CN113069494A - Agilawood leaf extract and health-preserving wine - Google Patents
Agilawood leaf extract and health-preserving wine Download PDFInfo
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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Abstract
The invention relates to the technical field of traditional Chinese medicines, and particularly discloses a agilawood leaf extract and health-preserving wine. The agilawood leaf extract is prepared by the following method: s01, crushing the agilawood leaves, and extracting the agilawood leaves with ethanol to obtain an ethanol extract of the agilawood leaves; s02, separating and purifying the ethanol extract of the agilawood leaves by using a silica gel column to obtain the agilawood leaf extract. Further, 0.5-10 g of agilawood leaf extract is added into every 1L of white spirit to obtain the health-preserving wine. The research shows that the agilawood leaf extract has the activity of resisting helicobacter pylori; therefore, the eaglewood leaf extract can be further used as an active ingredient to prepare medicines or health-care products with the anti-helicobacter pylori effect.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicines, and particularly relates to a agilawood leaf extract and health-preserving wine.
Background
Helicobacter pylori is a spiral, microaerophilic bacterium; helicobacter pylori infection in humans can cause gastritis, peptic ulcer, lymphoproliferative gastric lymphoma, etc. The world health organization international agency for research on cancer lists helicobacter pylori (infection) in a list of carcinogens. Therefore, the development of a Chinese medicinal material with the effect of resisting helicobacter pylori from the Chinese medicinal materials has important application value.
The agilawood leaf is leaf of Aquilaria sinensis (Lour.) Swingle of Aquilaria of Thymelaeaceae; the existing research shows that the agilawood leaf extract obtained by adopting different methods has the functions of resisting inflammation, easing pain, resisting tumors, reducing blood sugar, reducing blood fat and the like. However, the prior art does not report that the eaglewood leaves are adopted to resist helicobacter pylori independently; further, there is no report on the preparation of an extract having an anti-helicobacter pylori effect from Aquilaria sinensis. Therefore, the extract with the anti-helicobacter pylori effect is developed from the aquilaria sinensis leaves, and has important application value.
The health wine is a wine product with certain functions of preventing and treating diseases; is one of the wines popular with the people; however, the existing health wine does not report the effect of resisting helicobacter pylori, so that the health wine with the effect of resisting helicobacter pylori has important application value.
Disclosure of Invention
In view of the above, the present invention provides an extract of Aquilaria sinensis, which is prepared by a novel method and has an anti-helicobacter pylori effect.
The technical scheme of the invention is as follows:
the first aspect of the invention provides a agilawood leaf extract, which is prepared by the following method:
s01, crushing the agilawood leaves, and extracting the agilawood leaves with ethanol to obtain an ethanol extract of the agilawood leaves;
s02, separating and purifying the ethanol extract of the agilawood leaves by using a silica gel column to obtain the extract of the agilawood leaves;
the specific method for separating and purifying by using a silica gel column in the step S02 comprises the following steps:
loading the agilawood leaf ethanol extract to a silica gel column, eluting by using a mixed solvent consisting of chloroform and methanol in a volume ratio of 90-85: 10-15, and discarding the elution part; and then eluting with a mixed solvent consisting of chloroform and methanol in a volume ratio of 80-75: 20-25, collecting an eluted part eluted by the mixed solvent consisting of chloroform and methanol in a volume ratio of 80-75: 20-25, and concentrating and drying the eluted part to obtain a silica gel eluted part, namely the agilawood leaf extract.
In order to develop the eaglewood leaf extract with the anti-helicobacter pylori effect, a large number of experimental studies show that after the eaglewood leaf is extracted by ethanol, the eaglewood leaf is separated and purified by adopting a silica gel column under the conditions, and the obtained silica gel elution part is enriched with a large number of anti-helicobacter pylori effective components and has excellent anti-helicobacter pylori effect. The inventor needs to emphasize that the specific method for separating and purifying by using the silica gel column is the key point of the invention, the elution conditions of the method are different (the elution conditions of the invention are that the mixed solvent composed of chloroform and methanol with the volume ratio of 90-85: 10-15 is used for elution firstly, and then the mixed solvent composed of chloroform and methanol with the volume ratio of 80-75: 20-25 is used for elution later), and the obtained silica gel elution parts contain completely different effective components; the method also adopts silica gel column to separate and purify the eaglewood leaves, and if the elution conditions are different, the silica gel elution part with the anti-helicobacter pylori effect can not be obtained.
Preferably, the ethanol in the step S01 is ethanol with a volume fraction of 85-95%.
Preferably, the extraction in step S01 is a heating reflux extraction.
Preferably, the specific method of heating reflux extraction described in step S01 is:
crushing the agilawood leaves, and then adding ethanol to carry out heating reflux extraction for 1-2 h; wherein the dosage ratio of the agilawood leaves to the ethanol is 1g: 5-10 mL.
Preferably, the specific method of heating reflux extraction described in step S01 is:
crushing the agilawood leaves, and then adding ethanol for heating reflux extraction, wherein the extraction time is 1.5 h; wherein the dosage ratio of the agilawood leaves to the ethanol is 1g:8 mL.
Preferably, the specific method for separating and purifying by silica gel column in step S02 is:
loading the agilawood leaf ethanol extract to a silica gel column, eluting by using a mixed solvent which is 2-4 times of the column volume and consists of chloroform and methanol and has the volume ratio of 90-85: 10-15, and discarding the elution part; then eluting with a mixed solvent of chloroform and methanol in a volume ratio of 80-75: 20-25 of 3-5 times the volume of the column, collecting an eluted part eluted by the mixed solvent of chloroform and methanol in a volume ratio of 80-75: 20-25, and concentrating and drying the eluted part to obtain a silica gel eluted part, namely the agilawood leaf extract.
Most preferably, the specific method for separating and purifying by silica gel column in step S02 is:
loading the ethanol extract of the agilawood leaves on a silica gel column, eluting by using a mixed solvent consisting of chloroform and methanol with a volume ratio of 88:12 of 3 times of the column volume, and discarding the elution part; then eluting with a mixed solvent of chloroform and methanol with a volume ratio of 76:24 of 4 times of the column volume, collecting the eluted part eluted by the mixed solvent of chloroform and methanol with a volume ratio of 76:24, concentrating and drying the eluted part to obtain a silica gel eluted part, namely the eaglewood leaf extract.
Further preferably, the eaglewood leaf extract comprises a silica gel elution part and a macroporous resin elution part; the macroporous resin elution part is prepared by the following method:
s11, crushing the agilawood leaves, and extracting the agilawood leaves with ethanol to obtain an ethanol extract of the agilawood leaves;
s12, separating and purifying the ethanol extract of the agilawood leaves by using a macroporous resin column to obtain a macroporous resin elution part;
the specific conditions for separating and purifying by using the macroporous resin column in the step S12 are as follows:
loading the ethanol extract of the agilawood leaves on a macroporous resin column, and eluting and removing impurities by using ethanol with the volume fraction of 25% and 5 times of the volume of the column; eluting with 40% ethanol 5 times the column volume, collecting the effective fraction eluted with 40% ethanol, concentrating and drying to obtain the macroporous resin eluate.
The inventor further researches and discovers that after the silica gel elution part and the macroporous resin elution part are combined, the silica gel elution part and the macroporous resin elution part can generate a synergistic anti-helicobacter pylori effect; its anti-helicobacter pylori effect is further improved.
Preferably, in the agilawood leaf extract, the mass ratio of the silica gel elution part to the macroporous resin elution part is 2-4: 1.
Preferably, the ethanol in the step S11 is ethanol with a volume fraction of 85-95%.
Preferably, the extraction in step S11 is a heating reflux extraction.
Preferably, the specific method of heating reflux extraction described in step S11 is:
crushing the agilawood leaves, and then adding ethanol to carry out heating reflux extraction for 1-2 h; wherein the dosage ratio of the agilawood leaves to the ethanol is 1g: 5-10 mL.
Preferably, the specific method of heating reflux extraction described in step S11 is:
crushing the agilawood leaves, and then adding ethanol for heating reflux extraction, wherein the extraction time is 1.5 h; wherein the dosage ratio of the agilawood leaves to the ethanol is 1g:8 mL.
In a second aspect of the present invention, an application of the aforementioned eaglewood leaf extract in the preparation of a drug or health product with an anti-helicobacter pylori effect is provided.
In a third aspect of the invention, a health preserving wine is provided, which comprises the agaric leaf extract.
Preferably, 0.5-10 g of the agilawood leaf extract disclosed by any one of claims 1-6 is added into 1L of white spirit to obtain the health-preserving wine.
Preferably, the white spirit is Maotai-flavor white spirit, Luzhou-flavor white spirit, fen-flavor white spirit, drug-flavor white spirit or mixed-flavor white spirit.
Has the advantages that: the invention provides a Chinese eaglewood leaf extract with the function of resisting helicobacter pylori for the first time; researches show that the agilawood leaf extract prepared by the silica gel column chromatography method has excellent anti-helicobacter pylori effect. Further research shows that the agilawood leaf extract obtained by combining the silica gel elution part prepared by the method with the macroporous resin elution part can generate a synergistic anti-helicobacter pylori effect; its anti-helicobacter pylori effect is further improved. Because the eaglewood leaf extract has the effect of resisting helicobacter pylori, the active ingredients of the eaglewood leaf extract can be used for preparing medicines or health-care products with the effect of resisting helicobacter pylori; meanwhile, the active ingredients of the agilawood leaf extract are added into the white spirit, so that the white spirit has the effect of resisting helicobacter pylori.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the present invention is not limited to these examples in any way.
Example 1 preparation of extract of Aquilaria sinensis leaf
S01, crushing 500g of dried eaglewood leaves, then carrying out heating reflux extraction for 1.5h by using 4L of ethanol (the volume fraction is 95%), and concentrating and drying the extracting solution to obtain an eaglewood leaf ethanol extract;
s02, separating and purifying the ethanol extract of the agilawood leaves by using a silica gel column to obtain the extract of the agilawood leaves;
the specific method for separating and purifying by using a silica gel column in the step S02 comprises the following steps:
loading the agilawood leaf ethanol extract to a silica gel column (the weight of the silica gel is 30 times of that of the agilawood leaf ethanol extract, the mesh number of the silica gel is 200-300 meshes), eluting by using a mixed solvent which is 3 times of the column volume and consists of chloroform and methanol and has the volume ratio of 88:12, and discarding the elution part; then eluting with a mixed solvent of chloroform and methanol with a volume ratio of 76:24 of 4 times of the column volume, collecting the eluted part eluted by the mixed solvent of chloroform and methanol with a volume ratio of 76:24, concentrating and drying the eluted part to obtain a silica gel eluted part, namely the eaglewood leaf extract.
Example 2 preparation of extract of Aquilaria sinensis leaf
S01, crushing 500g of dried eaglewood leaves, then carrying out heating reflux extraction for 1.5h by using 4L of ethanol (the volume fraction is 95%), and concentrating and drying the extracting solution to obtain an eaglewood leaf ethanol extract;
s02, applying the linaloe leaf ethanol extract to a silica gel column (the weight of the silica gel is 30 times of that of the linaloe leaf ethanol extract, the mesh number of the silica gel is 200-300 meshes), eluting by using a mixed solvent composed of chloroform and methanol with the volume ratio of 88:12 and 3 times of the column volume, and discarding the elution part; then eluting with a mixed solvent of chloroform and methanol with a volume ratio of 76:24 of 4 times of the column volume, collecting the eluted part eluted by the mixed solvent of chloroform and methanol with a volume ratio of 76:24, concentrating and drying the eluted part to obtain a silica gel eluted part;
s11, crushing 500g of dried eaglewood leaves, then carrying out heating reflux extraction for 1.5h by using 4L of ethanol (the volume fraction is 95%), and concentrating and drying the extracting solution to obtain an eaglewood leaf ethanol extract;
s12, loading the ethanol extract of the agilawood leaves on a macroporous resin column (the weight of the macroporous resin is 50 times that of the ethanol extract of the agilawood leaves, and D101 type macroporous resin is selected as the macroporous resin), and eluting with ethanol with the volume fraction of 25% and 5 times of the column volume to remove impurities; eluting with 40% ethanol 5 times the column volume, collecting the effective fraction eluted with 40% ethanol, concentrating and drying to obtain the macroporous resin eluate;
and combining the prepared silica gel elution part with the macroporous resin elution part according to the mass ratio of 3:1 to obtain the agilawood leaf extract.
Comparative example 1 preparation of extract of eaglewood leaves
S01, crushing 500g of dried eaglewood leaves, then carrying out heating reflux extraction for 1.5h by using 4L of ethanol (the volume fraction is 95%), and concentrating and drying the extracting solution to obtain an eaglewood leaf ethanol extract;
s02, separating and purifying the ethanol extract of the agilawood leaves by using a silica gel column to obtain the extract of the agilawood leaves;
the specific method for separating and purifying by using a silica gel column in the step S02 comprises the following steps:
loading the agilawood leaf ethanol extract to a silica gel column (the weight of the silica gel is 30 times of that of the agilawood leaf ethanol extract, the mesh number of the silica gel is 200-300 meshes), eluting by using a mixed solvent which is 3 times of the column volume and consists of chloroform and methanol in a volume ratio of 95:5, and discarding the elution part; then eluting with a mixed solvent of chloroform and methanol with a volume ratio of 85:15 of 4 times of the column volume, collecting the eluted part eluted by the mixed solvent of chloroform and methanol with a volume ratio of 85:15, concentrating and drying the eluted part to obtain a silica gel eluted part, namely the eaglewood leaf extract.
Comparative example 1 is different from example 1 in that the elution conditions of the silica gel column are different; example 1 elution was carried out with a mixed solvent of chloroform and methanol at a volume ratio of 88: 12; then eluting by using a mixed solvent consisting of chloroform and methanol with the volume ratio of 76: 24; in contrast, in comparative example 1, the elution was carried out with a mixed solvent of chloroform and methanol at a volume ratio of 95: 5; then, the mixture was eluted with a mixed solvent of chloroform and methanol at a volume ratio of 85: 15.
Comparative example 2 preparation of extract of Aquilaria sinensis leaf
S01, crushing 500g of agilawood leaves, then carrying out heating reflux extraction for 1.5h by using 4L of ethanol (the volume fraction is 95%), and concentrating and drying the extracting solution to obtain an ethanol extract of the agilawood leaves;
s02, separating and purifying the ethanol extract of the agilawood leaves by using a silica gel column to obtain the extract of the agilawood leaves;
the specific method for separating and purifying by using a silica gel column in the step S02 comprises the following steps:
loading the agilawood leaf ethanol extract to a silica gel column (the weight of the silica gel is 30 times of that of the agilawood leaf ethanol extract, the mesh number of the silica gel is 200-300 meshes), eluting by using a mixed solvent which is 3 times of the column volume and consists of chloroform and methanol in a volume ratio of 75:25, and discarding the elution part; then eluting with a mixed solvent of chloroform and methanol with a volume ratio of 70:30 of 4 times of the column volume, collecting the eluted part eluted by the mixed solvent of chloroform and methanol with a volume ratio of 70:30, concentrating and drying the eluted part to obtain a silica gel eluted part, namely the eaglewood leaf extract.
Comparative example 2 is different from example 1 in that the elution conditions of the silica gel column are different; example 1 elution was carried out with a mixed solvent of chloroform and methanol at a volume ratio of 88: 12; then eluting by using a mixed solvent consisting of chloroform and methanol with the volume ratio of 76: 24; in contrast, in comparative example 2, the elution was carried out with a mixed solvent of chloroform and methanol at a volume ratio of 75: 25; then, the mixture was eluted with a mixed solvent of chloroform and methanol at a volume ratio of 70: 30.
Experimental example 1 Aquilaria sinensis leaf extract Activity test against helicobacter pylori
Testing the anti-helicobacter pylori activity of the agilawood leaf extract by adopting an oxford cup method; the specific method comprises the following steps:
under aseptic conditions, 0.3mL of a helicobacter pylori ATCC 700392-containing bacterial liquid (concentration 2.5X 10)7one/mL) is added into the culture medium, and then an L-shaped smearing rod (after sterilization) is used for smearing evenly; then drying for 8min at room temperature, and then putting the sterilized oxford cup into a culture dish by using sterile tweezers; then 100 mul of liquid medicine (containing the eaglewood leaf extract to be detected) with the concentration of 100mg/mL is added into the oxford cup; adding the liquid medicine, placing the culture dish into an incubator for culture, and measuring the diameter of the bacteriostatic circle after culturing for 16h at 37 ℃.
The eaglewood leaf extracts prepared in examples 1-2 and comparative examples 1-2 were tested by the above method; the eaglewood leaf extract prepared in each example is tested repeatedly for 10 times, the average value of the tests is taken, and the test results are shown in table 1; wherein the larger the inhibition zone, the better the anti-helicobacter pylori activity.
TABLE 1 results of anti-helicobacter pylori activity of extract of Aquilaria sinensis
| Diameter/mm of bacteriostatic circle | |
| Example 1 extract of Aquilaria sinensis leaf | 30.5 |
| Example 2 extract of Aquilaria sinensis leaf | 45.3 |
| Comparative example 1 extract of eaglewood leaves | 10.2 |
| Comparative example 2 extract of Aquilaria sinensis leaf | Without bacteriostatic zone |
As can be seen from the tests in Table 1, the diameter of the zone of inhibition of the eaglewood leaf extract prepared in example 1 is 30.5mm, which indicates that the silica gel elution part prepared by the method of the invention has a certain anti-helicobacter pylori effect; further, as can be seen from example 2, the inhibition zone of the extract is significantly greater than that of the eaglewood leaf extract prepared in example 1, which indicates that after the eaglewood leaf extract is combined by the silica gel elution part and the macroporous resin elution part, the anti-helicobacter pylori effect of the extract is further enhanced, and probably the two exert synergistic anti-helicobacter pylori effects.
In addition, the inhibition zone of the agilawood leaf extract prepared in the comparative example 1 is far smaller than that of the agilawood leaf extract prepared in the example 1, and the inhibition zone of the agilawood leaf extract prepared in the comparative example 2 is not higher than that of the agilawood leaf extract; this shows that the elution condition of the silica gel column is very critical for preparing the effective part with the function of resisting helicobacter pylori; the existing research shows that only mixed solvent composed of chloroform and methanol with the volume ratio of 90-85: 10-15 is used for elution; and then eluting with a mixed solvent consisting of chloroform and methanol in a volume ratio of 80-75: 20-25 to obtain a silica gel elution part which has the anti-helicobacter pylori activity, wherein the silica gel elution part prepared under other silica gel elution conditions does not have the anti-helicobacter pylori activity or has the insignificant anti-helicobacter pylori activity.
Example 3 preparation of health preserving wine
And (3) adding 0.5g of the agilawood leaf extract prepared in the example 1 into 1L of Maotai-flavor liquor to obtain the health-preserving liquor.
Example 4 preparation of health preserving wine
And 2g of the agilawood leaf extract prepared in the example 2 is added into 1L of Maotai-flavor liquor, so that the health-preserving liquor is obtained.
Example 5 preparation of health preserving wine
And 5g of the agilawood leaf extract prepared in the example 1 is added into 1L of Maotai-flavor liquor, so that the health-preserving liquor is obtained.
Example 6 preparation of health preserving wine
And (3) adding 0.5g of the agilawood leaf extract prepared in the example 2 into 1L of strong aromatic Chinese spirits to obtain the health-preserving wine.
Example 7 preparation of health preserving wine
And (3) adding 0.5g of the agilawood leaf extract prepared in the example 1 into 1L of fen-flavor liquor to obtain the health-preserving liquor.
Example 8 preparation of health preserving wine
And (3) adding 0.5g of the agilawood leaf extract prepared in the example 1 into 1L of the phoenix-flavor liquor to obtain the health-preserving liquor.
Example 9 preparation of health preserving wine
And (3) adding 0.5g of the agilawood leaf extract prepared in the example 1 into 1L of medicinal-flavor liquor to obtain the health-preserving liquor.
Example 10 preparation of health preserving wine
And (3) adding 0.5g of the agilawood leaf extract prepared in the example 1 into 1L of the mixed-flavor liquor to obtain the health-preserving liquor.
Claims (10)
1. The eaglewood leaf extract is characterized by being prepared by the following method:
s01, crushing the agilawood leaves, and extracting the agilawood leaves with ethanol to obtain an ethanol extract of the agilawood leaves;
s02, separating and purifying the ethanol extract of the agilawood leaves by using a silica gel column to obtain the extract of the agilawood leaves;
the specific method for separating and purifying by using a silica gel column in the step S02 comprises the following steps:
loading the agilawood leaf ethanol extract to a silica gel column, eluting by using a mixed solvent consisting of chloroform and methanol in a volume ratio of 90-85: 10-15, and discarding the elution part; and then eluting with a mixed solvent consisting of chloroform and methanol in a volume ratio of 80-75: 20-25, collecting an eluted part eluted by the mixed solvent consisting of chloroform and methanol in a volume ratio of 80-75: 20-25, and concentrating and drying the eluted part to obtain a silica gel eluted part, namely the agilawood leaf extract.
2. The eaglewood leaf extract according to claim 1, wherein the ethanol in step S01 is 85-95% by volume.
3. The eaglewood leaf extract according to claim 1, wherein the extraction in the step S01 is a heating reflux extraction.
4. The agilawood extract according to claim 3, wherein the specific method for heating reflux extraction in step S01 comprises:
crushing the agilawood leaves, and then adding ethanol to carry out heating reflux extraction for 1-2 h; wherein the dosage ratio of the agilawood leaves to the ethanol is 1g: 5-10 mL.
5. The eaglewood leaf extract according to claim 3, wherein the specific method of the heating reflux extraction in the step S01 is:
crushing the agilawood leaves, and then adding ethanol for heating reflux extraction, wherein the extraction time is 1.5 h; wherein the dosage ratio of the agilawood leaves to the ethanol is 1g:8 mL.
6. The eaglewood leaf extract according to claim 1, wherein the specific method for separation and purification by silica gel column in step S02 is:
loading the agilawood leaf ethanol extract to a silica gel column, eluting by using a mixed solvent which is 2-4 times of the column volume and consists of chloroform and methanol and has the volume ratio of 90-85: 10-15, and discarding the elution part; then eluting with a mixed solvent consisting of chloroform and methanol in a volume ratio of 80-75: 20-25 and 3-5 times the column volume, collecting an eluted part eluted by the mixed solvent consisting of chloroform and methanol in a volume ratio of 80-75: 20-25, and concentrating and drying the eluted part to obtain a silica gel eluted part, namely the agilawood leaf extract;
most preferably, the specific method for separating and purifying by using the silica gel column is as follows:
loading the ethanol extract of the agilawood leaves on a silica gel column, eluting by using a mixed solvent consisting of chloroform and methanol with a volume ratio of 88:12 of 3 times of the column volume, and discarding the elution part; then eluting with a mixed solvent of chloroform and methanol with a volume ratio of 76:24 of 4 times of the column volume, collecting the eluted part eluted by the mixed solvent of chloroform and methanol with a volume ratio of 76:24, concentrating and drying the eluted part to obtain a silica gel eluted part, namely the eaglewood leaf extract.
7. Use of the eaglewood leaf extract according to any one of claims 1 to 6 in the preparation of a medicine or health-care product with an anti-helicobacter pylori effect.
8. A health wine comprising the eaglewood leaf extract according to any one of claims 1 to 6.
9. The health wine according to claim 8, wherein 0.5-10 g of the eaglewood leaf extract according to any one of claims 1-6 is added into 1L of white wine to obtain the health wine.
10. The health preserving wine as claimed in claim 8, wherein the white spirit is Maotai-flavor, Luzhou-flavor, fen-flavor, drug-flavor or mixed-flavor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110373893.4A CN113069494B (en) | 2021-04-07 | 2021-04-07 | Application of agilawood leaf extract in preparation of medicine with anti-helicobacter pylori effect |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110373893.4A CN113069494B (en) | 2021-04-07 | 2021-04-07 | Application of agilawood leaf extract in preparation of medicine with anti-helicobacter pylori effect |
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| Publication Number | Publication Date |
|---|---|
| CN113069494A true CN113069494A (en) | 2021-07-06 |
| CN113069494B CN113069494B (en) | 2022-05-31 |
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