CN105250342A - Ligularia virgaurea extract and preparation method and application thereof in aspect of reducing blood glucose - Google Patents
Ligularia virgaurea extract and preparation method and application thereof in aspect of reducing blood glucose Download PDFInfo
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- CN105250342A CN105250342A CN201510861096.5A CN201510861096A CN105250342A CN 105250342 A CN105250342 A CN 105250342A CN 201510861096 A CN201510861096 A CN 201510861096A CN 105250342 A CN105250342 A CN 105250342A
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Abstract
The invention provides an extraction and preparation method of ligularia virgaurea components. The invention further provides application of prepared ligularia virgaurea extract in reducing the activity of glucosidase and treating diabetes. The prepared ligularia virgaurea extract serves as a plant source drug and can effectively reduce the activity of the glucosidase, and side effects are small; not only is a novel treatment means provided for diabetes treatment, but also the grassland poisonous weed resource of ligularia virgaurea is scientifically, reasonably and sustainably developed and utilized, a harmful thing is turned into treasure, and meanwhile important significance to protection of the ecological environment of the Tibet plateau is achieved. According to the ligularia virgaurea extract and the preparation method and application in the aspect of reducing the blood glucose, the preparation technology is simple, rapid and environmentally friendly, and due to the fact that the plant resources are rich, low in development cost and capable of being continuously developed, the economic benefit is obvious.
Description
Technical field
The invention belongs to biological pharmacy technical field, be specifically related to a kind of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract and preparation method thereof and the application in treating diabetes.
Background technology
Diabetes be a kind of produce to insulin and act on abnormal relevant, take hyperglycemia as the metabolic disease of principal character, there is no radical cure way.Recent research finds, diabetes disease is sent out process and often first occurred postprandial hyperglycemia, and then develop into diabetes, namely the former is the sign in advance of the latter.For diabetics, particularly II type patient, postprandial hyperglycemia to the harm of body considerably beyond hyperglycemia, a large amount of excreting insulin of appearance stimulating pancreas of postprandial hyperglycemia, multiple Developmental and Metabolic Disorder and vascular lesion can be caused, make pancreatic function suffer damage further simultaneously.Therefore, reduce post-prandial glycemia for treatment type Ⅱdiabetes mellitus and this disease of prevention, reduce one of complication and the important measures reducing mortality rate.
Alpha-glucosidase inhibitor is the antidiabetic medicine that a class of the appearance 1980s is new, it by the glycosidase on competitive inhibition intestinal villi film be used for reduce the degraded of saccharide, delay the absorption and digestion of saccharide, thus effectively reduce the peak value of after-dinner blood sugar of diabetes patients concentration.This inhibitor has been the first-line drug falling post-prandial glycemia by third time Asian-Pacific area Remedies for diabetes Guidelines recommend, is classified as the 3rd class oral antidiabetic drug by medical circle.But this product such as acarbose, the voglibose etc. that have gone on the market at present mostly are sintetics, though good drug efficacy is comparatively large to human body toxic and side effects, as gastrointestinal reaction, anaphylaxis etc.Taking Chinese herbal medicine as raw material, find and take safety for treating and preventing diabetes significant, is also a focus of research alpha-glucosidase inhibitor both at home and abroad at present.
Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski (Ligulariavirgaurea) is common in the herbaceos perennial on Qinghai-Tibet Alpine meadow.The effects such as this plant is Compositae Ligularia, greatly poison, and nature and flavor are sweet, bitter, is used as medicine and has place heat, removing toxic substances more skin ulcer clearly, emetic.Ligularia about has 112 kinds in China, and the overwhelming majority is distributed in Qinghai-xizang Plateau Region, and wherein multiple have medical value, but less to the blood sugar reducing function research of this platymiscium, and the correlational study had no about Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski and report.In recent years, along with the continuous degeneration on Qinghai-Tibet grassland, structure of community changes, and malicious weeds are increased, and Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski is exactly one of them, and resource is very abundant.This patent organizes the further investigation of blood sugar reducing function for Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski, changes harmful to treasure, and technique is simple, and production cost is low, is easy to industrialization, and for the comprehensive exploitation of Qinghai-Tibet ecological environment and Sustainable use significant.
Summary of the invention
The invention provides a kind of preparation method of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract, this extract has drawn from Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski tissue (blade or root or flower).Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski is widely distributed in Qinghai-Tibet Platean, aboundresources.The technology of the present invention raw material is sufficient, and technique is simple, can realize industrialization.
The open Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract of simultaneously the present invention, in treatment and the application in preventing diabetes, is provided fundamental basis for finding safe and reliable alpha-glucosidase inhibitor, and science integration is developed to Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski resource and had very important significance.Through checking, this extract can reduce the activity of alpha-glucosidase effectively, thus has positive meaning for the treatment of diabetes.
Below the preparation method of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract:
Described method gathers Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski period of maturation tissue, dries in the shade after pulverizing, and with 5-50 times of dehydrated alcohol or other organic solvent, ultrasonic or heating and refluxing extraction 10-100 minute, repeat to extract 2-4 time, concentrating under reduced pressure, dries in the shade, and obtains Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract.By the water dissolution of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract with 1-10 volume, first with petroleum ether extraction 1-10 time with aqueous solution 1-10 volume, petroleum ether part merged, concentrating under reduced pressure is air-dry, obtained petroleum ether part; Use extraction into ethyl acetate remaining aqueous solution 1-10 time of aqueous solution 1-10 volume again, merged by ethyl acetate portion, concentrating under reduced pressure is air-dry, obtained ethyl acetate extract.Remaining aqueous solution concentrating under reduced pressure, air-dry, obtained aqueous phase position.
Further, be Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski flower extract for the best position of diabetes effect.
Further, mainly ethyl acetate extract is retained in for diabetes useful effect position.
Further, organic solvent extraction be the Plant Secondary Metabolites of medium and small polarity, oxidation occurs portion of product in high temperature possibility inducing plant or other reacts and is transformed into other composition, and room temperature easily keeps the stable of composition, so extraction temperature should not higher than 40 DEG C.
Further, for the emulsion occurred in extraction process, can be solved by the mode adding NaCl.
Further, according to clinical needs, effective site can be processed into tablet or capsule.
Detailed description of the invention
Following embodiment is convenient to understand the present invention better, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is conventional method.Test material used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.
Embodiment 1
The preparation method of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski leaf extract of the present invention is as follows:
Be collected in the Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski period of maturation fresh blade in Southern Gansu Area August, pulverizing of drying in the shade, obtains fine powder by 100 mesh sieve.By blade fine powder 20 times of ethanol solutions, ultrasonic extraction, after 60 minutes, repeats extraction 3 times, lower than concentrating under reduced pressure at 40 DEG C of temperature.In dark low temperature and the place preferably that ventilates, dry, obtained extract.Extract is dark green solid.
Embodiment 2
The preparation method of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski root extract of the present invention is as follows:
Be collected in fresh of the Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski period of maturation in Southern Gansu Area August, pulverizing of drying in the shade, obtains fine powder by 100 mesh sieve.By blade fine powder 20 times of ethanol solutions, ultrasonic extraction, after 60 minutes, repeats extraction 3 times, lower than concentrating under reduced pressure at 40 DEG C of temperature.In dark low temperature and the place preferably that ventilates, dry, obtained extract.Extract is black solid.
Embodiment 3
The preparation method of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski flower extract of the present invention is as follows:
Be collected in Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski period of maturation fresh flowers (yellow) in Southern Gansu Area August, pulverizing of drying in the shade, obtains fine powder by 100 mesh sieve.By blade fine powder 20 times of ethanol solutions, ultrasonic extraction, after 60 minutes, repeats extraction 3 times, lower than concentrating under reduced pressure at 40 DEG C of temperature.In dark low temperature and the place preferably that ventilates, dry, obtained extract.Extract is dark brown viscous solid.
Embodiment 4
The extract preparation method of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski flower extract of the present invention is as follows:
By the water (30-40 DEG C of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski flower extract (2.76g) with 25 times of volumes, 50mL) dissolve, 3 times are extracted with the petroleum ether (100mL) with aqueous solution 2 times of volumes, by air-dry for petroleum ether part concentrating under reduced pressure at lower than 40 DEG C of temperature in upper strata, obtained petroleum ether part.If there is serious emulsification phenomenon in extraction process, add NaCl solid 1-3g.Flower extract petroleum ether part 0.19g, yield 6.71%.Remaining aqueous solution is extracted 3 times by the ethyl acetate (100mL) of aqueous solution 2 times of volumes, ethyl acetate portion is air-dry lower than concentrating under reduced pressure at 40 DEG C of temperature, obtained ethyl acetate extract.Flower extract ethyl acetate extract 0.38g, yield 13.64%.Remaining aqueous solution is lower than concentrating under reduced pressure at 40 DEG C of temperature, air-dry, obtained aqueous phase position.
Embodiment 5
Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract of the present invention and different extraction position are to alpha-glucosaccharase enzyme inhibition activity
5.1 experimental technique
Alpha-glucosaccharase enzyme inhibition activity is that alpha-glucosidase (0.1 unit/mL) and substrate (p-NPG, 1.0mM) are dissolved in phosphate buffer (0.1M, pH6.7).All test specimen DMSO dissolve.Sample cultivates 10min with alpha-glucosidase together with 37 DEG C, adds substrate reactions 30min.Add Na
2cO
3cessation reaction, is determined at the ultraviolet absorption value OD under 405nm.5 variable concentrations got by all samples, repeat 3 times respectively, and calculate IC
50value.With commercially available hypoglycemic medicine acarbose for contrast.
5.2 date processing
Suppression ratio is by following formulae discovery:
Suppression ratio %=(OD
contrast-OD
sample)/OD
contrast× 100
Table 1 different parts Inhibiting α-glucosidase IC
50value
Note: (>2 represent at 2mg/mL below do not have activity in concentration)
Carbohydrate in diet is under the effect of glucosidase, and release glucose also enters blood through intestinal absorption, is the main cause that post-prandial glycemia raises.Post-prandial glycemia raises and insulin sensitivity can be caused to reduce and cause serious complication.Therefore, the regulable control of alpha-glucosidase activity, can delay the absorption and digestion of carbohydrate, plays and reduces post-prandial glycemia and glycolated hemoglobin, reduces blood glucose fluctuation, the effect of control diabetic complication.
As can be known from Table 1, it is active that the Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski different tissues extractive part prepared by the present invention all has obvious Inhibiting α-glucosidase.Wherein root and flower extract activity are all higher than contrast acarbose, and are the strongest with flower extract activity, are better than contrast more than 20 times.Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski is in the wild in growth course, in 1 year 6-8 monthly have bloom, florescence is longer, and the Biomass of flower, and can sustainable utilization higher than root.In addition, gather in a large number and utilize the root of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski to ground environmental disruption comparatively greatly, to cause the plateau grassy marshland acceleration being originally just in degraded edge to worsen.Therefore, the present invention determine by for Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski flower utilization and exploitation.
Follow the tracks of to be separated by activity and find, flower extract activity mainly concentrates on ethyl acetate extract, and its activity is also much better than street drug acarbose.But extraction position activity is all weaker than flower extract, may be because of the synergistic result of opposed polarity composition in extract, and this phenomenon is also comparatively common in Natural products research with utilization.This also further simplify production technology for the present invention, without the need to carrying out extraction step.
Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski flower extract prepared by the present invention as antidiabetic drug or food additive etc., can be offered help to the prevention and therapy of hyperglycemia and diabetics.Though there is bibliographical information, Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski has certain toxicity, mainly concentrates in leaf, and without accumulative.Leaf is poisoning be occur the odd-numbered day edible in a large number after (>250g/ day), and as the consumption of food additive or antidiabetic drug at below 1g.Therefore institute of the present invention final choice is the flower extract that toxicity is very little, and suppress glucosidase activity strong, consumption is few, is very safe to people.
The present invention has taken into full account ecological protection and sustainable use on Process Route Planning, and not only technique is simple, and plant resources enriches, and development cost is low, has significantly ecological and economic benefit.
Last it is noted that the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although with reference to previous embodiment to invention has been detailed description, for a person skilled in the art, it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein portion of techniques feature.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (7)
1. the preparation method of a Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract, it is characterized in that, described method gathers Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski period of maturation tissue, dry in the shade after pulverizing, with 5-50 times of dehydrated alcohol or other organic solvent, ultrasonic or heating and refluxing extraction 10-100 minute, repeat to extract 2-4 time, concentrating under reduced pressure, dries in the shade, and obtains Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract; By the water dissolution of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract with 1-10 volume, first with petroleum ether extraction 1-10 time with aqueous solution 1-10 volume, petroleum ether part merged, concentrating under reduced pressure is air-dry, obtained petroleum ether part; Use extraction into ethyl acetate remaining aqueous solution 1-10 time of aqueous solution 1-10 volume again, merged by ethyl acetate portion, concentrating under reduced pressure is air-dry, obtained ethyl acetate extract; Remaining aqueous solution concentrating under reduced pressure, air-dry, obtained aqueous phase position.
2. the preparation method of a kind of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract according to claim 1, is characterized in that, the Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski period of maturation is organized as Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski flower portion.
3. the preparation method of a kind of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract according to claim 1, it is characterized in that, the extract after extraction mainly retains in ethyl acetate extract.
4. the preparation method of a kind of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract according to claim 1, is characterized in that, extraction process temperature is not higher than 40 DEG C.
5. the preparation method of a kind of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract according to claim 1, is characterized in that, add NaCl when emulsion appears in extraction process.
6. the application of a kind of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract according to claim 1, it is characterized in that, extract is mainly used in preventing and treating diabetes.
7. the application of a kind of Ligularia virgaurea (Maxim.) Mattf. Ex Rehd. Etkobuski extract according to claim 1, it is characterized in that, effective site can be processed as tablet or capsule.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106074770A (en) * | 2016-08-01 | 2016-11-09 | 吉林大学 | Multiple sequence Farfugium kaemferi application in preparing xanthine oxidase inhibitor |
| CN109777622A (en) * | 2019-03-14 | 2019-05-21 | 武汉理工大学 | A kind of technique of high efficiency extraction net vein Farfugium kaemferi volatile oil |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104336085A (en) * | 2014-10-11 | 2015-02-11 | 甘肃省科学院生物研究所 | Ligularia virgaurea extract insecticide and preparation method thereof |
Non-Patent Citations (3)
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| YA LI等: "Sesquiterpenoids and Lignans from Ligularia virgaurea spp. oligocephala", 《HELVETICA CHIMICA ACTA》 * |
| 唐艳丽等: "黄帚橐吾挥发油化学成分的研究", 《中国中药杂志》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106074770A (en) * | 2016-08-01 | 2016-11-09 | 吉林大学 | Multiple sequence Farfugium kaemferi application in preparing xanthine oxidase inhibitor |
| CN106074770B (en) * | 2016-08-01 | 2019-09-24 | 吉林大学 | Multiple sequence Farfugium kaemferi is preparing the application in xanthine oxidase inhibitor |
| CN109777622A (en) * | 2019-03-14 | 2019-05-21 | 武汉理工大学 | A kind of technique of high efficiency extraction net vein Farfugium kaemferi volatile oil |
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