CN107669708A - A kind of kelp base hypoglycemic product and preparation method thereof, application - Google Patents
A kind of kelp base hypoglycemic product and preparation method thereof, application Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The present invention relates to a kind of kelp base hypoglycemic product and preparation method thereof, application, comprise the following steps：Take kelp base multiple using ethanol extraction, extraction process adds antioxidant, obtains extraction concentrat；The extraction concentrat adds extract and extracted, and extraction process adds antioxidant, dries, obtains kelp base hypoglycemic product.Kelp base raw material used in the present invention is sufficient, and hypoglycemic effect is definite, is a kind of safely and effectively hypoglycemic product without toxic side effect；Hypoglycemic product obtained by the present invention has significant inhibitory action to alpha glucuroides, and IC50 reaches 140 μ g/ml, is a kind of noncompetitive inhibitor of alpha glucuroides；Antioxidant vitamin C is added, compared to the prior art, blood sugar decreasing effect IC50 improves more than 1 times, while the yield of product improves several times.
Description
Technical field
The present invention relates to field of medicaments, more particularly to a kind of kelp base hypoglycemic product and preparation method thereof, application.
Background technology
Diabetes are to seriously endanger the major disease of human health, and complication is more and serious, it is difficult to effects a radical cure.China's diabetes
Patient reaches tens of millions of people, is one of the country that diabetic is most in the world.The medicine of clinical treatment diabetes at present
Including sulfonylurea, biguanides etc., but there is serious side effect in these long term combined uses, as sulfonylureas has
The adverse reaction of hypoglycemia；Biguanides have gastrointestinal side effect.Insulin substitution medicine is relatively more safe and effective at this stage
Medicine, it is but expensive, it is restricted to preservation condition.Therefore safer and more effective, few side effects and small hypoglycemic are found
Product is significant.
Kelp base (the Root of Laminaria Japonica) is the byproduct of kelp processing, and in China, sea-tangle adds
Work area is very big including Shandong, Liaoning, Zhejiang, the kelp processing enterprise production of jiangsu coast, generally as soil fertility quality.Existing skill
There is technology to disclose the live part in kelp base with blood sugar reducing function in art, but blood sugar reducing component is active low.Such as China
Number of patent application is 200610070494.6 patent of invention, discloses blood sugar reducing component of kelp root and preparation method thereof, is made
Preparation Method is sea-tangle root herb water and/or ethanol or ethyl acetate extraction, and water extraction concentrat is obtained after concentration, and ethanol extraction is dense
Contracting thing, ethyl acetate extraction concentrat, or its corresponding concentrate；Corresponding concentrate is extracted with ethyl acetate, and obtains and mainly contains
The acetic acid ethyl ester extract of Polyphenols and ter penoidses, and from acetic acid ethyl ester extract, the group obtained by chromatography
Point.The component is inhibited to alpha-glucosidase, and IC50 is 380 μ g/ml.
The content of the invention
In view of this, the present invention is intended to provide a kind of kelp base hypoglycemic product and preparation method thereof, application, with solution
State the blood sugar reducing component mentioned it is active low the problem of.
The invention discloses a kind of preparation method of kelp base hypoglycemic product, comprise the following steps：Kelp base is taken to use
Ethanol extraction is multiple, and extraction process adds antioxidant, obtains extraction concentrat；The extraction concentrat adds extract and carried out
Extraction, extraction process add antioxidant, dry, obtain kelp base hypoglycemic product.
Further, the antioxidant is vitamin C.
Further, the amount that the extraction process or the extraction process add antioxidant is 0-1%.
Further, the addition that the extraction process or the extraction process add antioxidant is 0.1%.
Further, the extraction process is at least extracted three times, and temperature is more than or equal to 60 DEG C, and extraction conditions are vacuum decompression
Dry.
Further, the concentration of alcohol is at least more than 80%.
Further, the extraction process also includes：Room temperature extracts 24 hours, and extract is concentrated under reduced pressure.
Further, the amount of the extract is that 5 times of extracts of the extraction concentrat are ethyl acetate.
Another object of the present invention is to disclose a kind of kelp base hypoglycemic product, including a kind of above-mentioned kelp base drop
The preparation method of blood glucose product.
Third object of the present invention is to disclose a kind of kelp base hypoglycemic product answering in hypoglycemic drug is prepared
With.
A kind of kelp base hypoglycemic product provided by the invention and preparation method thereof, using having the advantages that：
Kelp base raw material used in the present invention is sufficient, and hypoglycemic effect is definite, is that one kind safely and effectively drops without toxic side effect
Blood glucose product；Hypoglycemic product obtained by the present invention has significant inhibitory action to alpha-glucosidase, and IC50 reaches
It is a kind of noncompetitive inhibitor of alpha-glucosidase to 140 μ g/ml；Antioxidant vitamin C is added, and it is existing
Technology is compared, and blood sugar decreasing effect IC50 improves more than 1 times, while the yield of product improves several times.
Sea-tangle root extract obtained by the present invention is a kind of alpha-glucosidase noncompetitive inhibitor.It is described non-
Competitive inhibitor refers to that active material contained in extract is not combined with the activated centre of enzyme, but can be with other portions of enzyme
Position combines, so as to inhibitory enzyme and Binding Capacity.Compared to competitive inhibitor, suppression of the noncompetitive inhibitor to enzymatic activity is made
With not suppressed by concentration of substrate.
For above-mentioned and related purpose, one or more embodiments include will be explained in below and in claim
In the feature that particularly points out.Other benefits and novel features will become obvious with following detailed description consideration, institute
Disclosed embodiment is to include all these aspects and they are equal.
Embodiment
In the following detailed description, a large amount of specific details are proposed, in order to provide thorough understanding of the present invention.But
It will be understood by those within the art that it can also implement the present invention even if without these specific details.
Below with reference to preferred embodiment, the present invention is described in detail.
A kind of preparation method of kelp base hypoglycemic product is disclosed, is comprised the following steps：Kelp base is taken to be carried using ethanol
Take repeatedly, extraction process adds antioxidant, obtains extraction concentrat；The extraction concentrat adds extract and extracted,
Extraction process adds antioxidant, dries, obtains kelp base hypoglycemic product.Preferably, the antioxidant is vitamin C.
Wherein, the active ingredient contained in the hypoglycemic material of kelp base is extracted, easily oxidation, adds in separation process is extracted
Antioxidant is added to significantly improve the content of hypoglycemic material, antioxidant can also be VitAVitE, gluathione
Peptide.Antioxidant vitamin C, IC50 raising is added in extraction process.
In some illustrative embodiments, the amount of the extraction process or extraction process addition antioxidant is 0-
1%.Preferably, the addition that the extraction process or the extraction process add antioxidant is 0.1%.
In some illustrative embodiments, the extraction process is at least extracted three times, and temperature is more than or equal to 60 DEG C, extracts bar
Part is dried for vacuum decompression.The concentration of alcohol is at least more than 80%.
In some illustrative embodiments, the extraction process also includes：Room temperature extracts 24 hours, and extract decompression is dense
Contracting.
In some illustrative embodiments, the amount of the extract is that 5 times of extracts of the extraction concentrat are
Ethyl acetate.
Embodiment 1
The kelp base 100kg of natural air drying is taken, 60 DEG C of drying 24h, is clayed into power after taking-up, 60 mesh sieves is crossed, according to solid-liquid ratio
1:5 add more than 80% ethanol, while add 0.1% vitamin C, 60 DEG C of refluxing extractions 2 hours, extract solution filtering, filter
Slag extracts twice according to the method described above, merging filtrate, is dried under reduced pressure, and obtains the extraction concentrat of kelp base.
The extraction concentrat for obtaining kelp base adds the ethyl acetate of 5 times of amounts, while adds 0.1% vitamin C, in room
Upper solution is collected in temperature stirring extraction 24h, siphon, and lower sediment is extracted 2 times with same method again, combining extraction liquid, and decompression is dry
It is dry, that is, obtain kelp base hypoglycemic product A.
Embodiment 2
The kelp base 100kg of natural air drying is taken, 60 DEG C of drying 24h, is clayed into power after taking-up, 60 mesh sieves is crossed, according to solid-liquid ratio
1:5 add more than 80% ethanol, 60 DEG C of refluxing extractions 2 hours, extract solution filtering, and filter residue extracts twice according to the method described above,
Merging filtrate, it is dried under reduced pressure, obtains the extraction concentrat of kelp base.
The extraction concentrat for obtaining kelp base adds the ethyl acetate of 5 times of amounts, while adds 0.1% vitamin C, in room
Upper solution is collected in temperature stirring extraction 24h, siphon, and lower sediment is extracted 2 times with same method again, combining extraction liquid, and decompression is dry
It is dry, that is, obtain kelp base hypoglycemic product B.
Embodiment 3
The kelp base 100kg of natural air drying is taken, 60 DEG C of drying 24h, is clayed into power after taking-up, 60 mesh sieves is crossed, according to solid-liquid ratio
1:5 add more than 80% ethanol, while add 0.1% vitamin C, 60 DEG C of refluxing extractions 2 hours, extract solution filtering, filter
Slag extracts twice according to the method described above, merging filtrate, is dried under reduced pressure, and obtains the extraction concentrat of kelp base.
The extraction concentrat for obtaining kelp base adds the ethyl acetate of 5 times of amounts, and extraction 24h is stirred at room temperature, and siphon is collected
Upper solution, lower sediment are extracted 2 times with same method, combining extraction liquid, are dried under reduced pressure, that is, obtain the hypoglycemic production of kelp base again
Product C.
Embodiment 4
The kelp base 100kg of natural air drying is taken, 60 DEG C of drying 24h, is clayed into power after taking-up, 60 mesh sieves is crossed, according to solid-liquid ratio
1:5 add more than 80% ethanol, 60 DEG C of refluxing extractions 2 hours, extract solution filtering, and filter residue extracts twice according to the method described above,
Merging filtrate, it is dried under reduced pressure, obtains the extraction concentrat of kelp base.
The extraction concentrat for obtaining kelp base adds the ethyl acetate of 5 times of amounts, and extraction 24h is stirred at room temperature, and siphon is collected
Upper solution, lower sediment are extracted 2 times with same method, combining extraction liquid, are dried under reduced pressure, that is, obtain the hypoglycemic production of kelp base again
Product D.
Embodiment 5
To the measure of alpha-glucosidase inhibitory activity：
Embodiment 1-4 is measured, is configured to testing sample solution respectively, concentration gradient is, 50,100,200,400,600,
800μg/ml.As steps described below, the inhibitory action to alpha-glucosidase is determined respectively.
1) solution is prepared：Solution A (K3PO4 0.1mol/L, MgCL, 3.2mmol/L, PH 6.8), solution B (K3PO4
0.5mol/L,MgCL,16mmol/L,PH 6.8)；Enzyme solutions：0.1unit/ml alpha-glucosidase (Sigma companies
Product, I types), it is dissolved in solution A, substrate solution：Substrate p- p-nitrophenyl-alpha-D- glucofuranoses (Sigma companies) is molten
In solution A.
2) 1.02ml solution Bs are taken, add 2.82ml distilled water, the above-mentioned substrate solutions prepared of 2ml is being added, is mixing
Obtain detecting liquid.
3) do not have the above-mentioned detection liquid that hole adds 58.4 μ l on 96 plates, while add in above-described embodiment 1 sample obtained,
If (one group is blank control group, not enzyme-added and testing sample for two groups of controls；Another group is negative control, enzyme-added but be not added with to be measured
Sample), incubate 5min in 37 DEG C of water-baths.
4) 96 orifice plates of above-mentioned water-bath are taken out, do not have the μ l of enzyme solutions 20 that hole adds above-mentioned preparation, control group adds 20 μ l solution
A, incubated 30 minutes in 37 DEG C.
5) 120 μ l glycine solutions (0.4mol/L), terminating reaction are respectively added in each hole.
6) using ELIASA (Thermo companies, Varioskan flash) measure absorbance A at 405nm.
7) inhibiting rate calculates：Inhibiting rate=(absorbances of absorbance-blank control wells of sample well)/negative control hole
Absorbance.
It the results are shown in Table 1.
Inhibitory action of the embodiment 1-4 kelp bases hypoglycemic product of table 1 to alpha-glucosidase
Measurement result shows that the IC50 values of the kelp base hypoglycemic product A suppression alpha-glucosidases of embodiment 1 are 140 μ
g/ml；The IC50 values that the kelp base hypoglycemic product B of embodiment 2 suppresses alpha-glucosidase are 395 μ g/ml；Embodiment 3
Kelp base hypoglycemic product C suppress alpha-glucosidase IC50 values be 382 μ g/ml；The kelp base drop blood of embodiment 4
The IC50 values that sugar product D suppresses alpha-glucosidase are 468 μ g/ml；Suppression type is Noncompetition inhibition.Kelp base drops
Blood glucose product A activity improves more than 2 times than kelp base hypoglycemic product B, kelp base hypoglycemic product C；Dropped than kelp base
Blood glucose product D activity improves more than 3 times.
The noncompetitive inhibitor refers to that active material contained in extract is not combined with the activated centre of enzyme, but
It can be combined with other positions of enzyme, so as to inhibitory enzyme and Binding Capacity.Compared to competitive inhibitor, noncompetitive inhibitor
The inhibitory action of enzymatic activity is not suppressed by concentration of substrate.
Embodiment 6
Add influence of the vitamin C to ocean hypoglycemic product yield：
Kelp base 1kg is taken, hypoglycemic product is prepared according to the step of above-described embodiment 1,2,3 and 4, after drying, is weighed.Implement
Example 1,2,3 and 4 obtain kelp base hypoglycemic product weight be respectively：5.38th, 3.85,3.72 and 2.78 grams, yield is respectively
0.538%th, 0.39%, 0.37% and 0.28%.Illustrate that extraction can make the hypoglycemic production in ocean with extraction process addition vitamin C
The yield of product significantly improves.
Embodiment 7
Kunming mouse 60 is taken, streptozotocin is injected intraperitoneally according to 100mg/kg body weight, control group gives isometric citric acid
Solution, 5 measure blood glucose are extracted after 48 hours immediately, determine whether modeling succeeds.After modeling success, random packet：It is negative right
According to group (only injecting normal saline)；Kelp base hypoglycemic product (by taking the kelp base hypoglycemic product A in embodiment 1 as an example)；Sun
Property control group (Acarbose), every group of mouse 10, gastric infusion, once a day, and successive administration 3 days, fasting 2h after last dose
(can't help water).After inframe takes blood, fasting blood-glucose is determined with automatic clinical chemistry analyzer.Each group blood sugar effects are shown in Table 2.
Influence of the kelp base hypoglycemic product of table 2 to model mice blood sugar concentration
(* * P ＜ 0.01)
Table 2 illustrates, kelp base hypoglycemic product can significantly reduce the blood of model mice in 100mg/kg body weight gastric infusions
Sugar, there is pole significant difference with negative control.Application of the kelp base hypoglycemic product of the present invention in hypoglycemic drug is prepared, it is
A kind of safely and effectively hypoglycemic product.
Embodiment 8
The kelp base 100kg of natural air drying is taken, 60 DEG C of drying 24h, is clayed into power after taking-up, 60 mesh sieves is crossed, according to solid-liquid ratio
1:5 add more than 80% ethanol, while add 0.1% vitamin A, 60 DEG C of refluxing extractions 2 hours, extract solution filtering, filter
Slag extracts twice according to the method described above, merging filtrate, is dried under reduced pressure, and obtains the extraction concentrat of kelp base.
The extraction concentrat for obtaining kelp base adds the ethyl acetate of 5 times of amounts, while adds 0.1% vitamin A, in room
Upper solution is collected in temperature stirring extraction 24h, siphon, and lower sediment is extracted 2 times with same method again, combining extraction liquid, and decompression is dry
It is dry, that is, obtain kelp base hypoglycemic product E.
Embodiment 9
The kelp base 100kg of natural air drying is taken, 60 DEG C of drying 24h, is clayed into power after taking-up, 60 mesh sieves is crossed, according to solid-liquid ratio
1:5 add more than 80% ethanol, while add 0.1% glutathione, 60 DEG C of refluxing extractions 2 hours, extract solution filtering, filter
Slag extracts twice according to the method described above, merging filtrate, is dried under reduced pressure, and obtains the extraction concentrat of kelp base.
The extraction concentrat for obtaining kelp base adds the ethyl acetate of 5 times of amounts, while adds 0.1% glutathione, in
Extraction 24h is stirred at room temperature, upper solution is collected in siphon, and lower sediment is extracted 2 times with same method again, combining extraction liquid, decompression
Dry, that is, obtain kelp base hypoglycemic product F.
Embodiment 10
To embodiment 1,8,9 carry out alpha-glucosidase inhibitory activity measure, specific method referring to embodiment 5 side
Method, it the results are shown in Table 3.
Inhibitory action of the kelp base hypoglycemic product of 3 embodiment of table 1,8,9 to alpha-glucosidase
Measurement result shows that kelp base hypoglycemic product A activity is than kelp base hypoglycemic product E, kelp base hypoglycemic product F
Improve more than 2 times.
Embodiment 11
The kelp base 100kg of natural air drying is taken, 60 DEG C of drying 24h, is clayed into power after taking-up, 60 mesh sieves is crossed, according to solid-liquid ratio
1:5 add more than 80% ethanol, while add 1% vitamin C, 60 DEG C of refluxing extractions 2 hours, extract solution filtering, filter residue
Extract twice, merging filtrate, be dried under reduced pressure according to the method described above, obtain the extraction concentrat of kelp base.
The extraction concentrat for obtaining kelp base adds the ethyl acetate of 5 times of amounts, while adds 0.1% vitamin C, in room
Upper solution is collected in temperature stirring extraction 24h, siphon, and lower sediment is extracted 2 times with same method again, combining extraction liquid, and decompression is dry
It is dry, that is, obtain kelp base hypoglycemic product G.
Embodiment 12
The kelp base 100kg of natural air drying is taken, 60 DEG C of drying 24h, is clayed into power after taking-up, 60 mesh sieves is crossed, according to solid-liquid ratio
1:5 add more than 80% ethanol, while add 0.1% vitamin C, 60 DEG C of refluxing extractions 2 hours, extract solution filtering, filter
Slag extracts twice according to the method described above, merging filtrate, is dried under reduced pressure, and obtains the extraction concentrat of kelp base.
The extraction concentrat for obtaining kelp base adds the ethyl acetate of 5 times of amounts, while adds 1% vitamin C, in room temperature
Upper solution is collected in stirring extraction 24h, siphon, and lower sediment is extracted 2 times with same method, combining extraction liquid, is dried under reduced pressure again,
Obtain kelp base hypoglycemic product H.
Embodiment 13
To embodiment 1,11,12 carry out alpha-glucosidase inhibitory activity measure, specific method referring to embodiment 5 side
Method, it the results are shown in Table 4.
Inhibitory action of the kelp base hypoglycemic product of 4 embodiment of table 1,11,12 to alpha-glucosidase
Measurement result shows that kelp base hypoglycemic product A activity is than kelp base hypoglycemic product G, kelp base hypoglycemic product H
Improve more than 1 times.Kelp base hypoglycemic product G extraction process vitamin Cs amount is 1%, and the ascorbic amount of extraction process is
0.1%；Kelp base hypoglycemic product H extraction process vitamin Cs amount is 0.1%, and the ascorbic amount of extraction process is 1%, activity
It is more or less the same.
In summary, embodiment 1 is optimum embodiment, and activity is better than other kelp base hypoglycemic products, and prior art
Compare, blood sugar decreasing effect IC50 improves more than 1 times, while the yield of product improves several times.And kelp base raw material is sufficient, drop
Blood glucose effect is definite, without toxic side effect, is a kind of safely and effectively hypoglycemic product, has to alpha-glucosidase aobvious
The inhibitory action of work.
The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention, for the skill of this area
For art personnel, the present invention can have various modifications and variations.Within the spirit and principles of the invention, that is made any repaiies
Change, equivalent substitution, improvement etc., should be included in the scope of the protection.
Claims (10)
1. a kind of preparation method of kelp base hypoglycemic product, it is characterised in that comprise the following steps：Kelp base is taken to use second
Alcohol extracting is multiple, and extraction process adds antioxidant, obtains extraction concentrat；The extraction concentrat adds extract and carried out
Extraction, extraction process add antioxidant, dry, obtain kelp base hypoglycemic product.
2. preparation method according to claim 1, it is characterised in that the antioxidant is vitamin C.
3. preparation method according to claim 1, it is characterised in that the extraction process or the extraction process add
The amount of antioxidant is 0-1%.
4. preparation method according to claim 2, it is characterised in that the extraction process or the extraction process add
The addition of antioxidant is 0.1%.
5. preparation method according to claim 1, it is characterised in that the extraction process is at least extracted three times, and temperature is big
In equal to 60 DEG C, extraction conditions are dried for vacuum decompression.
6. preparation method according to claim 1, it is characterised in that the concentration of alcohol is at least more than 80%.
7. preparation method according to claim 1, it is characterised in that the extraction process also includes：Room temperature extraction 24 is small
When, extract is concentrated under reduced pressure.
8. preparation method according to claim 1, it is characterised in that the amount of the extract is the extraction concentrat
5 times, the extract is ethyl acetate.
9. a kind of kelp base hypoglycemic product, it is characterised in that including the main laminaria any one of claim 1-8
The preparation method of root hypoglycemic product.
A kind of 10. application of kelp base hypoglycemic product in hypoglycemic drug is prepared.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109090512A (en) * | 2018-07-24 | 2018-12-28 | 哈尔滨工业大学（威海） | A kind of hypoglycemic Thallus Laminariae (Thallus Eckloniae) extract |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106539825A (en) * | 2015-09-17 | 2017-03-29 | 青岛佳日隆海洋食品有限公司 | A kind of Radix Laminariae hypoglycemic product and preparation method thereof, application |
-
2017
- 2017-11-06 CN CN201711075170.6A patent/CN107669708A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106539825A (en) * | 2015-09-17 | 2017-03-29 | 青岛佳日隆海洋食品有限公司 | A kind of Radix Laminariae hypoglycemic product and preparation method thereof, application |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109090512A (en) * | 2018-07-24 | 2018-12-28 | 哈尔滨工业大学（威海） | A kind of hypoglycemic Thallus Laminariae (Thallus Eckloniae) extract |
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