CN113663043B - Composition with alpha-glucosidase inhibition effect and preparation method and application thereof - Google Patents
Composition with alpha-glucosidase inhibition effect and preparation method and application thereof Download PDFInfo
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- CN113663043B CN113663043B CN202110977705.9A CN202110977705A CN113663043B CN 113663043 B CN113663043 B CN 113663043B CN 202110977705 A CN202110977705 A CN 202110977705A CN 113663043 B CN113663043 B CN 113663043B
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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
- A61K36/18—Magnoliophyta (angiosperms)
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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Abstract
The invention discloses a composition with alpha-glucosidase inhibition effect, a preparation method and application thereof, wherein the composition with alpha-glucosidase inhibition effect comprises the following components: mulberry leaf extract, licorice extract, turmeric extract and sea buckthorn extract. The composition obtained by matching the mulberry leaf extract, the liquorice extract, the turmeric extract and the sea buckthorn extract has good alpha-glucosidase inhibition effect, the drug effect is superior to that of the simple superposition of the single use of each component or the matching use of the two components, and the composition has obvious synergistic effect; the four raw materials of the composition are all pure natural traditional Chinese medicines, have low toxic and side effects, reduce the harm to human bodies, and have relatively low price. The composition can be used as an active ingredient for preparing medicines and health products for reducing blood sugar or food additives.
Description
Technical Field
The present invention belongs to the field of medicine, health product and food technology. More particularly, relates to a composition with alpha-glucosidase inhibition effect, and a preparation method and application thereof.
Background
Diabetes Mellitus (DM) is a multi-etiologic metabolic disease characterized by chronic hyperglycemia due to disorders of sugar, fat and protein metabolism caused by insufficient insulin secretion or defective insulin action or both. Diabetes can cause chronic damage, hypofunction and failure of tissues and organs such as eyes, kidneys, nerves, heart, blood vessels, and the like. Acute metabolic disorders such as diabetic ketoacidosis, hyperosmolar coma, etc. may also occur in severe or stress states.
The main means of the prior art for treating diabetes mellitus are oral medication therapy and insulin therapy, and the oral medication contains secretion promoting agents such as sulfonylureas and non-sulfonylureas, biguanides such as alpha-glucosidase inhibitors and thiazolidinediones and the like. Alpha-glucosidase inhibitors are a new class of antidiabetic drugs, and 3 drugs are currently on the market and widely used in clinical treatment. The medicine can obviously reduce the postprandial blood sugar level of diabetics, thereby reducing the occurrence of diabetic complications. The alpha-glucosidase inhibitor medicine can treat a plurality of diseases by regulating the activity of alpha-glucosidase in vivo, and has wide application prospect.
However, although the curative effect of western medicines for treating diabetes is determined, oral medicines for treating diabetes have toxic and side effects, and insulin for treating diabetes has adverse reactions such as hypoglycemia and the like. The traditional Chinese medicine for treating diabetes belongs to the category of diabetes, adopts pure natural plant extracts, has stable curative effect, small toxic and side effects and low price. For example, patent document CN102258570B discloses that the formula of selaginella tamariscina active extract and selfheal active extract is selected, the effect of inhibiting the activity of alpha-glucosidase is greater than the simple addition of the effects of the two drugs, and the composition can be applied to drugs, health products or food additives for preventing or treating diabetes, but the existing drugs for treating diabetes containing pure natural plant extracts are still few. Therefore, the research and development of more pure natural plant extract compositions with stable effect, less toxic and side effect and relatively low price and the alpha-glucosidase inhibition effect have important significance for preparing health products for reducing blood sugar.
Disclosure of Invention
The invention aims to solve the technical problems of the existing western medicine for treating diabetes, such as insufficient relieving effect, unstable curative effect and high cost, and provides a composition containing pure natural plant extracts and having an alpha-glucosidase inhibiting effect, and a preparation method thereof.
The first object of the present invention is to provide a composition having an alpha-glucosidase inhibitory effect.
The second purpose of the invention is to provide a preparation method of the composition with alpha-glucosidase inhibition effect.
The third purpose of the invention is to provide the application of the composition with alpha-glucosidase inhibition effect in preparing medicines, health products or foods with hypoglycemic effect.
The above purpose of the invention is realized by the following technical scheme:
the inventor researches and discovers that the composition obtained by compounding the four Chinese herbal medicine extracts of the mulberry leaf extract, the licorice extract, the turmeric extract and the sea buckthorn extract has a good alpha-glucosidase inhibiting effect, a good blood sugar reducing effect, a drug effect obviously superior to that of simple superposition of single use of each component or matching use of two components, and a remarkable synergistic effect. The four extracts are all extracted from pure natural traditional Chinese medicines, have low toxic and side effects, reduce the harm to human bodies and have relatively low price.
Therefore, the invention firstly provides a composition with alpha-glucosidase inhibition effect, and the composition comprises the following raw materials: mulberry leaf extract, licorice extract, turmeric extract and sea buckthorn extract.
Preferably, the mass ratio of the mulberry leaf extract to the licorice extract to the turmeric extract to the sea buckthorn extract is (1-5) to (1-4) to (1-3.5) to (1-5).
Preferably, the mass ratio of the mulberry leaf extract to the licorice extract to the turmeric extract to the sea buckthorn extract is (1-3) to (1-1.5) to (1-2).
More preferably, the mass ratio of the mulberry leaf extract to the licorice extract to the turmeric extract to the sea buckthorn extract is (1-3) to 1 to (1-2).
More preferably, the mass ratio of the mulberry leaf extract to the licorice extract to the turmeric extract to the sea buckthorn extract is (1-2) to 1:1 to (1-2).
Further preferably, the mass ratio of the mulberry leaf extract, the licorice extract, the turmeric extract and the sea buckthorn extract is 1: 1.
In the composition with the alpha-glucosidase inhibiting effect, mulberry leaves and liquorice are prepared into a mulberry leaf extract and a liquorice extract by a hot water extraction method, and turmeric and sea buckthorn are prepared into a turmeric extract and a sea buckthorn extract by an ethanol extraction method.
Preferably, the preparation method of the mulberry leaf extract comprises the following steps: crushing the dried mulberry leaves, sieving the crushed mulberry leaves with a 50-70-mesh sieve, adding 14-16 times of water, soaking for 0.5-2 h, boiling and extracting for 1.5-3 h, filtering, sieving with 90-110-mesh filter cloth, and collecting filtrate; adding 8-12 times of water into filter residues, boiling and extracting for 0.5-2 h, filtering through a filter cloth of 90-110 meshes, and collecting filtrate; and combining the filtrates, carrying out reduced pressure rotary evaporation and concentration, and freeze-drying for 36-72 h to finally obtain an extract, namely the mulberry leaf extract.
More preferably, the mulberry leaf extract is prepared by removing insects or deteriorated parts from sun-dried mulberry leaves, pulverizing the mulberry leaves with a pulverizer, and sieving the pulverized mulberry leaves with a 60-mesh sieve; soaking the medicinal powder in 15 times of water for 1 hr, boiling and extracting for 2 hr, filtering with 100 mesh filter cloth, and collecting filtrate; adding 10 times of water into the filter residue, boiling and extracting for 2h, filtering through 100-mesh filter cloth, and collecting the filtrate; mixing filtrates, concentrating under reduced pressure by rotary evaporation, and lyophilizing for 48 hr to obtain extract, i.e. folium Mori extract.
Preferably, the preparation method of the licorice extract comprises the following steps: crushing the dried liquorice, sieving the crushed liquorice with a 50-70-mesh sieve, adding 14-16 times of water, soaking for 0.5-2 hours, boiling and extracting for 1.5-3 hours, filtering, sieving with 90-110-mesh filter cloth, and collecting filtrate; adding 8-12 times of water into filter residues, boiling and extracting for 0.5-2 h, filtering through a filter cloth of 90-110 meshes, and collecting filtrate; and combining the filtrates, carrying out reduced pressure rotary evaporation and concentration, and freeze-drying for 36-72 h to finally obtain an extract, namely the mulberry liquorice extract.
More preferably, the preparation method of the licorice extract comprises the following steps: taking the dried liquorice, removing insect-containing or deteriorated parts, crushing by using a powder grinding machine, and sieving by using a 60-mesh sieve; soaking the medicinal powder in 15 times of water for 1 hr, boiling and extracting for 2 hr, filtering with 100 mesh filter cloth, and collecting filtrate; adding 10 times of water into the filter residue, boiling and extracting for 2h, filtering through 100-mesh filter cloth, and collecting the filtrate; mixing filtrates, concentrating under reduced pressure by rotary evaporation, and lyophilizing for 48 hr to obtain extract, i.e. Glycyrrhrizae radix extract.
Preferably, the turmeric extract is prepared by the following steps: crushing the dried turmeric, sieving with a 50-70-mesh sieve, adding 14-16 times of 50-75% ethanol, soaking for 0.5-1 h, performing reflux extraction at 70-85 ℃ for 1.5-3 h, sieving with 90-110-mesh filter cloth, and collecting filtrate; adding 9-12 times of 50-75% ethanol water into filter residues after suction filtration, performing reflux extraction at 70-85 ℃ for 0.5-2 h, filtering through filter cloth of 90-110 meshes, and collecting filtrate; and combining the filtrates, carrying out reduced pressure rotary evaporation and concentration, and freeze-drying for 36-72 h to finally obtain an extract, namely the turmeric extract.
More preferably, the turmeric extract is prepared by removing insects or deteriorated parts from sun-dried turmeric, pulverizing the turmeric with a pulverizer, and sieving the turmeric with a 60-mesh sieve. Soaking the medicinal powder in 15 times of 60% ethanol water for 0.5 hr, reflux extracting at 80 deg.C for 2 hr, filtering with 100 mesh filter cloth, and collecting filtrate; adding 10 times of 60% ethanol water into the filter residue after suction filtration, extracting under reflux at 80 deg.C for 1h, filtering through 100 mesh filter cloth, and collecting filtrate; mixing filtrates, concentrating under reduced pressure by rotary evaporation, and lyophilizing for 48 hr to obtain extract, namely Curcuma rhizome extract.
Preferably, the preparation method of the seabuckthorn extract comprises the following steps: crushing the dried sea buckthorn, sieving the crushed sea buckthorn with a 50-70-mesh sieve, adding 14-16 times of 50-75% ethanol, soaking for 0.5-1 h, performing reflux extraction at 70-85 ℃ for 1.5-3 h, sieving with 90-110-mesh filter cloth, and collecting filtrate; adding 9-12 times of 50-75% ethanol water into filter residues after suction filtration, performing reflux extraction at 70-85 ℃ for 0.5-2 h, filtering through filter cloth of 90-110 meshes, and collecting filtrate; and combining the filtrates, carrying out reduced pressure rotary evaporation and concentration, and freeze-drying for 36-72 h to finally obtain an extract, namely the sea buckthorn extract.
More preferably, the seabuckthorn extract is prepared by sun-drying seabuckthorn, removing insect-containing or deteriorated parts, pulverizing with a pulverizer, and sieving with a 60-mesh sieve. Soaking the medicinal powder in 15 times of 60% ethanol for 0.5 hr, reflux-extracting at 80 deg.C for 2 hr, filtering with 100 mesh filter cloth, and collecting filtrate; adding 10 times of 60% ethanol water into the filter residue after suction filtration, extracting under reflux at 80 deg.C for 1h, filtering through 100 mesh filter cloth, and collecting filtrate; mixing filtrates, rotary steaming under reduced pressure, concentrating, and lyophilizing for 48 hr to obtain extract, i.e. fructus Hippophae extract.
The invention also provides a preparation method of the composition with the alpha-glucosidase inhibition effect, which takes mulberry leaves, liquorice, turmeric and sea buckthorn as raw materials to prepare mulberry leaf extract, liquorice extract, turmeric extract and sea buckthorn extract; mixing the above extracts to obtain the composition with alpha-glucosidase inhibiting effect.
The invention also provides application of the composition with the alpha-glucosidase inhibiting effect in preparation of a preparation with the alpha-glucosidase inhibiting effect, and the conventional preparation is prepared according to a conventional process.
Preferably, the conventional formulation includes a tablet, a capsule, a pill, a powder, a granule, a soft extract, a solution, or a syrup.
The invention provides a medicine, health-care product or food with alpha-glucosidase inhibition effect, which contains the composition with alpha-glucosidase inhibition effect.
The invention has the following beneficial effects:
the composition obtained by compounding the mulberry leaf extract, the licorice extract, the turmeric extract and the sea buckthorn extract has good alpha-glucosidase inhibition effect and good blood sugar reduction effect, the drug effect is obviously better than that of the simple superposition of the single use of each component or the matching use of the two components, the inhibition rate can reach 79.34-87.73%, and the composition has obvious synergistic effect. The four extracts in the composition are all extracted from pure natural traditional Chinese medicines, have low toxic and side effects, reduce the harm to human bodies, have relatively low price and have wide application prospect.
Drawings
FIG. 1 is a graph showing the results of the test of the inhibition rate of the extracts of examples 1 to 3 and comparative examples 1 to 10 of the present invention against α -glucosidase.
Detailed Description
The invention is further described with reference to the drawings and the following detailed description, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
Example 1
1. The preparation method of the mulberry leaf extract comprises the following steps:
(1) pretreatment of mulberry leaves: taking dried folium Mori, removing insect or deterioration part, pulverizing with a pulverizer, and sieving with 60 mesh sieve;
(2) taking mulberry leaf powder, adding 15 times of water, soaking for 1h, boiling and extracting for 2h, filtering through 100-mesh filter cloth, and collecting filtrate;
(3) adding 10 times of water into the filter residue obtained in the step (2), boiling and extracting for 2h, filtering through 100-mesh filter cloth, and collecting the filtrate;
(4) combining the filtrates obtained in (2) and (3), performing rotary evaporation and concentration under reduced pressure, and freeze-drying for 48h to obtain the extract, namely the mulberry leaf extract.
2. The preparation method of the licorice extract comprises the following steps:
(1) pretreatment of liquorice: taking the dried liquorice, removing insect-containing or deteriorated parts, crushing by using a powder grinding machine, and sieving by using a 60-mesh sieve;
(2) soaking Glycyrrhrizae radix powder in 15 times of water for 1 hr, boiling and extracting for 2 hr, filtering with 100 mesh filter cloth, and collecting filtrate;
(3) adding 10 times of water into the filter residue obtained in the step (2), boiling and extracting for 2h, filtering through 100-mesh filter cloth, and collecting the filtrate;
(4) mixing the filtrates of (2) and (3), concentrating under reduced pressure by rotary evaporation, and lyophilizing for 48 hr to obtain extract, i.e. Glycyrrhrizae radix extract.
3. The preparation method of the turmeric extract comprises the following steps:
(1) pretreatment of turmeric: taking sun-dried Curcuma rhizome, removing pest or deterioration parts, pulverizing with a pulverizer, and sieving with 60 mesh sieve;
(2) soaking Curcuma rhizome powder in 15 times of 60% ethanol water for 0.5 hr, reflux-extracting at 80 deg.C for 2 hr, filtering with 100 mesh filter cloth, and collecting filtrate;
(3) adding 10 times of 60% ethanol water into the filter residue obtained in the step (2), carrying out reflux extraction at 80 ℃ for 1h, filtering through 100-mesh filter cloth, and collecting the filtrate;
(4) mixing the filtrates of (2) and (3), rotary-steaming under reduced pressure, removing ethanol, concentrating, and lyophilizing for 48 hr to obtain extract, namely Curcuma rhizome extract.
4. The preparation method of the sea buckthorn extract comprises the following steps:
(1) pretreatment of sea buckthorn: taking sun-dried sea buckthorn, removing insect or metamorphic parts, pulverizing with a pulverizer, and sieving with a 60-mesh sieve;
(2) soaking fructus Hippophae powder in 15 times of 60% ethanol water for 0.5 hr, reflux extracting at 80 deg.C for 2 hr, filtering with 100 mesh filter cloth, and collecting filtrate;
(3) adding 10 times of 60% ethanol water into the filter residue obtained in the step (2), carrying out reflux extraction at 80 ℃ for 1h, filtering through 100-mesh filter cloth, and collecting the filtrate;
(4) mixing the filtrates of (2) and (3), rotary steaming under reduced pressure, removing ethanol, concentrating, and lyophilizing for 48 hr to obtain extract, i.e. fructus Hippophae extract.
The present example provides a composition with α -glucosidase inhibitory effect, wherein the total dosage of the composition extracted according to the above steps is 0.1g/mL, and the mass ratio of the mulberry leaf extract, the licorice root extract, the turmeric root extract and the sea buckthorn extract is 1:1:1: 1.
Example 2
The extraction method of each extract is the same as that in example 1, this example provides a composition having an α -glucosidase inhibitory effect, wherein the total amount of the composition is 0.1g/mL, and the mass ratio of the mulberry leaf extract, the licorice extract, the turmeric extract and the sea buckthorn extract in the composition of this example is 2:1:1: 2.
Example 3
The extraction method of each extract is the same as that in example 1, this example provides a composition having an α -glucosidase inhibitory effect, wherein the total amount of the composition is 0.1g/mL, and the mass ratio of the mulberry leaf extract, the licorice extract, the turmeric extract and the sea buckthorn extract in the composition of this example is 3:1:1: 2.
Comparative example 1
The mulberry leaf extraction method is the same as example 1, and this comparative example provides an extract having an alpha-glucosidase inhibitory effect, i.e., a mulberry leaf extract, in an amount of 0.1 g/mL.
Comparative example 2
The same licorice extraction method as in example 1, this comparative example provides an extract having alpha-glucosidase inhibitory activity, i.e., licorice extract, in an amount of 0.1 g/mL.
Comparative example 3
Turmeric extraction method As in example 1, this comparative example provides an extract having alpha-glucosidase inhibitory activity, namely turmeric extract, in an amount of 0.1 g/mL.
Comparative example 4
The method for extracting seabuckthorn is the same as that of example 1, and the comparative example provides an extract with alpha-glucosidase inhibition, namely the seabuckthorn extract, and the dosage of the extract is 0.1 g/mL.
Comparative example 5
The extracts were extracted by the same method as in example 1, and this comparative example provides a composition having an α -glucosidase inhibitory effect, which contained only mulberry leaf extract and turmeric extract in a total amount of 0.1g/mL in a mass ratio of 1: 1.
Comparative example 6
The extraction method of each extract was the same as example 1, and this comparative example provides a composition having an α -glucosidase inhibitory effect, which contained only mulberry leaf extract and licorice extract in a total amount of 0.1g/mL in a mass ratio of 1: 1.
Comparative example 7
The extraction method of each extract is the same as that of example 1, and this comparative example provides a composition having an alpha-glucosidase inhibitory effect, and the composition of this example only contains a mulberry leaf extract and a sea buckthorn extract, the total amount of which is 0.1g/mL, and the mass ratio is 1: 1.
Comparative example 8
The extracts were extracted in the same manner as in example 1, and this comparative example provides a composition having an α -glucosidase inhibitory effect, which contained only turmeric extract and licorice extract in a total amount of 0.1g/mL in a mass ratio of 1: 1.
Comparative example 9
The same extraction method as that of example 1 this comparative example provides a composition having an α -glucosidase inhibitory effect, and the composition of this example contains only hippophae rhamnoides extract and licorice extract in an amount of 0.1g/mL, in a mass ratio of 1: 1.
Comparative example 10
The extraction methods of the extracts are the same as example 1, and this comparative example provides a composition having an α -glucosidase inhibitory effect, which contains only the hippophae rhamnoides extract and the turmeric extract in a total amount of 0.1g/mL in a mass ratio of 1: 1.
Example 4 alpha-glucosidase inhibition
1. Experimental methods
(1) The experiment was divided into blank, drug and drug blank (drug only), each reactant was dosed in 96-well plates with 3 parallel wells per group.
(2) Sequentially adding 250 mu L of PBS buffer solution, 0.01776mol/L of 50 mu L of PNPG substrate and 50 mu L of sample, uniformly mixing, carrying out water bath at 37 ℃ for heat preservation for 10min, and taking out after the heat preservation; adding 0.1U/mL enzyme solution in 37 deg.C water bath, mixing, and reacting in 37 deg.C water bath for 20 min; after the reaction was completed, 100. mu.L of 1mol/L sodium carbonate solution was added to stop the reaction. pNPG is hydrolyzed by alpha-glucosidase to yield glucose and PNP, wherein PNP has a maximum absorbance at 405nm and the absorbance is measured.
Calculating the formula:
(A1: blank enzyme activity; A2: enzyme activity after addition of drugs; A3: PNPG background; A4: drug background)
2. Results of the experiment
The detection shows that the compositions of examples 1-3 and the extracts and compositions of comparative examples 1-10 can inhibit the activity of alpha-glucosidase, but the inhibition rate of the alpha-glucosidase is different, and the total dosage of the extracts or the compositions in all the examples and the comparative examples is 0.1 g/mL. As shown in fig. 1, the inhibition ratios of the compositions of examples 1 to 3, which used the four components at different mass ratios of 1:1:1:1, 2:1:1:2 and 3:1:1:2, to α -glucosidase were 87.73%, 79.34% and 82.03%, respectively; in comparative examples 1-4, the inhibition rates of the mulberry leaf extract, the licorice extract, the turmeric extract and the sea buckthorn extract on the alpha-glucosidase are 40.34%, 7.08%, 9.12% and 11.73% respectively; in comparative examples 5 to 10, compositions in a ratio of 1:1 were used in pairs: the inhibition rates of the mulberry leaf and curcuma extract, the mulberry leaf and licorice extract, the mulberry leaf and seabuckthorn extract, the curcuma and licorice extract, the seabuckthorn extract and licorice extract and the curcuma and seabuckthorn extract on alpha-glucosidase are 47.29%, 50.51%, 66.11%, 22.19%, 20.03% and 25.37% respectively.
The inhibition effect of the mulberry leaf extract is found to be good in the inhibition of alpha-glucosidase by adopting the extract alone, and the inhibition effect of the mulberry leaf and sea buckthorn is good in the inhibition effect of the composition combined in pairs. The sum of the inhibition rates of the four extracts acting independently on the alpha-glucosidase is only 68.28%, which is not as good as the inhibition effect of the composition prepared by compounding the four components according to different qualities on the alpha-glucosidase in examples 1-3. In addition, the four compositions adopted in the example 1 have the best inhibition effect in terms of mass ratio, and the inhibition rate reaches 87.73%, which is obviously superior to the extracts or the extract compositions in the comparative examples 1-10.
The composition obtained by compounding the mulberry leaf extract, the liquorice extract, the turmeric extract and the sea buckthorn extract has good alpha-glucosidase inhibition effect and good blood sugar reduction effect, the drug effect is obviously superior to that of the simple superposition of the single use of each component or the matching use of the two components, and the composition has obvious synergistic effect.
Example 5
1. The preparation method of the mulberry leaf extract comprises the following steps:
(1) pretreatment of mulberry leaves: taking sun-dried mulberry leaves, removing worm or metamorphic parts, crushing by a powder grinding machine, and sieving by a 50-mesh sieve;
(2) taking mulberry leaf powder, adding 14 times of water, soaking for 0.5h, boiling and extracting for 1.5h, filtering through 90-mesh filter cloth, and collecting filtrate;
(3) adding 8 times of water into the filter residue obtained in the step (2), boiling and extracting for 0.5h, filtering through a 90-mesh filter cloth, and collecting the filtrate;
(4) and (3) combining the filtrates obtained in the steps (2) and (3), performing rotary evaporation and concentration under reduced pressure, and performing freeze-drying for 36h to finally obtain an extract, namely the mulberry leaf extract.
2. The preparation method of the licorice extract comprises the following steps:
(1) pretreatment of liquorice: taking the dried liquorice, removing insect-containing or deteriorated parts, crushing by using a powder grinding machine, and sieving by using a 50-mesh sieve;
(2) soaking Glycyrrhrizae radix powder in 14 times of water for 0.5 hr, boiling and extracting for 1.5 hr, filtering with 90 mesh filter cloth, and collecting filtrate;
(3) adding 8 times of water into the filter residue obtained in the step (2), boiling and extracting for 0.5h, filtering through a 90-mesh filter cloth, and collecting the filtrate;
(4) mixing the filtrates of (2) and (3), concentrating under reduced pressure by rotary evaporation, and lyophilizing for 36h to obtain extract, i.e. Glycyrrhrizae radix extract.
3. The preparation method of the turmeric extract comprises the following steps:
(1) pretreatment of turmeric: taking sun-dried Curcuma rhizome, removing worm or deteriorated part, pulverizing with a pulverizer, and sieving with 50 mesh sieve;
(2) soaking Curcuma rhizome powder in 14 times of 50% ethanol water for 0.5 hr, reflux-extracting at 70 deg.C for 1.5 hr, filtering with 90 mesh filter cloth, and collecting filtrate;
(3) adding 9 times of 50% ethanol water into the filter residue obtained in the step (2), carrying out reflux extraction at 70 ℃ for 0.5h, filtering through a 90-mesh filter cloth, and collecting the filtrate;
(4) mixing the filtrates of (2) and (3), rotary-steaming under reduced pressure, removing ethanol, concentrating, and lyophilizing for 48 hr to obtain extract, namely Curcuma rhizome extract.
4. The preparation method of the sea buckthorn extract comprises the following steps:
(1) pretreatment of sea buckthorn: taking sun-dried sea buckthorn, removing insect or metamorphic parts, pulverizing with a pulverizer, and sieving with a 50-mesh sieve;
(2) soaking fructus Hippophae powder in 14 times of 50% ethanol water for 1 hr, reflux extracting at 70 deg.C for 1.5 hr, filtering with 90 mesh filter cloth, and collecting filtrate;
(3) adding 9 times of 50% ethanol water into the filter residue obtained in the step (2), carrying out reflux extraction at 70 ℃ for 0.5h, filtering through a 90-mesh filter cloth, and collecting the filtrate;
(4) mixing the filtrates of (2) and (3), rotary-steaming under reduced pressure, removing ethanol, concentrating, and lyophilizing for 36h to obtain extract, i.e. fructus Hippophae extract.
The present example provides a composition with α -glucosidase inhibitory effect, the total dosage of the composition extracted according to the above steps is 0.1g/mL, and the mass ratio of the mulberry leaf extract, the licorice root extract, the turmeric root extract and the sea buckthorn extract is 1:1:1: 1.
The method for preparing the composition provided in this example is different from that of example 1 in specific parameters, and the composition extracted from mulberry leaves, licorice roots, turmeric and sea buckthorn according to the above method has a less α -glucosidase inhibition rate than that of example 1, so that the method is not the most preferable scheme, but still has a significant synergistic effect compared with the extracts and compositions of comparative examples 1 to 10.
Example 6
1. The preparation method of the mulberry leaf extract comprises the following steps:
(1) pretreatment of mulberry leaves: taking dried folium Mori, removing insect or deterioration part, pulverizing with a pulverizer, and sieving with 70 mesh sieve;
(2) taking mulberry leaf powder, adding 16 times of water, soaking for 2h, boiling and extracting for 3h, filtering through 110-mesh filter cloth, and collecting filtrate;
(3) adding 12 times of water into the filter residue obtained in the step (2), boiling and extracting for 2h, filtering through 110-mesh filter cloth, and collecting the filtrate;
(4) combining the filtrates obtained in (2) and (3), performing rotary evaporation and concentration under reduced pressure, and freeze-drying for 72h to obtain the extract, namely the mulberry leaf extract.
2. The preparation method of the licorice extract comprises the following steps:
(1) pretreatment of liquorice: taking the dried liquorice, removing insect-containing or deteriorated parts, crushing by using a powder grinding machine, and sieving by using a 70-mesh sieve;
(2) soaking Glycyrrhrizae radix powder in 16 times of water for 2 hr, boiling and extracting for 3 hr, filtering with 110 mesh filter cloth, and collecting filtrate;
(3) adding 12 times of water into the filter residue obtained in the step (2), boiling and extracting for 2h, filtering through 110-mesh filter cloth, and collecting the filtrate;
(4) mixing the filtrates of (2) and (3), concentrating under reduced pressure by rotary evaporation, and lyophilizing for 72h to obtain extract, i.e. Glycyrrhrizae radix extract.
3. The preparation method of the turmeric extract comprises the following steps:
(1) pretreatment of turmeric: taking sun-dried Curcuma rhizome, removing pest or deterioration parts, pulverizing with a pulverizer, and sieving with a 70 mesh sieve;
(2) soaking Curcuma rhizome powder in 16 times of 75% ethanol water for 1 hr, reflux extracting at 85 deg.C for 3 hr, filtering with 110 mesh filter cloth, and collecting filtrate;
(3) adding 12 times of 75% ethanol water into the filter residue obtained in the step (2), carrying out reflux extraction at 85 ℃ for 2h, filtering through 110-mesh filter cloth, and collecting the filtrate;
(4) mixing the filtrates of (2) and (3), rotary steaming under reduced pressure, removing ethanol, concentrating, and lyophilizing for 72 hr to obtain Curcuma rhizome extract.
4. The preparation method of the sea buckthorn extract comprises the following steps:
(1) pretreatment of sea buckthorn: taking sun-dried sea buckthorn, removing insects or deteriorated parts, crushing by using a powder grinding machine, and sieving by using a 70-mesh sieve;
(2) soaking fructus Hippophae powder in 16 times of 75% ethanol water for 1 hr, reflux-extracting at 85 deg.C for 3 hr, filtering with 110 mesh filter cloth, and collecting filtrate;
(3) adding 12 times of 75% ethanol water into the filter residue obtained in the step (2), carrying out reflux extraction at 85 ℃ for 2h, filtering through 110-mesh filter cloth, and collecting the filtrate;
(4) mixing the filtrates of (2) and (3), rotary steaming under reduced pressure, removing ethanol, concentrating, and lyophilizing for 72 hr to obtain extract, i.e. fructus Hippophae extract.
The present example provides a composition with α -glucosidase inhibitory effect, the total dosage of the composition extracted according to the above steps is 0.1g/mL, and the mass ratio of the mulberry leaf extract, the licorice root extract, the turmeric root extract and the sea buckthorn extract is 1:1:1: 1.
The preparation method of the composition provided in this example is different from that of example 1 in specific parameters, and the extract composition extracted according to the above method has a poor α -glucosidase inhibition rate compared to that of example 1, so that the method is not the most preferable scheme, but still has a significant synergistic effect compared to the extracts and compositions of comparative examples 1 to 10.
Example 7
The extraction methods of the extracts are the same as example 1, and this example provides a composition having an α -glucosidase inhibitory effect, wherein the total dosage of the composition is 0.1g/mL, and the mass ratio of the mulberry leaf extract, the licorice extract, the turmeric extract and the sea buckthorn extract in the composition of this example is 5:4:3.5: 5.
The difference between the present example and example 1 is that the mass ratio of the prepared mixture is different, and the extracts mixed according to the above mass ratio have less effect on the α -glucosidase than example 1, so that the most preferable scheme is not adopted, but still have significant synergistic effect compared with the extracts and compositions of comparative examples 1 to 10.
Example 8
The extraction methods of the extracts are the same as example 1, and this example provides a composition having an α -glucosidase inhibitory effect, wherein the total dosage of the composition is 0.1g/mL, and the mass ratio of the mulberry leaf extract, the licorice extract, the turmeric extract and the sea buckthorn extract in the composition of this example is 3:1.5:1.5: 2.
The difference between the present example and example 1 is that the mass ratio of the prepared mixture is different, and the extracts mixed according to the above mass ratio have less effect on the α -glucosidase than example 1, so that the most preferable scheme is not adopted, but still have significant synergistic effect compared with the extracts and compositions of comparative examples 1 to 10.
The preparation methods under different conditions and the mass ratios of different compositions are also detected to inhibit the alpha-glucosidase, and the results show that the detection shows that the extraction compositions prepared by the preparation methods under different conditions have different inhibition effects on the alpha-glucosidase. The best inhibition was obtained with the composition prepared under the conditions of example 1; the compositions of examples 7 and 8 had a less effective inhibition than the compositions of examples 1 to 3, but still had a significant synergistic effect compared to the extracts and compositions of comparative examples 1 to 10.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (7)
1. A composition with alpha-glucosidase inhibiting effect is characterized by being prepared from mulberry leaf extract, licorice extract, turmeric extract and sea buckthorn extract; the mass ratio of the mulberry leaf extract to the licorice extract to the turmeric extract to the sea buckthorn extract is (1-5) to (1-4) to (1-3.5) to (1-5);
the preparation method of the mulberry leaf extract comprises the following steps: crushing the dried mulberry leaves, sieving the crushed mulberry leaves with a 50-70-mesh sieve, adding 14-16 times of water, soaking for 0.5-2 h, boiling and extracting for 1.5-3 h, filtering, sieving with 90-110-mesh filter cloth, and collecting filtrate; adding 8-12 times of water into filter residues, boiling and extracting for 0.5-2 h, filtering through a filter cloth of 90-110 meshes, and collecting filtrate; mixing the filtrates, carrying out reduced pressure rotary evaporation and concentration, and freeze-drying for 36-72 h to finally obtain an extract, namely a mulberry leaf extract;
the preparation method of the licorice extract comprises the following steps: crushing the dried liquorice, sieving the crushed liquorice with a 50-70-mesh sieve, adding 14-16 times of water, soaking for 0.5-2 hours, boiling and extracting for 1.5-3 hours, filtering, sieving with 90-110-mesh filter cloth, and collecting filtrate; adding 8-12 times of water into filter residues, boiling and extracting for 0.5-2 h, filtering through a filter cloth of 90-110 meshes, and collecting filtrate; mixing the filtrates, carrying out reduced pressure rotary evaporation and concentration, and freeze-drying for 36-72 h to finally obtain an extract, namely a licorice extract;
the preparation method of the turmeric extract comprises the following steps: crushing the dried turmeric, sieving with a 50-70-mesh sieve, adding 14-16 times of 50-75% ethanol water, soaking for 0.5-1 h, performing reflux extraction at 70-85 ℃ for 1.5-3 h, sieving with 90-110-mesh filter cloth, and collecting filtrate; adding 9-12 times of 50-75% ethanol water into the filter residue after suction filtration, performing reflux extraction at 70-85 ℃ for 0.5-2 h, filtering through filter cloth of 90-110 meshes, and collecting filtrate; mixing the filtrates, carrying out reduced pressure rotary evaporation and concentration, and freeze-drying for 36-72 h to finally obtain an extract, namely the turmeric extract;
the preparation method of the sea buckthorn extract comprises the following steps: crushing the dried sea buckthorn, sieving the crushed sea buckthorn with a 50-70-mesh sieve, adding 14-16 times of 50-75% ethanol water, soaking for 0.5-1 h, performing reflux extraction at 70-85 ℃ for 1.5-3 h, sieving with 90-110-mesh filter cloth, and collecting filtrate; adding 9-12 times of 50-75% ethanol water into filter residues after suction filtration, performing reflux extraction at 70-85 ℃ for 0.5-2 h, filtering through filter cloth of 90-110 meshes, and collecting filtrate; and combining the filtrates, carrying out reduced pressure rotary evaporation and concentration, and freeze-drying for 36-72 h to finally obtain an extract, namely the sea buckthorn extract.
2. The composition with alpha-glucosidase inhibitory effect according to claim 1, wherein the mass ratio of the mulberry leaf extract, licorice extract, turmeric extract and sea buckthorn extract is (1-3) to (1-1.5) to (1-2).
3. The composition having an alpha-glucosidase inhibitory effect according to claim 2, wherein the mass ratio of the mulberry leaf extract, licorice extract, turmeric extract and sea buckthorn extract is (1-3) to 1:1 to (1-2).
4. The composition having an α -glucosidase inhibitory effect according to claim 3, wherein the mulberry leaf extract, licorice extract, turmeric extract and sea buckthorn extract are in a mass ratio of 1: 1.
5. The method for preparing a composition having an α -glucosidase inhibitory effect as defined in any one of claims 1 to 4, wherein the mulberry leaf extract, licorice root extract, turmeric root extract and sea buckthorn extract are prepared from mulberry leaf, licorice root, turmeric root and sea buckthorn as raw materials; mixing the above extracts to obtain the composition with alpha-glucosidase inhibiting effect.
6. The composition with alpha-glucosidase inhibitory effect according to any of claims 1-4, wherein the composition is in a dosage form selected from the group consisting of tablets, capsules, pills, powders, granules, electuary and solution.
7. A pharmaceutical product having an α -glucosidase inhibitory effect, which is characterized by being prepared from the composition having an α -glucosidase inhibitory effect according to any one of claims 1 to 4.
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| CN102258570A (en) * | 2011-07-18 | 2011-11-30 | 贵州省中国科学院天然产物化学重点实验室 | Composition for inhibiting activity of alpha-glycuronide and preparation method of composition |
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| CN101797312B (en) * | 2010-01-19 | 2011-11-16 | 南昌大学 | Method for preparing Alisma extract capable of inhibiting alpha-glucuroide activity |
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