CN111437236B - Dendrobium extract, preparation method and application thereof - Google Patents
Dendrobium extract, preparation method and application thereof Download PDFInfo
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- A61K8/00—Cosmetics or similar toiletry preparations
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A61K36/898—Orchidaceae (Orchid family)
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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
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Abstract
The invention discloses a dendrobe extracting solution, a preparation method and application thereof. The preparation method comprises the steps of flash extraction, high-pH high-concentration ethanol soaking, activated carbon decolorization, filtration to obtain a dendrobe ethanol extract, concentration at 50 ℃ and evaporation to remove ethanol to obtain an ethanol extract part; extracting the filter residue with water, performing enzymolysis, and filtering to obtain a water extract part; mixing the ethanol extract and water extract, and adding propylene glycol to obtain herba Dendrobii extractive solution. The preparation conditions are mild, the effective components of the dendrobium are protected to the greatest extent, and the dendrobium can be used as a full-component raw material liquid to be added into cosmetics to moisten skin, effectively inhibit bacteria, remove acnes, diminish inflammation and resist allergy.
Description
Technical Field
The invention relates to the technical field of extraction and processing of active ingredients of medicinal plants, and particularly relates to a dendrobium extract, a preparation method and application thereof.
Background
Dendrobii is called "Qianjin grass" and is called "panda of medicine world" by the international medicinal plant world. In recent years, scholars at home and abroad carry out systematic research on chemical components of dendrobium and find that the dendrobium contains various chemical components such as alkaloids, polysaccharides, flavonoids, phenanthrenes, sesquiterpenes, bibenzenes and the like, and the existing research shows that: the dendrobine has antibacterial, antiinflammatory, tumor inhibiting, immunity regulating and antioxidant effects; the dendrobe polysaccharide can stimulate macrophage action, enhance immunity and regulate biological reaction; the flavonoids and phenolic compounds have antibacterial, antiviral, antiinflammatory, antitumor and antioxidant activities; the phenanthrene and bibenzyl compounds have the effects of resisting tumor, oxidation and mutation, and the like; the sesquiterpene compound can effectively enhance the immunity of organisms and inhibit the growth of staphylococcus aureus, escherichia coli and candida albicans. Health foods, medicines and cosmetics related to the dendrobium concept are popular among consumers.
The extraction technology of the effective components of the dendrobium has become mature day by day, and a plurality of preparation processes and methods of the dendrobium extract exist, but the problems of high energy consumption, large amount of used solvent, large loss of target effective components, long extraction time and the like generally exist in the traditional extraction technology, and although the new extraction technology such as the microwave-assisted extraction method, the ultrasonic-assisted extraction method and other methods make up the defects of the traditional extraction method to a certain extent, reduce the consumption of solvent and energy, and improve the extraction efficiency and yield, the process operation is more complex, the equipment requirement and the extraction technical requirement are higher, and the method is difficult to be applied to mass production.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention provides the preparation method which has the advantages of small investment of production equipment, simple process operation, easy realization of conditions and low energy consumption of the preparation process, and the dendrobium extract with all components is obtained. The preparation method comprises the steps of comprehensively adopting flash extraction, high-pH high-concentration ethanol soaking and activated carbon decoloration, filtering to obtain dendrobe ethanol extract, and concentrating and steaming at 50 ℃ to remove ethanol to obtain an ethanol extract part; extracting the filter residue with water, performing enzymolysis, and filtering to obtain a water extract part; mixing the ethanol extract and water extract, and adding propylene glycol to obtain herba Dendrobii extractive solution. The preparation conditions are mild, the effective components of the dendrobium are protected to the greatest extent, and the dendrobium can be used as a full-component raw material liquid to be added into cosmetics, so that the dendrobium can moisten skin, effectively inhibit bacteria, remove acnes, diminish inflammation and resist allergy.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides a preparation method of a dendrobium extract, which comprises the following steps:
(1) selecting and processing raw materials: taking fresh stem segments of herba Dendrobii, cleaning with water, and cutting into small segments;
(2) flash extraction: mixing the small sections obtained in step (1) with ethanol solution, extracting in a flash extractor at room temperature, centrifuging, adding ethanol solution into the precipitate, repeatedly extracting in the flash extractor, centrifuging, mixing the supernatants, and precipitating;
(3) activated carbon decolorization: adding activated carbon into the supernatant obtained in the step (2), stirring, and then removing the activated carbon;
(4) and (3) evaporation and concentration: concentrating and evaporating the combined solution obtained in the step (3) in vacuum, removing residual ethanol in the solution, and collecting the concentrated solution to obtain the dendrobe ethanol extract;
(5) and (3) protease hydrolysis: adding purified water into the precipitate obtained in the step (2), stirring and heating, adding trypsin, carrying out enzymolysis, stirring and heating, centrifuging, and filtering supernatant to obtain a dendrobium water extract;
(6) mixing and compounding: and (4) mixing the dendrobe alcohol extract obtained in the step (4) and the dendrobe water extract obtained in the step (5), and adding propylene glycol to obtain a dendrobe extract.
As a further improvement of the invention, the dendrobium comprises dendrobium nobile, dendrobium loddigesii, dendrobium verbena, dendrobium loddigesii and the like, preferably dendrobium nobile.
As a further improvement of the invention, the step (1) is cut into small sections with the length of 1-2 cm.
As a further improvement of the invention, the flash extraction in the step (2) comprises the following specific processes: and (2) uniformly mixing the small sections obtained in the step (1) with ethanol solution with the volume 5-10 times that of the small sections, placing the small sections in a flash extractor for extraction for 5-10 min at room temperature, centrifuging for 5-10 min, adding ethanol solution with the volume 5-10 times that of the precipitate into the flash extractor for repeated extraction once, centrifuging for 5-10 min, combining supernate, and precipitating for later use. Preferably 75-85% ethanol solution with the volume fraction of the pH value of 8-10.
As a further improvement of the invention, the adding amount of the activated carbon in the step (3) is that 2-4 g of activated carbon is added into every 100mL of supernatant, the mixture is stirred for 30-60 min, and the activated carbon is removed by centrifugation or filtration.
As a further improvement of the invention, the vacuum degree of the step (4) is-0.04 to-0.08 MPa, and the concentration and evaporation are carried out at 45 to 55 ℃, and the purpose of the concentration and evaporation is to remove the residual ethanol in the solution.
As a further improvement of the invention, in the step (5), the adding amount of the purified water is 5-10 times of the mass of the precipitate, the purified water is added, the mixture is stirred and heated to 50-60 ℃, preferably to 55 ℃, food-grade trypsin powder with the enzyme activity of 10000U/g is added, the adding amount of the enzyme is 0.05-0.1 percent of the adding amount of the purified water, the enzymolysis is carried out for 20-40 min, the mixture is stirred and heated to 90-100 ℃, preferably to 95 ℃, and the enzymolysis is maintained for 20-40 min, preferably for 30 min.
As a further improvement of the invention, the addition amount of the propylene glycol in the step (6) is 25-30%.
The invention also provides the dendrobe extracting solution prepared by the preparation method of the dendrobe extracting solution.
The invention also provides application of the dendrobium extract in preparing cosmetics.
As a further improvement of the invention, the dendrobium extract is used as a main effective component of a cosmetic raw material.
The invention discloses the following technical effects:
the method adopts two solvents of water and alcohol for extraction, can extract the effective components of the dendrobium to the maximum extent, combines the flash crushing extraction and the proteolysis method during extraction, can extract the water-soluble and alcohol-soluble components in the dendrobium to the maximum extent under the condition of not destroying the effective components as far as possible, and has mild reaction conditions, simple operation, easy realization of conditions, low cost and environmental protection. The dendrobium extract obtained by the method protects the effective components in the plant body to the maximum extent, and the obtained dendrobium extract as a raw material solution is added into cosmetics, so that the skin can be moistened, and the effects of inhibiting bacteria, removing acnes, diminishing inflammation and resisting allergy can be achieved effectively.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without inventive exercise.
FIG. 1 is a flow chart of the preparation method of the present invention.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
Example 1
(1) Taking 500g of fresh stem segments of the two-year-old dendrobium nobile, cleaning the stem segments with water, draining the stem segments, and cutting the stem segments into small segments with the length of 1 cm;
(2) adding 3L of 80% ethanol solution with pH of 9, mixing, extracting in a flash extractor at room temperature for 8min, centrifuging at room temperature 4000r/min for 5min, collecting supernatant, extracting precipitate with 3L of 80% ethanol solution with pH of 9, centrifuging at room temperature 4000r/min for 5min, mixing supernatants, and precipitating;
(3) weighing 180g of activated carbon, adding into the combined supernatant, continuously stirring for 40min, centrifuging for 5min at the room temperature of 4000r/min, and taking the supernatant;
(4) evaporating and concentrating the supernatant obtained after the activated carbon is decolorized under reduced pressure at 50 ℃, removing residual ethanol in the supernatant, and collecting the concentrated solution;
(5) adding 3L of purified water into the precipitate obtained in the step (2), stirring and heating to 55 ℃, adding 0.15g of trypsin powder, carrying out enzymolysis at a constant temperature for 30min, stirring and heating to 95 ℃, maintaining for 30min, centrifuging for 5min at the room temperature of 4000r/min, filtering the supernatant with a 3 mu filter membrane, and collecting the filtrate;
(6) and (3) mixing the concentrated solution collected in the step (4) with the filtrate collected in the step (5), and adding propylene glycol according to the proportion of 30% by volume to obtain the dendrobium nobile extract.
Example 2
(1) Taking 500g of fresh stem segments of the two-year-old dendrobium candidum, cleaning the stem segments with water, draining the stem segments, and cutting the stem segments into small segments with the length of 1 cm;
(2) adding 5L of 80% ethanol solution with pH of 9, mixing, extracting in a flash extractor at room temperature for 8min, centrifuging at room temperature 4000r/min for 5min, collecting supernatant, precipitating, extracting with 5L of 80% ethanol solution with pH of 9 again, centrifuging at room temperature 4000r/min for 5min, mixing supernatants, and precipitating;
(3) weighing 260g of activated carbon, adding into the combined supernatant, continuously stirring for 40min, centrifuging for 5min at the room temperature of 4000r/min, and taking the supernatant;
(4) evaporating and concentrating the supernatant obtained after the activated carbon is decolorized under reduced pressure at 50 ℃, removing residual ethanol in the supernatant, and collecting the concentrated solution;
(5) adding 5L of purified water into the precipitate obtained in the step (2), stirring and heating to 55 ℃, adding 0.3g of trypsin powder, carrying out enzymolysis at a constant temperature for 30min, stirring and heating to 95 ℃, maintaining for 30min, centrifuging for 5min at the room temperature of 4000r/min, filtering the supernatant with a 3 mu filter membrane, and collecting the filtrate;
(6) and (3) mixing the concentrated solution collected in the step (4) with the filtrate collected in the step (5), and adding propylene glycol according to the proportion of 30% to obtain the dendrobium candidum extract.
Example 3
(1) Taking 500g of fresh stem segments of the two-year-old dendrobium nobile, cleaning the stem segments with water, draining the stem segments, and cutting the stem segments into small segments with the length of 2 cm;
(2) adding 3L of 80% ethanol solution with pH of 9, mixing, extracting in a flash extractor at room temperature for 10min, centrifuging at room temperature 4000r/min for 5min, collecting supernatant, extracting precipitate with 3L of 80% ethanol solution with pH of 9, centrifuging at room temperature 4000r/min for 5min, mixing supernatants, and precipitating;
(3) weighing 180g of activated carbon, adding into the combined supernatant, continuously stirring for 50min, centrifuging for 5min at the room temperature of 4000r/min, and taking the supernatant;
(4) evaporating and concentrating the supernatant obtained after the activated carbon is decolorized under reduced pressure at 55 ℃, removing residual ethanol in the supernatant, and collecting the concentrated solution;
(5) adding 3L of purified water into the precipitate obtained in the step (2), stirring and heating to 55 ℃, adding 0.15g of trypsin powder, carrying out enzymolysis at a constant temperature for 30min, stirring and heating to 95 ℃, maintaining for 30min, centrifuging for 5min at the room temperature of 4000r/min, filtering the supernatant with a 3 mu filter membrane, and collecting the filtrate;
(6) and (3) mixing the concentrated solution collected in the step (4) with the filtrate collected in the step (5), and adding propylene glycol according to a proportion of 25% to obtain the dendrobium nobile extracting solution.
Example 4
(1) Taking 1000g of fresh stem segments of dendrobium nobile lindl of three years old, cleaning with water, draining, and cutting into small segments with the length of 2 cm;
(2) adding 10L 85% ethanol solution with pH of 10, mixing, extracting in a flash extractor at room temperature for 8min, centrifuging at room temperature 4000r/min for 5min, collecting supernatant, extracting precipitate with 10L 85% ethanol solution with pH of 10, centrifuging at room temperature 4000r/min for 5min, mixing supernatants, and precipitating;
(3) weighing 560g of activated carbon, adding into the combined supernatant, continuously stirring for 40min, centrifuging for 5min at the room temperature of 4000r/min, and taking the supernatant;
(4) evaporating and concentrating the supernatant obtained after the activated carbon is decolorized under reduced pressure at 45 ℃, removing residual ethanol in the supernatant, and collecting the concentrated solution;
(5) adding 10L of purified water into the precipitate obtained in the step (2), stirring and heating to 55 ℃, adding 0.5g of trypsin powder, carrying out enzymolysis at a constant temperature for 30min, stirring and heating to 95 ℃, maintaining for 30min, centrifuging for 5min at the room temperature of 4000r/min, filtering the supernatant with a 3 mu filter membrane, and collecting the filtrate;
(6) and (3) mixing the concentrated solution collected in the step (4) with the filtrate collected in the step (5), and adding propylene glycol according to a proportion of 28% to obtain the dendrobium nobile extracting solution.
The bacteriostatic activity of the dendrobium nobile extract prepared according to the method on propionibacterium acnes and staphylococcus aureus is investigated by a bacteriostatic ring method (table 1) to evaluate the bacteriostatic and anti-acne effects of the dendrobium nobile extract; the anti-inflammatory and anti-allergic effects of the extracts were evaluated by examining their effects on the in vitro inhibition of hyaluronidase (Table 2) and on the swelling of mouse pinna caused by xylene and the increase in the permeability of mouse capillaries caused by acetic acid (Table 3).
The results in table 1 show that 2%, 5% and 10% of the dendrobium nobile extract has obvious inhibition effect on propionibacterium acnes and staphylococcus aureus, and the diameter of the bacteriostatic ring is obviously increased along with the increase of the amount.
The results in table 2 show that 0.5%, 1%, 2% and 5% of the dendrobium nobile lindl extract has obvious inhibition effect on the in vitro activity of the hyaluronidase, and the inhibition effect is increasingly obvious along with the increase of the dosage within a certain range.
The results in table 3 show that 1% and 2% of the dendrobium nobile extract solutions significantly decrease the swelling degree of auricles and the exudation amount of evans blue compared with the control group, which indicates that the dendrobium nobile extract solutions can significantly inhibit the swelling of ears of mice caused by xylene and inhibit the increase of capillary permeability of mice caused by acetic acid. TABLE 1 bacteriostatic activity of Dendrobium nobile extract on Propionibacterium acnes and Staphylococcus aureus
TABLE 2 influence of Dendrobium nobile extract on hyaluronidase inhibition in vitro
TABLE 3 influence of Dendrobium nobile extract on mouse auricle swelling caused by xylene and mouse capillary permeability increase caused by acetic acid
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (8)
1. A preparation method of a dendrobium extract is characterized by comprising the following steps:
(1) selecting and processing raw materials: taking fresh stem segments of herba Dendrobii, cleaning with water, and cutting into small segments;
(2) flash extraction: uniformly mixing the small sections obtained in the step (1) with an ethanol solution, placing the mixture in a flash extractor for extraction at room temperature, centrifuging, adding the ethanol solution into the precipitate, placing the precipitate in the flash extractor for repeated extraction once, centrifuging, combining supernate, and using the ethanol solution with the volume fraction of 75-85% and the pH of 8-10 for precipitation for later use;
(3) activated carbon decolorization: adding activated carbon into the supernatant obtained in the step (2), stirring, and then removing the activated carbon;
(4) and (3) evaporation and concentration: concentrating and evaporating the combined solution obtained in the step (3) in vacuum, removing residual ethanol in the solution, and collecting the concentrated solution to obtain the dendrobe ethanol extract;
(5) and (3) protease hydrolysis: adding purified water into the precipitate obtained in the step (2), stirring and heating, adding trypsin, carrying out enzymolysis, stirring and heating, centrifuging, and filtering supernatant to obtain a dendrobium water extract; adding purified water in an amount which is 5-10 times of the mass of the precipitate in the step (5), stirring and heating to 50-60 ℃ after adding the purified water, adding food-grade trypsin powder with the enzyme activity of 10000U/g, wherein the adding amount of the enzyme is 0.05-0.1% of the adding amount of the purified water, performing enzymolysis for 20-40 min, stirring and heating to 90-100 ℃, and maintaining for 20-40 min;
(6) mixing and compounding: mixing the alcohol extract of dendrobe obtained in the step (4) and the water extract of dendrobe obtained in the step (5), and adding propylene glycol to obtain a dendrobe extract; in the step (6), the addition amount of the propylene glycol is 25-30% of the total volume of the mixed solution.
2. The preparation method of the dendrobe extracting solution according to claim 1, wherein the dendrobe extracting solution is cut into small sections with the length of 1-2 cm in the step (1).
3. The method for preparing the dendrobe extracting solution according to claim 1, wherein the flash extraction in the step (2) comprises the following specific steps: and (2) uniformly mixing the small sections obtained in the step (1) with ethanol solution with the volume 5-10 times that of the small sections, placing the small sections in a flash extractor for extraction for 5-10 min at room temperature, centrifuging for 5-10 min, adding ethanol solution with the volume 5-10 times that of the precipitate into the flash extractor for repeated extraction once, centrifuging for 5-10 min, combining supernate, and precipitating for later use.
4. The method for preparing the dendrobe extracting solution according to claim 1, wherein the adding amount of the activated carbon in the step (3) is 2-4 g of activated carbon per 100mL of supernatant.
5. The method for preparing the dendrobe extracting solution according to claim 1, wherein the vacuum degree in the step (4) is-0.04 to-0.08 MPa, and the dendrobe extracting solution is concentrated and evaporated at 45-55 ℃.
6. A dendrobe extracting solution, which is characterized by being prepared according to the preparation method of the dendrobe extracting solution of any one of claims 1 to 5.
7. The application of the dendrobium extract liquid of claim 6 in preparing cosmetics.
8. The use of claim 7, wherein the extract of Dendrobium nobile is used as a main effective ingredient of cosmetic raw material.
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