CN118178298A - Bletilla striata extract and preparation method and application thereof - Google Patents
Bletilla striata extract and preparation method and application thereof Download PDFInfo
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- CN118178298A CN118178298A CN202410605589.1A CN202410605589A CN118178298A CN 118178298 A CN118178298 A CN 118178298A CN 202410605589 A CN202410605589 A CN 202410605589A CN 118178298 A CN118178298 A CN 118178298A
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- bletilla striata
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- bletilla
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- 241001313857 Bletilla striata Species 0.000 title claims abstract description 98
- 238000002360 preparation method Methods 0.000 title claims abstract description 58
- 238000000605 extraction Methods 0.000 title claims description 52
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- 241001313855 Bletilla Species 0.000 claims abstract description 40
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 30
- 239000000843 powder Substances 0.000 claims abstract description 26
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- 229940035437 1,3-propanediol Drugs 0.000 claims description 24
- FHKSXSQHXQEMOK-UHFFFAOYSA-N hexane-1,2-diol Chemical compound CCCCC(O)CO FHKSXSQHXQEMOK-UHFFFAOYSA-N 0.000 claims description 24
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- VQECOUFHPVSSCD-UHFFFAOYSA-N 1-phenanthren-1-ylphenanthrene Chemical compound C1=CC2=CC=CC=C2C2=C1C(C1=C3C=CC=4C(C3=CC=C1)=CC=CC=4)=CC=C2 VQECOUFHPVSSCD-UHFFFAOYSA-N 0.000 description 1
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- OQUKIQWCVTZJAF-UHFFFAOYSA-N phenol;sulfuric acid Chemical compound OS(O)(=O)=O.OC1=CC=CC=C1 OQUKIQWCVTZJAF-UHFFFAOYSA-N 0.000 description 1
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- 150000003505 terpenes Chemical class 0.000 description 1
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- 238000001291 vacuum drying Methods 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- 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
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9794—Liliopsida [monocotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/028—Flow sheets
- B01D11/0284—Multistage extraction
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0288—Applications, solvents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Dermatology (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of cosmetics, and discloses a bletilla striata extract and a preparation method and application thereof. The preparation method comprises the following steps: adding ethanol into the bletilla striata powder, desorbing at normal temperature, placing the desorbed bletilla striata liquid in a water bath at the vacuum degree of-0.1 Mpa to-0.08 Mpa and the temperature of 45-55 ℃, heating in the water bath, adding deionized water when bubbles appear in the liquid, keeping the vacuum degree and the temperature, continuously extracting, treating the extracted liquid by high-pressure microjet, filtering, and concentrating to obtain the bletilla striata extract. The obtained extract contains polysaccharide and bletilla glycoside, and has effects of keeping moisture, relieving inflammation, resisting aging, and caring skin.
Description
Technical Field
The invention belongs to the technical field of cosmetics, and particularly relates to a bletilla striata extract and a preparation method and application thereof.
Background
Rhizoma bletillae, rhizoma bletilla powder and the like belong to perennial land orchid herbaceous plants, are mostly distributed in relatively wet places such as mountain areas, valleys and the like in three provinces of southwest in China, and are also distributed in small amounts in places such as Anhui, zhejiang, jiangsu and the like in coastal areas of southeast. The main chemical components of rhizoma bletilla comprise bibenzyl, biphenanthrene and derivatives thereof, glycosides, terpenes, polysaccharides, etc., wherein the polysaccharides are main active components of rhizoma bletilla, and have antibacterial, antitumor, blood coagulation promoting, and antioxidant effects.
The functional special effects of the bletilla striata are increasingly developed by people, the bletilla striata can be used for medical clinical application, can be applied to daily cosmetics, and has the effects of resisting oxidation, resisting inflammation, preserving moisture and the like.
The existing extraction method of rhizoma bletilla extract comprises heating extraction (heating reflux extraction, leaching method, decoction method), ultrasonic extraction, microwave extraction, enzyme-assisted extraction, etc. The heating extraction method has the defect that the structure of an extracted product is easily damaged under a continuous high-temperature environment; the ultrasonic extraction method has the defects that the glycosidic bond is broken after the ultrasonic time is too long, and the extraction efficiency of polysaccharide products is affected; the microwave extraction method has the defect that the gelatinization of polysaccharide is caused by slightly high microwave power; enzyme-assisted extraction has the disadvantage that the reaction conditions are easily deactivated if they are controlled for a short time; some processes have the disadvantages of high toxicity and safety problems due to the adoption of organic reagents.
The bletilla striata extract prepared by the prior art often uses a single substance such as polysaccharide or glycoside as a target product, so that the skin care effect is not rich enough, and the application range of the bletilla striata extract in the effect type skin care product is narrow. In addition, the existing bletilla striata extract is mostly in a powder state, but the powder needs to be dissolved in advance before use, the process is complex, the cost is high, and the problems of unstable cosmetic formula, precipitation and the like can be caused in the follow-up process; the preparation of powders typically requires vacuum or freeze drying, consuming large amounts of refrigerant and electrical energy. The existing bletilla striata extract (in a liquid state) has poor corrosion resistance and is easy to hydrolyze due to high polysaccharide content, so that the bletilla striata extract has the problems of a large number of microorganisms, reduced viscosity, increased turbidity and the like in a long-term storage process.
Disclosure of Invention
In order to overcome the drawbacks and disadvantages of the prior art, a primary object of the present invention is to provide a method for preparing rhizoma bletillae extract; the method sequentially applies two processes of a decompression internal boiling method and a high-pressure micro-jet method to the preparation of rhizoma bletillae extract.
The invention also aims to provide the bletilla striata extract prepared by the preparation method.
Still another object of the present invention is to provide a formulation of rhizoma bletilla extract prepared from the above rhizoma bletilla extract.
It is still another object of the present invention to provide a use of the above-mentioned bletilla striata extract and bletilla striata extract preparation.
The aim of the invention is achieved by the following technical scheme:
a preparation method of rhizoma bletillae extract comprises the following steps:
(1) And (3) desorption: adding 70-80% ethanol in volume percentage concentration into dried and crushed bletilla striata powder with 200 meshes according to a feed liquid ratio of 1 g (0.8-1.0) mL, and desorbing at normal temperature for 10-20 min to obtain bletilla striata feed liquid;
(2) Decompression internal boiling extraction: heating the desorbed rhizoma bletillae feed liquid in a water bath at the vacuum degree of-0.1 Mpa to-0.08 Mpa and the temperature of 45-55 ℃, rapidly adding deionized water at the temperature of 45-55 ℃ which is 100 times the weight of the rhizoma bletillae powder when bubbles rise in the feed liquid, and maintaining the vacuum degree and the temperature for extraction for 30-50 min;
(3) High-pressure micro-jet treatment: treating the material liquid extracted in the step (2) with high-pressure micro-jet equipment at 25 ℃, wherein the treatment pressure is 80-120 MPa, and the treatment times are 2-4 times;
(4) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (3) at 12000r/min for 10min, passing through a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to obtain rhizoma bletillae concentrated solution which is 3-4 times of the weight of the rhizoma bletillae powder, namely the rhizoma bletillae extract.
Rhizoma bletilla extract prepared by the above preparation method.
The bletilla striata extract preparation is prepared from the bletilla striata extract, wherein the bletilla striata extract preparation comprises the following components in percentage by mass of 15-20: 50-70: 15-25: 0.5-2 parts of bletilla striata extract, water, 1, 3-propanediol and 1, 2-hexanediol; the rhizoma bletilla extract preparation contains rhizoma bletilla polysaccharide more than 13 mg/ml and rhizoma bletilla glycoside more than 0.5mg/ml.
The total weight of the bletilla striata extract preparation is 20 times of the weight of the bletilla striata powder in the step (1).
The application of rhizoma bletilla extract or rhizoma bletilla extract preparation in preparing skin care product with moisturizing, antiinflammatory and antiaging effects is provided.
The principle of the invention is as follows:
In the step of decompression internal boiling, the high-concentration ethanol has better permeability, and can quickly permeate into cells when contacted with rhizoma bletillae powder; then adding a hot solvent (water with the temperature of 45-55 ℃) and simultaneously decompressing, boiling an ethanol solution in the material, and converting the extraction process of active ingredients (polysaccharide and bletilla glycoside) from molecular diffusion to convection diffusion, so that the mass transfer efficiency of the active ingredients can be improved; the method has the advantages of less organic solvent consumption, short time consumption and low temperature, can reduce degradation of polysaccharide and bletilla striata glycoside to the maximum extent, and has extraction effect obviously superior to that of the conventional method. In the high-pressure micro-jet treatment process, the residual intact cell walls are broken under the actions of instantaneous high-speed collision, strong shearing, instantaneous pressure release and the like of the bletilla striata feed liquid, so that the mass transfer rate is obviously improved, and the yields of polysaccharide and bletilla striata glycoside are further improved. Therefore, the invention uses decompression internal boiling and high pressure micro-jet extraction successively, plant cells which are not destroyed and fully swelled in the first step decompression internal boiling method are more easily crushed under the high pressure micro-jet condition, the dissolution of active ingredients is accelerated, and the two extraction processes can play a synergistic role when used successively.
Compared with the prior art, the invention has the following advantages and effects:
(1) The invention applies two processes of a decompression internal boiling method and a high-pressure micro-jet method to the preparation of the bletilla striata extract according to a specific sequence, and the obtained extract contains polysaccharide and bletilla striata glycoside and has the effects of moisturizing, anti-inflammatory and anti-aging various skin care.
(2) The invention can improve the mass transfer speed of the active ingredient by an internal boiling method and improve the extraction rate; the high-pressure micro-jet treatment can make the feed liquid more uniform, form a stable system, reduce the risks of precipitation and turbidity return, and synchronously improve the extraction effect of the active ingredients.
(3) The bletilla striata extract and the preparation thereof prepared by the invention are in transparent liquid state, and the appearance and the active matter content are kept stable under extreme conditions such as high temperature, illumination, low temperature and the like, so that the application range of the bletilla striata extract in skin care products is remarkably widened.
Detailed Description
The present invention is further illustrated below in conjunction with specific examples, but should not be construed as limiting the invention.
Example 1:
(1) And (3) desorption: drying and crushing 10g of rhizoma bletillae powder with 200 meshes, adding ethanol with the volume percentage concentration of 75% according to the feed liquid ratio of 1 g:0.9 mL, and desorbing 15 and min at normal temperature to obtain the rhizoma bletillae feed liquid;
(2) Internal boiling extraction: heating desorbed rhizoma bletilla feed liquid in water bath under vacuum degree of-0.09 Mpa at 50deg.C, rapidly adding 1000g deionized water at 50deg.C when slight boiling of the surface of feed liquid occurs (i.e. when bubbles rise in feed liquid), and extracting for 40min under vacuum degree and temperature conditions;
(3) High-pressure micro-jet treatment: all the feed liquid extracted in the step (2) is transferred to high-pressure micro-jet equipment for treatment at 25 ℃, the pressure is 100MPa, and the treatment times are 3 times;
(4) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (3) for 10min at 12000r/min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 40g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(5) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (4) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 20:54:25:1.
Example 2
(1) And (3) desorption: drying and crushing 10g of rhizoma bletillae powder with 200 meshes, adding 70% ethanol in volume percentage concentration according to a feed liquid ratio of 1 g:0.8 mL, and desorbing 20 min at normal temperature to obtain rhizoma bletillae feed liquid;
(2) Internal boiling extraction: heating desorbed rhizoma bletilla feed liquid in water bath at-0.1 Mpa and 55deg.C under vacuum degree, rapidly adding 1000g deionized water at 55deg.C when the surface of feed liquid is slightly boiled (i.e. bubbles rise in feed liquid), and extracting for 30min under vacuum degree and temperature conditions;
(3) High-pressure micro-jet treatment: all the feed liquid extracted in the step (2) is transferred to high-pressure micro-jet equipment for treatment at 25 ℃, the pressure is 120MPa, and the treatment times are 2 times;
(4) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (3) for 10min at 12000r/min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 36g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(5) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (4) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 18:61:20:1.
Example 3
(1) And (3) desorption: drying and crushing 10g of rhizoma bletillae powder with 200 meshes, adding 80% ethanol in volume percentage concentration according to a feed liquid ratio of 1 g:1 mL, and desorbing at normal temperature for 10min to obtain rhizoma bletillae feed liquid;
(2) Internal boiling extraction: heating desorbed rhizoma bletilla feed liquid in water bath at-0.08 Mpa and 45deg.C under vacuum, rapidly adding 1000g deionized water at 45deg.C when slight boiling of the feed liquid surface occurs (i.e. when rising of bubbles in the feed liquid occurs), and extracting for 50min under vacuum and temperature conditions;
(3) High-pressure micro-jet treatment: all the feed liquid extracted in the step (2) is transferred to high-pressure micro-jet equipment for treatment at 25 ℃, the pressure is 80MPa, and the treatment times are 4 times;
(4) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (3) for 10min at 12000r/min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 30g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(5) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (4) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 15:69:15:1.
Comparative example 1
The other steps are the same as in example 1, except that the high pressure micro-jet extraction is performed first, and then the internal boiling extraction is performed, specifically the steps are as follows:
(1) Adding 10g and 1000g of deionized water into high-pressure micro-jet equipment for treatment at 25 ℃ under the pressure of 100MPa for 2 times to obtain feed liquid;
(2) And (3) desorption: centrifugally separating the feed liquid to obtain filter residues and filtrate, adding ethanol with the volume percentage concentration of 75% into the dried filter residues according to the feed liquid ratio of 1 g:0.9 mL, and desorbing 15: 15 min at normal temperature to obtain bletilla striata feed liquid;
(3) Internal boiling extraction: heating the bletilla striata feed liquid desorbed in the step (2) in a water bath at the vacuum degree of-0.09 Mpa and 50 ℃, rapidly adding the filtrate obtained in the step (2) (preheated to 50 ℃ if less than 1000g is required to be filled with deionized water) when the surface of the feed liquid is slightly boiled (namely, when bubbles in the feed liquid rise), and extracting for 40min under the vacuum degree and temperature conditions;
(4) Concentrating: centrifuging the material liquid extracted in the step (3) at 12000r/min for 10min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 36g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(5) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (4) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 20:54:25:1.
Comparative example 2
The other steps are the same as in example 1, except that the internal boiling extraction is changed to conventional extraction, and the specific steps are as follows:
(1) Conventional extraction: adding 10g of rhizoma bletillae powder and 1000g of deionized water which are dried and crushed to 200 meshes into an extraction pot, and extracting at 50 ℃ for 55min at 40 r/min;
(2) High-pressure micro-jet treatment: all the feed liquid extracted in the step (1) is transferred to high-pressure micro-jet equipment for treatment at 25 ℃, the pressure is 100MPa, and the treatment times are 2 times;
(3) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (2) for 10min at 12000r/min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 40g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(4) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (3) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 20:54:25:1.
Comparative example 3
The other steps are the same as in example 1, except that the high pressure microfluidic extraction is replaced by conventional extraction, and the specific steps are as follows:
(1) And (3) desorption: drying and crushing 10g of rhizoma bletillae powder with 200 meshes, adding ethanol with the volume percentage concentration of 75% according to the feed liquid ratio of 1 g:0.9 mL, and desorbing 15 and min at normal temperature to obtain the rhizoma bletillae feed liquid;
(2) Internal boiling extraction: heating desorbed rhizoma bletilla feed liquid in water bath under vacuum degree of-0.09 Mpa at 50deg.C, rapidly adding 1000g deionized water at 50deg.C when slight boiling of the surface of feed liquid occurs (i.e. when bubbles rise in feed liquid), and extracting for 40min under vacuum degree and temperature conditions;
(3) Conventional extraction: transferring all the extracted feed liquid in the step (2) into an extraction pot, and extracting at 50 ℃ for 55min at 40 r/min;
(4) Concentrating: centrifuging the feed liquid subjected to conventional extraction in the step (3) at 12000r/min for 10min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 40g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(5) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (4) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 20:54:25:1.
Comparative example 4
The other steps are the same as in example 1, except that the internal boiling extraction is replaced by enzymatic extraction, and the specific steps are as follows:
(1) Enzymatic extraction: placing 10g and 1000g deionized water of rhizoma bletilla powder dried and crushed to 200 meshes into an extraction pot, adding pectase to 6000U/mL of enzyme activity, and extracting at 50 ℃ for 55min at 40 r/min;
(2) High-pressure micro-jet treatment: all the feed liquid extracted by the enzyme method in the step (1) is transferred to high-pressure micro-jet equipment for treatment at 25 ℃, the pressure is 100MPa, and the treatment times are 2 times;
(3) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (2) for 10min at 12000r/min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 40g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(4) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (3) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 20:54:25:1.
Comparative example 5
The other steps are the same as in example 1, except that the high pressure microfluidic extraction is replaced by ultrasonic extraction, the specific steps are as follows:
(1) And (3) desorption: drying and crushing 10g of rhizoma bletillae powder with 200 meshes, adding ethanol with the volume percentage concentration of 75% according to the feed liquid ratio of 1 g:0.9 mL, and desorbing 15 and min at normal temperature to obtain the rhizoma bletillae feed liquid;
(2) Internal boiling extraction: heating desorbed rhizoma bletilla feed liquid in water bath under vacuum degree of-0.09 Mpa at 50deg.C, rapidly adding 1000g deionized water at 50deg.C when slight boiling of the surface of feed liquid occurs (i.e. when bubbles rise in feed liquid), and extracting for 40min under vacuum degree and temperature conditions;
(3) Ultrasonic extraction: transferring all the extracted feed liquid in the step (2) into ultrasonic extraction equipment, and extracting for 20min at the power of 400 w;
(4) Concentrating: centrifuging the material liquid subjected to ultrasonic extraction in the step (3) at 12000r/min for 10min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 40g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(5) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (4) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 20:54:25:1.
Comparative example 6
The other steps are the same as in example 1, except that ethanol with a volume percentage concentration of 60% is used for desorption, and the specific steps are as follows:
(1) And (3) desorption: drying and crushing 10g of rhizoma bletillae powder with 200 meshes, adding ethanol with the volume percentage concentration of 60% according to the feed liquid ratio of 1 g:0.9 mL, and desorbing 15 and min at normal temperature to obtain the rhizoma bletillae feed liquid;
(2) Internal boiling extraction: heating desorbed rhizoma bletilla feed liquid in water bath of vacuum degree-0.11 Mpa at 50deg.C, rapidly adding 1000g deionized water at 50deg.C when slight boiling of the feed liquid surface occurs (i.e. when rising of bubbles in the feed liquid occurs), and extracting for 40min under the vacuum degree and temperature conditions;
(3) High-pressure micro-jet treatment: all the feed liquid extracted in the step (2) is transferred to high-pressure micro-jet equipment for treatment at 25 ℃, the pressure is 100MPa, and the treatment times are 2 times;
(4) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (3) for 10min at 12000r/min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 40g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(5) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (4) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 20:54:25:1.
Comparative example 7
The other steps are the same as in example 1, except that the high pressure micro-jet extraction pressure is 60Mpa, and the specific steps are as follows:
(1) And (3) desorption: drying and crushing 10g of rhizoma bletillae powder with 200 meshes, adding ethanol with the volume percentage concentration of 75% into the powder according to the proportion of 1 g:0.9: 0.9 mL, and desorbing the powder at normal temperature of 15: 15min to obtain rhizoma bletillae feed liquid;
(2) Internal boiling extraction: heating desorbed rhizoma bletilla feed liquid in water bath under vacuum degree of-0.09 Mpa at 50deg.C, rapidly adding 1000g deionized water at 50deg.C when slight boiling of the surface of feed liquid occurs (i.e. when bubbles rise in feed liquid), and extracting for 40min under vacuum degree and temperature conditions;
(3) High-pressure micro-jet treatment: all the feed liquid extracted in the step (2) is transferred to high-pressure micro-jet equipment for treatment at 25 ℃, the pressure is 60MPa, and the treatment times are 2 times;
(4) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (3) at 12000r/min for 10min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 40g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(5) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (4) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 20:54:25:1.
Comparative example 8: the difference from example 1 is that the high pressure microjet extraction pressure was 140Mpa, specifically as follows:
(1) And (3) desorption: drying and crushing 10g of rhizoma bletillae powder with 200 meshes, adding ethanol with the volume percentage concentration of 75% according to the feed liquid ratio of 1 g:0.9 mL, and desorbing 15 and min at normal temperature to obtain the rhizoma bletillae feed liquid;
(2) Internal boiling extraction: heating desorbed rhizoma bletilla feed liquid in water bath under vacuum degree of-0.09 Mpa at 50deg.C, rapidly adding 1000g deionized water at 50deg.C when slight boiling of the surface of feed liquid occurs (i.e. when bubbles rise in feed liquid), and extracting for 40min under vacuum degree and temperature conditions;
(3) High-pressure micro-jet treatment: all the feed liquid extracted in the step (2) is transferred to high-pressure micro-jet equipment for treatment at 25 ℃, the pressure is 140MPa, and the treatment times are 2 times;
(4) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (3) for 10min at 12000r/min, filtering with a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to 40g to obtain rhizoma bletillae concentrated solution, namely rhizoma bletillae extract;
(5) Adding water and preservative which is 1, 3-propanediol and 1, 2-hexanediol into the bletilla striata extract obtained in the step (4) to obtain 200g of bletilla striata extract preparation, wherein the mass ratio of the bletilla striata extract to the water to the 1, 3-propanediol to the 1, 2-hexanediol is 20:54:25:1.
Test example 1: determination of active ingredient in bletilla tuber extract preparation
Determination of polysaccharide content in bletilla extract preparation
The polysaccharide content of the bletilla striata extract preparations finally obtained in examples 1 to 3 and comparative examples 1 to 8 was measured by a phenol-sulfuric acid method using glucose as a reference substance.
Determination of the content of bletilla glycoside in bletilla extract preparation
Determining contents of bletilla glycoside in the bletilla extract preparations finally obtained in examples 1-3 and comparative examples 1-8 by adopting an HPLC method; HPLC test method: c18 chromatographic column, acetonitrile-0.1% phosphoric acid solution (22:78) is used as mobile phase, column temperature box is 30 ℃, detection wavelength is: 223nm.
Third, determination of active substance content in rhizoma Bletillae extract preparation
TABLE 1 active content in bletilla extract formulations
From the above table, the content of active substances (bletilla polysaccharide and bletilla glycoside) in the bletilla striata extract preparation obtained in examples 1-3 is significantly higher than that in comparative examples 2-5, which shows that the extraction efficiency is high after the two technologies of internal boiling extraction and high-pressure micro-jet extraction are combined, and the extraction efficiency of the active substances is reduced by replacing other extraction modes (such as conventional water extraction, enzyme method and ultrasonic).
Test example 2: moisture retention efficacy test of bletilla striata extract preparation
110 Healthy skin-free volunteers aged 20-40 were selected to test the moisturizing effect of the bletilla extract preparation. The test samples are bletilla striata extract preparations finally obtained in examples 1-3 and comparative examples 1-8, and each group of 10 people has a test environment temperature of 24-26 ℃ and a humidity of 40% -60%; the testing instrument is a CM825 type skin oil-water acid-base tester; the inner side of the arm of the test part is 2 cm zone; the results are shown in Table 2.
Table 2 moisture efficacy test of bletilla striata extract formulations
Polysaccharide in the bletilla striata extract preparation contains a large number of hydroxyl groups, can be combined with water molecules in a hydrogen bond mode, and can be combined with each other to form a net-shaped compact protective film so as to achieve the effects of moisture absorption and moisture preservation. From the above table, it can be seen that the skin still has a high moisture content after using the bletilla striata extract preparation of examples 1-3 for 6 hours; and the bletilla striata extract preparation with the extraction process changed in comparative examples 2-5 has low polysaccharide content, has weaker moisturizing effect than the examples, and is basically recovered before use after 6 hours of use.
Test example 3: anti-inflammatory efficacy test of bletilla striata extract preparation
An inflammation model is established by inducing RAW 264.7 cells by lipopolysaccharide LPS, and the effects of bletilla striata extract preparations obtained in examples 1-3 and comparative examples 1-8 of the present invention on the secretion of IL-6 and NO induced by LPS are detected by ELISA method.
The testing method comprises the following steps: selecting RAW 264.7 cells with good morphology in a logarithmic growth phase, inoculating the RAW 264.7 cells into a 24-well plate, and incubating the RAW 264.7 cells in an incubator for 24 hours. A blank control group (BC), an LPS-induced stimulation group (PC), a positive control group (100 mug/mL dexamethasone, NC) and a bletilla extract preparation (50 mg/mL, the cell viability is greater than 90% under the pre-test of the administration concentration and is a safe concentration) are set, incubated for 2 hours in a 5% CO 2 incubator at 37 ℃, LPS is added to each hole except the blank control group, and incubated for 24 hours in the incubator. Cell supernatants were assayed according to ELISA kit and nitric oxide assay kit protocol, and the amounts of NO and IL-6 in the collected cell supernatants were measured separately. And (3) analyzing an experimental result by software in data processing, and calculating the relative expression quantity of the NO and the IL-6 in the sample group by taking the relative expression quantity of the LPS positive control group as 1.
Table 3 anti-inflammatory efficacy test of bletilla striata extract formulations
As can be seen from the data in the table, compared with the BC group, the two inflammatory factors in the PC group are significantly increased, which indicates that the experimental stimulation conditions are effective; after dexamethasone is added into the NC group, the secretion of two inflammatory factors is obviously reduced, which proves that the experimental modeling is effective. The bletilla striata extract preparation of examples 1-3 has good anti-inflammatory effect on RAW 264.7 cell inflammatory factors IL-6 and NO secretion inhibition rate reaching about 40% under LPS induction. The bletilla striata extract preparation prepared in comparative examples 2-5 has low bletilla striata glycoside content and poor anti-inflammatory effect.
Test example 4: anti-aging test
Tricine-buffer (50 mmol/L, pH 7.5.5) containing 400 mmol/L NaCl and 10mmol/L CaCl 2 was prepared, and 0.8U/mL collagenase solution and 2mmol/L FALGPA solution were prepared using Tricine-buffer. 40. Mu.L of a sample solution (bletilla striata extract preparation obtained in examples 1-3 and comparative examples 1-8) with a mass concentration of 50mg/mL was mixed with 100. Mu.L of Tricine-buffer, then 20. Mu.L of a collagenase solution (0.8U/mL) was added, after mixing, incubated at a constant temperature of 25℃for 15min, then 40. Mu.L of a solution (2 mmol/L) of FALGPA was added, and after 15 minutes the absorbance was measured at 335 nm. The inhibition rate of the sample to collagenase is calculated according to the formula:
Inhibition ratio =
Wherein: a is the absorbance of the reaction solution without the sample; b is the absorbance of the reaction solution without sample and enzyme; c is the absorbance of the reaction solution containing the sample and the enzyme; d is the absorbance of the reaction solution without enzyme.
Table 4 anti-aging efficacy test of bletilla striata extract formulations
From the above table, the inhibition ratio of collagenase activity was found: examples 1-3 > comparative examples 2-5 show that the bletilla striata extract preparation prepared by combining the two processes has better collagenase inhibition effect than the bletilla striata extract preparation prepared by combining other processes, so that the degradation of collagen in skin can be obviously reduced, and skin relaxation caused by aging can be resisted.
Test example 5: stability test
The final bletilla striata extract preparations obtained in examples 1 to 3 and comparative examples 1 to 8 were directly placed in environments of 4 ℃ and normal temperature (25 ℃) and 50 ℃ respectively, the appearance stability was examined for 3 months, the degree of change in appearance of the samples before and after the stability test was scored, the scoring was 10 minutes, the no difference in color before and after the test was 10 minutes, the complete discoloration was 0 minutes, and the middle was analogized in order, and the occurrence of precipitation was recorded, and the experimental results are shown in table 5.
Table 5 stability test of bletilla extract formulation
From the data, the rhizoma bletillae extract preparation obtained in examples 1-3 has no change in color and no precipitation after being placed in high, low and normal temperature environments for a long time. The method shows that substances in the feed liquid can be dispersed more uniformly through the combined process of internal boiling and high-pressure micro-jet, collision and agglomeration among particles are not easy to occur, and the risk of precipitation or sedimentation in the subsequent long-term use process is reduced.
Application example 1: bai skin care essence
The skin care essence containing bletilla striata extract was prepared as shown in table 6 below:
table 6 skin care essence formulation containing bletilla striata extract
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (5)
1. A preparation method of rhizoma bletillae extract is characterized by comprising the following steps:
(1) And (3) desorption: adding 70-80% ethanol in volume percentage concentration into dried and crushed bletilla striata powder with 200 meshes according to a feed liquid ratio of 1 g (0.8-1.0) mL, and desorbing at normal temperature for 10-20 min to obtain bletilla striata feed liquid;
(2) Decompression internal boiling extraction: heating the desorbed rhizoma bletillae feed liquid in a water bath at the vacuum degree of-0.1 Mpa to-0.08 Mpa and the temperature of 45-55 ℃, rapidly adding deionized water at the temperature of 45-55 ℃ which is 100 times the weight of the rhizoma bletillae powder when bubbles rise in the feed liquid, and maintaining the vacuum degree and the temperature for extraction for 30-50 min;
(3) High-pressure micro-jet treatment: treating the material liquid extracted in the step (2) with high-pressure micro-jet equipment at 25 ℃, wherein the treatment pressure is 80-120 MPa, and the treatment times are 2-4 times;
(4) Concentrating: centrifuging the feed liquid subjected to the high-pressure microjet treatment in the step (3) at 12000r/min for 10min, passing through a 0.4 mu m filter membrane to obtain filtrate, and concentrating the filtrate to obtain rhizoma bletillae concentrated solution which is 3-4 times of the weight of the rhizoma bletillae powder, namely the rhizoma bletillae extract.
2. An extract of bletilla striata prepared by the method of claim 1.
3. A formulation of rhizoma bletilla extract prepared from the rhizoma bletilla extract of claim 2, wherein: the bletilla striata extract preparation comprises the following components in percentage by mass: 50-70: 15-25: 0.5-2 parts of bletilla striata extract, water, 1, 3-propanediol and 1, 2-hexanediol; the rhizoma bletilla extract preparation contains rhizoma bletilla polysaccharide more than 13 mg/ml and rhizoma bletilla glycoside more than 0.5mg/ml.
4. A formulation of bletilla striata extract according to claim 3, characterized in that: the total weight of the bletilla striata extract preparation is 20 times of the weight of the bletilla striata powder in the step (1) of claim 1.
5. Use of the bletilla striata extract according to claim 2 or the bletilla striata extract preparation according to claim 3 for the preparation of skin care products with moisturizing, anti-inflammatory and anti-ageing effects.
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