 1-20 agent, as: 1-20 bag or grain or sheet, every dose of 1mg-1000mg.
Following data declaration beneficial effect of the present invention by experiment.
Folium camelliae assamicae blood sugar lowering, weight losing function are investigated experiment
1, test material
1.1 experimental animal: 40 of KKAy mices, female, age in 7-8 week, rank SPF level, body weight 33.5 ± 1.9g is provided by Chinese Academy of Medical Sciences's laboratory animal.
1.2 reagent and medicine
Pu'er tea (abbreviation tea powder): the embodiment of the invention 1, the dark brown powder, water-soluble, dissolubility is good, lucifuge.
Positive drug: rosiglitazone maleate sheet
1.3 experimental apparatus:
BIOSEN5030 type blood glucose/lactic acid analyser
Hitachi's 7080 full automatic biochemical apparatus
2.1 experimental technique
2.1 fasting glucose assay method:
All animals all in 9 beginnings of commercial affairs fasting, are got blood behind the fasting 4h, get preceding each the treated animal gastric infusion of 1h of blood, cut tail when getting blood and get tip blood 10ul, with BIOSEN5030 type blood glucose/lactose analysis-e/or determining fasting glucose.
2.2 grouping and administration
The animal adaptability is measured fasting glucose according to 2.1 methods after feeding a week, according to the fasting blood sugar stratified random grouping of measuring.40 KKAy are divided into 4 groups, model group, positive controls, experimental group according to the grouping of fasting blood sugar stratified random.
Crowd's operational version, it is 3g/ people/day that the crowd intends using dosage, i.e. 0.05g/kg/d, the use approach brews to be drunk.
Positive controls, administration every day 1 time, dosage 1.33mg/kg/ day, oral administration gavage administration.
Test group: the animal test sample amount of drinking reaches the dosage requirement of 1.5g/kgBW, and replaces drinking water every day with the Folium camelliae assamicae of this concentration, gives for 4 weeks continuously.
2.3 test procedure
2.3.1 the weight of animals and feed situation, feces situation of change: observe animal spirit, activity, hair color, food-intake, amount of drinking water and body weight change
2.3.2 fasting glucose is measured: the set time is measured fasting glucose according to 2.1 methods weekly before the administration and after the administration.
2.3.3 carbohydrate tolerance experiment
Tested the 21st day, animal gavages glucose 2.5g/kg after measuring fasting glucose according to 2.1 methods, and respectively at 0.5h, blood glucose value, area under the calculated curve (AUC) AUC=0.5 * fasting glucose+0.5h blood glucose+1.5 * 1h blood glucose+2h blood glucose are measured in 1h, 2h blood sampling.
2.3.4 serum electrolyte is measured
When testing the 22nd day, pluck eyeball behind the animal fasting 16h and get blood, separation of serum is surveyed mensuration serum K ion, Na ion, Ca ion concentration with the XD685 electrolyte analyser.
2.3.5 triglyceride, T-CHOL are measured;
When testing the 29th day, pluck eyeball behind the animal fasting 16h and get blood, separation of serum is measured serum triglycerides, total cholesterol level with full automatic biochemical apparatus.
3, result of the test
3.1 the weight of animals, food-intake changes result of the test
The experimental session experimental animal is in good condition, and food-intake, amount of drinking water are stablized.
Experimental group was freely drunk for two weeks continuously according to dosage 1.5g/kg/ day, and food-intake and body weight all change not quite, each treated animal body weight, and the food-intake result of variations sees Table 1, table 2.
Table 1 the weight of animals situation of change (g)
| Grouping | The animal number of elements | Dosage | Before the administration | First week | Second week |
| The KKAy model group | 10 | 33.0±1.1 | 35.1±2.4 | 35.8±2.9 | |
| Positive controls | 10 | 1.33g/kg/ day | 33.7±2.0 | 34.8±1.7 | 34.9±1.3 |
| Tea powder | 10 | 1.5g/kg/ day | 32.9±2.2 | 33.2±1.4* | 34.3±1.3 |
Annotate: compare * p＜0.05 with model.
Each treated animal food-intake situation of change (g) of table 2
| Grouping | The animal number of elements | Dosage | Before the administration | First week | Second week |
| The KKAy model group | 10 | 4.5±1.1 | 5.2±1.1 | 5.4±1.7 | |
| Positive controls | 10 | 1.33g/kg/ day | 4.0±1.3 | 3.9±0.4 | 4.0±1.3 |
| Tea powder | 10 | 1.5g/kg/ day | 4.2±1.4 | 4.7±0.7 | 4.4±0.3 |
3.2 fasting glucose determination test result
The results are shown in Table 3, after 7 days, fasting glucose all is starkly lower than model group (p＜0.01) to the experimental group experimental animal in administration, positive controls, and after 14 days, fasting glucose all is starkly lower than model group (p＜0.01, p＜0.01) to the experimental group animal in administration.As seen, Folium camelliae assamicae has obvious hypoglycemic activity.
Table 3 Folium camelliae assamicae is to the influence of fasting glucose (nmol/l)
| Grouping | Dosage | Before the administration | First week | Second week |
| The KKAy model group | 19.86±5.11 | 24.77±6.62 | 21.23±5.29 | |
| Positive controls | 1.33g/kg/ day | 19.57±4.67 | 20.34±4.60** | 13.91±3.32** |
| Tea powder | 1.5g/kg/ day | 19.62±4.71 | 16.21±2.62** | 13.27±2.00*** |
Annotate: compare material p＜0.01 with model, * * * p＜0.01.
3.3 carbohydrate tolerance test result:
By table 4 as seen, in the test of anti-sugar amount, the blood glucose value of each time point of model group and AUC value all are higher than matched group, the blood glucose value and the AUC of each time point of positive drug group all are lower than model group, experimental group is in 0h, 0.5h, 1h and AUC value and model group there were significant differences p＜0.01, p＜0.001., visible Pu'er tea can suppress the rising of blood glucose value after the oral sucrose of mice.
Table 4 Pu'er tea is to the influence of carbohydrate tolerance
| Grouping | ?n | 0h | 0.5h | 1h | 2h | AUC |
| The KKAy model group | 10 | 22.83±2.17 | 30.91±2.18 | 29.59±3.26 | 27.46±4.89 | 114.58±11.76 |
| Positive controls | 10 | 13.03±3.86** | 18.38±5.34*** | 18.58±5.63*** | 13.74±3.82*** | 66.12±18.93*** |
| Tea powder | 10 | 15.32±5.39*** | 24.34±3.37*** | 24.06±4.26** | 25.08±5.14 | 9363±15.51** |
Annotate: compare * * p＜0.01 with model group, * * * p＜0.001.
3.4 serum electrolyte measurement result
By table 5 as seen, the positive drug group diabetic mice K that can obviously raise
+,Na
+,Cl
-Concentration (p＜0.001), the experimental group Na that can obviously raise
+,Cl
-Concentration (p＜0.01, p＜0.001)
Table 5 Pu'er tea is to the influence of serum electrolyte
| Grouping | n | K + | Na + | Cl - | Ca 2+ |
| The KKAy model group | 10 | 6.07±0.61 | 151.38±2.68 | 113.90±1.84 | 1.10±0.11 |
| Positive controls | 10 | 7.02±0.57** | 156.19±2.07*** | 118.45±1.89*** | 1.10±0.06 |
| Tea powder | 10 | 6.35±0.33 | 155.03±2.14** | 117.19±1.28*** | 1.06±0.08 |
3.6 triglyceride, T-CHOL measurement result
By table 6 as seen, in the time of 28 days in triglyceride (TG), T-CHOL (CHOL) measurement result, each experimental group does not have influence to total cholesterol level in administration.The positive drug group, the Pu'er tea high dose group, the low dose group content of triglyceride has been compared the reduction effect with model group, and significant difference (p＜0.05) is arranged.
Table 6 Pu'er tea triglyceride, T-CHOL result
Annotate: compare with model group
*P＜0.05
Compare with normal group
△ △P＜0.01,
△ △ △P＜0.001
Experiment conclusion
To each treated animal food-intake, the continuous observation of amount of drinking water and body weight gain finds that positive drug and experimental group are to the animal food-intake by experiment, and the body weight influence is little, and is not only smaller than model group on the numerical value, and not statistically significant.From fasting glucose determination test result, Pu'er tea all has the effect of tangible reduction fasting glucose.The beta-oxybutyria acidosis is the severe complication of diabetes, such patient is because relative or absolute shortage of insulin, tissue utilizes the glucose ability drop, catabolism of fat and glyconeogenesis strengthen, liver generation ketoboidies increases and causes hyperglycemia and ketosis, thereby brings out a series of water, electrolyte and acid base imbalance.Mensuration by serum electrolyte Pu'er tea as can be seen has certain regulating action to the diabetes electrolyte disturbance.
In the investigation experiment of effect for reducing fat, Pu'er tea shows can triglyceride reducing content.
The invention has the advantages that: the effective ingredient of Folium Camelliae sinensis and material are easy to stripping especially after the micronizing, extracting all more conventional Folium Camelliae sinensis extraction of water and extraction time reduces, reduced production cost, the membrane filtration product can ensure the clarity behind the cold-water solution of finished product, the film trapped fluid that is difficult to continue membrane filtration operation after simultaneously concentration being increased carries out centrifugal treating, not only line has reduced effective ingredient and the material base that centrifugal workload of later stage has also at utmost kept Folium camelliae assamicae, the membrane filtration process of conventional report is not handled the film trapped fluid, this process route can be realized industrialization, and is stable and controllable for quality.
Relevant test shows, Pu'er tea of the present invention is than prior art curative effect height, the purity height, and good absorbing, steady quality, the purposes novelty, preparation method technology is simple, easy to operate, with low cost simultaneously, is fit to suitability for industrialized production.
The specific embodiment
Below with bright the present invention specifically, embodiment is for the ease of understanding the present invention, and claim that does not limit the present invention in any way and core content.
The preparation of embodiment 1 Pu'er tea, wherein the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 18.97%
Abrownin: 15.87%
Caffeine: 44.83%
Tea polysaccharide: 12.45%
Get Folium camelliae assamicae superfine powder flour, add water, add 14 times of water multiples, extraction time 2 hours in 95 ℃ of lixiviates 3 times; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and the centrifugal liquid nature is to 50 ℃, and 200,000 molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, it is 10% concentrated solution that the film trapped fluid is evaporated to solid content, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, membrane filtration liquid and the merging of secondary centrifuging liquid are concentrated into 1.14/60 ℃ of proportion, and the condensed cream microwave drying gets final product.
The preparation of embodiment 2 Pu'er teas
Get Folium camelliae assamicae superfine powder flour, add water, add 16 times of water multiples, extraction time 2 hours in 95 ℃ of lixiviates 3 times; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and the centrifugal liquid nature is to 40 ℃, and 300,000 molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, it is 10% concentrated solution that the film trapped fluid is evaporated to solid content, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, membrane filtration liquid and the merging of secondary centrifuging liquid are concentrated into 1.13/55 ℃ of proportion, and the condensed cream microwave drying gets final product.
Detect by following method, wherein the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 21.07%
Abrownin: 16.67%
Caffeine: 8.96%
Tea polysaccharide: 32.27%
The method of inspection is as follows:
Content Determination Method of Green Tea Polyphenol
The tartaric acid ferrous solution
Take by weighing ferrous sulfate (GB 664) 1.0g, sodium potassium tartrate tetrahydrate (GB 1288) 5.0g is dissolved in water and is settled to IL,
The phosphate buffer of pH7.5
The disodium phosphate soln of a liquid 1/15mol/L: take by weighing sodium hydrogen phosphate (GB 1263) 23.877g, be dissolved in water and be diluted to IL.
The potassium dihydrogen phosphate of b liquid 1/15mol/L: take by weighing potassium dihydrogen phosphate (GB1274) 9.078g, be dissolved in water to IL through 110 ℃ of oven dry 2h.
Get a liquid 85mL and b liquid 15mL mixing, promptly get the slow juice of pH7.5.
The preparation of standard solution
Accurately take by weighing progallin A 250mg, be dissolved in the 100mL water, draw mother solution 2,4 respectively as mother solution, 6,8,10mL is mixed with standardize solution not and contains progallin A 50 among the 100mL in the 10mL volumetric flask, 100,150,200, the standard solution of five kinds of variable concentrations of 250mg.
The making of standard working curve
Accurately the progallin A standard solution ImL and the tartaric acid ferron 5mL of the variable concentrations of drawing place the volumetric flask of a series of 25mL, with the buffer standardize solution of pH7.5.Water replaces progallin A in contrast, with the cuvette of 1cm, measures absorbance at the 540nm place.The absorbance of being surveyed is depicted as standard working curve with corresponding progallin A concentration.
The preparation of test liquid and mensuration
Preparation: accurately take by weighing tea extract 200mg sample, place the 100mL beaker, add the boiling water dissolving more than 20～30mL90 ℃, cooling moves in the 100mL volumetric flask, and standardize solution, filtration discard the about 20mL of initial filtrate, and last filtrate is test liquid.
Measure: accurately draw test liquid ImL, put in the 25mL volumetric flask, tartarize ferrous solution 5ml, abundant mixing is with the phosphate buffer standardize solution of pH7.5.Make reference with reagent blank liquid, measure absorbance in the 540nm place.
The result calculates
According to standard working curve, obtain the corresponding content of the progallin A that is equivalent to the sample absorbance;
The tea polysaccharide method of inspection
The preparation of need testing solution
Get the about 0.2g of this product powder (being accurate to 0.001g), put in the 10ml measuring bottle, it is an amount of to add water, and dissolving in ultrasonic 20 minutes is cooled off and added water to scale, shakes up.Precision is measured 1ml, puts in the 100ml measuring bottle, and thin up shakes up to scale, as need testing solution.
The preparation of reference substance solution
It is an amount of to get D-anhydrous glucose reference substance, accurate claims fixed (being accurate to 0.001g), is dissolved in water and dilutes and make the solution that contains 0.2mg among every 1ml, in contrast product solution.
The drafting of standard curve
Accurate respectively absorption reference substance solution 0.0,0.1,0.2,0.3,0.4,0.5ml put respectively in the 10ml tool plug test tube, add water to 1.0ml respectively, each is accurate to add 5% phenol solution and (takes by weighing the 2.5g re-distilled phenol, be dissolved in water and be diluted to 50ml, shake up, promptly.) 1.0ml, the jolting mixing, add 5.0ml sulphuric acid, with the rapid jolting mixing of miniature whirlpool mixed instrument, placed 30 minutes under the room temperature, with the 1st pipe is blank, according to ultraviolet visible spectrophotometry (appendix VB of Chinese Pharmacopoeia version in 2005), measures absorbance (measured and finish) at 487nm wavelength place in 1 hour.Make standard curve with absorbance (Y) and quality (X), regression equation is:
The mensuration of need testing solution
The accurate 1ml need testing solution of drawing under the drafting item according to standard curve, from " the phenol solution 1.0ml of accurate adding 5% ", with the method operation, is measured absorbance respectively at the 487nm place.
Calculate content;
Theaflavin, thearubigins and abrownin content assaying method
Test liquid preparation: accurately take by weighing the 0.05-1g powder, add boiling water 125ml, shake up back lixiviate 10 minutes in boiling water bath, shake bottle in the lixiviate once, after lixiviate finished, taking-up shook up, and filtered in exsiccant conical flask (residue does not need the water flushing) with Cotton Gossypii while hot, filtrate can extract and spectrophotometry after soaking and be chilled to room temperature in cold water.
Extraction: shake up test liquid, draw 25ml and be placed in the 60ml separatory funnel, add the 25ml ethyl acetate, speed jolting 5 minutes with secondary each second roughly, static layering is emitted water layer respectively and is poured the ethyl acetate layer into stand-by in the tool plug triangular flask (after the layering, intermediary opacifying layer discards).
Absorption ethyl acetate layer solution 2ml is placed in the 25ml volumetric flask, adds 95% ethanol dilution to 25ml, shakes up to be solution A.Draw ethyl acetate layer solution 10ml and be placed in the 30ml separatory funnel, add 2.5%NaHCO3 aqueous solution 10ml, 30 seconds of jolting.Behind the standing demix, emit NaHCO3 water layer solution immediately and discard, carefully ethyl acetate layer solution is poured in the tool plug test tube, get this ethyl acetate layer solution 4ml, be placed in the 25ml volumetric flask, add 95% ethanol dilution, shake up and be solution C to 25ml.
Draw the water layer solution 2ml that tells with ethyl acetate extraction for the first time, be placed in the 25ml volumetric flask, add 2ml saturated oxalic acid solution and 6ml distilled water, and with 95% ethanol dilution to 25ml, shake up solution D afterwards.
Draw millet paste test liquid 10ml, be placed in the 30ml separatory funnel, add the 10ml n-butyl alcohol, jolting 3 minutes, leave standstill slowly layering after, emit following water layer.Draw this water layer solution 2ml, be placed in the 25ml volumetric flask, add 2ml saturated oxalic acid and 6ml distilled water, and with 95% ethanol dilution to 25ml, shake up and be solution B.
Colorimetric determination
Select the 380nm wavelength, use the 1cm cuvette, make blank, measure the optical density E of A, B, C, D solution respectively with 95% ethanol.Calculate content.
The content of caffeine assay method:
1), chromatographic condition: mobile phase (methanol: water=30: 70), detect wavelength 273nm, sample size (reference substance 10ul, test sample 5ul), column temperature (25 ℃), flow velocity (1ml/min)
2), reference substance solution preparation: get the about 10mg of caffeine reference substance, the accurate title, decide, and puts in the 100ml volumetric flask, adds methanol 20ml and make dissolving, adds water and be settled to scale, filters, promptly.
3), need testing solution preparation: accurate claim decide extract be 0.5g in the 100ml conical flask, accurately add purified water 100ml, weigh, boiling water bath 1h weighs, add purified water and supply weight, filtration, promptly.
The preparation of embodiment 3, Pu'er tea
Get Folium camelliae assamicae superfine powder flour, add water, add 10 times of water multiples, extraction time 1 hour in 75 ℃ of lixiviates 1 time; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and the centrifugal liquid nature is to 30 ℃, and 100,000 molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, it is 5% concentrated solution that the film trapped fluid is evaporated to solid content, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, membrane filtration liquid and the merging of secondary centrifuging liquid are concentrated into 1.01/55 ℃ of proportion, and the condensed cream spray drying gets final product.
The preparation of embodiment 4, Pu'er tea
Get Folium camelliae assamicae superfine powder flour, add water, add 20 times of water multiples, extraction time 3 hours in 98 ℃ of lixiviates 3 times; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and the centrifugal liquid nature is to 55 ℃, and 300,000 molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, it is 30% concentrated solution that the film trapped fluid is evaporated to solid content, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, membrane filtration liquid and the merging of secondary centrifuging liquid are concentrated into 1.4/65 ℃ of proportion, and the condensed cream vacuum belt type drying gets final product.
The preparation of embodiment 5, Pu'er tea
Get Folium camelliae assamicae superfine powder flour, add water, add 16 times of water multiples, extraction time 2 hours in 80 ℃ of lixiviates 2 times; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and the centrifugal liquid nature is to 38 ℃, and 200,000 molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, it is 30% concentrated solution that the film trapped fluid is evaporated to solid content, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, membrane filtration liquid and the merging of secondary centrifuging liquid are concentrated into 1.1/60 ℃ of proportion, and the condensed cream vacuum belt type drying gets final product.
The preparation of embodiment 6, Pu'er tea
Get Folium camelliae assamicae superfine powder flour, add water, add 12 times of water multiples, extraction time 3 hours in 85 ℃ of lixiviates 3 times; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and the centrifugal liquid nature is to 38 ℃, and 300,000 molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, it is 30% concentrated solution that the film trapped fluid is evaporated to solid content, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, membrane filtration liquid and the merging of secondary centrifuging liquid are concentrated into 1.2/60 ℃ of proportion, and the condensed cream spray drying gets final product.
The preparation of embodiment 7, Pu'er tea
Get Folium camelliae assamicae superfine powder flour, add water, add 30 times of water multiples, extraction time 3 hours in 85 ℃ of lixiviates 4 times; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and the centrifugal liquid nature is to 50 ℃, and 200,000 molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, it is 30% concentrated solution that the film trapped fluid is evaporated to solid content, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, membrane filtration liquid and secondary centrifuging liquid are concentrated into 1.15/60 ℃ of proportion separately, and the condensed cream spray drying gets final product.
Embodiment 8,
The extract of embodiment 3 detects according to the method for embodiment 2
Wherein the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 40.46%
Abrownin: 16.51%
Caffeine: 6.79%
Tea polysaccharide: 28.33%.
Embodiment 9,
The extract of embodiment 4 detects according to the method for embodiment 2
Wherein the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 28.05%
Abrownin: 16.58%
Caffeine: 4.40%
Tea polysaccharide: 40.73%.
Embodiment 10,
The extract of embodiment 5 detects according to the method for embodiment 2
Wherein the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 22.15%
Abrownin: 20.82%
Caffeine: 8.9%
Tea polysaccharide: 32.43%.
Embodiment 11,
The extract of embodiment 6 detects according to the method for embodiment 2
Wherein the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 40.55%
Abrownin: 25.56%
Caffeine: 4.79%
Tea polysaccharide: 15.46%.
Embodiment 12,
The extract of embodiment 6 detects according to the method for embodiment 2
Wherein the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 20.45%
Abrownin: 18.71%
Caffeine: 7.8%
Tea polysaccharide: 43.49%.
Embodiment 13,
The extract of embodiment 7 detects according to the method for embodiment 2
Wherein the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 20.12%
Abrownin: 16.10%
Caffeine: 9.04%
Tea polysaccharide: 23.71%.
Embodiment 14, Folium camelliae assamicae preparation of compositions
With Pu'er tea as active constituents of medicine, the medicine acceptable carrier be can contain as required, tablet, capsule, oral liquid, granule, pill, powder, unguentum, sublimed preparation, injection, suppository, cream, spray, drop pill, patch are prepared into according to the galenic pharmacy routine techniques.
Embodiment 15
The Pu'er tea of Pu'er tea that has gone on the market in the prior art and the most preferred embodiment of the present invention carries out content relatively, the result shows Pu'er tea tea polyphenols of the present invention, abrownin, the more approaching former tea of the portfolio ratio of composition such as caffeine or tea polysaccharide brews the proportioning of each component in the tea, and mouthfeel is more natural, true.
Claims (10)
1. a Pu'er tea is characterized in that, the main active that described extract contains is a tea polyphenols, tea polysaccharide, and abrownin, caffeine, the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols 19-41%, tea polysaccharide 15-45%, abrownin 16-25%, caffeine 4-13%, wherein four percentage by weight sums are less than 100%.
2. the extract of claim 1 is characterized in that, preparation method is as follows:
Get Folium camelliae assamicae superfine powder flour, add water, add water multiple 8-30 doubly, extraction time 1-3 hour in 75-98 ℃ of lixiviate 1-4 time; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with centrifuge, merges centrifugal liquid, and centrifugal liquid nature or heat exchange are cooled to 30-55 ℃, and 10-30 ten thousand molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; The film trapped fluid is concentrated into the concentrated solution that solid content is 5-30%, and concentrated solution is once centrifugal, a centrifugal liquid secondary centrifuging, with membrane filtration liquid and secondary centrifuging liquid separately or merge and be concentrated into proportion 1.01-1.4/55-65 ℃, the condensed cream drying.
3. the extract of claim 1 is characterized in that, preparation method is as follows:
Get Folium camelliae assamicae superfine powder flour, add water, add water multiple 12-18 doubly, extraction time 1-3 hour in 75-98 ℃ of lixiviate 2-3 time; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and centrifugal liquid nature or heat exchange are cooled to 30-55 ℃, and 10-30 ten thousand molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, the film trapped fluid is evaporated to the concentrated solution that solid content is 5-30%, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, with membrane filtration liquid and secondary centrifuging liquid separately or merge and be concentrated into proportion 1.13-1.15/55-65 ℃, condensed cream microwave drying, vacuum belt type drying, or spray drying gets final product.
4. the extract of claim 1 is characterized in that, preparation method is as follows:
Get Folium camelliae assamicae superfine powder flour, add water, add 14 times of water multiples, extraction time 2 hours in 95 ℃ of lixiviates 3 times; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and the centrifugal liquid nature is to 50 ℃, and 200,000 molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, it is 10% concentrated solution that the film trapped fluid is evaporated to solid content, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, membrane filtration liquid and the merging of secondary centrifuging liquid are concentrated into 1.14/60 ℃ of proportion, and the condensed cream microwave drying gets final product.
5. the pharmaceutical composition that contains the extract of claim 1.
6. the pharmaceutical composition of claim 5 can contain the medicine acceptable carrier as required, and wherein Pu'er tea is as active constituents of medicine, and its shared percentage by weight in preparation can be 0.1-99.9%, and all the other are the medicine acceptable carrier.
7. the pharmaceutical composition of claim 5 is any pharmaceutically useful dosage forms.
The compositions of the extract of claim 1 and claim 5 preparation treatment diabetes with slimming medicine in application.
9. content of effective assay method in the Pu'er tea of claim 1, the following Content Determination Method of Green Tea Polyphenol of step
The tartaric acid ferrous solution
Take by weighing ferrous sulfate 1.0g, sodium potassium tartrate tetrahydrate 5.0g is dissolved in water and is settled to IL,
The phosphate buffer of pH7.5
The disodium phosphate soln of a liquid 1/15mol/L: take by weighing sodium hydrogen phosphate 23.877g, be dissolved in water and be diluted to IL;
The potassium dihydrogen phosphate of b liquid 1/15mol/L: take by weighing potassium dihydrogen phosphate 9.078g, be dissolved in water to IL through 110 ℃ of oven dry 2h;
Get a liquid 85mL and b liquid 15mL mixing, promptly get the slow juice of pH7.5;
The preparation of standard solution
Accurately take by weighing progallin A 250mg, be dissolved in the 100mL water, draw mother solution 2,4 respectively as mother solution, 6,8,10mL is mixed with standardize solution not and contains progallin A 50 among the 100mL in the 10mL volumetric flask, 100,150,200, the standard solution of five kinds of variable concentrations of 250mg;
The making of standard working curve
Accurately the progallin A standard solution ImL and the tartaric acid ferron 5mL of the variable concentrations of drawing place the volumetric flask of a series of 25mL, with the buffer standardize solution of pH7.5; Water replaces progallin A in contrast, with the cuvette of 1cm, measures absorbance at the 540nm place; The absorbance of being surveyed is depicted as standard working curve with corresponding progallin A concentration;
The preparation of test liquid and mensuration
Preparation: accurately take by weighing tea extract 200mg sample and place the 100mL beaker, add the boiling water dissolving more than 20～30mL90 ℃, cooling moves into 100mL
In the volumetric flask, standardize solution, filtration discard the about 20mL of initial filtrate, and last filtrate is test liquid;
Measure: accurately draw test liquid ImL, put in the 25mL volumetric flask, tartarize ferrous solution 5ml, abundant mixing is with the phosphate buffer standardize solution of pH7.5; Make reference with reagent blank liquid, measure absorbance in the 540nm place;
The result calculates:
According to standard working curve, obtain the corresponding content of the progallin A that is equivalent to the sample absorbance; The tea polysaccharide method of inspection
The preparation of need testing solution
Get the about 0.2g of this product powder, put in the 10ml measuring bottle, it is an amount of to add water, and dissolving in ultrasonic 20 minutes is cooled off and added water to scale, shakes up; Precision is measured 1ml, puts in the 100ml measuring bottle, and thin up shakes up to scale, as need testing solution;
The preparation of reference substance solution
It is an amount of to get D-anhydrous glucose reference substance, accurate claims surely, is dissolved in water and dilutes and make the solution that contains 0.2mg among every 1ml, in contrast product solution;
The drafting of standard curve
Accurate respectively absorption reference substance solution 0.0,0.1,0.2,0.3,0.4,0.5ml put respectively in the 10ml tool plug test tube, add water to 1.0ml respectively, the phenol solution 1.0ml of each accurate adding 5%, the jolting mixing adds 5.0ml sulphuric acid, with the rapid jolting mixing of miniature whirlpool mixed instrument, placed 30 minutes under the room temperature, with the 1st pipe is blank, according to ultraviolet visible spectrophotometry, measures absorbance at 487nm wavelength place; Make standard curve with absorbance (Y) and quality (X), regression equation is:
The mensuration of need testing solution
The accurate 1ml need testing solution of drawing under the drafting item according to standard curve, from " the phenol solution 1.0ml of accurate adding 5% ", with the method operation, is measured absorbance respectively at the 487nm place;
Calculate content;
Theaflavin, thearubigins and abrownin content assaying method
Test liquid preparation: accurately take by weighing the 0.05-1g powder, add boiling water 125ml, shake up back lixiviate 10 minutes in boiling water bath, shake bottle in the lixiviate once, after lixiviate finished, taking-up shook up, and filtered in exsiccant conical flask with Cotton Gossypii while hot, filtrate can extract and spectrophotometry after soaking and be chilled to room temperature in cold water;
Extraction: shake up test liquid, draw 25ml and be placed in the 60ml separatory funnel, add the 25ml ethyl acetate, with the speed jolting 5 minutes of secondary each second roughly, static layering is emitted water layer respectively and is poured in the tool plug triangular flask ethyl acetate layer stand-by;
Absorption ethyl acetate layer solution 2ml is placed in the 25ml volumetric flask, adds 95% ethanol dilution to 25ml, shakes up to be solution A; Draw ethyl acetate layer solution 10ml and be placed in the 30ml separatory funnel, add 2.5%NaHCO3 aqueous solution 10ml, 30 seconds of jolting; Behind the standing demix, emit NaHCO3 water layer solution immediately and discard, carefully ethyl acetate layer solution is poured in the tool plug test tube, get this ethyl acetate layer solution 4ml, be placed in the 25ml volumetric flask, add 95% ethanol dilution, shake up and be solution C to 25ml;
Draw the water layer solution 2ml that tells with ethyl acetate extraction for the first time, be placed in the 25ml volumetric flask, add 2ml saturated oxalic acid solution and 6ml distilled water, and with 95% ethanol dilution to 25ml, shake up solution D afterwards;
Draw millet paste test liquid 10ml, be placed in the 30ml separatory funnel, add the 10ml n-butyl alcohol, jolting 3 minutes, leave standstill slowly layering after, emit following water layer; Draw this water layer solution 2ml, be placed in the 25ml volumetric flask, add 2ml saturated oxalic acid and 6ml distilled water, and with 95% ethanol dilution to 25ml, shake up and be solution B;
Colorimetric determination
Select the 380nm wavelength, use the 1cm cuvette, make blank, measure the optical density E of A, B, C, D solution respectively with 95% ethanol; Calculate content.
10. the preparation method of the Pu'er tea of claim 1, step is as follows
Get Folium camelliae assamicae superfine powder flour, add water, add 14 times of water multiples, extraction time 2 hours in 95 ℃ of lixiviates 3 times; Continuous stirring in extraction and the blowing process, each extraction slurry is centrifugal with tripod pendulum type batch centrifugal, merges centrifugal liquid, and the centrifugal liquid nature is to 40 ℃, and 300,000 molecular weight membrane filtrations get membrane filtration liquid and film trapped fluid; Temperature≤70 ℃, it is 10% concentrated solution that the film trapped fluid is evaporated to solid content, concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, the centrifugal liquid centrifugal secondary centrifuging of tube centrifuge, temperature≤70 ℃, membrane filtration liquid and the merging of secondary centrifuging liquid are concentrated into 1.13/55 ℃ of proportion, and the condensed cream microwave drying gets final product.
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| CN103315101A (en) * | 2013-05-07 | 2013-09-25 | 江苏丹枫神源生物科技有限公司 | Mono maple bud tea and preparation process thereof |
| CN104397291A (en) * | 2014-12-04 | 2015-03-11 | 广州王老吉大健康产业有限公司 | Preparation method of concentrated juice of herbal tea |
| CN104623097A (en) * | 2013-11-07 | 2015-05-20 | 云南天士力帝泊洱生物茶集团有限公司 | Application of Pu'er tea extract in preparation of fat reducing medicines or health foods |
| CN104688934A (en) * | 2013-12-04 | 2015-06-10 | 云南天士力帝泊洱生物茶集团有限公司 | Composition of Pu'er tea effective component and application of composition in preparation of medicine or health food for reducing fat |
| CN106819288A (en) * | 2017-01-19 | 2017-06-13 | 合肥观云阁商贸有限公司 | The preparation method of Longtube Ground Ivy Herb tealeaves Tea Polyphenols |
| CN106820144A (en) * | 2017-01-19 | 2017-06-13 | 合肥观云阁商贸有限公司 | The preparation method of sweet tea polyphenols |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103315101A (en) * | 2013-05-07 | 2013-09-25 | 江苏丹枫神源生物科技有限公司 | Mono maple bud tea and preparation process thereof |
| CN103315101B (en) * | 2013-05-07 | 2014-10-15 | 江苏丹枫神源生物科技有限公司 | Mono maple bud tea and preparation process thereof |
| CN104623097A (en) * | 2013-11-07 | 2015-05-20 | 云南天士力帝泊洱生物茶集团有限公司 | Application of Pu'er tea extract in preparation of fat reducing medicines or health foods |
| CN104688934A (en) * | 2013-12-04 | 2015-06-10 | 云南天士力帝泊洱生物茶集团有限公司 | Composition of Pu'er tea effective component and application of composition in preparation of medicine or health food for reducing fat |
| CN104397291A (en) * | 2014-12-04 | 2015-03-11 | 广州王老吉大健康产业有限公司 | Preparation method of concentrated juice of herbal tea |
| CN106819288A (en) * | 2017-01-19 | 2017-06-13 | 合肥观云阁商贸有限公司 | The preparation method of Longtube Ground Ivy Herb tealeaves Tea Polyphenols |
| CN106820144A (en) * | 2017-01-19 | 2017-06-13 | 合肥观云阁商贸有限公司 | The preparation method of sweet tea polyphenols |
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