CN101401829A - Wild Jinchai liveness extract, preparation and uses thereof - Google Patents
Wild Jinchai liveness extract, preparation and uses thereof Download PDFInfo
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- CN101401829A CN101401829A CNA2008100264288A CN200810026428A CN101401829A CN 101401829 A CN101401829 A CN 101401829A CN A2008100264288 A CNA2008100264288 A CN A2008100264288A CN 200810026428 A CN200810026428 A CN 200810026428A CN 101401829 A CN101401829 A CN 101401829A
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Abstract
The invention relates to a wild Jinchai extract, as well as a preparation method and application thereof. In the wild Jinchai active extract, the Jiangxi wild Jinchai beech family plant, namely the Lithocarpus polystachyus (wall) Rehd or L.litseifolius (Hamce) Chun is used as a raw material; one of a low-temperature differential pressure type extraction method, a solvent extraction method, a macropore adsorptive resin method, a column chromatography method, a membrane separation concentration technique, a spray drying technique and a freeze drying technique or the free combination of the methods is adopted to prepare the extract. The invention is characterized in that a modernly advanced low-temperature differential pressure type extraction technique is matched with the membrane separation concentration technique and the spray drying technique to prepare the wild Jinchai active extract; and the macropore adsorptive resin technique and the polyamide separation enrichment technique are matched with the freeze drying technique to prepare a wild Jinchai total flavone extract. The content of the total flavone in the wild Jinchai active extract is between 10 and 90 percent, and the pharmacological activity of the wild Jinchai active extract on protecting liver, reducing fat and resisting type 2 diabetes and diabetic complication is higher than that of the prior aqueous extract and alcohol extract.
Description
Technical field
The present invention relates to the field of Chinese medicines, be specifically related to a kind of activity extract of YEJINCHAI and application.
Technical background
The YEJINCHAI source mainly contains the young tender tip, the leaf of plants such as Rosaceae YEJINCHAI, Hydrangeaceae Radix Hydrangeae Strigosae, Fagaceae multiple-ear rock Ke, Vitaceae Ampelopsis grossedentata.Indication YEJINCHAI of the present invention, also claim YEJINCHAI to be mainly derived from Jiangxi Fagaceae plant multiple-ear rock Ke (Lithocarpus polystachyus (wall) Rehd or L.litseifolius (Hamce) Chun.) all plant or any position, RUGEN, rhizome, stem, leaf, spica etc.Multiple-ear rock Ke is distributed on the south the Changjiang river in the low mountain thick forest of each provinces and regions height above sea level more than 400 meters with wild state. and abundant and concentrate with resource-savings such as Jiangxi, Guangxi, Hunan, Anhui especially, provinces such as Guangdong, Yunnan, Sichuan, Fujian take second place, the widest with the Wuyi Mountain Range distribution that Jiangxi, Fujian have a common boundary, resource is very abundant.The YEJINCHAI extract is mainly used in preparation sweeting agent and coloring agent, also is used for clearing away heat and promoting diuresis, control hygropyretic dysentery, skin pruritus, carbuncle malignant boil.This tea of long-term drink, energy liver nourishing nourishing the stomach, clearing heat and moistening lung, promoting the production of body fluid to quench thirst is refreshed oneself and is recovered from fatigue, and the blood pressure lowering fat-reducing is effective in cure to hypertension and coronary heart disease.
Research for the Pasania cuspidata leaf focuses mostly in the raw material as tea and extraction sweeting agent and pigment, and it gives off a strong fragrance, lovely luster, and sweet in the mouth is sweet lasting, unique flavor.(the development and utilization of wild YEJINCHAI such as Yang Yongyao, the tea machinery magazine, 2000 02 phases) water extract to wild plant Pasania cuspidata tender leaf has carried out the physical and chemical composition analysis, the result shows, this lixiviating solution is nutritious, is rich in 17 seed amino acids and various trace elements, vitamin and the multiple flavones ingredient of needed by human body.Wang Yaqian (research of Pasania cuspidata and application, Chinese wild plant resource, 1999 the 18th the 4th phases of volume) report separation from the Pasania cuspidata extract obtains three kinds of dihydrochalcone glycosides, phlorhizin, trilobatin and 3-hydroxyl phlorhizin.Wherein the highest with trilobatin content, reach 95%, its sugariness is 300 times of sucrose.And from the YEJINCHAI extract of Hunan, be separated to two kinds of natural glucins that content is high, total content is 12.6%, be accredited as dihydrochalcone-glycoside through chemical constitution, point out also to exist in the Pasania cuspidata extract eight kinds of triterpenoid compound, be oleanane, dammarane, class materials such as cycloartane, this compounds may be YEJINCHAI cough-relieving effective ingredient.In addition, the Pasania cuspidata lixiviating solution also contain K, Na, Ca, etc. the trace element of macroelement and multiple needed by human, aminoacid, vitamin.(the irritated effective ingredient preliminary study of Yunnan YEJINCHAI such as Li Wanyi, Yunnan University's journal, the 23rd the 6th phase of volume of calendar year 2001) from the ethyl acetate extract of Yunnan YEJINCHAI (Pasania cuspidata), separates 2 chemical compounds that obtain having hyaluronidase inhibitor, through spectrum analysis, methods such as chemical reaction and document contrast, Preliminary Identification confirm that effective ingredient is dihydrochalcone-2 '-β-D-glucopyanoside (dihydrochareone-2 '-β-D-glu-copyranoside) and dihydrochalcone-4 '-β-D-glucopyanoside (dihydrocharcone4 '-β-D-glucopyranoside).Fringe Ke brown pigment is to be extracted, separated by leaf, its productive rate can reach cured leaf heavy about 10%, belong to phenolic acid compound.This pigment can be used for the painted of confection, cold drink, beverage, assembled alcoholic drinks and cake etc., and the sugar-coat that also can be used for medicine is painted, has been GB2760-36 by Ministry of Public Health approval implementation criteria.
Many about Folium Lithocarpi Polystachyi chemical constituent and Pharmacological action study thereof, it uses the research that also is confined to health tea, beverage and sweet ingredient thereof, tool tea, sugar, three kinds of economic worths of medicine are YEJINCHAI all over the body, wait to develop to the effect of its " medicine " aspect is anxious, and Jiangxi YEJINCHAI research report in this regard still less, and do not have the report of preparation aspect.The present invention carries out comprehensive research of pharmacy, pharmacodynamics to the Jiangxi YEJINCHAI, adopt modern advanced low-temperature differential pressure type extractive technique to cooperate the concentrated technology of membrance separation, spray drying technology to prepare Wild Jinchai liveness extract, utilize macroporous adsorbent resin, polyamide separation and concentration technology to cooperate Freeze Drying Technique to prepare highly purified YEJINCHAI extractive of general flavone, and pharmacological actions such as its liver-protecting and blood fat-reducing, anti-type 2 diabetes mellitus and diabetic complication are better than conventional water extract, ethanol extract; Made dosage forms such as capsule, tablet, pill as required, method is simple, and is easy and simple to handle, is beneficial to commercial production.
Summary of the invention
The purpose of this invention is to provide a kind of Wild Jinchai liveness extract with clear and definite index.
Another object of the present invention provides the preparation method of this kind Wild Jinchai liveness extract.
The YEJINCHAI of indication of the present invention, refer in particular to and be the whole plant of Jiangxi Fagaceae plant multiple-ear rock Ke (Lithocarpuspolystachyus (wall) Rehd or L.litseifolius (Hamce) Chun.) or any position, RUGEN, rhizome, stem, leaf, spica etc. also comprise the processed product through various concocting methods.
Wild Jinchai liveness extract of the present invention contains 10%～90% total flavones, and its color is faint yellow to brown.The qualitative identification experiment shows: hydrochloric acid-magnesium powder reaction is positive.Further, this Wild Jinchai liveness extract contains 45%～60% total flavones.
Wild Jinchai liveness extract high-efficiency liquid-phase fingerprint 285nm of the present invention detects 26 characteristic peaks, and wherein 6 peaks are main peaks, account for more than 89% of total peak area; Extract high-efficiency liquid-phase fingerprint 345nm detects 26 characteristic peaks, and wherein 14 peaks are main peaks, account for more than 95% of total peak area.
The Wild Jinchai liveness extract of the present invention hydrochloric acid hydrolysis of concentration 2%～6% (volume ratio), 90 ℃ of hydrolysis temperatures, hydrolysis time is 60 minutes, high-efficiency liquid-phase fingerprint has 4 characteristic peaks, when concentration of hydrochloric acid reaches 6% but tiny peak showed increased in the chromatogram when being lower than 12%, concentration of hydrochloric acid to increase the fragment that chemical compound is hydrolyzed into many more, when concentration reaches 12%, do not have chromatographic peak in the high-efficiency liquid-phase fingerprint, it is complete that chemical compound is hydrolyzed oxidation substantially.
It is reference substance that general flavone content detection method of the present invention adopts with the naringin, ultraviolet spectrophotometry; The general flavone content detection method is with reference to pharmacopeia version in 2005, appendix VA ultraviolet visible spectrophotometry.
Flavones ingredient in the Wild Jinchai liveness extract of the present invention, can with any and slaine sodium carbonate in alkali sodium hydroxide, the potassium hydroxide, potassium carbonate, calcium carbonate, zinc acetate in any, metallic salt derivant that forms and the metal ion complex that forms with any metal ion in ferrum, aluminum, zinc, copper, barium, chromium, the strontium, these derivants have pharmacologically active or the purposes identical or close with above-mentioned YEJINCHAI extractive of general flavone with complex.
Described YEJINCHAI method for preparing extractive of the present invention: the combination in any of any one method or these methods is prepared in solvent extraction method, low-temperature differential pressure type extraction method, solvent extraction, macroporous adsorbent resin method, column chromatography, the concentrated technology of membrance separation, spray drying technology, the Freeze Drying Technique.Wherein be preferably low-temperature differential pressure type extraction method, macroporous adsorbent resin method, polyamide column chromatography, the concentrated technology of membrance separation, spray drying technology, Freeze Drying Technique.Preferably, the initiative low-temperature differential pressure type extraction method of using of the present invention is extracted YEJINCHAI, and finds that the extract that obtains has extraordinary character.
When these methods of use are prepared, comprise following step:
(1) extract: solvent for use can be water or any one alcohols, ketone and esters solvent, or the mixed solvent formed by a certain percentage of these solvents, or the acidity or the basic solvent that are made into by these solvents and acid, alkali, salt; Extracting method can be decoction, reflux, supersound extraction, merceration, percolation, microwave extraction, low-temperature differential pressure type extraction etc.
Preferred extraction process is:
The YEJINCHAI medical material adds 0～95% ethanol, and reflux, extract, 2～3 times was extracted 1～2 hour at every turn, and solvent load is 5～20 times of amounts (ml/g).
The YEJINCHAI medical material through size reduction machinery separate broken for granularity be granule below the 10mm, add 10～50 times of amounts of 0～80% ethanol, stir and keep dynamically, at-35 ℃～40 ℃, " normal pressure-vacuum-Jia solvent-normal pressure-pressurization-normal pressure " constantly circulation carries out the differential pressure extraction, vacuum remains on 13.33Pa～1.00MPa as required, and pressure remains on 1MPa～25.00MPa as required, and cycle-index is selected 4～8 times as required.
(2) filter: comprise methods such as centrifugal, sucking filtration, ultrafiltration, microfiltration, use or do not use following any clarifier or its combination: any in precipitate with ethanol liquor, gelatin, Kaolin, silicon bath soil, various resin, Polyethylene Glycol, poly-second triol, chitosan and natural clarifying agent finished product 101 fruit juice clarifiers, the ZTC+1 natural clarifying agent.
(3) concentrate: comprise that thin film evaporation, rotary evaporation, decocting and concentrating, membrance concentration technology, suspension freeze concentration, progressive freeze under normal pressure or the reduced pressure concentrates, natural outer circulation two phase flow concentrates, in concentrating any of outer circulation type freeze concentration, spray chilling.
(4) drying: comprise in vacuum drying, spray drying, the lyophilization any.
When adopting solvent extraction to be prepared, earlier extract mixture is suspended from the water, then with low polar esters, ether solvent extraction weeding of grease solubility impurity, use suitable solvent then, can be a kind of in chloroform, ethyl acetate, acetone, the n-butyl alcohol, or the mixture of these solvents, extraction obtains YEJINCHAI total flavonoid composition.
When using the macroporous adsorbent resin method to be prepared, used macroporous resin can be nonpolar, low pole, middle polarity, polarity, the any type of alkalescence or faintly acid, can be D101, X-5, NKA, S-8, NKA-9, HPD-600, ADS-7, AB-8, LSA-5B, HPD-722, ADS-17, DM130, HPD-826, DN-300, DN-603, FL-1, FL-2, among the FL-3 any, low pole preferably wherein, middle polarity or polar resin, comprise AB-8, ADS-17, among the HPD-600 any, used eluant can be water, aquiferous ethanol, methanol, in the acetone any wherein is preferably 0～100% ethanol.
When using column chromatography with polyamide during as immobile phase, used chromatography polyamide is 30-60 order, 60-120 order, 100-120 order, wherein be preferably 30-60 purpose resin, used eluant is any in water, aquiferous ethanol, methanol, the acetone, wherein is preferably 0～100% ethanol.
The preferred pure metallization processes of YEJINCHAI total flavonoid is: select for use to comprise AB-8, ADS-17, any low pole among the HPD-600, middle polarity or polar macroporous adsorbent resin and 30～60 purpose chromatography polyamides are as the purification resin, the YEJINCHAI extract is 1: 5～1: 10 in crude drug amount sample solution extension rate, absorption flow velocity 2～5BV/h, resin column blade diameter length ratio 1: 5～1: 10, in crude drug amount applied sample amount is 0.1～1g/ml, purified water remove impurity with 1～4 times of resin volume, the remove impurity flow velocity is 2～5BV/h, with 2～8 times of resin volumes of 10%～95% ethanol elution, elution flow rate is 2～5BV/h.
When adopting column chromatography to be prepared, its process object can be the product that the said extracted step is obtained, and also can be the product behind above-mentioned solvent extraction method, solvent extraction or the macroporous adsorbent resin method preliminary purification.Used immobile phase can be any in silica gel, polyamide, aluminium oxide, glucosan, C-8, C-18, active carbon, the cellulose, used eluent is different because of the difference of immobile phase, generally by any or several mixed solvent of forming in water, methanol, ethanol, acetone, chloroform, ethyl acetate, acetone, the petroleum ether.
When using membrance separation to concentrate technology, its microfiltration is held back the material of diameter more than 0.1 μ m; The solable matter of ultrafiltration molecular cut off between 1000～500000; The nanofiltration molecular cut off more than 150, the material of diameter about 1nm.The material of its use can be that organic class comprises that cellulose acetate, polypropylene, Merlon, polysulfones, polyamide and mineral-type cover pottery, metal, metal-oxide, charcoal, cellular glass.
This extract can be pressed the arbitrary proportion compatibility separately or with other any Chinese and western drugs or food, be used to prepare medicine or functional food, prepared medicine or functional food can be any in capsule, tablet, pill, granule, oral liquid, syrup, electuary, medicated wine, injection, unguentum, powder, beverage, the cataplasma.
Multiple of the present invention is meant that solvent volume consumption (ml) is the multiple of quality of medicinal material consumption (g).
Concentration of alcohol of the present invention is meant volumetric concentration (v/v).
Wild Jinchai liveness extract preparation method abundant raw material of the present invention, the processing technology simple and convenient.
The application of Wild Jinchai liveness extract of the present invention in preparing cardiovascular system diseases such as treating hyperlipidemia, coronary heart disease and disease medicaments such as fatty liver, diabetes and diabetic complication.
Description of drawings
Fig. 1 is a YEJINCHAI extract UV scanning collection of illustrative plates.
Fig. 2 a, b are the HLPC finger printing (285nm and 345nm) of YEJINCHAI total extract.
Fig. 3 a, b are the HPLC collection of illustrative plates (285nm and 345nm) behind 2% hydrochloric acid hydrolysis of YEJINCHAI extract.
Fig. 4 a, b are the HPLC collection of illustrative plates (285nm and 345nm) behind 4% hydrochloric acid hydrolysis of YEJINCHAI extract.
Fig. 5 a, b are the HPLC collection of illustrative plates (285nm and 345nm) behind 6% hydrochloric acid hydrolysis of YEJINCHAI extract.
Fig. 6 is and normal group model group diabetes rat serum saccharifying serum albumin (GSP), triglyceride (TG), cholesterol (TC), nephridial tissue peroxidating product (MDA) level relatively.
The specific embodiment
Related method is the technological means that those skilled in the art can grasp and use, but following exemplifying embodiment must not be interpreted as the restriction to claim of the present invention of going up in all senses.
YEJINCHAI medical material 5kg, through size reduction machinery separate broken for granularity be granule below the 10mm, the water that adds 20 times of amounts, stir and keep dynamically, under 0 ℃～5 ℃ condition, to carry out differential pressure by " normal pressure-vacuum-Jia solvent-normal pressure-pressurization-normal pressure " continuous circulation and extract, vacuum remains on 14Pa, pressure keeps 10.00MPa, carries out 4 circulations and extracts.Extracting solution is carried out solid-liquid separation to clarification through solid-liquid separation mechanics, get clarifying extracting solution, clarifying extracting solution is carried out microfiltration and ultrafiltration continuously get concentrated solution, spray drying gets the YEJINCHAI total extract.With the naringin is reference substance, and general flavone content is about 60% in the determined by ultraviolet spectrophotometry Wild Jinchai liveness extract.
Get YEJINCHAI medical material 5kg, through size reduction machinery separate broken for granularity be granule below the 10mm, the process water that adds 20 times of amounts, stir and keep dynamically, at 0 ℃～5 ℃, " normal pressure-vacuum-Jia solvent-normal pressure-pressurization-normal pressure " constantly circulation carries out the differential pressure extraction, and vacuum remains on 14Pa, pressure keeps 10.00MPa, carries out 4 circulations and extracts.Extracting solution is carried out solid-liquid separation to clarification through solid-liquid separation mechanics, get extracting solution.Get extracting solution 200ml, add equimultiple water dilution, cross AB-8 type macroporous adsorbent resin, be washed to colourlessly, 70% ethanol elution is collected eluent to colourless, and being evaporated to does not have the alcohol flavor, dry Wild Jinchai liveness extract.With the naringin is reference substance, and general flavone content is about 48% in the determined by ultraviolet spectrophotometry Wild Jinchai liveness extract.
Get YEJINCHAI medical material 5kg, through size reduction machinery separate broken for granularity be granule below the 10mm, the process water that adds 20 times of amounts, stir and keep dynamically, at 0 ℃～5 ℃, " normal pressure-vacuum-Jia solvent-normal pressure-pressurization-normal pressure " constantly circulation carries out the differential pressure extraction, and vacuum remains on 14Pa, pressure keeps 10.00MPa, carries out 4 circulations and extracts.Extracting solution is carried out solid-liquid separation to clarification through solid-liquid separation mechanics, get extracting solution.Get extracting solution 200ml, add the water dilution of equimultiple, cross polyamide column (30～60 order), be washed to colourlessly, 80% ethanol elution is collected eluent to colourless, and being evaporated to does not have the alcohol flavor, dry Wild Jinchai liveness extract.With the naringin is reference substance, and general flavone content is about 52% in the determined by ultraviolet spectrophotometry Wild Jinchai liveness extract.
Get the spray drying medicated powder 15g of YEJINCHAI low-temperature differential pressure type extract, adding distil water 200ml makes dissolving, crosses the AB-8 macroporous adsorbent resin, be washed to colourless, 70% ethanol elution is collected eluent then, and being evaporated to does not have the alcohol flavor, cross polyamide column (30～60 order), be washed to colourlessly, reuse 70% ethanol elution is collected eluent, being evaporated to does not have the alcohol flavor, the dry Wild Jinchai liveness extract that gets.With the naringin is reference substance, and general flavone content is about 45% in the determined by ultraviolet spectrophotometry YEJINCHAI extract.
Get the spray drying medicated powder 100g of YEJINCHAI low-temperature differential pressure type extract, add the equimultiple carboxymethyl starch sodium, mixing, dry-pressing is granulated, and is encapsulated, gets capsule.
Get the spray drying medicated powder 100g of YEJINCHAI low-temperature differential pressure type extract, add equimultiple sucrose, equimultiple dextrin and appropriate amount of starch, make granule, drying, granulate is made 1000g, is distributed into the every bag of 10 grams.
Embodiment 7
Get the spray drying medicated powder 100g of YEJINCHAI low-temperature differential pressure type extract, adding distil water 1000ml makes dissolving, adding ethanol is 80% to containing the alcohol amount, standing over night is got supernatant concentration to about 500ml, adds 0.1% active carbon and boils half an hour, filter, filtrate is crossed 0.2 μ m microporous filter membrane, obtains activity extract, and wherein general flavone content is about 39%.Above-mentioned Wild Jinchai liveness extract is added the injection water to 1000ml, packing (every bottle of 100ml), sterilization promptly gets the YEJINCHAI injection.
Embodiment 8
Get the spray drying medicated powder 100g of YEJINCHAI low-temperature differential pressure type extract, starch 100g, mix homogeneously, it is an amount of to add Pulvis Talci, is pressed into 1000.
Embodiment 9
Get three parts of the spray drying medicated powder 10g of YEJINCHAI low-temperature differential pressure type extract, add 2%, 4%, 6% hydrochloric acid solution 200ml respectively, 90 ℃ of heating in water bath were hydrolyzed in 1 hour, took out, and put cold, with water saturation ethyl acetate extraction 5 times, each 100ml, combined ethyl acetate liquid, low-temperature reduced-pressure reclaims solvent, get residue, with methanol 100ml dissolving.The HPLC collection of illustrative plates of its hydrolysis afterproduct such as Fig. 4 ab, 5ab, 6ab.
Embodiment 10
Get the spray drying medicated powder 10g of YEJINCHAI low-temperature differential pressure type extract, add distilled water 200ml and dissolve, use water saturation n-butanol extraction 5 times, each 100ml merges n-butyl alcohol liquid, and low-temperature reduced-pressure reclaims solvent, gets residue, uses methanol 100ml dissolving.With the naringin is reference substance, and determined by ultraviolet spectrophotometry Wild Jinchai liveness extract hydrolysate general flavone content is about 55%.
Get YEJINCHAI medical material 5kg, through size reduction machinery separate broken for granularity be granule below the 10mm, the process water that adds 30 times of amounts, stir and keep dynamically, at 5 ℃～10 ℃, " normal pressure-vacuum-Jia solvent-normal pressure-pressurization-normal pressure " constantly circulation carries out the differential pressure extraction, and vacuum remains on 0.4MPa, pressure keeps 15.00MPa, carries out 4 circulations and extracts.Extracting solution is carried out solid-liquid separation to clarification through solid-liquid separation mechanics, get extracting solution, extracting solution carries out microfiltration and ultrafiltration gets concentrated solution, and spray drying gets the YEJINCHAI extract.With the naringin is reference substance, and general flavone content is about 62% in the determined by ultraviolet spectrophotometry Wild Jinchai liveness extract.
Embodiment 12
Get YEJINCHAI medical material 5kg, through size reduction machinery separate broken for granularity be granule below the 10mm, the process water that adds 40 times of amounts, stir and keep dynamically, at 10 ℃～15 ℃, " normal pressure-vacuum-Jia solvent-normal pressure-pressurization-normal pressure " constantly circulation carries out the differential pressure extraction, and vacuum remains on 0.8MPa, pressure keeps 20.00MPa, carries out 4 circulations and extracts.Extracting solution is carried out solid-liquid separation to clarification through solid-liquid separation mechanics, get extracting solution, extracting solution carries out microfiltration and ultrafiltration gets concentrated solution, and spray drying gets the YEJINCHAI extract.With the naringin is reference substance, and general flavone content is about 55% in the determined by ultraviolet spectrophotometry Wild Jinchai liveness extract.
The pharmacodynamics data
One, experiment material
1, is subjected to reagent thing and control drug
The YEJINCHAI low-temperature differential pressure type extract that the method for embodiment 1 makes (being called for short " activity extract "), the conventional water extract of YEJINCHAI (being called for short " conventional water extract "), the conventional ethanol extract of YEJINCHAI (being called for short " conventional ethanol extract ") provides by Traditional Chinese Medicine University Of Guangzhou new drug development research center.
Positive control drug: diamicron, Tianjin Hua Jin pharmaceutical factory, lot number: 070813.Novolin, lot number: SVG0426, Denmark Novo Nordisk Co.,Ltd produces.Sodium chloride injection (500ml, Hunan Cologne Pharmaceutical Co., Ltd, 041221).
2, laboratory animal
The SD rat, the SPF level, male and female half and half are provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center, normally raise after 3 days for examination.
3, key instrument and reagent
The BS110S electronic balance, Sartorius company produces; 722 grating spectrophotometers, Shanghai Precision Scientific Apparatus Co., Ltd produces; SN-695B type intelligence is put and is exempted from the r measuring instrument, and day ring one factory of Shanghai nuclear research institute produces.
Serum biochemistry test kit (AST, ALT, TG, TC, BUN, CREA) is provided by Beijing Zhong Shengbei control bio tech ltd; The MDA test kit, building up biological company limited by Nanjing provides.(streptozotocin, STZ), Sigma company faces with preceding that to be made into concentration with citrate buffer solution be 3% STZ solution to streptozotocin.(lot number: 449497), all company of Switzerland Roche Group provides for the full blood glucose meter of Luo Kang, blood sugar test paper.The insulin test kit, Kemei Dongya Biological Technology Co., Ltd., Beijing provides, lot number: 20071025.(lot number: 396494), sigma company provides GER-11..The saccharifying serum albumin is measured test kit, and (lot number: 20070621), building up bio-engineering research by Nanjing is provided.(lot number: 20070804), the sharp glad biotechnology research in Shanghai provides glucose kit.(lot number: 20070410), Guangzhou city flower all Gao Erbao Bioisystech Co., Ltd provides Tes-Tape.
4, statistical method
Use SPSS12.0 to carry out data analysis.Data are all used mean ± standard deviation (x ± s) expression.Each index adopts one factor analysis of variance and t check; The data Kruskal-Wallis H check and q check to heterogeneity of variance
Two, method and result
1. to the protective effect of alcoholic fatty liver
Get SD female rats 160～200g, be divided into normal control group, model group, silymarin positive controls at random, join this dibasic acid esters positive controls, Wild Jinchai liveness extract, the conventional water extract of YEJINCHAI, the conventional ethanol extract of YEJINCHAI.Normal group every day at the upper and lower noon gavages 10ml/kg, 20ml/kg drinking water respectively; Model group gavages 55 ° of alcoholic solution 10ml/kg the morning, gavages the 20ml/kg drinking water afternoon; Each administration group every morning gavages 55 ° of alcoholic solution 10ml/kg, gavages the medicinal liquid 20ml/kg of variable concentrations afternoon.Normal group is raised with normal diet, freely drinks water; Model group, each administration group are raised with the high fat low-nutrition feed of making 20% by oneself, freely drink water.After 4 weeks, behind the animal fasting 12h, eye socket is got blood, and separation of serum is measured T-CHOL (TG), total triglyceride (TC), glutamate pyruvate transaminase (ALT), and each treated animal is put to death in anesthesia.Survey body weight, get liver and weigh, calculate organ coefficient.The results are shown in Table 1.
Table 1 YEJINCHAI extract is to the therapeutical effect of alcoholic fatty liver rat (x ± s)
Annotate: model group and normal group be #p＜0.05 relatively, ##p＜0.01; Administration group and model group compare, * p＜0.05, * * p＜0.01.
The result shows: compare with normal group, gavage ethanol and raise with behind the high lipid food, the obvious swelling of model group liver, liver coefficient extremely significantly raise (p＜0.01), simultaneously blood fat T-CHOL, total triglyceride, gpt level significantly raise (p＜0.05 or p＜0.01).Compare with model group, the positive control drug silymarin can significantly alleviate liver enlargement degree and suppress the rising (p＜0.05 or p＜0.01) of blood fat T-CHOL, total triglyceride; The positive control drug bifendate also can extremely significantly suppress the rising (p＜0.01) of blood fat T-CHOL, total triglyceride; Wild Jinchai liveness extract can be shown the rising (p＜0.05 or p＜0.01) that suppresses blood fat T-CHOL, total triglyceride; The conventional extract of YEJINCHAI can be shown the rising (p＜0.05) that suppresses blood fat T-CHOL, total triglyceride, but effect is weaker than activity extract.Each treatment group all alleviates the liver coefficient and reduces the gpt level effect except that bifendate, but no difference of science of statistics.Results suggest: the chronic hepatic injury that Wild Jinchai liveness extract is increased due to the fat feedstuff ethanol has therapeutical effect, and obviously blood fat reducing improves liver function, and effect is better than conventional water, ethanol extract.
2. to the therapeutical effect of type 2 diabetes mellitus rat
2.1 therapeutical effect to the type 2 diabetes mellitus rat
Randomly drawing 9 rats feeds to normal diet as the normal control group.All the other rats are as model group, the model group rat gives high lipid food (1% cholesterol, 10% Adeps Sus domestica, 10% yolk powder, 2% sucrose, 77% normal feedstuff) feeds, feed after 4 weeks animal fasting (can't help water) with model group after 12 hours, the 4%STZ solution 30mg/kg of intravenous injection citrate buffer solution (PH4.6) configuration, solution is used filtering with microporous membrane earlier.Normal control group lumbar injection is with the citrate buffer solution of volume.Behind the 3d, the tail vein is got blood with blood taking needle and is surveyed blood glucose, surveys glucose in urine simultaneously.Whole blood blood glucose 〉=11mmol/L, glucose in urine +++~ ++ ++, keep and be defined as the modeling success more than 1 week.The diabetes rat of modeling success is divided into 4 groups, be respectively model group (9), Wild Jinchai liveness extract beverage group (9), the conventional water extract filling of YEJINCHAI stomach group (9), the conventional ethanol extract filling of YEJINCHAI stomach group (9), reach the interior animal of group between group and mark respectively.The normal control group gives normal feedstuff, drinking public water supply.Model group gives high lipid food, drinking public water supply.Other each administration group all gives high lipid food, corresponding YEJINCHAI extract, changes/administration 1 time 4 weeks of successive administration every day.Glucose in urine, urine protein and the body weight that detects each treated animal after the administration weekly once and note down.Body is skinny, body weight gain is slow, under the glucose in urine, urine protein strong positive animal per perithelium insulin injection 5u/kg in case the death of stop thing.Medicinal liquid is drunk preparation: get the 10ml medicinal liquid and directly drink with distilled water diluting to 250mL rat.Successive administration is after 4 weeks, animal fasting 12h, and the urethane intraperitoneal injection of anesthesia is weighed and record, after the ventral aorta blood sampling, the centrifugal 10min separation of serum of 3000 commentaries on classics/min (surveying fasting blood sugar with blood glucose meter when getting blood).Get part serum and survey SOD, MDA, GSP level; All the other serum are surveyed insulin (FNS), triglyceride (TG), cholesterol (TC) level.Dissect rat and get liver and kidney, weigh, calculate organ coefficient; Get a side kidney and hepatomegaly leaf, 5% formalin is fixed, and does the routine pathology histological examination.The results are shown in Table 2,3,4 and Fig. 1.
Table 11 is respectively organized relatively (x ± s) of 4 week of rat treatment back FBG, FNS, Ln (ISI) result
Annotate: compare #p＜0.05, ##p＜0.01 with the blank group; Compare with model group: * p＜0.05, * * p＜0.01.
Behind the injection STZ, the rat model hair color is uninteresting loose, tarnish, and spontaneous activity reduces, and irritating to external world sensitivity descends, and the normal treated animal of stool is rare half congealed, polyuria obviously occurs, polydipsia, polyphagia symptom.Symptom more than three all has to a certain degree improvement after each treatment group rat administration, and hair color is moist than the modeling group, and spontaneous activity increases, and stimulation to external world is responsive.When weekly the urine glucose being detected, not treatment group rat significantly is strong positive.As can be seen from Table 2, compare with the blank group, after model group treated for 4 weeks, the fasting glucose utmost point is significantly higher than blank group (P＜0.01), and the fasting insulin serum levels raises, but no difference of science of statistics (P＞0.05), and insulin sensitivity index obviously descends, and remarkable significant difference (P＜0.01) is arranged; Compare with model group, after the Wild Jinchai liveness extract group treated for 4 weeks, fasting glucose reduced, significant difference (P＜0.05) is arranged, and the fasting insulin serum levels reduces, no difference of science of statistics (P＞0.05), insulin sensitivity index obviously raises, and remarkable significant difference (P＜0.01) is arranged; Compare with model group, after the conventional water extract of YEJINCHAI, ethanol extract group treated for 4 weeks, fasting glucose reduced, and insulin sensitivity index slightly raises to some extent, but the rising of fasting insulin serum levels, all no difference of science of statistics (p＞0.05).Prompting: the Wild Jinchai liveness extract type 2 diabetes mellitus rat insulin Sensitivity Index that can raise again in the time of to a certain extent can blood sugar lowering, promptly improve insulin resistant; Effect is better than conventional water, ethanol extract.
Table 3 is respectively organized relatively (x ± s) of dirty, the kidney coefficient results in 4 week of rat treatment back
Annotate: compare #p＜0.05, ##p＜0.01 with the blank group; Compare with model group: * p＜0.05, * * p＜0.01.
Normal rats liver, scarlet, the smooth surface of kidney perusal color; Fed with high is after 8 weeks, and model group diabetes rat liver is ochre yellow, rough surface tarnish a little, and the kidney volume increases; Each administration group liver of YEJINCHAI, the color of kidney, lustrous surface degree have improvement slightly than model group.Can also find out from table 12: model group liver, kidney coefficient have significant difference (P＜0.01) all greater than normal group, and the modeling of hints model group rat is fed can cause liver, injury of kidney after the fed with high, causes its enlargement.Compare with model group, the Wild Jinchai liveness extract group has the effect that suppresses liver, renomegaly in various degree, and effect is better than conventional water, ethanol extract, but equal no difference of science of statistics.The prompting Wild Jinchai liveness extract can improve the degree of injury of type 2 diabetes mellitus rat Liver and kidney to a certain extent, may be with its blood circulation promoting and blood stasis dispelling, to increase body relevant to the effect of insulin sensitivity.
Table 4 is respectively organized relatively (x ± s) of 4 week of rat treatment back GSP, TG, TC and nephridial tissue MDA, SOD result
Annotate: compare #p＜0.05, ##p＜0.01 with the blank group; Compare with model group: * p＜0.05, * * p＜0.01.
Table 4 shows: compare with normal group, model group diabetes rat serum saccharifying serum albumin (GSP), triglyceride (TG), cholesterol (TC), nephridial tissue peroxidating product (MDA) level extremely significantly raise (P＜0.01), and nephridial tissue superoxide dismutase (SOD) extremely significantly descends (P＜0.01), point out higher with the diabetes rat blood lipid level of streptozotocin and high lipid food modeling, hyperglycemia brings out glycation product and the peroxidating product increases.Compare with model group, Wild Jinchai liveness extract group rat blood serum GSP, TG and MDA water half descends, and SOD raises, and significant difference (P＜0.05 or P＜0.01) is all arranged, cholesterol (TC) level has decline to a certain degree, but no difference of science of statistics (p＞0.05); Conventional water, ethanol extract group GSP, TG, TC, the horizontal no difference of science of statistics of MDA, SOD, effect is weaker than activity extract.Results suggest: Wild Jinchai liveness extract is drunk the generation that can reduce type 2 diabetes mellitus rat hyperlipidemia degree, glycation product, organize peroxide, improve the ability organize peroxide injury due to the hyperglycemia, have the effect that control system diabetic complication produces or increases the weight of.
Fig. 1 different dosing group diabetes rat liver, kidney pathology Histological change
The result of histopathologic examination: the serious edema of hepatocyte appears in model group rat 9 examples, the endochylema puffing, and obviously fat change appears in 5 examples; Part animal renal tubular epithelial tissue distribution is inhomogeneous; Wild Jinchai liveness extract is drunk 9 examples and the slight edema of hepatocyte occurred, and 1 example fat becomes, and kidney is not found bright typical cytopathic; 6 routine hepatocyte edema appear in YEJINCHAI extract water extract group, the slight edema of 3 examples, and 2 example fat become, and degree all is lighter than model group, and kidney is not found bright typical cytopathic; 5 routine hepatocyte edema appear in YEJINCHAI extract ethanol extract group, the slight edema of 4 examples, and 1 example fat becomes, and degree all is lighter than model group, and kidney is not found bright typical cytopathic.
26.2 to the glycometabolic influence of diabetes rat
Randomly drawing 6 rats feeds to normal diet as the normal control group.All the other rats are as model group, the model group rat gives high lipid food (1% cholesterol, 10% Adeps Sus domestica, 10% yolk powder, 10% sucrose, 69% normal feedstuff) feeds, after 4 weeks the animal fasting (can't help water) of model group is spent the night, the STZ solution 30mg/kg of lumbar injection citrate buffer solution (PH4.4) configuration.Normal control group lumbar injection is with the citrate buffer solution of volume.Overnight fasting behind the 3d, the tail vein is surveyed blood glucose with the blood taking needle blood sampling.Whole blood blood glucose 〉=11mmol/L, glucose in urine +++~ ++ ++, be defined as diabetes model more than keeping for 1 week.Diabetes rat is divided into 3 groups, is respectively model group, diamicron administration group, Wild Jinchai liveness extract group, the conventional water extract group of YEJINCHAI, the conventional ethanol extract group of YEJINCHAI, 10 every group, reach the interior animal of group between group and mark respectively.The normal control group gives normal feedstuff, and all the other each groups all give high-sugar-fat-diet.Each administration group dosage sees the following form, and model control group and normal control group such as gavage every day at the distilled water of capacity, administration every day 1 time, 5 weeks of successive administration.2 weeks before administration, after the administration, animal fasting 12h, the urethane intraperitoneal injection of anesthesia, the eyeball blood sampling, separation of serum is measured blood glucose (FBG) and insulin level (INS) before and after the administration, calculates insulin sensitivity index (ISI), the results are shown in Table 5.In administration the 21st day, animal fasting 3h, adopt 0min blood after, gavage glucose solution 2.0g/kg, measure blood glucose respectively in gavaging back 30min, 1h, 2h.In administration the 25th day, do not get hematometry 0min blood glucose value under the fasting state, subcutaneous injection novolin (40 times of normal saline dilutions) 0.5u/kg measures blood glucose in injection back 40min, 80min, 120min immediately, the results are shown in Table 6.
Table 5 YEJINCHAI extract is to the influence of diabetes rat insulin sensitivity index (x ± s)
Annotate: model group and normal group be #p＜0.05 relatively, ##p＜0.01; Compare * p＜0.05, * * p＜0.01 before the administration with after the administration
Table 5 result shows: compare with normal rats, insulin sensitivity index all extremely significantly is lower than normal rat before and after the administration of model group rat, and prompting diabetes rat insulin sensitivity index descends.With comparison before the administration, no change before and after the insulin sensitivity index administration of normal group, the insulin sensitivity index of model group slightly descends but no difference of science of statistics (p＞0.05), the insulin sensitivity index of positive control diamicron, Wild Jinchai liveness extract group all be significantly increased (p＜0.05); The insulin sensitivity index of the conventional water of YEJINCHAI, ethanol extract group is improved, but no difference of science of statistics.The same insulin sensitivity index that can improve diabetes rat with diamicron of prompting Wild Jinchai liveness extract improves carbohydrate metabolism.
Table 6 YEJINCHAI extract is to the influence of diabetes rat insulin tolerance, carbohydrate tolerance (x ± s)
In insulin tolerance test, behind the rat skin lower injection insulin, blood glucose begins to descend, and different dosing group rat blood sugar reaches the minimum time and has nothing in common with each other.Table 6 result shows: during insulin injection 40min, the blood glucose of normal rat, Wild Jinchai liveness extract, the conventional water extract of YEJINCHAI, the conventional ethanol extract treated animal of YEJINCHAI drops to minimum, model group, positive control drug diamicron group blood glucose then just reach minimum when 80min, prompting normal group, each extract administration group rat of YEJINCHAI is big to the sensitivity of exogenous insulin than model group and diamicron administration group rat, the YEJINCHAI extract can strengthen the sensitivity of diabetes rat to exogenous insulin, wherein with the best results of activity extract.In carbohydrate tolerance test, behind the glucose of rats gavaged doses, blood glucose begins to rise, different dosing group rat blood sugar reaches the time to peak difference: normal group, diamicron administration group, the conventional water extract of YEJINCHAI, the time that peak value appears in the conventional ethanol extract group of YEJINCHAI group blood glucose is 30min, blood glucose begins to descend subsequently, and model group blood glucose continues to improve, when 60min, peak, just begin subsequently to descend, hints model group rat carbohydrate tolerance is poorer than normal group and administration group, each extract of YEJINCHAI can improve the carbohydrate tolerance of diabetes rat, improve carbohydrate metabolism, wherein the blood sugar increasing amplitude of activity extract group is less than conventional water, the ethanol extract group.Comprehensive above two result of the tests are found: Wild Jinchai liveness extract can not only strengthen the sensitivity of diabetes rat to exogenous insulin, can improve simultaneously carbohydrate tolerance, promote the blood glucose metabolism, suppress to take the photograph sugar back blood glucose and continue to raise, show that Wild Jinchai liveness extract has the good curing effect to the type 2 diabetes mellitus rat.
The preparation that the extract that the inventor further makes with embodiment 2-4 and all the other embodiment make carries out various pharmacodynamics tests and controlled trial, obtains and the similar effect of extract that makes with embodiment 1, here no longer repetition.Those skilled in the art should be appreciated that the extract that extracting method of the present invention obtains all can reach " extract contains 10～90 quality % total flavones; and the extract color is faint yellow to brown " effect, and infer that thus learning these extracts has better drug effect character with respect to conventional water extract that makes according to prior art or ethanol extract.These extracting method all are that the inventor passes through that a large amount of tests obtain and proposition first, there is no record in the prior art, and these extracting method are owing to the effect that can obtain to be close makes to have unicity between these extracting method.
Claims (14)
1, a kind of Wild Jinchai liveness extract, the combination that it is characterized in that adopting macroporous adsorbent resin method, column chromatography, membrance separation to concentrate in technology, spray drying technology, the Freeze Drying Technique any one or several different methods and low-temperature differential pressure type extraction method prepares, this extract contains 10～90 quality % total flavones, and the extract color is faint yellow to brown.
2, Wild Jinchai liveness extract as claimed in claim 1 is characterized in that this extract high-efficiency liquid-phase fingerprint 285nm detection has 26 characteristic peaks, and wherein 6 peaks are main peaks, account for more than 89% of total peak area; Extract high-efficiency liquid-phase fingerprint 345nm detects 26 characteristic peaks, and wherein 14 peaks are main peaks, account for more than 95% of total peak area.
3, Wild Jinchai liveness extract as claimed in claim 1, it is characterized in that the hydrochloric acid hydrolysis that this extract volume ratio is a concentration 2%～6%, 90 ℃ of hydrolysis temperatures, hydrolysis time 60 minutes, high-efficiency liquid-phase fingerprint has 4 characteristic peaks, when concentration of hydrochloric acid reaches 6% but tiny peak showed increased in the chromatogram when being lower than 12%, concentration of hydrochloric acid to increase the fragment that chemical compound is hydrolyzed into many more, when concentration reaches 12%, do not have chromatographic peak in the high-efficiency liquid-phase fingerprint, it is complete that chemical compound is hydrolyzed oxidation substantially.
4, the described Wild Jinchai liveness extract of claim 1, it is characterized in that YEJINCHAI be Jiangxi Fagaceae plant multiple-ear rock Ke (Lithocarpus polystachyus (wall) Rehd or L.litseifolius (Hamce) Chun.) whole plant or any position with and the processed product that makes through various concocting methods.
5, the described Wild Jinchai liveness extract of claim 1 is characterized in that described flavones ingredient comprises metallic salt derivant that forms with alkali and slaine and the metal ion complex that forms with metal ion.
6, the preparation method of the described Wild Jinchai liveness extract of claim 1 is characterized in that comprising following one or several step:
Extract: solvent for use is water or any one alcohols, ketone and esters solvent, or the mixed solvent formed by a certain percentage of these solvents, or the acidity or the basic solvent that are made into by these solvents and acid, alkali, salt; Extracting method is one or more in low-temperature differential pressure type extraction, supersound extraction, merceration, percolation, the microwave extraction;
Filter: comprise centrifugal, sucking filtration, ultrafiltration, microfiltration, use or do not use following any clarifier or its combination: precipitate with ethanol liquor, gelatin, Kaolin, silicon bath soil, various resin, Polyethylene Glycol, poly-second triol, chitosan and natural clarifying agent finished product;
Concentrate: comprise that thin film evaporation, rotary evaporation, decocting and concentrating, membrance separation under normal pressure or the reduced pressure concentrates that technology, suspension freeze concentration, progressive freeze concentrate, natural outer circulation two phase flow concentrates, outer circulation type freeze concentration, spray chilling concentrate;
Dry: as to comprise vacuum drying, spray drying, lyophilization.
7, the preparation method of the described Wild Jinchai liveness extract of claim 1, it is characterized in that when using the macroporous adsorbent resin method, used macroporous resin is nonpolar, low pole, middle polarity, polarity, any type in alkalescence or the faintly acid, as D101, X-5, NKA, S-8, NKA-9, HPD-600, ADS-7, AB-8, LSA-5B, HPD-722, ADS-17, DM130, HPD-826, DN-300, DN-603, FL-1, FL-2, FL-3 etc., low pole preferably wherein, middle polarity or polar resin, as AB-8, ADS-17, HPD-600 etc., used eluant are water, aquiferous ethanol, methanol, acetone etc.
8, the preparation method of the described Wild Jinchai liveness extract of claim 1, it is characterized in that when using column chromatography with polyamide during as immobile phase, used chromatography polyamide is 30-60 order, 60-120 order, 100-120 order, wherein be preferably 30-60 purpose resin, used eluant is water, aquiferous ethanol, methanol, acetone.
9, the preparation method of the described Wild Jinchai liveness extract of claim 1, it is characterized in that when using the low-temperature differential pressure type extraction method, unite simultaneously and use membrance separation to concentrate technology, spray drying technology prepares Wild Jinchai liveness extract, this method is: at-35 ℃～40 ℃ temperature conditions, by " normal pressure-vacuum-Jia solvent-normal pressure-pressurization-normal pressure " constantly circulation pressure change and extract chemical constituent, vacuum is selected in 13.33Pa～0.99MPa scope as required, pressure is selected in 0.99MPa～25.00MPa scope as required, and cycle-index is selected in 1～100 underrange as required; Solvent is chosen water, organic solvent or their mixture as required, and addition is pressed the w/v of extract and solvent as required and selected 1: 1～1: 500 scope.
10, the preparation method of the described Wild Jinchai liveness extract of claim 1, it is characterized in that when using membrance separation to concentrate technology, can adopt microfiltration to hold back the material of diameter more than 0.1 μ m, the solable matter of ultrafiltration molecular cut off between 1000～500000, the nanofiltration molecular cut off separates concentrated in the combination that reaches any two modes in the material of diameter about 1nm more than 150, the material of use is organic class cellulose acetate, polypropylene, Merlon, polysulfones, polyamide or mineral-type pottery, metal, metal-oxide, charcoal, in the cellular glass any.
11, the preparation method of claim 7 and 8 each described Wild Jinchai liveness extracts, it is characterized in that: select AB-8 for use, ADS-17, the low pole of HPD-600, middle polarity or polar macroporous adsorbent resin and 30～60 purpose chromatography polyamides are as the purification resin, YEJINCHAI extract sample solution is 1: 1～1: 20 in crude drug amount extension rate, absorption flow velocity 0.5～10BV/h, resin column blade diameter length ratio 1: 1～1: 15, in crude drug amount applied sample amount is 0.1～1g/ml, purified water remove impurity with 1～10 times of resin volume, the remove impurity flow velocity is 1～10BV/h, with 1～10 times of resin volume of 10%～95% ethanol elution, elution flow rate is 1～10BV/h.
12, the preparation method of the described Wild Jinchai liveness extract of claim 1, it is characterized in that getting the YEJINCHAI dry product, be ground into 100 order powder, adding 10 times of water gaging soak at room temperature extracted with low-temperature differential pressure type after 1.5 hours, 20 ℃ of temperature, pressure 10.00MPa, cycle-index 30 times, use Merlon membrance separation concentration technique to concentrate, can adopt microfiltration to hold back the material of diameter more than 0.1 μ m, the solable matter of ultrafiltration molecular cut off between 1000～500000, the nanofiltration molecular cut off separates concentrated in the combination that reaches any two modes in the material of diameter about 1nm more than 150, with gained concentrated extracting solution AB-8 middle polarity purification with macroreticular resin, in crude drug amount sample solution extension rate is 1: 10, absorption flow velocity 6BV/h, resin column blade diameter length ratio 1: 12, in crude drug amount applied sample amount is 0.5g/ml, purified water remove impurity with 6 times of resin volumes, the remove impurity flow velocity is 8BV/h, with 5 times of resin volumes of 10%～95% ethanol elution, elution flow rate is 6BV/h, extracting solution is through vacuum drying, and gained extractum is the YEJINCHAI extract.
13, the application of the described Wild Jinchai liveness extract of claim 1, it is characterized in that this extract can be separately or with other any Chinese and western drugs or food by the arbitrary proportion compatibility, be used to prepare medicine or functional food.
14, the described application of claim 13 is characterized in that prepared medicine or functional food are capsule, tablet, pill, granule, oral liquid, syrup, electuary, medicated wine, injection, unguentum, powder, beverage, cataplasma.
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| CN101904882A (en) * | 2010-07-28 | 2010-12-08 | 广西壮族自治区中医药研究院 | Preparation method of lithocarpus litseifolius total flavone |
| CN102441291A (en) * | 2011-12-01 | 2012-05-09 | 吉林省包装工程研究中心 | New method for extracting or purifying effective component of traditional Chinese medicine |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101904882A (en) * | 2010-07-28 | 2010-12-08 | 广西壮族自治区中医药研究院 | Preparation method of lithocarpus litseifolius total flavone |
| CN102441291A (en) * | 2011-12-01 | 2012-05-09 | 吉林省包装工程研究中心 | New method for extracting or purifying effective component of traditional Chinese medicine |
| CN107581306A (en) * | 2017-08-22 | 2018-01-16 | 深圳市瑞世兴科技有限公司 | It is a kind of that there is hypoglycemic, anti-trioxypurine leaf beet health protection tea and preparation method thereof |
| CN107467307A (en) * | 2017-09-22 | 2017-12-15 | 王红波 | A kind of method for making Jin Chaigong tea using wild golden bavin |
| CN108771112A (en) * | 2018-05-17 | 2018-11-09 | 哈尔滨医科大学 | Multiple-ear rock Ke's compound rice flour and preparation method thereof |
| CN109134557A (en) * | 2018-07-16 | 2019-01-04 | 华北理工大学 | The method of phloridzin is extracted from manyspike tanoak leaf piece |
| CN109134557B (en) * | 2018-07-16 | 2020-07-28 | 华北理工大学 | Method for extracting phlorizin from lithocarpus polystachyus rehd leaves |
| CN109432200A (en) * | 2018-12-13 | 2019-03-08 | 吉林化工学院 | A kind of preparation method of rose hip flavones calcium ion complexes |
| CN109432200B (en) * | 2018-12-13 | 2021-04-13 | 吉林化工学院 | Preparation method of rose hip flavone calcium ion complex |
| CN113633674A (en) * | 2021-09-08 | 2021-11-12 | 吉林大学 | Lithocarpus polystachyus (rehd.) Rehd oral liquid for livestock and preparation method thereof |
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