CN115634187B - Dendrobium officinale flower extract, preparation method and application thereof, dendrobium officinale flower extract, application thereof, soothing cream and preparation method thereof - Google Patents
Dendrobium officinale flower extract, preparation method and application thereof, dendrobium officinale flower extract, application thereof, soothing cream and preparation method thereof Download PDFInfo
- Publication number
- CN115634187B CN115634187B CN202211670137.9A CN202211670137A CN115634187B CN 115634187 B CN115634187 B CN 115634187B CN 202211670137 A CN202211670137 A CN 202211670137A CN 115634187 B CN115634187 B CN 115634187B
- Authority
- CN
- China
- Prior art keywords
- dendrobium officinale
- alcohol
- flower extract
- officinale flower
- extract
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Medicines Containing Plant Substances (AREA)
- Cosmetics (AREA)
Abstract
The invention provides a dendrobium officinale flower extract, a preparation method and application thereof, a dendrobium officinale flower extract, an application thereof, a soothing cream and a preparation method thereof, and belongs to the technical field of deep processing of plants. The method adopts an alcohol extraction mode to extract the flavonoid compounds in the dendrobium officinale flower to the maximum degree, and adopts HPD100, D101, AB-8 or amide adsorption resin to improve the yield of the flavonoid compounds, so that the dendrobium officinale flower extract prepared by the method has obvious inhibition effect on inflammatory factors TNF-alpha and IL-6, and has excellent anti-inflammatory and relieving effects; the dendrobium officinale flower extract prepared by the invention has no irritation to eyes, low cytotoxicity, high safety concentration, good water solubility, lighter color and wide application range, and has good application prospect in cosmetics, especially cosmetics suitable for sensitive skin. The preparation method adopts only alcohol and water as solvents, has low cost and simple and convenient operation, and is suitable for industrial production.
Description
Technical Field
The invention relates to the technical field of plant deep processing, and particularly relates to a dendrobium officinale flower extract, a preparation method and application thereof, a dendrobium officinale flower extract, an application thereof, soothing cream and a preparation method thereof.
Background
Sensitive skin is characterized in that skin cells are damaged to reduce the immunity of the skin, the skin moistening degree is insufficient due to the fact that the cuticle is thinned, the barrier function of the skin is too weak finally, external stimulation cannot be resisted, and when nerve fibers of the skin are frequently stimulated and excited, the phenomena of redness, fever, pruritus, stabbing pain, red rash and the like are easily caused. The problem of sensitive skin can be solved to a great extent by using the flower-shaped product of natural raw materials with anti-inflammatory and soothing effects.
Dendrobium officinale (Dendrobium officinale) is a perennial herb of the genus Dendrobium of the family Orchidaceae, is attached to trees or rocks of cliff in semi-yin and damp conditions, has important medical and health care values, and is a traditional and famous and precious traditional Chinese medicine. The current chemical composition and pharmacological research shows that the dendrobium officinale contains various alkaloids, various water-soluble polysaccharides, mucoids and trace elements, has high medicinal value and obvious effects of preventing aging, enhancing immunity and the like, and is called 'lifesaving grass herb' by folks. Wherein the Dendrobium officinale stem has effects of benefiting stomach, promoting fluid production, nourishing yin, clearing heat, moistening lung, relieving cough, invigorating kidney and improving eyesight; the dendrobium officinale flower contains flavonoid, amino acid, polysaccharide, volatile components and the like, and has the effects of nourishing yin, protecting liver and relieving mental stress.
Chinese patent CN110477397A discloses a method for preparing a dendrobium flower extract, which comprises the steps of crushing dendrobium flowers, soaking in deionized water, performing steam distillation extraction, respectively collecting distillate and water extract, and filtering to obtain the dendrobium flower extract. Chinese patent CN109172748A discloses a preparation method of a dendrobium officinale flower extract with tyrosinase activation effect, wherein dried dendrobium officinale flower is subjected to water or ethanol extraction, filtration, reduced-pressure concentration of filtrate, and freeze drying to obtain the dendrobium officinale flower extract with tyrosinase activation effect, so that the whitening effect of cosmetics can be improved. Chinese patent CN107158240A discloses a dendrobium officinale flower extract with whitening effect, which comprises 80-90% of dendrobium officinale flower essence and 10-20% of rice bran oil, wherein the dendrobium flower essence is prepared by combining a flash extraction technology (95% ethanol) with a subcritical extraction method (propane, butane, dimethyl ether, tetrafluoroethane and ethanol), and the whitening effect of cosmetics can be improved. Chinese patent CN108606209A is a method for breaking cell wall of Dendrobium officinale flower, which comprises the steps of sequentially performing physical crushing, chemical alcohol dissolving, biological enzymolysis and ultrasonic cell wall breaking to obtain a cell wall breaking extract of Dendrobium officinale flower with polysaccharide content of more than 15wt% and flavone content of more than 2mg/g, wherein the cell wall breaking extract of Dendrobium officinale flower is used as a main component of the cell wall breaking beverage of Dendrobium officinale flower, and the cell wall breaking beverage has the effects of promoting body absorption, strengthening spleen and nourishing stomach, protecting liver and improving eyesight, nourishing and tonifying kidney, improving physical strength, resisting oxidation and resisting fatigue.
At present, no report exists on a preparation method of the dendrobium flower extract with anti-inflammatory and relieving effects.
Disclosure of Invention
In view of the above, the invention aims to provide a dendrobium officinale flower extract, a preparation method and an application thereof, a dendrobium officinale flower extracting solution and an application thereof, and a soothing cream and a preparation method thereof.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of a dendrobium officinale flower extract, which comprises the following steps:
performing alcohol extraction on dendrobium officinale flowers, and then performing first concentration to obtain an alcohol-extracted crude extract;
loading the alcohol-extracted crude extract into an adsorption resin column, sequentially carrying out water elution and alcohol-water solution desorption, and carrying out second concentration on the obtained desorption solution to obtain a dendrobium officinale flower extract;
the adsorption resin in the adsorption resin column comprises at least one of HPD100 type macroporous adsorption resin, D101 type macroporous adsorption resin, AB-8 macroporous adsorption resin and polyamide resin;
the mass fraction of the alcohol in the alcohol-water solution is 50 to 80 percent.
Preferably, the alcohol extraction comprises at least one of reflux alcohol extraction, ultrasonic-reflux alcohol extraction, and flash alcohol extraction;
the alcohol extraction agent for alcohol extraction is an alcohol-water mixed solvent, and the mass fraction of alcohol in the alcohol-water mixed solvent is 40-80%;
the volume ratio of the dry weight of the dendrobium officinale flowers to the alcohol extract is 1kg:15 to 30L.
Preferably, the water consumption of the water elution is 1.5 to 5BV.
Preferably, the dosage of the alcohol-water solution desorbed by the alcohol-water solution is 1.5 to 5BV.
Preferably, the second concentrating further comprises: freeze-drying the dendrobium officinale flower extract obtained by the second concentration; the temperature of the freeze-drying is-50 to-40 ℃, the vacuum degree is 5 to 30Pa, and the time is 24 to 72h.
The invention provides the dendrobium officinale flower extract prepared by the preparation method in the technical scheme, and the active ingredients of the dendrobium officinale flower extract comprise flavonoid compounds, phenolic compounds and polysaccharide compounds.
The invention provides a dendrobium officinale flower extract, which comprises the dendrobium officinale flower extract and an alcohol dispersant.
The invention provides an application of the dendrobium officinale flower extract or the dendrobium officinale flower extracting solution in the technical scheme in cosmetics.
The invention provides a soothing cream which comprises the following components in percentage by mass: 0.8 to 2.5 percent of cetearyl alcohol, 1.2 to 3.0 percent of cetyl glucoside, 0.1 to 0.5 percent of acryloyl dimethyl sodium taurate copolymer, 3236 zxft Of caprylic/capric triglyceride, 3236 percent of tocopherol, 0.15 to 0.5 percent of tocopherol, 0.1 to 0.5 percent of dendrobium officinale flower extract in the technical scheme and/or dendrobium officinale flower extract liquid in the technical scheme, 0.1 to 0.3 percent of carbomer, 1~5 percent of butanediol, 2~5 percent of glycerin, 0.2 to 0.8 percent of preservative, 0.05 to 0.3 percent of triethanolamine and the balance of water; the mass of the dendrobium officinale flower extracting solution is based on the dry weight of the dendrobium officinale flower extract.
The invention provides a preparation method of a soothing cream in the technical scheme, which comprises the following steps:
(1) Mixing cetostearyl alcohol, cetyl glucoside, sodium acryloyldimethyl taurate copolymer, caprylic/capric triglyceride and tocopherol to obtain phase A;
(2) Mixing the Dendrobium officinale flower extract and/or Dendrobium officinale flower extract solution, carbomer resin, butanediol, glycerol, preservative and water to obtain phase B;
(3) Mixing the phase A and the phase B with triethanolamine to obtain a soothing cream;
the steps (1) and (2) have no time sequence.
The invention provides a preparation method of a dendrobium officinale flower extract, which comprises the following steps: performing alcohol extraction on dendrobium officinale flowers, and then performing first concentration to obtain an alcohol-extracted crude extract; loading the alcohol-extracted crude extract into an adsorption resin column, sequentially carrying out water elution and alcohol-water solution desorption, and carrying out second concentration on the obtained desorption solution to obtain a dendrobium officinale flower extract; the adsorption resin in the adsorption resin column comprises at least one of HPD100 type macroporous adsorption resin, D101 type macroporous adsorption resin, AB-8 macroporous adsorption resin and polyamide resin; the mass fraction of the alcohol in the alcohol-water solution is 50 to 80 percent. According to the invention, active ingredients (flavonoids, phenolic compounds and polysaccharides) in the dendrobium officinale flower can be extracted to the maximum extent through alcohol extraction, and the yield of the flavonoids can be improved by adopting an adsorption resin column for purification; moreover, the dendrobium officinale flower extract prepared by the method disclosed by the invention is good in water solubility, lighter in color and wide in application range. As shown in the test results of the examples, the dendrobium officinale flower extract prepared by the invention has obvious inhibition effect on inflammatory factors (TNF-alpha and IL-6) secreted by macrophage Raw264.7 cell inflammation caused by LPS, and the dendrobium officinale flower extract prepared by the invention has excellent anti-inflammatory and relieving effects; meanwhile, the dendrobium officinale flower extract prepared by the method has no irritation to eyes within the concentration of 10wt%, and the maximum safe concentration is 10wt%, which shows that the dendrobium officinale flower extract prepared by the method has low cytotoxicity and high safe concentration, and has good application prospect in cosmetics, especially cosmetics suitable for sensitive skin. The solvent adopted in the preparation process is only alcohol and water, so that the method is low in toxicity, low in cost, simple and convenient to operate, short in process time consumption and suitable for industrial production.
The invention provides a dendrobium officinale flower extract, which comprises the dendrobium officinale flower extract and an alcohol dispersant. In the dendrobium officinale flower extract provided by the invention, the dendrobium officinale flower extract has anti-inflammatory and soothing effects and low toxicity, and the dendrobium officinale flower extract has high solubility in alcohol dispersants, so that the dendrobium officinale flower extract has a good application prospect in cosmetics, especially in cosmetics suitable for sensitive skin.
The invention provides a soothing cream which comprises the following components in percentage by mass: 1.2% of cetearyl alcohol, 2.5% of cetyl glucoside, 0.3% of sodium acryloyldimethyl taurate copolymer, 6% of caprylic/capric triglyceride, 0.5% of tocopherol, 0.3% of the dendrobium officinale flower extract and/or the dendrobium officinale flower extract in the technical scheme, 0.1% of carbomer resin, 2% of butanediol, 5% of glycerol, 0.7% of preservative, 0.1% of triethanolamine and 81.2% of water. The dendrobium officinale flower extract has anti-inflammatory and soothing effects and low toxicity, improves the anti-inflammatory and soothing effects of the soothing cream, and is suitable for sensitive skin.
Drawings
FIG. 1 is a color development result chart of the Dendrobium officinale flower extract prepared in example 7;
FIG. 2 is a graph showing the results of the test of the activity of the flower extract of Dendrobii ferruginea on mouse macrophage cells in test example 2;
FIG. 3 is the standard curve TNF-. Alpha.in test example 3;
FIG. 4 is a graph showing the results of the TNF-. Alpha.inhibition by the Dendrobium officinale flower extract of test example 3;
FIG. 5 shows the standard curve IL-6 in test example 4;
FIG. 6 is a graph showing the result of the IL-6 inhibitory effect of the Dendrobium officinale flower extract in test example 4;
FIG. 7 is a graph showing the results of skin reaction experiments in 3 volunteers (No. 1, no. 2 and No. 3) in test example 5.
Detailed Description
The invention provides a preparation method of a dendrobium officinale flower extract, which comprises the following steps:
the invention provides a preparation method of a dendrobium officinale flower extract, which comprises the following steps:
extracting dendrobium officinale flowers with alcohol, and then carrying out first concentration to obtain an alcohol-extracted crude extract;
loading the alcohol-extracted crude extract into an adsorption resin column, sequentially performing water elution and alcohol-water solution desorption, and performing second concentration on the obtained desorption solution to obtain the dendrobium officinale flower extract;
the adsorption resin in the adsorption resin column comprises at least one of HPD100 type macroporous adsorption resin, D101 type macroporous adsorption resin, AB-8 macroporous adsorption resin and polyamide resin;
the mass fraction of the alcohol in the alcohol-water solution is 50 to 80 percent.
In the present invention, all the raw material components are commercially available products well known to those skilled in the art unless otherwise specified.
The invention carries out alcohol extraction on dendrobium officinale flowers, and then carries out first concentration to obtain alcohol extraction crude extract.
In the invention, the alcohol extraction agent for alcohol extraction is preferably an alcohol-water mixed solvent, and the mass fraction of alcohol in the alcohol-water mixed solvent is preferably 40 to 80%, more preferably 45 to 70%, and still more preferably 50 to 60%; the alcohol preferably comprises methanol and/or ethanol, more preferably ethanol. In the invention, the volume ratio of the dry weight of the dendrobium officinale flowers to the single alcohol extraction alcohol extractant is preferably 1kg:15 to 30L, more preferably 1kg:20 to 25L.
In the present invention, the alcohol extraction preferably includes at least one of reflux alcohol extraction, ultrasonic-reflux alcohol extraction, and flash alcohol extraction, more preferably reflux alcohol extraction, ultrasonic-reflux alcohol extraction, or flash alcohol extraction, and further preferably reflux alcohol extraction; the alcohol extraction frequency is preferably 1~3 times, and more preferably 1~2 times.
In the invention, the temperature of the reflux alcohol extraction is preferably 80 to 90 ℃, and more preferably 85 to 90 ℃; the time of single reflux alcohol extraction is preferably 1 to 3h, and more preferably 2h.
In the present invention, the ultrasonic-reflux alcohol extraction preferably comprises sequentially performing ultrasonic alcohol extraction and reflux alcohol extraction; the temperature of the ultrasonic alcohol extraction is preferably 15-50 ℃, more preferably 35 ℃, the power of the ultrasonic alcohol extraction is preferably 600-3500W, more preferably 2500W, the frequency of the ultrasonic alcohol extraction is preferably 20-40kHz, more preferably 35kHz, and the time of single ultrasonic alcohol extraction is preferably 20-120min, more preferably 30min; the temperature of the reflux alcohol extraction is preferably 80 to 90 ℃, and more preferably 85 to 90 ℃; the time for refluxing and alcohol extraction is preferably 1.0 to 3.0 hours, and more preferably 1.5 hours.
In the invention, the flash alcohol extraction is preferably carried out by using a flash extractor, the rotation speed of the flash extractor is preferably 1000 to 3000r/min, more preferably 2000r/min, and the time of single flash alcohol extraction is preferably 2 to 10min, more preferably 2 to 5min.
In the present invention, in the specific embodiment of the present invention, taking the number of times of alcohol extraction as an example, the alcohol extraction specifically is: performing first alcohol extraction on the dendrobium officinale flowers, and then performing first solid-liquid separation to obtain first alcohol extract and residues respectively; carrying out second alcohol extraction on the residues, and then carrying out second solid-liquid separation to obtain a second alcohol extract; and combining the first alcohol extract and the second alcohol extract to obtain the alcohol extract. The first solid-liquid separation and the second solid-liquid separation are not particularly limited in the present invention, and a solid-liquid separation method known to those skilled in the art may be adopted, specifically, filtration, suction filtration or centrifugal separation.
In the present invention, the temperature of the first concentration is preferably 60 to 80 ℃, and more preferably 70 ℃; the pressure of the first concentration is preferably-0.05 to-0.07 MPa, and more preferably-0.06 MPa; the first concentration time is not specially limited, and the first concentration is carried out until the mass ratio of the dry weight of the dendrobium officinale flowers to the alcohol extraction crude extract is 1:0.5 to 3, and more preferably 1:1~2.
After the alcohol-extracted crude extract is obtained, the alcohol-extracted crude extract is loaded into an adsorption resin column, water elution and alcohol-water solution desorption are sequentially carried out, and the obtained desorption solution is subjected to second concentration to obtain the dendrobium officinale flower extract.
In the present invention, the adsorption resin in the adsorption resin column preferably comprises at least one of an HPD100 type macroporous adsorption resin, a D101 type macroporous adsorption resin, an AB-8 macroporous adsorption resin and a polyamide resin, more preferably comprises an HPD100 type macroporous adsorption resin, a D101 type macroporous adsorption resin, an AB-8 macroporous adsorption resin or a polyamide resin, and further preferably an AB-8 macroporous adsorption resin. In the invention, the mass ratio of the volume of the adsorption resin in the adsorption resin column to the alcohol extraction crude extract (calculated by the dry weight of dendrobium officinale flowers) is preferably 0.5-3L: 1kg, more preferably 1 to 2L:1kg.
In the invention, the water consumption for water elution is preferably 1.5 to 5BV, more preferably 2 to 4BV, and further preferably 2.5 to 3BV; the flow rate of the water elution is preferably 2 to 5BV/h, more preferably 2.5 to 4.5BV/h, and still more preferably 3 to 4BV/h.
In the invention, the mass fraction of the alcohol in the alcohol-water solution is 50 to 80%, preferably 50 to 70%, and more preferably 55 to 60%; the using amount of the alcohol-water solution is preferably 1.5 to 5BV, more preferably 2 to 4BV, and further preferably 2.5 to 3BV; the flow rate of the alcohol-water solution desorption is preferably 2 to 5BV/h, more preferably 2.5 to 4.5BV/h, and still more preferably 3 to 4BV/h.
In the present invention, the temperature of the second concentration is preferably 60 to 80 ℃, and more preferably 70 ℃; the pressure of the second concentration is preferably-0.05 to-0.07 MPa, and more preferably-0.06 MPa; the time for the second concentration is not particularly limited, and the concentration is carried out until the solid content of the dendrobium officinale flower extract is 20 to 45wt%, and the solid content of the dendrobium officinale flower extract is more preferably 30 to 40wt%.
After the second concentration, the present invention preferably further comprises: and freeze-drying the dendrobium officinale flower extract obtained by the second concentration to obtain the dendrobium officinale flower extract (namely the dendrobium officinale flower extract freeze-dried powder). In the invention, the temperature of freeze-drying is-50 to-40 ℃, and the preferable temperature is-45 ℃; the vacuum degree of the freeze-drying is 5 to 30Pa, preferably 10 to 20Pa; the freeze-drying time is 24 to 72h, and preferably 48 to 60h.
The invention provides the dendrobium officinale flower extract prepared by the preparation method in the technical scheme, and the active ingredients comprise flavone. In the invention, the dendrobium officinale flower extract is preferably in the form of a dendrobium officinale flower extract or a dendrobium officinale flower extract freeze-dried powder.
The invention provides a dendrobium officinale flower extract, which comprises the dendrobium officinale flower extract and an alcohol dispersant. In the invention, the solid content of the dendrobium officinale flower extracting solution is preferably 0.1 to 99wt%, and more preferably 20 to 50wt%. In the present invention, the alcohol dispersant preferably includes at least one of 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, and glycerol, more preferably 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, or glycerol.
The invention provides an application of the dendrobium officinale flower extract or the dendrobium officinale flower extracting solution in the technical scheme in cosmetics. In the present invention, the cosmetic preferably includes a cream, essence, water cream, or pack. In the invention, the usage amount of the dendrobium officinale flower extract in the cosmetic is preferably 0.05 to 99wt%, more preferably 0.1 to 50wt%, and further preferably 0.2 to 5wt%; the dendrobium officinale flower extract is preferably used in the form of at least one of a dendrobium officinale flower extract, a dendrobium officinale flower extract freeze-dried powder and a dendrobium officinale flower extracting solution, and more preferably used in the form of the dendrobium officinale flower extract freeze-dried powder and/or the dendrobium officinale flower extracting solution. The dendrobium officinale flower extract adopted by the invention has the effects of resisting inflammation and relieving, has low toxicity, and has high solubility in alcohol dispersants, so that the dendrobium officinale flower extract has a good application prospect in cosmetics, especially in cosmetics suitable for sensitive skin.
The invention provides a soothing cream which comprises the following components in percentage by mass: 0.8 to 2.5 percent of cetearyl alcohol, 1.2 to 3.0 percent of cetyl glucoside, 0.1 to 0.5 percent of acryloyl dimethyl sodium taurate copolymer, 3236 zxft Of caprylic/capric triglyceride, 3236 percent of tocopherol, 0.15 to 0.5 percent of tocopherol, 0.1 to 0.5 percent of dendrobium officinale flower extract in the technical scheme and/or dendrobium officinale flower extract liquid in the technical scheme, 0.1 to 0.3 percent of carbomer, 1~5 percent of butanediol, 2~5 percent of glycerin, 0.2 to 0.8 percent of preservative, 0.05 to 0.3 percent of triethanolamine and the balance of water; the mass of the dendrobium officinale flower extracting solution is based on the dry weight of the dendrobium officinale flower extract.
The components of the soothing cream provided by the invention comprise, by mass, 0.8 to 2.5% of cetearyl alcohol, preferably 1~2%, and more preferably 1.2 to 1.5%.
The components of the soothing cream provided by the invention comprise 1.2 to 3.0% of cetyl glucoside by mass, preferably 1.5 to 2.8% of cetyl glucoside, and more preferably 2 to 2.5% of cetyl glucoside by mass.
The components of the soothing cream provided by the invention comprise 0.1 to 0.5% of acryloyl dimethyl sodium taurate copolymer, preferably 0.1 to 0.4%, and more preferably 0.2 to 0.3% in percentage by mass.
The components of the soothing cream provided by the invention comprise caprylic/capric triglyceride 1~8%, preferably 3~7%, more preferably 5~6% in percentage by mass.
The components of the soothing cream provided by the invention comprise, by mass, 0.15 to 0.5% of tocopherol, preferably 0.3 to 0.5%, and more preferably 0.4 to 0.5%.
The components of the soothing cream provided by the invention comprise, by mass percentage, 0.1 to 0.5% of the dendrobium officinale flower extract in the technical scheme and/or 0.2 to 0.4% of the dendrobium officinale flower extract in the technical scheme, preferably 0.3 to 0.4%, and more preferably 0.3 to 0.4%, wherein the mass of the dendrobium officinale flower extract and the mass of the dendrobium officinale flower extract are calculated by the dry weight of the dendrobium officinale flower extract.
The components of the soothing cream provided by the invention comprise 0.1-0.3% of carbomer, preferably 0.1-0.2%, and more preferably 0.1-0.15% in percentage by mass.
The components of the soothing cream provided by the invention comprise 1~5% of butanediol, preferably 1.5 to 4%, and more preferably 2~3% in percentage by mass.
The components of the soothing cream provided by the invention comprise 2~5%, preferably 3~5%, more preferably 4~5% by mass.
The components of the soothing cream provided by the invention comprise 0.2-0.8% of preservative, preferably 0.4-0.8%, and more preferably 0.5-0.7% by mass. In the invention, the preservative preferably comprises p-hydroxyacetophenone and 1,2-hexanediol, and the mass ratio of the p-hydroxyacetophenone to 1,2-hexanediol is preferably 1:1~5, more preferably 1:2~3.
The components of the soothing cream provided by the invention comprise 0.05 to 0.3% of triethanolamine, preferably 0.1 to 0.2%, and more preferably 0.1 to 0.15% by mass.
The components of the soothing cream provided by the invention comprise the balance of water in percentage by mass.
The invention provides a preparation method of the soothing cream in the technical scheme, which comprises the following steps:
(1) Mixing cetostearyl alcohol, cetyl glucoside, sodium acryloyldimethyl taurate copolymer, caprylic/capric triglyceride and tocopherol to obtain phase A;
(2) Mixing the Dendrobium officinale flower extract and/or Dendrobium officinale flower extract solution, carbomer resin, butanediol, glycerol, preservative and water to obtain phase B;
(3) Mixing the phase A and the phase B with triethanolamine to obtain a soothing cream;
the steps (1) and (2) have no time sequence. In the invention, cetostearyl alcohol, cetyl glucoside, sodium acryloyldimethyl taurate copolymer, caprylic/capric triglyceride and tocopherol are mixed to obtain phase A. In the present invention, the temperature of the mixing is preferably room temperature, the mixing is preferably stirring mixing, and the speed and time of the stirring mixing are not particularly limited, and the raw materials may be uniformly mixed.
The invention mixes the dendrobium officinale flower extract and/or dendrobium officinale flower extract liquid, carbomer resin, butanediol, glycerol, preservative and water to obtain phase B. In the present invention, the mixing temperature is preferably room temperature, the mixing is preferably stirring mixing, and the speed and time of the stirring mixing are not particularly limited, and the raw materials may be uniformly mixed.
After the phase A and the phase B are obtained, the phase A, the phase B and triethanolamine are mixed to obtain the soothing cream. In the present invention, the temperature of the mixing is preferably room temperature, the mixing is preferably stirring mixing, and the speed and time of the stirring mixing are not particularly limited, and the raw materials may be uniformly mixed.
The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Adding 600mL of 50wt% ethanol solution into 30g of dendrobium officinale flower, carrying out reflux ethanol extraction for 2h under the condition of 90 ℃ water bath, filtering, concentrating the obtained extracting solution to 30mL under the condition of 60 ℃ and-0.06 MPa under reduced pressure, loading the obtained ethanol extraction crude extract into a polyamide resin column, eluting with water (the water washing flow rate is 4BV/h and the water elution amount is 2.5 BV), desorbing with 60wt% ethanol solution (the desorption flow rate is 2BV/h and the desorption elution amount is 2.5 BV), respectively collecting water washing solution and desorption solution, concentrating the obtained desorption solution to 30mL under the condition of 60 ℃ and-0.06 MPa under reduced pressure, and freeze-drying the obtained dendrobium officinale flower extract (the temperature is-45 ℃, the vacuum degree is 20Pa and the time is 48 h) to obtain the dendrobium officinale flower extract; wherein the volume of the polyamide resin in the polyamide resin column: the mass of the dendrobium officinale flower =2mL:1g of the total weight of the composition.
Example 2
The dendrobium officinale flower extract was prepared according to the method of example 1, differing from example 1 only in that the polyamide resin was replaced with D101 resin.
Example 3
The dendrobium officinale flower extract was prepared according to the method of example 1, differing from example 1 only in that the polyamide resin was replaced with an AB-8 resin.
Example 4
The dendrobium officinale flower extract was prepared according to the method of example 1, differing from example 1 only in that the polyamide resin was replaced with HPD-100 resin.
The content of flavone in the water-washed solution and the desorbed solution obtained in example 1~4 was measured by an ultraviolet spectrophotometer method, and the results are shown in table 1, with 1mL of sample per sample and 3 times of parallel tests.
TABLE 1 flavone content of aqueous washes and desorbents from example 1~4
As shown in Table 1, the adsorption effect of AB-8 resin on flavone is best, and the yield of flavone is highest.
Example 5
Screening of extractant
(1) Adding 600mL of water into 30g of dendrobium officinale flowers respectively, performing reflux extraction for 2h under the condition of water bath at 90 ℃, filtering, concentrating the obtained extracting solution, and freeze-drying the obtained water-extracted crude extract (the temperature is-45 ℃, the vacuum degree is 20Pa, and the time is 48 h) to obtain the water extract of the dendrobium officinale flowers.
(2) Replacing the water in the step (1) with 50wt% ethanol solution, and freeze-drying the obtained 50% ethanol extract crude extract to obtain the 50% ethanol extract of the dendrobium officinale flowers.
(3) Replacing the water in the step (1) with 80wt% ethanol solution, and freeze-drying the obtained 80% ethanol extract crude extract to obtain the 80% ethanol extract of dendrobium officinale flowers.
The content of the flavonoid in the aqueous extract, 50% alcohol extract and 80% alcohol extract of the dendrobium officinale flower is tested by an ultraviolet spectrophotometer method, the yellow ketone content is tested for 3 times in parallel, and the test results are shown in table 2:
TABLE 2 flavone content in Dendrobium officinale flower extract
As can be seen from Table 2, the yield of flavone was the highest when 50wt% ethanol solution was used as the extractant.
Example 6
Ultrasonic-reflux alcohol extraction
Adding 600mL of 50wt% ethanol solution into 30g of dendrobium officinale flowers, performing ultrasonic-reflux alcohol extraction (ultrasonic extraction is performed for 30min under the conditions of 35 ℃, 750W and 35kHz, and then reflux alcohol extraction is performed for 1.5h under the condition of 90 ℃ water bath), filtering to obtain a primary alcohol extract and residues, repeating the ultrasonic-reflux alcohol extraction on the obtained residues for 1 time, filtering to obtain a secondary alcohol extract, combining the primary alcohol extract and the secondary alcohol extract, performing reduced pressure concentration to 30mL under the conditions of 60 ℃ and 0.06MPa, loading the obtained alcohol extract crude extract into an AB-8 resin column, performing water elution (water washing flow rate is 4BV/h and water elution amount is 2.5 BV), then performing desorption by using 60wt% ethanol solution (desorption flow rate is 2BV/h and desorption elution amount is 2.5 BV), collecting desorption solution, performing reduced pressure concentration to 30mL under the conditions of 60 ℃ and 0.06MPa, freeze-drying the obtained dendrobium officinale flower extract (temperature is-45 ℃, vacuum degree is 20 h and time is 48 h), and obtaining dendrobium officinale flower extract; wherein, the volume of the AB-8 resin in the AB-8 resin column: the mass of the dendrobium officinale flower =2mL:1g.
Example 7
Reflux alcohol extraction
Adding 600mL of 50wt% ethanol solution into 30g of dendrobium officinale flowers, performing reflux alcohol extraction for 2h under the condition of 90 ℃ water bath, filtering to obtain primary alcohol extract and residues, repeating the reflux alcohol extraction for 1 time on the obtained residues, filtering to obtain secondary alcohol extract, combining the primary alcohol extract and the secondary alcohol extract, performing reduced pressure concentration to 30mL under the conditions of 60 ℃ and 0.06MPa, loading the obtained alcohol extract into an AB-8 resin column, performing water elution (the water washing flow rate is 4BV/h and the water elution amount is 2.5 BV), then performing desorption (the desorption flow rate is 2BV/h and the desorption elution amount is 2.5 BV) by using 60wt% ethanol solution, collecting desorption solution, performing reduced pressure concentration to 30mL under the conditions of 60 ℃ and 0.06MPa on the desorption solution, and freeze-drying the obtained dendrobium officinale flower extract (the temperature is-45 ℃, the vacuum degree is 20Pa and the time is 48 h) to obtain the dendrobium officinale flower extract; wherein, the volume of the AB-8 resin in the AB-8 resin column: the mass of the dendrobium officinale flower =2mL:1g.
Example 8
Flash extraction
Adding 600mL of 50wt% ethanol solution into 30g of dendrobium officinale flowers, carrying out flash extraction at 2000r/min and 21 ℃ for 5min, filtering to obtain primary alcohol extract and residue, carrying out flash extraction on the residue for 1 time, filtering to obtain secondary alcohol extract, combining the primary alcohol extract and the secondary alcohol extract, carrying out reduced pressure concentration at 60 ℃ and 0.06MPa to 30mL, loading the obtained alcohol extract crude extract into an AB-8 resin column, carrying out water elution (the water washing flow rate is 4BV/h and the water elution amount is 2.5 BV), desorbing by using 60wt% ethanol solution (the desorption flow rate is 2BV/h and the desorption elution amount is 2.5 BV), collecting desorption solution, carrying out reduced pressure concentration on the desorption solution at 60 ℃ and 0.06MPa to 30mL, freeze-drying the obtained dendrobium officinale flower extract (the temperature is-45 ℃, the vacuum degree is 20Pa and the vacuum degree is 48 h) to obtain the dendrobium officinale flower extract; wherein, the volume of the AB-8 resin in the AB-8 resin column: the mass of the dendrobium officinale flower =2mL:1g.
The content of flavonoid in the dendrobium officinale flower extract prepared in example 6~8 was tested in parallel 3 times by using an ultraviolet spectrophotometer method, and the test results are shown in table 3:
TABLE 3 content of flavonoid in Dendrobium officinale flower extract from example 6~8
As shown in Table 3, the yield of flavone was the highest when the reflux-alcohol extraction was used.
Example 9
Example 7 qualitative identification of the components of the Dendrobium officinale flower extract prepared
The test principle and the steps are as follows: (1) identifying phenolic substances: adding ferric trichloride into the sample solution for reaction, wherein the solution is dark green; (2) identifying flavonoids: putting magnesium powder into a test tube, adding a sample solution, inclining the test tube, and slowly adding concentrated hydrochloric acid along the wall to obtain a red solution; (3) polysaccharide substance identification: concentrated sulfuric acid is slowly added into the sample solution to produce strong acid environment, and phenol is added for reaction to produce orange derivatives. The color development result of the dendrobium officinale flower extract prepared in example 7 is shown in fig. 1, and as can be seen from fig. 1, the dendrobium officinale flower extract prepared in the invention contains phenols, flavonoids and polysaccharides.
Example 10
Example 7 quantitative analysis of flavone component of Dendrobium officinale flower extract
(1) Control test materials: the standards were purchased from Shanghai leaf Biotech, inc. Adding 50% alcohol into rutin (HPLC is more than or equal to 98%), catechin (HPLC is more than or equal to 98%), quercetin (HPLC is more than or equal to 98%), naringin (HPLC is more than or equal to 98%), quercetin (HPLC is more than or equal to 98%) and naringenin (HPLC is more than or equal to 98%) to prepare the rutin with the concentration of 179.34 mug/mL; the catechin concentration is 45.668 mug/mL; the concentration of the quercetin is 52.234 mug/mL; the concentration of naringin is 76.44 mug/mL; the concentration of quercetin is 126.42 mug/mL; a control solution with naringenin concentration of 96.432 μ g/mL.
(2) Test solution: precisely taking 2mg of the dendrobium officinale flower extract, placing the dendrobium officinale flower extract in a 25mL colorimetric tube, diluting the dendrobium officinale flower extract with 50% methanol, and fixing the volume to 25mL.
(3) HPLC chromatographic conditions: mobile phase A:0.1% aqueous formic acid; mobile phase B: acetonitrile; a chromatographic column: hypersil BDS C18 μm (4.6mm. Times.250mm); detection wavelength: 270 nm; column temperature: 30 ℃; flow rate: 1 mL/min; sample introduction amount: 10. mu L; elution gradient: 0min (2 v/v% mobile phase B) -5 min (2 v/v% mobile phase B) -10 min (10 v/v% mobile phase B) -15 min (15 v/v% mobile phase B) -30 min (15 v/v% mobile phase B) -35 min (20 v/v% mobile phase B) -45 min (20 v/v% mobile phase B) -46 min (27.5 v/v% mobile phase B) -65 min (30 v/v% mobile phase B) -70 min (50 v/v% mobile phase B) -75 min (50 v/v% mobile phase B) -90 min (60 v/v% mobile phase B) -100 min (80 v/v% mobile phase B) -110 min (80 v/v% mobile phase B) -120 min (2 v/v% mobile phase B);
(4) HPLC assay results are shown in Table 4~6.
TABLE 4 HPLC detection chromatography results of control
TABLE 5 HPLC detection chromatogram result of herba Dendrobii flower extract (test sample)
TABLE 6 HPLC assay results of dendrobe flower extract (test article)
As shown in Table 4~6, the flavonoids in the dendrobium officinale flower extract prepared by the invention comprise rutin, quercetin and naringenin, wherein the rutin content in the test solution is 7.26mg/mL, the quercetin content is 0.45mg/mL, and the naringenin content is 5.25mg/mL.
Test example 1
Safety test
The dendrobium officinale flower extract prepared in example 8 is subjected to eye irritation measurement according to OECD TG492, and the test results are shown in table 7:
table 7 result of eye prick test of dendrobium officinale flower extract prepared in example 8
As can be seen from Table 7, the extract of Dendrobium officinale flowers prepared by the present invention has no irritation to eyes at 10 wt%.
Test example 2
MTT method for determining maximum safe concentration of dendrobium officinale flower extract prepared in example 8
Cell plating: the density of mouse RAW264.7 macrophage was adjusted to 1X 10 5 cell/mL, 200. Mu.L (1X 10) per well 4 cell/Wells) were inoculated in 96-well plates, the basal medium was added around the 96-well plates one round to prevent edge effects, a null set (200. Mu.L of basal medium without cells) was additionally provided, and CO was% 2 ) Medium culture;
sample preparation: diluting the dendrobium officinale flower extract into a solution with the concentration of 1.5wt% by utilizing a DMEM basic culture medium, filtering and sterilizing by using a 0.22 mu m filter membrane, taking out the solution before use, and diluting the solution by utilizing the DMEM basic culture medium in a gradient manner into sample solutions to be detected with the concentrations of 0.75wt%, 0.375wt%, 0.188wt%, 0.094wt% and 0.047wt% respectively for later use.
MTT: accurately weighing appropriate amount of MTT powder, adding PBS to dissolve and prepare into 5mg/mL concentration, filtering with 0.22 μm filter membrane, storing at 4 deg.C in dark place, and keeping effective period for 15 days.
Administration: after 24h of culture, taking out the 96-well plate, discarding the old culture medium, and distributing the plate according to 3 multiple wells per concentration in each group; adding 200 μ L DMEM basal medium to each well of the zeroing group and the blank control group, adding 200 μ L of each solution of the sample to be tested at different concentrations to the test group, and returning to the incubator for culturing (37 deg.C, 5% CO) 2 ）；
MTT assay cell viability: after 24h of dosing, the 96-well plate was removed, 20. Mu.L of MTT (5 mg/mL) was added to each well, the plate was returned to the incubator for further incubation for 4h, the well contents were discarded, 150. Mu.L of DMSO was added to each well again, and the absorbance (OD value) was measured at a wavelength of 560nm after shaking for 10 min.
And (3) processing an experimental result: cell viability% = (experimental group OD value-zero adjustment hole OD value)/(blank control group OD value-zero adjustment group OD value) × 100%, when cell viability is greater than or equal to 90%, the corresponding sample concentration is the maximum safe concentration, and the test results are shown in table 8 and fig. 2.
TABLE 8 influence of Dendrobium officinale flower extract on macrophage activity test results
As can be seen from Table 8 and FIG. 2, the cell viability was 89.31% when the concentration of the Dendrobium officinale flower extract was 1.5wt%, which is used as a reference for screening the concentration of the sample to be tested.
Test example 3
The effect of the dendrobium officinale flower extract prepared in example 8 on TNF-alpha secretion of mouse RAW264.7 macrophage caused by LPS
Cell plating: adjusting the cell density to 1X 10 5 cells/mL were inoculated into 96-well plates at 200. Mu.L per well, basal medium was added around the plate to prevent edge effects, and a zero set (200. Mu.L basal medium without cells) was returned to the incubator (37 ℃,5% CO) 2 ) Culturing;
preparing a reagent: positive drug (pomalidomide): taking out the solution with a concentration of 1X 10 6 Diluting pomalidomide with nM concentration with DMEM to obtain positive control solution with 5000nM concentration for later use;
the dendrobium officinale flower extract: preparing the dendrobium officinale flower extract into a sample solution to be detected with the concentration of 1% by using a DMEM culture medium, filtering and sterilizing by using a 0.22 mu m filter membrane, and then diluting the solution in a gradient manner to obtain the sample solution with the concentrations of 0.5%, 0.5% and 0.25%.
LPS: preparing LPS into 200 mug/mL concentration by PBS, and subpackaging in 1.5mL centrifuge tubes according to 100 mug/tube, and storing at-20 ℃ for later use; the solution is taken out before use and diluted to 3ng/mL concentration by a DMEM basic culture medium, and is prepared for use.
Administration: culturing for 24h, taking out 96-well plate, removing old culture medium, and adding 180 μ L DMEM basal medium into each well of zero setting group (3 multiple wells) and blank control group (BC group, 3 multiple wells); LPS stimulation group (NC group, 3 multiple wells) 180. Mu.L DMEM basal medium per well; positive control group (PC group, 3 duplicate wells) 160 μ L DMEM basal medium and 20 μ L pomalidomide at 5000nM concentration per well; 160. Mu.L of DMEM basal medium and 20. Mu.L of each of the test sample solutions with different concentrations were added to each well of the experimental group, and then returned to the incubator for culture (37 ℃,5% CO) 2 ) (ii) a Taking out the 96-well plate after 1h, and adding 20 mu L of DMEM basal medium into each well of the zero adjustment group and the blank control group; adding 20 mu L of LPS solution with the concentration of 3ng/mL into each hole of the positive control group, the LPS stimulation group and the experimental group, and then putting the holes back to the incubator for incubation;
collecting cell supernatants: taking out the 96-well plate after 24h of administration, respectively collecting cell supernatants corresponding to each sample group into a 1.5mL centrifuge tube, and storing at-20 ℃ for later use;
ELISA kit determination of TNF-alpha concentration in cell supernatant: 1) Sample preparation: taking out a sample to be measured, centrifuging the sample in a centrifuge (1000rpm, 5 min) after the sample is melted, directly measuring cell supernatant of a blank control group sample, and measuring other groups of samples after the supernatant is diluted by 40 times; 2) Standard solution: taking out the standard substance, adding 650 mu L of distilled water, dissolving to prepare a standard substance solution of 1000pg/mL, diluting according to 2-fold gradient to prepare a reference substance solution with the concentration of 500-3.906pg/mL, and preparing for use; 3) TNF-alpha concentration determination: the procedures were performed exactly as described in the kit instructions.
The standard curve TNF-. Alpha.is shown in FIG. 3 and Table 9, and the results of absorbance measurement (TNF-. Alpha.) are shown in Table 10.
TABLE 9 Standard Curve TNF-. Alpha.
TABLE 10 Absorbance assay results (TNF-. Alpha.)
And (3) analyzing an experimental result: according to a formula obtained by standard curve: y =0.0031x +0.0439, and the OD value (y) of each group is substituted into a formula to obtain the concentration (x, pg/mL) of TNF-alpha of each group, and then the concentration is multiplied by the dilution multiple (40 times), so that the final concentration of TNF-alpha of each group is obtained.
Statistical analysis was performed using T-Test, and the significance was expressed in x for the NC group compared to the BC group, and P values < 0.001 were expressed in x; compared with the PC group and the sample group to be tested, the significance is represented by #, and the P value is less than 0.001 is represented by # # #. The difference was not statistically significant and was expressed as P > 0.05.
The test results are shown in table 11 and fig. 4.
TABLE 11 TNF-. Alpha.concentration in cell supernatants of various groups of samples
As can be seen from table 11 and fig. 4, the LPS-stimulated group (NC) showed a significant increase in TNF- α concentration (P < 0.001) compared to the blank control group (BC); the positive control group has obviously lower TNF-alpha concentration than LPS stimulation group (NC); the concentration of TNF-alpha is obviously lower than that of LPS stimulation group (NC) under the action of 1.0%,0.5% and 0.25% concentration of dendrobium officinale flower extract; the dendrobium officinale flower extract prepared by the invention has obvious inhibition effect on TNF-alpha secretion generated by macrophage Raw264.7 cell inflammation caused by LPS.
Test example 4
Example 8 Effect of the Dendrobium officinale flower extract on IL-6 secretion by RAW264.7 macrophage of LPS-induced mouse
Cell plating: adjusting the cell density to 1X 10 5 cell/mL was inoculated into a 96-well plate at 200. Mu.L/well, the basal medium was added around the plate to prevent edge effects, and a zeroing group (200. Mu.L of basal medium without cells) was placed back in the incubator (37 ℃,5% 2 ) Culturing;
preparing a reagent: positive drug (pomalidomide): taking out the solution with a concentration of 1X 10 6 Diluting pomalidomide with nM concentration with DMEM to obtain positive control solution with 5000nM concentration for later use;
the dendrobium officinale flower extract: preparing the dendrobium officinale flower extract into a sample solution to be detected with the concentration of 1% by using a DMEM medium, filtering and sterilizing by using a 0.22-micron filter membrane, and diluting the sample solution into the sample solution to be detected with the concentrations of 0.5%, 0.5% and 0.25% in a gradient manner by using the DMEM medium.
LPS: preparing LPS into 200 mug/mL concentration by PBS, and subpackaging in 1.5mL centrifuge tubes according to 100 mug/tube, and storing at-20 ℃ for later use; the solution is taken out before use and diluted to a concentration of 10 mu g/mL by a DMEM basic culture medium, and is used as it is.
Administration: culturing for 24h, taking out 96-well plate, removing old culture medium, and adding 180 μ L DMEM basal medium into each well of zero setting group (3 multiple wells) and blank control group (BC group, 3 multiple wells); LPS stimulation group (NC group, 3 multiple wells) 180. Mu.L DMEM basal medium per well; positive control group (PC group, 3 multiple wells) 160. Mu.L DMEM basal medium per wellAnd 20 μ L pomalidomide at a concentration of 5000 nM; adding 160 μ L of sample solution with corresponding concentration to the basal medium 160 μ L of LDMEM and the extract of Dendrobium officinale flower into each well of the experimental group, and placing the mixture in an incubator for culturing (37 deg.C, 5% 2 ) (ii) a Taking out the 96-well plate after 1h, and adding 20 mu L of DMEM basal medium into each well of the zero adjustment group and the blank control group; adding 20 mu L of LPS solution with the concentration of 10 mu g/mL into each hole of the positive control group, the LPS stimulation group and the experimental group, and then putting the positive control group, the LPS stimulation group and the experimental group back into the incubator for incubation;
collecting cell supernatants: taking out a 96-well plate after administration for 24h, respectively collecting cell supernatants corresponding to the sample groups into a 1.5mL centrifuge tube, and storing at-20 ℃ for later use;
the kit is used for measuring the concentration of IL-6 in cell supernatant: 1) Sample preparation: taking out a sample to be detected, centrifuging the sample in a centrifuge (1000rpm, 5 min) after the sample is melted, directly measuring cell supernatant taken from a blank control group sample, and measuring the cell supernatant taken from other groups of samples after the cell supernatant is diluted by 4 times; 2) Standard solution: taking out the standard substance, adding 650 mu L of distilled water, dissolving to prepare a standard substance solution of 500pg/mL, diluting according to a 2-time gradient to prepare reference substance solutions with the concentrations of 250, 125, 62.5, 31.25, 15.625, 7.8125, 3.90625 and 1.953pg/mL, and preparing for use; 3) Determination of IL-6 concentration: the procedures were performed exactly as described in the kit instructions.
The standard curve IL-6 is shown in FIG. 5 and Table 12, and the results of absorbance measurement (IL-6) are shown in Table 13.
TABLE 12 Standard Curve-IL-6
TABLE 13 Absorbance measurement results (IL-6)
And (3) analyzing an experimental result: according to a formula obtained by marking the music: y =0.013x +0.021, and the OD value (y) of each group is substituted into the formula to calculate the IL-6 concentration (x, pg/mL) of each group, and then the concentration is multiplied by the dilution multiple (4 times), so that the final IL-6 concentration of each group is obtained, and the test results are shown in Table 14 and FIG. 6.
Statistical analysis was performed using T-Test, and the significance was expressed in x for the NC group compared to the BC group, and P values < 0.001 were expressed in x; compared with the PC group and the NC group, the significance is represented by #, and the P value is less than 0.001 is represented by # # #. The difference was not statistically significant and was expressed as P > 0.05.
TABLE 14 IL-6 concentration in cell supernatants of various groups of samples
As can be seen from table 14 and fig. 6, the concentration of IL-6 was significantly increased in the LPS-stimulated group (NC) compared to the blank control group (BC) (. X.p < 0.001); the positive control group had significantly lower IL-6 concentrations than the LPS-stimulated group (NC); under the action of the dendrobium officinale flower extract with the concentration of 1.0% and 0.5%, the concentration of IL-6 is obviously lower than that of LPS stimulation group (NC); the dendrobium officinale flower extract prepared by the invention has obvious inhibition effect on IL-6 secretion generated by macrophage Raw264.7 cell inflammation caused by LPS.
Example 9
A soothing cream containing dendrobium officinale flowers was prepared from the dendrobium officinale flower extract prepared in example 8, and the formula of the soothing cream is shown in table 15:
TABLE 15 formula of soothing cream containing dendrobium officinale flower extract
The preparation method of the soothing cream comprises the following steps:
heating and dissolving carbomer, butanediol, glycerol, herba Dendrobii flower extract, p-hydroxyacetophenone and 1,2-hexanediol at 80 deg.C to obtain phase B;
heating and stirring cetostearyl alcohol, M68, EMT10, GTCC and VE oil until completely dissolving, and stirring at 80 deg.C for 30min to obtain phase A;
adding the phase A into the phase B under stirring, uniformly mixing, keeping the temperature and stirring for 10min at 80 ℃ after the addition is finished, homogenizing for 3min at 6000rpm, cooling to 50 ℃, adding triethanolamine, and uniformly stirring to obtain the soothing cream.
Comparative example 1
A cream was prepared according to the method of example 9, except that DENDROBIUM officinale (DENDROBIUM CANDIDUM) flower extract was not added to the cream of example 9, to obtain a base cream.
Test example 5
Human body class label experiment
The experimental method comprises the following steps: cutting the patch paper into small blocks of 3cm × 3cm, applying 20 μ L of histamine solution (anaphylactic agent, concentration of 2 wt%) on the patch paper, applying the patch paper to 2 different positions on the inner side of forearm of arm, taking off the patch paper after 30min, and taking pictures to record skin redness, swelling, pimple, edema, etc. in the patch area; then, a sample is smeared on a blank control area, the sample to be detected is smeared on the sample area to be detected, pictures are taken again after 30min and 60min, the fading conditions of redness, pimples and the like in a patch area are compared, the change condition of the itching pain feeling is subjectively evaluated by a volunteer, and the skin reaction grading standard of the histamine skin patch test is shown in a table 16.
TABLE 16 Histamine Patch test skin response grading Standard
Sensitization test: dividing the left forearm of 9 subjects into 2 small square areas (respectively marked as area 1 and area 2) of 3cm × 3cm, respectively, applying 20 μ L of histamine solution (anaphylactic agent, concentration of 2wt%, solvent water) onto patch cloth, sticking to each area for 30min, tearing off patch cloth, taking pictures with camera, and recording anaphylaxis;
and (3) anti-allergy experiment: area 1 as a sensitization control group, a blank group (matrix cream) was applied; zone 2 application experimental group (soothing cream prepared in example 9); the coating amount is 0.2g, pictures are taken after coating for 30min and 60min respectively, and the sensitivity phenomenon of each area is observed.
The results of the skin reaction experiments of 3 volunteers (1, 2 and 3) are shown in fig. 7, and it can be seen from fig. 7 that the skin of the volunteer shows different degrees of red swelling 30min after histamine spot-pasting, and the red swelling is obviously relieved compared with the blank control after the sample is smeared.
Skin reaction scoring: the skin redness and swelling status of the volunteers was scored according to the skin reaction evaluation criteria and statistically analyzed, and the test results are shown in table 17:
TABLE 17 volunteer skin redness and swelling status scoring results
From table 17, it can be seen that erythema of different degrees appears on the skin after histamine is applied (30 min), the score is 1.0 to 2.5 minutes, and erythema on the skin after a sample to be detected and a blank solvent are applied (60 min) is dissipated, but compared with the blank group, the soothing cream containing the dendrobium officinale flower extract has an obvious improvement effect on the erythema on the skin caused by histamine.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and amendments can be made without departing from the principle of the present invention, and these modifications and amendments should also be considered as the protection scope of the present invention.
Claims (10)
1. A preparation method of a dendrobium officinale flower extract is characterized by comprising the following steps:
performing alcohol extraction on dendrobium officinale flowers, and then performing first concentration to obtain an alcohol-extracted crude extract; the alcohol extraction alcohol extract is an alcohol-water mixed solvent, and the mass fraction of alcohol in the alcohol-water mixed solvent is 40-80%;
loading the alcohol-extracted crude extract into an adsorption resin column, sequentially carrying out water elution and alcohol-water solution desorption, and carrying out second concentration on the obtained desorption solution to obtain a dendrobium officinale flower extract;
the adsorption resin in the adsorption resin column is AB-8 macroporous adsorption resin;
the alcohol extraction is reflux alcohol extraction;
the mass fraction of alcohol in the alcohol-water solution is 50 to 80 percent;
the active ingredients of the dendrobium officinale flower extract comprise flavonoids, and the flavonoids comprise rutin, quercetin and naringenin.
2. The preparation method of claim 1, wherein the ratio of the dry weight of the dendrobium officinale flowers to the volume of the alcohol extract is 1kg:15 to 30L.
3. The method of claim 1, wherein the water is used in an amount of 1.5 to 5BV.
4. The method according to claim 1, wherein the alcohol-water solution is desorbed at 1.5 to 5BV.
5. The method of claim 1, further comprising, after the second concentrating: freeze-drying the dendrobium officinale flower extract obtained by the second concentration; the temperature of the freeze-drying is-50 to-40 ℃, the vacuum degree is 5 to 30Pa, and the time is 24 to 72h.
6. The dendrobium officinale flower extract prepared by the preparation method of any one of claim 1~5, wherein the active ingredients comprise flavonoids, phenolic compounds and polysaccharides.
7. An extract solution of dendrobium officinale flowers, which is characterized by comprising the extract of dendrobium officinale flowers of claim 6 and an alcohol dispersant.
8. The use of the dendrobium officinale flower extract of claim 6 or the dendrobium officinale flower extract of claim 7 in cosmetics.
9. The soothing cream is characterized by comprising the following components in percentage by mass: 0.8 to 2.5 percent of cetearyl alcohol, 1.2 to 3.0 percent of cetyl glucoside, 0.1 to 0.5 percent of acryloyl dimethyl sodium taurate copolymer, 3236 zxft Of caprylic/capric triglyceride, 3236, 0.15 to 0.5 percent of tocopherol, 0.1 to 0.5 percent of the dendrobium officinale flower extract of claim 6 and/or the dendrobium officinale flower extract of claim 7, 0.1 to 0.3 percent of carbomer, 1~5 percent of butanediol, 2~5 percent of glycerin, 0.2 to 0.8 percent of preservative, 0.05 to 0.3 percent of triethanolamine and the balance of water; the mass of the dendrobium officinale flower extracting solution is based on the dry weight of the dendrobium officinale flower extract.
10. A method of preparing a soothing cream as claimed in claim 9, comprising the steps of:
(1) Mixing cetostearyl alcohol, cetyl glucoside, sodium acryloyldimethyl taurate copolymer, caprylic/capric triglyceride and tocopherol to obtain phase A;
(2) Mixing the Dendrobium officinale flower extract and/or Dendrobium officinale flower extract solution, carbomer resin, butanediol, glycerol, preservative and water to obtain phase B;
(3) Mixing the phase A and the phase B with triethanolamine to obtain soothing cream;
the steps (1) and (2) have no time sequence.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211670137.9A CN115634187B (en) | 2022-12-26 | 2022-12-26 | Dendrobium officinale flower extract, preparation method and application thereof, dendrobium officinale flower extract, application thereof, soothing cream and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211670137.9A CN115634187B (en) | 2022-12-26 | 2022-12-26 | Dendrobium officinale flower extract, preparation method and application thereof, dendrobium officinale flower extract, application thereof, soothing cream and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115634187A CN115634187A (en) | 2023-01-24 |
| CN115634187B true CN115634187B (en) | 2023-03-31 |
Family
ID=84950009
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211670137.9A Active CN115634187B (en) | 2022-12-26 | 2022-12-26 | Dendrobium officinale flower extract, preparation method and application thereof, dendrobium officinale flower extract, application thereof, soothing cream and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115634187B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116211783B (en) * | 2023-05-04 | 2023-07-25 | 云南凌垭生物科技有限公司 | Cosmetic composition containing dendrobium candidum extract and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106420557A (en) * | 2016-10-09 | 2017-02-22 | 上海兰葹生物科技有限公司 | Herba dendrobii combined skin cream capable of promoting skin regeneration |
| WO2018090805A1 (en) * | 2016-11-15 | 2018-05-24 | Hong Kong Baptist University | Skin-protection composition containing dendrobium-based ingredients |
| WO2019056908A1 (en) * | 2017-09-20 | 2019-03-28 | 杭州丹鹤医药有限公司 | Application method of dendrobium candidum in preparing medicine for treating hypertension |
| CN114767787A (en) * | 2022-05-09 | 2022-07-22 | 中国药科大学 | Application of dendrobium officinale flower extract in preparation of medicine or health-care product for preventing cerebral ischemia |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102126469B1 (en) * | 2017-09-04 | 2020-06-24 | 주식회사 엘지생활건강 | Cosmetic Compositions comprising extract of Dendrobium candidum flower extract |
| CN108991331A (en) * | 2018-06-27 | 2018-12-14 | 广西驰胜农业科技有限公司 | A kind of dendrobium candidum medicine materical crude slice and its preparation method and application |
| CN109820959B (en) * | 2019-03-25 | 2022-03-18 | 浙江中医药大学 | Application of dendrobium officinale extract in preparation of medicines for preventing and treating dyslipidemia |
| CN114796073A (en) * | 2022-05-05 | 2022-07-29 | 浙江工业大学 | Anti-aging and anti-wrinkle dendrobium officinale essence and preparation method and application thereof |
-
2022
- 2022-12-26 CN CN202211670137.9A patent/CN115634187B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106420557A (en) * | 2016-10-09 | 2017-02-22 | 上海兰葹生物科技有限公司 | Herba dendrobii combined skin cream capable of promoting skin regeneration |
| WO2018090805A1 (en) * | 2016-11-15 | 2018-05-24 | Hong Kong Baptist University | Skin-protection composition containing dendrobium-based ingredients |
| WO2019056908A1 (en) * | 2017-09-20 | 2019-03-28 | 杭州丹鹤医药有限公司 | Application method of dendrobium candidum in preparing medicine for treating hypertension |
| CN114767787A (en) * | 2022-05-09 | 2022-07-22 | 中国药科大学 | Application of dendrobium officinale flower extract in preparation of medicine or health-care product for preventing cerebral ischemia |
Non-Patent Citations (2)
| Title |
|---|
| Q Zeng等.Dendrobium officinale kimura et migo improved dry eye symptoms via promoting tear production in an experimental dry eye rat model.《Pharmacognosy Magazine》.2020,第69卷(第16期),全文. * |
| 刘倩葶 ; 刘琨毅 ; 李崇萍 ; 焦文文 ; 念波 ; 蒋宾 ; 施扬 ; 程建胜 ; 赵菊润 ; 张新凤 ; 赵明 ; .铁皮石斛花固体饮料工艺初探及功能性物质鉴定.美食研究.2020,(第02期),全文. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115634187A (en) | 2023-01-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ohsugi et al. | Active-oxygen scavenging activity of traditional nourishing-tonic herbal medicines and active constituents of Rhodiola sacra | |
| AU2002348988B2 (en) | Composition comprising wenguanguo extracts, methods for preparing same and uses thereof | |
| Zainol et al. | Profiling of Centella asiatica (L.) Urban extract | |
| Yalavarthi et al. | A review on identification strategy of phyto constituents present in herbal plants | |
| KR102304498B1 (en) | Composition for anti-transmissible gastroenteritis virus (tgev) and pharmaceutical composition including the same | |
| CN115634187B (en) | Dendrobium officinale flower extract, preparation method and application thereof, dendrobium officinale flower extract, application thereof, soothing cream and preparation method thereof | |
| Attard et al. | The phytochemical and in vitro pharmacological testing of Maltese medicinal plants | |
| Karan et al. | Antihyperglycaemic effect of Vetiveria zizanioides (L.) Nash root extract in alloxan induced diabetic rats | |
| CN112545926A (en) | Comprehensive extraction method of folium artemisiae argyi volatile oil and folium artemisiae argyi total flavonoids and oxidation resistance evaluation of folium artemisiae argyi volatile oil | |
| Chand et al. | In vitro antioxidant activity of alcoholic extract of seed of Cucumis callosus (Rottl.) cogn | |
| CN101547701B (en) | Enhancement of antitubercular activity of extracts of salicornia brachiata | |
| KR102115669B1 (en) | Preparation method of black Platycodon grandiflorus having increased crude saponin and reduced Platycodin D, inhibiting production of benzopyrene | |
| CN110664865A (en) | Method for separating and preparing extract of cinnamon steaming oil residues and application of extract | |
| Kumar et al. | Hepatoprotective effects of 50% ethanolic extract of Ficus hispida Linn against CCl4 induced hepatotoxicity in rats | |
| Afroze et al. | Proximate analysis, phytochemical screening and antioxidant activity of Psidium guajava leaves growing in coastal area of Bangladesh | |
| Padmalochana | Anticancer (liver cancer cell lines) and antioxidant activity of Cassia auriculata flower extract from acetone and methanol solvents | |
| CN108497246A (en) | A kind of plant weight losing function beverage and preparation method thereof | |
| CN1943647B (en) | The method for preparing triterpenic acid extract from the loquat leaves | |
| CN100488543C (en) | Local externally applied itch stopping garlic liniments | |
| Sutar et al. | Phytochemical investigation, anthelmintic and antioxidant activities of Quisqualis indica | |
| Marliani | A review: Aquilaria species as potential of pharmacological | |
| Pandurangan et al. | Evaluation of anti-inflammatory and antioxidant activity of Ichnocarpus frutescens root | |
| CN111973666B (en) | Preparation method and application of total alkaloids of aerial part of radix Aconiti lateralis | |
| KR20110040024A (en) | Mixture extract of natural plants and anti-atopy cosmetic composition containing the same | |
| Kabir et al. | In vitro Study of Antioxidant Potentials, and Thrombolytic Activity of the Leaves of Delonix regia (Family: Fabaceae) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |