CN101961423B - Pu'er tea extract and preparation method thereof - Google Patents
Pu'er tea extract and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine extractive and a preparation method thereof, especially to a Pu'er tea extractive and a preparation method thereof, wherein the preparation method comprises the technical means of water extracting, concentrating, centrifuging, etc.
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 two types of bulk tea and compressed teas on the type system again; All also continue behind the finished product carrying out natural ageing process, have the fragrant unique quality of Chen Yue more.Folium camelliae assamicae is to process 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, and harmful material such as its theophylline, tea polyphenols is divided to have melted in secular sweat, so moral character is gentle; 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
The preparation process is following
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. the temperature of 3-5 times of volume of filtering residue use is 30 minutes after-filtration of distilled water immersion of 80～90 ℃;
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 filtrating 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 carries 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 method for preparing consumption alcohol amount is big, and hold facility is big, and the power consumption of extracting solution Direct Filtration is consuming time again, and the inventor is carrying out in the research process Pu'er tea; Adopt new isolation technics, test out one group of new Pu'er tea and preparation method thereof, these Pu'er teas have curative effect height, good absorbing; Steady quality, the method for preparing separating effect is good simultaneously, and technology is simple; Easy to operate, with low cost, be fit to suitability for industrialized production.
Summary of the invention
The present invention provides a kind of Pu'er tea.
The main active that said extract contains is a tea polyphenols, tea polysaccharide, abrownin, caffeine.
The percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 12-50%, abrownin: 12-30%, caffeine: 6-11%, tea polysaccharide: 15-45%, wherein four percentage by weight sums are less than 100%.
The effective ingredient of preferred weight percentage composition:
Tea polyphenols 20-50%
Abrownin 15-25%
Caffeine 6-10%
Tea polysaccharide 17-30%
Pu'er tea of the present invention, method for preparing is following:
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and is made up of the 2-5 jar for every group; Every jar of boiling decocts extracts 3-8 time, adds water 3-10 doubly at every turn, each extraction time 0.5-1.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish and get into circulation after the jar that extracts adds new raw material; Begin new extraction, extracting solution 40-300 order filters, and filtrating is concentrated into Folium Camelliae sinensis (weight): concentrated solution (volume)=1: 0.5-10; Concentrated solution is once centrifugal; One time centrifugal liquid is passed through secondary centrifuging again, and secondary centrifuging liquid is concentrated into proportion 1.01-1.4/55-65 ℃, and condensed cream is dry.
Preferred manufacturing procedure is following:
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and is made up of the 3-4 jar for every group; Every jar of boiling decocts extracts 3-5 time, adds water 5-10 doubly at every turn, each extraction time 0.5-1.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 60-300 order filters; Filtrate decompression concentrates, and temperature≤70 ℃ are to Folium Camelliae sinensis (weight): concentrated solution (volume)=1: 2-3; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated into proportion 1.01-1.4/55-65 ℃, and condensed cream is with microwave drying, vacuum belt type drying or spray drying.
The most preferred method for preparing of the present invention in embodiments of the present invention.
The present invention comprises that also said 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 said 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 through 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, can this chemical compound be suspended or dissolving.The preparation of solution is normally through 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; Said medicine acceptable carrier is selected from: mannitol, sorbitol, sodium pyrosulfite, sodium sulfite, sodium thiosulfate, cysteine hydrochloride, TGA, methionine, vitamin C, 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-, phospholipid material, Kaolin, Pulvis Talci, calcium stearate, magnesium stearate etc.
Pharmaceutical composition of the present invention is confirmed 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.
Below through experimental data beneficial effect of the present invention is described.
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 divide into groups and administration
The animal adaptability is measured fasting glucose according to 2.1 methods after feeding a week, divides into groups according to the fasting blood sugar stratified random of measuring.40 KKAy divide into groups according to the fasting blood sugar stratified random, are divided into 4 groups, model group, positive controls, experimental group.
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, and after animal is measured fasting glucose according to 2.1 methods, irritated clothes glucose 2.5g/kg, 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 according to dosage 1.5g/kg/ day continuously, and food-intake and body weight all change not quite, each treated animal body weight, and the food-intake result of variations is seen table 1, table 2.
Table 1 the weight of animals situation of change (g)
| Divide into groups | The animal number of elements | Dosage | Before the administration | First week | Second week |
| The KKAy model group | 10 | 33.0±1.8 | 35.7±2.5 | 35.4±2.7 | |
| Positive controls | 10 | 1.33g/kg/ day | 33.9±2.4 | 34.7±1.7 | 34.9±1.6 |
| Tea powder | 10 | 1.5g/kg/ day | 32.4±2.3 | 33.2±1.1* | 34.1±1.2 |
Annotate: compare with model
*P＜0.05.
Each treated animal food-intake situation of change (g) of table 2
| Divide into groups | The animal number of elements | Dosage | Before the administration | First week | Second week |
| The KKAy model group | 10 | 4.5±1.1 | 5.2±1.7 | 5.4±1.1 | |
| Positive controls | 10 | 1.33g/kg/ day | 4.0±1.5 | 3.9±0.7 | 4.1±1.2 |
| Tea powder | 10 | 1.5g/kg/ day | 4.1±1.3 | 4.7±0.6 | 4.3±0.8 |
3.2 fasting glucose determination test result
The result sees table 3, and 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.It is thus clear that Folium camelliae assamicae has obvious hypoglycemic activity.
Table 3 Folium camelliae assamicae is to the influence of fasting glucose (nmol/l)
| Divide into groups | Dosage | Before the administration | First week | Second week |
| The KKAy model group | 19.86±5.18 | 24.88±6.18 | 21.48±5.29 | |
| Positive controls | 1.33g/kg/ day | 19.58±4.57 | 20.30±4.63** | 13.91±3.57** |
| Tea powder | 1.5g/kg/ day | 19.67±4.63 | 16.25±2.67** | 13.41±2.17*** |
Annotate: compare with model
*P＜0.01,
* *P＜0.01.
3.3 carbohydrate tolerance test result:
Visible by table 4; 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, and the blood glucose value and the AUC of each time point of positive drug group all are lower than model group, and 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
| Divide into groups | n | 0h | 0.5h | 1h | 2h | AUC |
| The KKAy model group | 10 | 22.83±2.17 | 30.92±2.17 | 29.47±3.27 | 27.36±4.99 | 114.58±11.76 |
| Positive controls | 10 | 13.03±3.86** | 18.38±5.29*** | 18.43±5.63*** | 13.87±3.81*** | 66.48±18.88*** |
| Tea powder | 10 | 15.3±5.39*** | 24.61±3.71*** | 24.07±4.15** | 25.16±5.03 | 9333±15.52** |
Annotate: compare with model group
*P＜0.01,
* *P＜0.001.
3.4 serum electrolyte is measured the result
Visible by table 5, 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
| Divide into groups | n | K + | Na + | Cl - | Ca 2+ |
| The KKAy model group | 10 | 6.07±0.61 | 151.38±2.87 | 113.90±1.76 | 1.04±0.11 |
| Positive controls | 10 | 7.02±0.77** | 156.19±2.17*** | 118.48±1.84*** | 1.10±0.17 |
| Tea powder | 10 | 6.31±0.34 | 155.14±2.14** | 117.18±1.22*** | 1.05±0.04 |
3.6 triglyceride, T-CHOL are measured the result
Visible by table 6, administration in the time of 28 days triglyceride (TG), T-CHOL (CHOL) measure among the result, each experimental group does not have influence to total cholesterol level.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 through experimental session, 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, and liver generation ketoboidies increases and causes hyperglycemia and ketosis, thereby brings out a series of water, electrolyte and acid base imbalance.Can find out that through the mensuration of serum electrolyte Pu'er tea 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: violent boiling helps the conversion and the stripping of effective ingredient; Filling group countercurrent extraction technology can reduce the extraction water consumption greatly when carrying out continuous production in enormous quantities; Alleviating the later stage concentrates cost, reduces energy loss, and this process route can ensure the clarity behind the cold-water solution of finished product; This process route has reduced centrifugal workload of later stage, but the lower industrialization of this process route cost, stable and controllable for quality.Relevant test shows that Pu'er tea of the present invention is higher than prior art curative effect, and purity is high, good absorbing, and steady quality, purposes is novel, and method for preparing 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: 49.95%
Abrownin: 24.10%
Caffeine: 9.10%
Tea polysaccharide: 15.00%.
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and forms by 3 jars for every group; Every jar of boiling decocts extracts 4 times, adds 6 times in water at every turn, each extraction time 1h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 80 orders filter; Filtrate decompression concentrates, and temperature≤70 ℃ are to Folium Camelliae sinensis (weight): concentrated solution (volume)=1: 2; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated to 60 ℃ and measure proportions 1.14, and condensed cream is used microwave drying.
The preparation of embodiment 2 Pu'er teas
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and forms by 3 jars for every group; Every jar of boiling decocts extracts 3 times, adds 5 times in water at every turn, each extraction time 0.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 60 orders filter; Filtrate decompression concentrates, and temperature≤70 ℃ are to Folium Camelliae sinensis (weight): concentrated solution (volume)=1: 0.5; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated to 55 ℃ and measure proportions 1.01, condensed cream spray drying.
Detect through following method, wherein the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 54.35%
Abrownin: 12.47%
Caffeine: 5.53%
Tea polysaccharide: 25.12%.
The method of inspection is following:
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
The progallin A standard solution ImL and the tartaric acid ferron 5mL of the variable concentrations of accurately drawing place the volumetric flask of a series of 25mL, with the buffer standardize solution of pH7.5.Water replaces progallin A as 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 filtrating, and last filtrating 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
The about 0.2g of these article of getting powder (being accurate to 0.001g) puts in the 10ml measuring bottle, and 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 process the solution that contains 0.2mg among every 1ml, as reference substance 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 get.) 1.0ml; The jolting mixing adds 5.0ml sulphuric acid, with the rapid jolting mixing of miniature whirlpool mixed instrument; Room temperature held 30 minutes; With the 1st pipe is blank, according to ultraviolet visible spectrophotometry (appendix VB of Chinese Pharmacopoeia version in 2005), measures absorbance (in 1 hour, measure and finish) in the 487nm wavelength.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; Filtrating can extract and spectrophotometry after soaking and in cold water, being chilled to room temperature.
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 (after the layering, intermediary opacifying layer discards) for use in the tool plug triangular flask.
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, pour ethyl acetate layer solution in the tool plug test tube into carefully, 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, and promptly gets.
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, and adds purified water and supplies weight, filtration promptly gets.
The preparation of embodiment 3, Pu'er tea
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and forms by 5 jars for every group; Every jar of boiling decocts extracts 8 times, adds 10 times in water at every turn, each extraction time 1.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 300 orders filter; Filtrate decompression concentrates, and temperature≤70 ℃ are concentrated into Folium Camelliae sinensis (weight): concentrated solution (volume)=1: 10; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated into 65 ℃ and measure proportions 1.4, and condensed cream is used vacuum belt type drying.
The preparation of embodiment 4, Pu'er tea
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and forms by 2 jars for every group; Every jar of boiling decocts extracts 3 times, adds 3 times in water at every turn, each extraction time 1h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 60-300 order filters; Filtrate decompression concentrates, and temperature≤70 ℃ are concentrated into Folium Camelliae sinensis (weight): concentrated solution (volume)=1: 3; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated into 65 ℃ and measure proportions 1.14, and condensed cream is used vacuum belt type drying.
The preparation of embodiment 5, Pu'er tea
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and forms by 4 jars for every group; Every jar of boiling decocts extracts 6 times, adds 7 times in water at every turn, each extraction time 1.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 150 orders filter; Filtrate decompression concentrates, and temperature≤70 ℃ are concentrated into Folium Camelliae sinensis (weight): concentrated solution (volume)=1: 0.5; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated into 65 ℃ and measure proportions 1.13, and condensed cream is used spray drying.
The preparation of embodiment 6, Pu'er tea
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and forms by 3 jars for every group; Every jar of boiling decocts extracts 5 times, adds 8 times in water at every turn, each extraction time 1.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 150 orders filter; Filtrate decompression concentrates, and temperature≤70 ℃ are concentrated into Folium Camelliae sinensis (weight): concentrated solution (volume)=1: 3; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated into 65 ℃ and measure proportions 1.15, and condensed cream is used vacuum belt type drying.
The preparation of embodiment 7, Pu'er tea
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and forms by 4 jars for every group; Every jar of boiling decocts extracts 5 times, adds 6 times in water at every turn, each extraction time 1.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 100 orders filter; Filtrate decompression concentrates, and temperature≤70 ℃ are concentrated into Folium Camelliae sinensis (weight): concentrated solution (volume)=1: 6; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated into 60 ℃ and measure proportions 1.01, and condensed cream is used spray drying.
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: 35.46%
Abrownin: 16.51%
Caffeine: 6.79%
Tea polysaccharide: 18%.
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: 20.95%
Abrownin: 16.51%
Caffeine: 7.70%
Tea polysaccharide: 34.73%.
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: 17.98%
Abrownin: 15.52%
Caffeine: 6.7%
Tea polysaccharide: 45.23%.
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: 30.95%
Abrownin: 16.51%
Caffeine: 8.2%
Tea polysaccharide: 35.46%.
Embodiment 12,
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: 36.75%
Abrownin: 13.71%
Caffeine: 9.25%
Tea polysaccharide: 37.31%.
Embodiment 13, 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 14
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 portfolio ratio of composition such as caffeine or tea polysaccharide more brews the proportioning of each component in the tea near former tea, mouthfeel is more natural, true.
Claims (9)
1. a Pu'er tea is characterized in that, is extracted by following method to obtain:
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and is made up of the 2-5 jar for every group; Every jar of boiling decocts extracts 3-8 time, adds water 3-10 doubly at every turn, each extraction time 0.5-1.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 40-300 order filters; The weight/volume that filtrating is concentrated into Folium Camelliae sinensis and concentrated solution is=1:0.5-10; Concentrated solution is once centrifugal, and one time centrifugal liquid is passed through secondary centrifuging again, and secondary centrifuging liquid is concentrated into proportion 1.01-1.4/55-65 ℃; Condensed cream is dry
The main active that Pu'er tea contains is a tea polyphenols, tea polysaccharide, and abrownin, caffeine, the percentage by weight of each ingredients constitute extract gross weight is:
Tea polyphenols: 12-50%, abrownin: 12-30%, caffeine: 6-11%, tea polysaccharide: 15-45%, wherein four percentage by weight sums are less than 100%.
2. the extract of claim 1 is characterized in that, method for preparing is following:
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and is made up of the 3-4 jar for every group; Every jar of boiling decocts extracts 3-5 time, adds water 5-10 doubly at every turn, each extraction time 0.5-1.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 60-300 order filters; Filtrate decompression concentrates, and temperature≤70 ℃ to the weight/volume of Folium Camelliae sinensis and concentrated solution are=1:2-3; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated into proportion 1.01-1.4/55-65 ℃, and condensed cream is with microwave drying, vacuum belt type drying or spray drying.
3. the extract of claim 1 is characterized in that, method for preparing is following:
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and forms by 3 jars for every group; Every jar of boiling decocts extracts 4 times, adds 6 times in water at every turn, each extraction time 1h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 80 orders filter; Filtrate decompression concentrates, and temperature≤70 ℃ to the weight/volume of Folium Camelliae sinensis and concentrated solution are=1:2; The concentrated solution tripod pendulum type batch centrifugal is once centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated to 60 ℃ and measure proportions 1.14, and condensed cream is used microwave drying.
4. the pharmaceutical composition that contains the extract of claim 1.
5. the pharmaceutical composition of claim 4 contains 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 is 0.1-99.9%, and all the other are the medicine acceptable carrier.
6. the pharmaceutical composition of claim 4 is any pharmaceutically useful dosage forms.
The compositions of the extract of claim 1 and claim 4 preparation treatment diabetes with slimming medicine in application.
8. content of effective assay method in the Pu'er tea of claim 1, step is following
Content Determination Method of Green Tea Polyphenol
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 l00mL water, draw mother solution 2,4 respectively as mother solution; 6,8, l0mL is mixed with standardize solution not and contains progallin A 50 among the l00mL in the l0mL volumetric flask; 100,150,200, the standard solution of five kinds of variable concentrations of 250mg;
The making of standard working curve
The progallin A standard solution ImL and the tartaric acid ferron 5mL of the variable concentrations of accurately drawing place the volumetric flask of a series of 25mL, with the buffer standardize solution of pH7.5; Water replaces progallin A as 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 l00mL beaker, add the boiling water dissolving more than 20～30mL90 ℃, cooling moves into l00mL
In the volumetric flask, standardize solution, filtration discard the about 20mL of initial filtrating, and last filtrating 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
The about 0.2g of these article of getting powder puts in the 10ml measuring bottle, and 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 process the solution that contains 0.2mg among every 1ml, as reference substance 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; Room temperature held 30 minutes; With the 1st pipe is blank, according to ultraviolet visible spectrophotometry, measures absorbance in the 487nm wavelength; 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; Filtrating can extract and spectrophotometry after soaking and in cold water, being chilled to room temperature;
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 for use;
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, pour ethyl acetate layer solution in the tool plug test tube into carefully, 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.
9. the method for preparing of the Pu'er tea of claim 1, step is following
Get Folium camelliae assamicae, filling group circulated in countercurrent is extracted, and forms by 3 jars for every group; Every jar of boiling decocts extracts 3 times, adds 5 times in water at every turn, each extraction time 0.5h; Every jar the extracting solution first time gets into fluid reservoir, and every jar subsequent extracted liquid is successively as the extraction solvent of next jar; Accomplish getting into circulation after the jar that extracts adds new raw material, begin new extraction, extracting solution 60 orders filter; Filtrate decompression concentrates, and temperature≤70 ℃ to the weight/volume of Folium Camelliae sinensis and concentrated solution are=1:0.5; Concentrated solution is once centrifugal with tripod pendulum type batch centrifugal, and one time centrifugal liquid is used the tube centrifuge secondary centrifuging, secondary centrifuging liquid concentrating under reduced pressure; Temperature≤70 ℃ are concentrated to 55 ℃ and measure proportions 1.01, and condensed cream is used spray drying.
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| CN104688933B (en) * | 2013-12-04 | 2021-01-08 | 云南天士力帝泊洱生物茶集团有限公司 | Composition of Pu' er tea effective components and application thereof in preparing hypoglycemic drug or health food |
| 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 |
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| 李宝才等.腐植酸与普洱茶茶色素.《第六届全国绿色环保肥料新技术、新产品交流会》.2006, |
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