CN111494500B - Black tiger pericarp extract and preparation method and application thereof - Google Patents
Black tiger pericarp extract and preparation method and application thereof Download PDFInfo
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- CN111494500B CN111494500B CN202010415281.2A CN202010415281A CN111494500B CN 111494500 B CN111494500 B CN 111494500B CN 202010415281 A CN202010415281 A CN 202010415281A CN 111494500 B CN111494500 B CN 111494500B
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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
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/57—Magnoliaceae (Magnolia family)
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- A—HUMAN NECESSITIES
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- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/20—Antivirals for DNA viruses
- A61P31/22—Antivirals for DNA viruses for herpes viruses
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- 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
- A61K2236/30—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
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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The invention relates to the technical field of natural medicinal chemistry, and particularly discloses a black tiger pericarp extract and a preparation method and application thereof. The preparation method of the black tiger pericarp extract comprises the following steps: taking black tiger pericarp, adding ethanol, carrying out ultrasonic extraction at 50-70 ℃, concentrating and drying an extracting solution to obtain the black tiger pericarp alcohol extract, namely the black tiger pericarp extract. The extract of the pericarp of Kadsura coccinea prepared by the method has certain antiviral activity.
Description
Technical Field
The invention relates to the technical field of natural medicinal chemistry, in particular to a black tiger pericarp extract and a preparation method and application thereof.
Background
Kadsura coccinea is a perennial evergreen climbing vine of Schisandra of Schisandraceae, also known as Kadsura coccinea, caulis Endadae Phaseoloidis, Buforna, Zuguifeng, etc., and is widely distributed in Hunan, Guizhou, Fujian, Yunnan, etc. The black cutworm is an edible and medicinal dual-purpose wild fruit with high development potential, has high medicinal value commonly called sargentgloryvine stem and is an important Chinese medicinal material; the fruit is large and purple red, is polymerized berry, has sour and sweet taste, can be eaten raw, and contains abundant amino acids, mineral elements and polyphenol substances. At present, the chemical components, the pharmacological effects, the volatile oil components and the like of the tiger rhizome are researched more, and the active components such as polyphenols, lignins, terpenes, volatile oil and the like have the effects of antioxidation, anti-inflammation, bacteriostasis, tumor resistance, liver protection, HIV resistance, anticoagulation, blood fat regulation and the like.
At present, few antiviral research reports on the black tiger pericarp exist, so that whether the black tiger pericarp contains an antiviral component or not, which viruses can be resisted by the effective component, and how to prepare the antiviral component from the black tiger pericarp exist; to be further investigated by those skilled in the art.
Disclosure of Invention
In order to overcome the defects of the prior art in researching the black tiger pericarp, a preparation method of the black tiger pericarp extract is provided.
The technical scheme of the invention is as follows:
a preparation method of a black tiger pericarp extract comprises the following steps:
taking black tiger pericarp, adding ethanol, carrying out ultrasonic extraction at 50-70 ℃, concentrating and drying an extracting solution to obtain the black tiger pericarp alcohol extract, namely the black tiger pericarp extract.
Preferably, the ethanol is ethanol with the volume fraction of 70-95%.
Most preferably, the ethanol is 80% ethanol by volume fraction.
Preferably, the feed-liquid ratio of the black tiger skin to the ethanol is 1 g: 20-30 mL.
Most preferably, the feed-liquid ratio of the black tiger skin to the ethanol is 1 g: 25 mL.
Preferably, the extraction time is 30-50 min.
Most preferably, the extraction time is 35 min.
Preferably, the power of ultrasonic extraction is 150-200W.
Most preferably, the power of the ultrasound extraction is 180W.
Preferably, the preparation method of the black tiger pericarp extract further comprises a silica gel column chromatography step; the silica gel column chromatography step specifically comprises:
putting the black tiger pericarp alcohol extract on a silica gel column, and firstly, mixing the extract with a solvent with a volume ratio of 92-90: eluting and removing impurities by using a mixed solvent consisting of 8-10 parts of chloroform and methanol; and the reuse volume ratio is 85-87: and (3) eluting with a mixed solvent consisting of chloroform and methanol of 15-13, collecting the eluted part, concentrating and drying to obtain the black tiger pericarp extract.
Most preferably, the silica gel column chromatography step specifically comprises:
putting the black tiger pericarp alcohol extract on a silica gel column, and firstly mixing the extract with a solvent with a volume ratio of 90: 10, eluting with a mixed solvent consisting of chloroform and methanol to remove impurities; reuse volume ratio 85: 15, eluting with a mixed solvent of chloroform and methanol, collecting the eluted part, concentrating and drying to obtain the black tiger pericarp extract.
Most preferably, the black cutworm pericarp alcohol extract is taken and put on a silica gel column, and the volume ratio of 3-5 times of column volume is firstly 92-90: eluting and removing impurities by using a mixed solvent consisting of 8-10 parts of chloroform and methanol; and then, the volume ratio of 3-5 times of column volume is 85-87: and (3) eluting with a mixed solvent consisting of chloroform and methanol of 15-13, collecting the eluted part, concentrating and drying to obtain the black tiger pericarp extract.
Preferably, the black tiger pericarp extract is a black tiger pericarp extract with antiviral effect.
The invention also provides the black tiger pericarp extract prepared by the preparation method.
The invention also provides an application of the black tiger pericarp extract in preparation of antiviral drugs.
Preferably, the virus is coxsackie virus and herpes simplex virus.
Has the advantages that: the invention researches the effective components in the black tiger pericarp for the first time; the black tiger pericarp extract is found to have the effect of resisting coxsackie virus and herpes simplex virus for the first time; the extract particularly shows stronger inhibitory activity to coxsackie virus; in addition, the black tiger pericarp alcohol extract further enhances the inhibition effect of the black tiger pericarp extract obtained after the silica gel column chromatography technology of the invention on the coxsackie virus, because the silica gel column chromatography technology of the invention enriches a large amount of active ingredients for resisting the coxsackie virus.
Detailed Description
The present invention is further explained below with reference to specific examples, which are not intended to limit the present invention in any way.
Example 1 preparation of extract from pericarp of Kadsura coccinea
Taking fresh black tiger pericarp, and mixing the materials according to a material-liquid ratio of 1 g: adding ethanol with volume fraction of 80% into 25mL, performing ultrasonic extraction at 60 deg.C for 35min (power of ultrasonic extraction is 180W), concentrating the extractive solution under reduced pressure to obtain extract, and drying the extract to obtain the extract of black tiger pericarp.
Example 2 preparation of extract from the pericarp of Kadsura coccinea
(1) Taking fresh black tiger pericarp, and mixing the materials according to a material-liquid ratio of 1 g: adding ethanol with volume fraction of 80% into 25mL, performing ultrasonic extraction at 60 deg.C for 35min (power of ultrasonic extraction is 180W), concentrating the extractive solution under reduced pressure to obtain extract, and drying the extract to obtain the extract;
(2) putting the black tiger pericarp alcohol extract into a silica gel column (the silica gel column is filled with 200-mesh 300-mesh silica gel, the weight of the silica gel is 30 times of the weight of the black tiger pericarp alcohol extract), and mixing the extract with a solvent with the volume ratio of 5 times of the column volume of 90: 10, eluting with a mixed solvent consisting of chloroform and methanol to remove impurities; then the volume ratio of 5 times of column volume is 85: 15, eluting with a mixed solvent of chloroform and methanol, collecting the eluted part, concentrating and drying to obtain the black tiger pericarp extract.
Comparative example 1 preparation of extract from pericarp of black tiger
(1) Taking fresh black tiger pericarp, and mixing the materials according to a material-liquid ratio of 1 g: adding ethanol with volume fraction of 80% into 25mL, performing ultrasonic extraction at 60 deg.C for 35min (power of ultrasonic extraction is 180W), concentrating the extractive solution under reduced pressure to obtain extract, and drying the extract to obtain the extract;
(2) putting the black tiger pericarp alcohol extract into a silica gel column (the silica gel column is filled with 200-mesh 300-mesh silica gel, the weight of the silica gel is 30 times of the weight of the black tiger pericarp alcohol extract), and firstly, using a silica gel column with the volume ratio of 5 times of the column volume of 85: 15, eluting with a mixed solvent consisting of chloroform and methanol to remove impurities; then the volume ratio of 5 times of column volume is 80: 20, eluting with a mixed solvent of chloroform and methanol, collecting the eluted part, concentrating and drying to obtain the black tiger pericarp extract.
The difference between the comparative example 1 and the example 2 is that the effective components in the obtained black tiger pericarp extract are different under different silica gel elution conditions.
Example 3 black tiger pericarp extract antiviral activity assay
The test method comprises the following steps: after MDCK cells were grown in a monolayer in a 96-well cell culture plate, the supernatant was discarded by washing with Eagle's solution, and then 100. mu.L of 100TCID was added to each well50The virus solution of (4); then, 100. mu.L of a MEM-diluted chemical solution was added to each well, and the mixture was incubated at 37 ℃ with 5% CO2And incubating for 48h in an incubator with saturated humidity, removing old liquid in a 96-well plate, and adding 10 mu L of CCK8 solution into each well; incubating in incubator for 1h, and using enzyme-labeling instrumentAbsorbance at 450nm was measured. Setting cell control group and virus control group at the same time, and calculating IC50The value is obtained.
The test viruses are coxsackievirus B3(VR30 strain) and herpes simplex virus type I (SM44 strain);
the test drugs were the extracts of the pericarp of black tiger prepared in examples 1 and 2 and comparative example 1.
IC50The calculation method is as follows: the concentration of the drug is taken as the X axis and the cell survival rate is taken as the Y axis, and the concentration of the drug corresponding to the 50 percent cell survival rate is calculated and is IC when the graph is drawn on Origin50(ii) a The cell survival rate is (drug-added well OD-virus control OD)/(cell control OD-virus control OD) × 100%.
TABLE 1 test results of the antiviral activity of the extract of Kadsura coccinea
As can be seen from the data in Table 1, the black tiger pericarp extract of the invention has the effect of resisting coxsackie virus and herpes simplex virus; wherein the extract has better anti-coxsackie virus activity than herpes simplex virus.
Wherein, the black tiger peel extract prepared by the silica gel column chromatography technology of the invention in the embodiment 2 has better coxsackie virus resistant function; however, the effect of the medicine against herpes simplex virus is not improved obviously. This shows that the significant anti-coxsackie virus active ingredient is obtained by the enrichment of the silica gel column chromatography method of the invention; however, the silica gel column chromatography method does not enrich a large amount of anti-herpes simplex virus active ingredients.
As can be seen from comparative example 1, the black tiger pericarp extract prepared under other silica gel column chromatography conditions has obviously reduced activity against coxsackie virus and herpes simplex virus; the result shows that the specific conditions of the silica gel column chromatography play a decisive factor for the enrichment of the antiviral active ingredient, and different conditions of the silica gel column chromatography can lead to the obtainment of active ingredients with different activities. The silica gel column chromatography condition of the invention is the best condition for obtaining the antiviral active ingredient by enrichment.
Claims (7)
1. A preparation method of a black tiger pericarp extract with antiviral effect is characterized by comprising the following steps:
taking black tiger pericarp, adding ethanol, carrying out ultrasonic extraction at 50-70 ℃, and concentrating and drying an extracting solution to obtain the black tiger pericarp alcohol extract; wherein the ethanol is 70-95% ethanol by volume fraction; the material-liquid ratio of the black tiger skin to the ethanol is 1 g: 20-30 mL; the extraction time is 30-50 min, and the ultrasonic extraction power is 150-200W;
putting the black tiger pericarp alcohol extract on a silica gel column, and firstly using a silica gel column with the volume ratio of 5 times of the column volume of 90: 10, eluting with a mixed solvent consisting of chloroform and methanol to remove impurities; then the volume ratio of 5 times of column volume is 85: 15, eluting with a mixed solvent of chloroform and methanol, collecting the eluted part, concentrating and drying to obtain the black tiger pericarp extract.
2. The method of claim 1, wherein the ethanol is 80% ethanol by volume.
3. The method for preparing the extract of pericarp of black tiger having antiviral effect as claimed in claim 1, wherein the ratio of the material to the liquid of the skin of black tiger to the ethanol is 1 g: 25 mL.
4. The method of claim 1, wherein the extraction time is 35 min.
5. The method of claim 1, wherein the ultrasonic extraction power is 180W.
6. An antiviral extract of the pericarp of black tiger prepared by the method of any one of claims 1 to 5.
7. The use of the extract of pericarp of black tiger as claimed in claim 6 for preparing antiviral drugs.
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| CN113439812A (en) * | 2020-10-26 | 2021-09-28 | 通道黑老虎电子商务有限公司 | Black tiger acetic acid beverage and preparation method thereof |
| CN114259444B (en) * | 2022-01-18 | 2023-06-02 | 湖南环境生物职业技术学院 | A black tiger fruit extract with whitening effect, its preparation method and application in preparing bath lotion with whitening effect |
| CN114467981B (en) * | 2022-01-21 | 2023-03-21 | 湖南环境生物职业技术学院 | Black tiger fruit extract with antibacterial and antiviral effects, preparation method thereof and application thereof in preparing handmade soap or cleaning agent |
| CN115057770B (en) * | 2022-06-09 | 2023-06-13 | 湖南农业大学 | Preparation method of triterpenic acid compound in black tiger peel |
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