CN1742803A - Chinese medicine extract with function of reducing blood-sugar and preparing method - Google Patents
Chinese medicine extract with function of reducing blood-sugar and preparing method Download PDFInfo
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Abstract
The present invention relates to a Chinese medicine extract with action of reducing blood sugar. It is composed of polysaccharides, flavones and alkaloids obtained by separating and extracting mulberry leaf, mulberry twig and mulberry root bark. Their contents respectively are 24-27%, 53-61% and 15-20%. Said invention also provides the concrete steps of the above-mentioned separation and extraction process.
Description
Technical field
The invention belongs to the field of Chinese medicines, disclose a kind of from Chinese medicine the method for effective component extracting, more particularly, the present invention relates to extract the component with treatment diabetes and complication thereof from moraceae plants Folium Mori, Ramulus Mori, Cortex Mori, these components can be applicable to the treatment of diabetes and complication thereof.
Background technology
Folium Mori, Ramulus Mori and Cortex Mori are the leaf of mulberry, and the skin of twig and tree root is the different parts of same plant, one of Chinese medicine commonly used.Folium Mori are the leaves of (Morus alba L.) on the Moraceae Morus plant.The medicinal head of Folium Mori is stated from Shennong's Herbal, Folium Mori bitter but sweet flavor, cold, thus sweet beneficial blood, so cold removing heat from blood.Sweet cold being harmonious, thus the therapeutic method to keep the adverse QI flowing downwards and tonifying YIN, again can cough-relieving, the merit of tonification is arranged.Ramulus Mori is the dry twig of Moraceae Morus plant Mulberry, and bitter in the mouth is flat, goes into Liver Channel.The function curing rheumatism, sharp joint, diuretic.Cortex Mori is the root bark of removing cork of Moraceae Morus plant Mulberry.Nature and flavor are sweet, and are cold.Go into lung, spleen channel.The function eliminating pathogen from the lung for relieving asthma, inducing diuresis to remove edema.Treatment lung-heat cough with asthma is spitted blood edema, beriberi, dysuria.All there is Mulberry can treat the record of quenching one's thirst in the successive dynasties Chinese medicine book.Carry as " Bencao Jingshu ", the Folium Mori bitter in the mouth, sweet, cold, so sweet beneficial blood, so cold removing heat from blood, sweet cold being harmonious is so the therapeutic method to keep the adverse QI flowing downwards and tonifying YIN can make eye bright again and quench the thirst; Compendium of Material Medica is carried, and Folium Mori are the medicine of brothers Yang Ming, fries in shallow oil juice for tea, can only quench one's thirst.
The main component of moraceae plants has flavone and flavonoid glycoside, alkaloids, steroid constituents, volatile oil component, polysaccharide, amino acids, vitamins and trace element etc.The lot of domestic and international result of study shows Folium Mori in recent years, and Ramulus Mori and Cortex Mori have good function of blood sugar reduction, the mechanism research of its hypoglycemic activity think with its effective ingredient in alkaloid have closely related.
About extraction separation alkaloid in Folium Mori, Ramulus Mori and the Cortex Mori with and the patent of glucoside inhibiting activity more, as Chinese patent publication number CN1338275A, CN13094A, CN14394623A, CN1466961A, CN1471934A, CN1559539A etc., only relate to and utilize organic solvent or cation exchange resin layer to analyse method to separate total alkaloids in Folium Mori, the Cortex Mori, and do not relate to the method that macroporous resin and ion exchange resin continuous chromatography separate different components.And for example relate to the separation of polysaccharides of Folium Mori among the Chinese patent publication number CN1351085A, extraction separation method adopts organic solvent and ion exchange resin, does not all relate to macroporous resin and ion exchange resin continuous separation method.
The present invention adopts the column chromatography continuous separation method first, with Folium Mori, the water of Ramulus Mori and Cortex Mori or aqueous ethanol extract continuous adsorption are in macroporous resin and ion exchange resin, solvent such as water with opposed polarity, the pure and mild ammonia eluting of variable concentrations, separate simultaneously and obtain flavonoid, alkaloids and polysaccharide component, its yield and content all reach expected result, and the column chromatography continuous separation method that adopts among the present invention is compared with traditional organic solvent continuous extraction partition method, has safety non-toxic, advantage such as pollution-free, feasible and reliable, can promote large-scale production.
Summary of the invention
An object of the present invention is to disclose a kind of Chinese medicine extract with hypoglycemic activity, it is formed by separating the polysaccharide, flavonoid and the alkaloids component that obtain in moraceae plants Folium Mori, Ramulus Mori, the Cortex Mori.
Another object of the present invention is the method for the main component of this Chinese medicine extract of preparation.
Another object of the present invention is to point out the purposes of this this Chinese medicine extract.
In a first aspect of the present invention, the invention provides a kind of Chinese medicine extract with hypoglycemic activity, by moraceae plants Folium Mori, Ramulus Mori, Cortex Mori is arbitrary or its combination in separate polysaccharide, flavonoid and the alkaloids component obtain and form, it is characterized in that, separate in the component that obtains that each component is according to mass percent, polysaccharide component content is 24-27%; Flavonoid component content is 53-61%; Alkaloids component content is 15-20%.
The present invention separates the impurity that minute quantity can be arranged in the component that obtains.
The Quercetin mass percent is 30～50% in the flavonoid component of the present invention.
Polyhydroxylated alkaloid 1-deoxynojirimycin (DNJ) mass percent is 30～45% in the alkaloids component of the present invention.
In a second aspect of the present invention, the invention provides the preparation method of the main component of above-mentioned Chinese medicine extract:
(1) with moraceae plants Folium Mori, Ramulus Mori, after Cortex Mori is arbitrary or its combination pulverizes, adds the water of 5-10 times of volume, alcohol or aqueous alcohol solvent, at 25-100 ℃, extract 2-3 time, be no less than 20 minutes at every turn, get extracting solution, the extracting liquid filtering slag is got filtrate 1, remove precipitation again, get filtrate 2, filtrate 2 is crossed macroporous resin, use the deionized water eluting, eluent is concentrated the back precipitate with ethanol, precipitate gets component 1 through the decolouring after drying, that is, and and the polysaccharide component;
(2) the pure eluting macroporous resin of reuse variable concentrations, each several part collect eluent colourless till, respectively behind the concentrate drying component 2, that is, and the flavonoid component;
(3) with macroporous resin penetrating component liquid after ion exchange resin, washing with 0.2-1N ammonia eluting, is collected eluent, concentrate drying and component 3, that is, and the alkaloids component.
Further, preparation method step of the present invention (1) is described can be methanol, ethanol, isopropyl alcohol as the alcohol that extracts solvent.
Further, the described removal precipitation process of preparation method step of the present invention (1) has adopted flocculation process, that is, filtrate 1 is concentrated to about 1/10 of original volume adds chitin or clarifier, and after 5-6 hour, centrifugal going precipitated 70-80 ℃ of insulation.
Further, the described alcohol precipitation process of preparation method step of the present invention (1) is that eluent is concentrated to and adds methanol behind 1/5～1/2 small size or ethanol to total concentration reaches according to volume ratio 70-80%, and 4 ℃ of placements are spent the night, and is centrifugal and obtain precipitate.More the volume ratio of You Huaing is 70%.
Further, the described depigmenting agent of preparation method step of the present invention (1) is ethanol and acetone.
Further, the described macroporous resin of preparation method of the present invention can be selected from the AB-8 type, D101 type, Amberlite 252 or Diaion HP-20.
Further, the described ion exchange resin of preparation method of the present invention can adopt styrene resin, is selected from 001 * 1.1 type, 001 * 4 type, 001 * 7 type, Amberlite IR 120, Dowex 50 or Diaion SK-IA.
Further, the described macroporous resin eluent of preparation method of the present invention is a methanol or alcoholic acid a kind of, and concentration is 30-95%.
Further, the described ammonia eluent of preparation method of the present invention concentration is 0.5N.
In preparation method step of the present invention (1), the condition of extracting method is in order to extract the ingredient in moraceae plants Folium Mori, Ramulus Mori, the Cortex Mori comparatively fully, in this area the condition of extracting method is done suitably rather than material change's method also should be a content of the present invention, for example extract each 15 minutes 5 times.
In preparation method step of the present invention (1), concentrate and exsiccant method can realize according to the method for the many routines in this area, evaporation and concentration for example, vacuum drying, atomization drying, or the like method.
Extraction component in the preparation method of the present invention is identified and control of quality by the following method:
(1) polysaccharide:
Test tube method (naphthols-sulphate reagent method (Molish reaction)): respectively get component 1, component 2, component 3 a little, test respectively, that is, it is centrifugal to add 80% ethanol jolting dissolving back, and the precipitation heating is dissolved in the ethanol, add isopyknic 10% alpha-Naphthol-ethanol liquid again, shake up, drip concentrated sulphuric acid along test tube wall, in the test of component 1, purplish red colour circle appears in two liquid at the interface, shows that component 1 contains polysaccharide.
Thin layer chromatography: get each component 0.1g, test respectively, promptly, add in the 2mL methanol, firmly get supernatant 10 μ L points on silica gel g thin-layer plate after the jolting, with methylene chloride-methanol=expansion in 8: 2, in the test of component 1, observing to show under uviol lamp has the purple fluorescence speckle, shows that component 1 contains polysaccharide.
Determination of polysaccharide: precision takes by weighing component 1, and 0.1g adds 80% ethanol 10mL, and is after 10 minutes after-filtration of supersound process, insoluble matter divide 4 washings with 80% ethanol 20mL, with hot water dissolving's precipitation, to 100mL, standby.Precision takes by weighing above-mentioned solution 0.5mL, places in the 10mL tool plug test tube, adds water to 1mL.Add 1mL 5% phenol solution, shake up, slowly add concentrated sulphuric acid 5mL, shake up, put in the boiling water bath cooling 5 minutes along tube wall.Adding reagent with 1mL water is blank, measures absorbance at 490nm wavelength place.Calculate polyoses content by calibration trace.
(2) flavonoid:
Test tube method (hydrochloric acid magnesium powder reduction chromogenic reaction): get each component and be made into 10mgmL respectively
-1Aqueous solution, test promptly, adds a small amount of magnesium powder and 1mL concentrated hydrochloric acid respectively, observes the foam color.In the test of component 2, sample solution produces pink foam rapidly after adding magnesium powder and concentrated hydrochloric acid, show that component 2 contains Flavonoid substances.
Thin layer chromatography: get each component a little, test respectively, that is, add 60% dissolve with ethanol, get 10 μ L points on polyamide film, to launch in 90% ethanol, with the colour developing of aluminum chloride ethanol liquid, observe the fluorescence speckle down in uv analyzer 365nm, in the test of component 2, the fluorescence speckle shows as purple dot, shows that component 2 contains Flavonoid substances.
The assay of flavone: with 60% ethanol configuration 0.098mgmL
-1The Quercetin standard solution, get 0.0lmL respectively, 1.0mL, 2.0mL, 3.0mL, 4.0mL, 5.0mL adds 5% sodium nitrite solution 0.3mL and shakes up the back and place 6min; Add 10% sodium nitrite solution 0.3ml and shake up back placement 6 minutes; Add 1mol sodium hydroxide solution 4mL and add 60% ethanol dilution, shake up, place 15min to 10mL.Measure absorbance in 510nm, ask the basis of calculation curve, measure the content of the Quercetin in the extract then respectively.Get flavone component 1g and add 10ml 60% ethanol, supersound extraction is 30 minutes then, gets supernatant 1ml and places the 10ml measuring bottle respectively, and working sample quercetin content is as stated above measured the content of the Quercetin in the extract then respectively.
(3) alkaloidal qualitative identification:
Test tube method (the silico-tungstic acid sedimentation method): get each component and be mixed with 1mg/ml solution with water respectively, test respectively, promptly, drip concentrated hydrochloric acid number droplet, dropwise drip the silico-tungstic acid test solution again, if any alkaloid then the Lycoperdon polymorphum Vitt insoluble matter occur, with the adding of silico-tungstic acid test solution, insoluble matter increases.Test shows that component 3 contains alkaloids substance.
Thin layer chromatography: get each component point sample on lamellae respectively, developing solvent is an isopropyl alcohol: acetic acid: water=4: 2: 1, connection aminotoluene-chlorine development process develops the color.It is alkaloid that the egg yellow spotting is arranged.Test shows that component 3 contains alkaloids substance.
Alkaloid detects: the HPLC method is measured content: with the 1-deoxynojirimycin is standard reference material, is the inserts agent with amino bonded phase silica gel, and acetonitrile-water is a mobile phase, detects on the parallax refraction detector.Precision is drawn certain density reference substance solution and each 10 μ L of component 3 sample solutions respectively, injects chromatograph of liquid, measures in accordance with the law, presses external standard method with calculated by peak area, thereby measures the content of the 1-deoxynojirimycin in the component 3.
In a third aspect of the present invention, the present invention has pointed out the application of this Chinese medicine extract aspect the preparation alpha-glucosidase inhibitor, and the application in the medicine of preparation treatment diabetes and complication thereof.
Preparation method beneficial effect of the present invention: the present invention adopts the column chromatography continuous separation method first, with Folium Mori, the water of Ramulus Mori and Cortex Mori or aqueous ethanol extract continuous adsorption are in macroporous resin and ion exchange resin, solvent such as water with opposed polarity, the pure and mild ammonia eluting of variable concentrations, separate continuously respectively and obtain flavonoid, alkaloids and polysaccharide component, its yield and content all reach expected result, and the column chromatography continuous separation method that adopts among the present invention is compared with traditional organic solvent continuous extraction partition method, has safety non-toxic, advantage such as pollution-free, feasible and reliable, can promote large-scale production.
Description of drawings
Fig. 1 has shown the extraction and separation process flow process of blood-sugar decreasing active.
Fig. 2 has shown and has respectively extracted influence and the glycemic peaks relative inhibition of component to the normal mouse carbohydrate tolerance.
Fig. 3 has shown and has respectively extracted influence and the glycemic peaks relative inhibition of component to the inductive diabetic mice carbohydrate tolerance of STZ.
The specific embodiment
Below with the present invention is further illustrated.
One, the extraction of effective ingredient in Folium Mori, Ramulus Mori or the Cortex Mori
Embodiment 1
Folium Mori 1kg, the deionized water of 5-10 times of volume of adding, 100 ℃ are extracted 3 times, be no less than 20 minutes, merge extractive liquid, gets filtrate 1 with the extracting liquid filtering slag at every turn, remove precipitation again, described removal precipitation process has adopted flocculation process, promptly, when the filtrate 1 of extracting is concentrated to 1/10 original volume, add chitin or clarifier, 70-80 ℃ of insulation after 5-6 hour, the centrifugal precipitation of going, get filtrate 2, filtrate 2 is crossed AB-8 macroporous resin, flow velocity 5mlmin
-1With deionized water be eluted to do not have precipitation when adding 95% ethanol in the eluent till, eluent is concentrated to adds 95% ethanol behind 1/4 volume, make the ethanol final concentration reach 70%, the static after-filtration that spends the night, precipitation with ethanol and acetone be eluted to repeatedly eluent colourless after, cold drying and must component 1,9g comprises that with the qualitative identification experiment test tube method and thin layer chromatography prove that this component is the polysaccharide component; Then then with 30%, 50%, 70%, 90% ethanol elution macroporous resin, each several part collect eluent colourless till, concentrate drying gets component 2 respectively, 11g, 5.5g, 0.9g and 0.3g, comprise that with the qualitative identification experiment test tube method and thin layer chromatography prove that this component is the flavonoid component, and show that with the assay that colorimetry is carried out the flavonoid component Quercetin quality accounts for 30%～50% of flavonoid component quality.In addition, after Amberlite IR 120 cation exchange resiies, 0.2molL is used in washing with macroporous resin penetrating component liquid
-1The ammonia eluting, collect eluent, cryoconcentration, vacuum drying gets component 3,6.7g, comprise that with the qualitative identification experiment test tube method and this component of thin layer chromatography are the alkaloids component, and show that with the assay that the HPLC method is carried out the alkaloids component 1-deoxynojirimycin quality accounts for 35%～45% of alkaloids component quality.
The leaching process raw material and the technical parameter of the embodiment of the invention 1～3 see Table 1.
The mass percent of the each component of the embodiment of the invention 1～3 Chinese medicine extract that extracts sees Table 2.
Each 500g of Folium Mori and Ramulus Mori common 1kg that mixes adds 7 times of volume 50% methanol, 25 ℃.Totally 3 times, merging filtrate concentrates, centrifugal, removal precipitation, and supernatant is crossed the AB-8 macroporous resin, flow velocity 5mlmin
-1With deionized water be eluted to do not have precipitation when adding 95% ethanol in the eluent till, eluent is concentrated to adds 95% ethanol behind the certain volume, make the ethanol final concentration reach 75%, the static after-filtration that spends the night, precipitation with ethanol and acetone be eluted to repeatedly eluent colourless after, cold drying and must component 1,8.4g, comprise that with the qualitative identification experiment test tube method and thin layer chromatography prove that this component is the polysaccharide component; Then then with 30%, 50%, 70%, 90% ethanol elution macroporous resin, each several part collect eluent colourless till, concentrate drying gets component 2 respectively, 12g, 6g, 0.92g and 0.3g, comprise that with the qualitative identification experiment test tube method and thin layer chromatography prove that this component is the flavonoid component, and show that with the assay that colorimetry is carried out the flavonoid component Quercetin quality accounts for 32%～41% of flavonoid component quality.In addition, after 732 ion exchange resin, 0.5molL is used in washing with macroporous resin penetrating component liquid
-1The ammonia eluting, collect eluent, cryoconcentration, vacuum drying gets component 3,6.3g, comprise that with the qualitative identification experiment test tube method and this component of thin layer chromatography are the alkaloids component, and show that with the assay that the HPLC method is carried out the alkaloids component 1-deoxynojirimycin quality accounts for 31%～40% of alkaloids component quality.
Each 500g of Ramulus Mori and Cortex Mori is 1kg altogether, and 10 times of volumes remove 30% ethanol, 70 ℃.Totally 3 times, merging filtrate concentrates, centrifugal, removal precipitation, and supernatant is crossed the D101 macroporous resin, flow velocity 5mlmin
-1With deionized water be eluted to do not have precipitation when adding 95% ethanol in the eluent till, eluent is concentrated to adds 95% ethanol behind the certain volume, make the ethanol final concentration reach 80%, the static after-filtration that spends the night, precipitation with ethanol and acetone be eluted to repeatedly eluent colourless after, cold drying and must component 1,9.4g, comprise that with the qualitative identification experiment test tube method and thin layer chromatography prove that this component is the polysaccharide component; Then then with 30%, 50%, 70%, 90% ethanol elution macroporous resin, each several part collect eluent colourless till, concentrate drying gets component 2 respectively, 15g, 8g, 0.9g and 0.2g, comprise that with the qualitative identification experiment test tube method and thin layer chromatography prove that this component is the flavonoid component, and show that with the assay that colorimetry is carried out the flavonoid component Quercetin quality accounts for 31%～45% of flavonoid component quality.In addition, after 732 ion exchange resin, 1molL is used in washing with macroporous resin penetrating component liquid
-1The ammonia eluting, collect eluent, cryoconcentration, vacuum drying gets component 3,6g comprises that with the qualitative identification experiment test tube method and this component of thin layer chromatography are the alkaloids component, and shows that with the assay that the HPLC method is carried out the alkaloids component 1-deoxynojirimycin quality accounts for 30%～43% of alkaloids component quality.
Table 1 leaching process raw material of the present invention and technical parameter
| Embodiment 1 | | | |||
| Material | Technological parameter | Material | Technological parameter | Material | Technological parameter |
| Folium Mori | Folium Mori and Ramulus Mori | Ramulus Mori and Cortex Mori | |||
| The extraction solvent is | 5 times of volumes | The extraction solvent is | 7 times of volumes | The extraction solvent is | 10 times of volumes |
| Water | 50% | 30% | ||||
| Extract | ||||||
| 100 ℃ of temperature | Extract 25 ℃ of temperature | Extract 70 ℃ of temperature | ||||
| Determining alcohol 70% in the alcohol precipitation process | Determining alcohol 75% in the alcohol precipitation | Determining alcohol | 80% in the alcohol precipitation process | |||
| The AB-8 macroporous resin | The AB-8 macroporous resin | D101 type macroporous | ||||
| Amberlite IR | ||||||
| 120 cation exchange resiies | 0.2mol·L -1The ammonia eluting | 732 ion exchange resin | 0.5mol·L -1The ammonia eluting | 732 ion exchange resin | 1mol·L -1The ammonia eluting |
The mass percent of each component in table 23 kinds of Chinese medicine extraction groups that leaching process obtained of the present invention
| Percentage composition | |||
| Polysaccharide | Flavonoid | Alkaloids | |
| Embodiment 1 | 27％ | 53％ | 20％ |
| | 25％ | 56％ | 19％ |
| | 24％ | 61％ | 15％ |
Two, external glycosidase suppresses experiment
Embodiment 4
(1) extraction of the big intestinal mucosa glycosidase of rat
Frozen rat small intestine (jejunum) is thawed, take mucosa with the pincet extruding.What add 5 times of amounts in this mucosa contains 5 * 10
-3MolL
-1The 0.1molL of ethylenediaminetetraacetic acid
-1Kaliumphosphate buffer (pH value 7.0) is while cool off homogenate.Centrifugalize then (4 ℃, 21000rmin
-1, 60min), in the gained precipitate, add the 0.1molL that contains 1%TritonX-100
-1Kaliumphosphate buffer (pH value 7.0) carries out dissolving processing (4 ℃ 60min), are carried out ultracentrifugation (110000rmin with it to 5 times of amounts
-1, 90min), with its supernatant 0.1molL
-1Kaliumphosphate buffer (pH value 7.0) dialyse (4 ℃, 24h), make enzyme liquid, protein content is pressed the Lowry method and is measured, resulting zyme extract is kept at-70 ℃.
(2) detect
This detection is carried out on 96 orifice plates, establishes 4 every group multiple holes of 3 groups, is respectively: a. matched group (buffer+substrate+enzyme liquid); B. blank group (buffer); C. sample determination group (sample+substrate+enzyme liquid).Operational approach is: add 80 μ L sample solutions in advance in the plate hole and (use 0.1molL
-1The pH7.0 phosphate buffer is diluted to 6 Concentraton gradient (20-1000 μ g.mL respectively
-1)), add 40 μ L 20 * 10 then
-3MolL
-1Substrate (PNPG), 37 ℃ of insulation 5min add enzymatic solution 40 μ L, and 37 ℃ of insulation 15min add 160 μ L 0.2molL
-1Na
2CO
3Stop enzyme reaction.The 405nm place measures absorbance on microplate reader at last, calculates suppression ratio by following method, suppression ratio (%)=[(a-b)-(c-b)/(a-b)] * 100%, and calculate corresponding IC
50The results are shown in Table 3.
Table 3 Chinese medicine extraction component is to glucoside inhibiting activity
| Component | Suppression ratio/% | IC 50/μg·mL -1 |
| Alpha-glucosidase rat small intestine glycosidase | Alpha-glucosidase rat small intestine glycosidase | |
| Polysaccharide flavonoid alkaloids | 63 65 79 80 95 90 | 80.1 91.3 50.4 67.4 12.7 10.6 |
In the table 3, external glucoside inhibiting activity testing result shows that each component concentrations is 20 ~ 1000 μ gmL
-1The time glycosidase activity suppression ratio be 32% ~ 95%, what suppression ratio was the highest is alkaloids, is flavonoid then, and polysaccharide.
Three, normal mouse carbohydrate tolerance test
ICR mice adaptability was raised 2-3 days, 20 hours on an empty stomach.Mice is divided into model group, acarbose group, Chinese medicine sample sets (comprising alkaloid group, flavone group and polysaccharide group) at random.Model group is irritated stomach 0.5mL soluble starch and (is pressed 2.5gkg with normal saline
-1Body weight dosage is prepared) and the 0.5mL normal saline; The BAITANGPING group is irritated stomach 0.5mL soluble starch and 0.5mL BAITANGPING solution (is pressed 10mgkg with normal saline
-1Body weight dosage faces uses preceding preparation).Chinese drug-treated group is irritated stomach 0.5mL soluble starch and the 0.5mL sample solution (is pressed 150mgkg with normal saline
-1Body weight dosage faces uses preceding preparation).Each treated animal is given 0min behind the starch, 30min, and 60min, 90min, 120min eye socket venous plexus is got blood, measures the blood glucose value of each time point and calculates the glycemic peaks relative inhibition.With the variation of the total serum glucose of glucose total in serum increment (TIG) expression (with 0 constantly as baseline correction), glycemic peaks relative inhibition (%)=(matched group TIG-administration group TIG)/matched group TIG * 100%.
Each extracts component the influence and the glycemic peaks relative inhibition of normal mouse carbohydrate tolerance is seen Fig. 2, the normal mouse carbohydrate tolerance test shows that each component all has remarkable inhibition normal mouse to irritate to feed the rising (P＜0.01 or P＜0.05) of each time point blood glucose in the 2h behind the starch, suppresses effect the most obvious (P＜0.05) when wherein giving starch 30min.The glycemic peaks relative inhibition is the highest with glucobay (acarbose), secondly alkaloid, flavone and polysaccharide.
Four, the inductive mice carbohydrate tolerance test of STZ
Embodiment 6
ICR mice adaptability was raised 2-3 days, 20 hours on an empty stomach.Lumbar injection STZ 200mgkg
-1(under 4 ℃ of conditions face with the citrate buffer solution of pH value 4.5 use preceding configuration), after 3 days, measuring blood sugar of blood extracting＞11mmolL
-1The selected experiment of person.Injection STZ one Zhou Houke experimentizes, and grouping is the same with experimental technique.
The results are shown in Figure 3, show in the inductive diabetic mice carbohydrate tolerance test of STZ, diabetic mice is given during the blood glucose value of each time point changes in the 2h behind the starch obvious inhibition effect (P＜0.01 or P＜0.05) being arranged wherein for the blood sugar increasing behind the starch 30min, all the other time periods do not have obviously influence (P＜0.01) except acarbose, the glycemic peaks relative inhibition is the highest with glucobay (acarbose), next alkaloid, flavone and polysaccharide.
Claims (15)
1. Chinese medicine extract with hypoglycemic activity, by moraceae plants Folium Mori, Ramulus Mori, Cortex Mori is arbitrary or its combination in separate polysaccharide, flavonoid and the alkaloids component obtain and form, it is characterized in that, separate in the component that obtains that each component is according to mass percent, polysaccharide component content is 24-27%; Flavonoid component content is 53-61%; Alkaloids component content is 15-20%.
2. Chinese medicine extract according to claim 1 is characterized in that the Quercetin mass percent is 30～50% in the described flavonoid component.
3. Chinese medicine extract according to claim 1 is characterized in that polyhydroxylated alkaloid 1-deoxynojirimycin (DNJ) mass percent is 30～45% in the described alkaloids component.
4. the preparation method of the main component of claims 1 described Chinese medicine extract is characterized in that, may further comprise the steps:
(1) with moraceae plants Folium Mori, Ramulus Mori, after Cortex Mori is arbitrary or its combination pulverizes, adds the water of 5-10 times of volume, alcohol or aqueous alcohol solvent, extract 2-3 time at 25-100 ℃, be no less than 20 minutes at every turn, get extracting solution, the extracting liquid filtering slag is got filtrate 1, remove precipitation again, get filtrate 2, filtrate 2 is crossed macroporous resin, use the deionized water eluting, eluent is concentrated the back precipitate with ethanol, precipitate gets component 1 through the decolouring after drying, that is polysaccharide component;
(2) the pure eluting macroporous resin of reuse variable concentrations, each several part collect eluent colourless till, respectively behind the concentrate drying component 2, that is, and the flavonoid component;
(3) with macroporous resin penetrating component liquid after ion exchange resin, washing with 0.2-1N ammonia eluting, is collected eluent, concentrate drying and component 3, that is, and the alkaloids component.
5. according to the preparation method of the main component of claims 4 described Chinese medicine extract, it is characterized in that step (1) is described can be methanol, ethanol, isopropyl alcohol as the alcohol that extracts solvent.
6. according to the preparation method of the main component of claims 4 described Chinese medicine extract, it is characterized in that, the described removal precipitation process of step (1) has adopted flocculation process, promptly, filtrate 1 is concentrated to 1/10 o'clock of original volume adds chitin or clarifier, after 5-6 hour, centrifugal going precipitated 70-80 ℃ of insulation.
7. according to the preparation method of the main component of claims 4 described Chinese medicine extract, it is characterized in that, described alcohol precipitation process is that eluent is concentrated to and adds methanol behind 1/5～1/2 small size or ethanol to total concentration reaches according to volume ratio 70-80%, 4 ℃ of placements are spent the night, and are centrifugal and obtain precipitate.
8. according to the preparation method of the main component of claims 7 described Chinese medicine extract, it is characterized in that adding methanol or ethanol to total concentration in the described alcohol precipitation process, to reach according to volume ratio be 70%.
9. according to the preparation method of the main component of claims 4 described Chinese medicine extract, it is characterized in that the described depigmenting agent of step (1) is ethanol and acetone.
10. according to the preparation method of the main component of claims 4 described Chinese medicine extract, it is characterized in that described macroporous resin can be selected from the AB-8 type, D101 type, Amberlite 252 or Diaion HP-20.
11. the preparation method according to the main component of claims 4 described Chinese medicine extract is characterized in that described ion exchange resin can adopt styrene resin, be selected from 001 * 1.1 type, 001 * 4 type, 001 * 7 type, Amberlite IR 120, Dowex 50 or Diaion SK-IA.
12. the preparation method according to the main component of claims 4 described Chinese medicine extract is characterized in that, the described macroporous resin eluent of step (2) is a methanol or alcoholic acid a kind of, and concentration is 30-95%.
13. the preparation method according to the main component of claims 5 described Chinese medicine extract is characterized in that described ammonia concn is 0.5N.
14. according to the arbitrary described Chinese medicine extract of claim 1 to 3 in the application of preparation aspect the alpha-glucosidase inhibitor.
15. according to the application of the arbitrary described Chinese medicine extract of claim 1 to 3 in the medicine of preparation treatment diabetes and complication thereof.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100287093A CN100361674C (en) | 2005-08-11 | 2005-08-11 | Chinese medicine extract with function of reducing blood-sugar and preparing method |
Applications Claiming Priority (1)
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