CN115040564A - Application of tartary buckwheat seed coat extract in reducing number or area of red spots on skin - Google Patents
Application of tartary buckwheat seed coat extract in reducing number or area of red spots on skin Download PDFInfo
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- CN115040564A CN115040564A CN202210896717.3A CN202210896717A CN115040564A CN 115040564 A CN115040564 A CN 115040564A CN 202210896717 A CN202210896717 A CN 202210896717A CN 115040564 A CN115040564 A CN 115040564A
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- China
- Prior art keywords
- seed coat
- extract
- skin
- tartary buckwheat
- agents
- Prior art date
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Classifications
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- 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]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K2236/50—Methods involving additional extraction steps
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Abstract
The invention provides application of tartary buckwheat seed coat extract in preparing a composition for reducing the number or area of red spots on skin.
Description
The application is a divisional application, the application number of a parent application is 202010533517.2, the application is Baiyuet biotechnology (Shanghai) limited company, the application date is 2020 and 06 and 11 months, and the invention name is the application of the tartary buckwheat seed coat extract for slowing down skin aging and the preparation method thereof.
Technical Field
The invention relates to a use of a plant extract, in particular to a use of a tartary buckwheat seed coat extract for preparing a composition for reducing the number or area of red spots on skin.
Background
The skin provides a first stage of protection for human subjects against environmental factors such as Ultraviolet (UV) radiation, pathogens, friction in the sun. The skin comprises, in order from the outside to the inside, an epidermal layer, a dermal layer mainly composed of connective tissue, and a subcutaneous tissue. The dermis layer contains molecules such as collagen (collagen), elastin (elastin), and hyaluronic acid (hyaluronic acid), the collagen is "scaffold" in the skin, and the elastin is "latch" in the skin, and the elastin is linked by the elastin to provide skin with support and elasticity.
Collagen is an important matrix protein in skin, has a triple-helix long-chain structure, occupies a high weight ratio in a dermis layer, and provides structural stability for the skin. According to recent studies, one of the possible factors of collagen chain cleavage is the glycosylation of collagen (Non-enzymatic hydrolysis, the addition of carbohydrate molecules to protein molecules). Since the glycosylation of protein can result in the loss of protein function, the collagen protein chain is broken to accelerate the loss rate, thereby promoting the skin aging or even other related diseases.
In addition to collagen, which has been the most important of the previous research, elastin also has an important effect on the general condition of the skin. Elastin is a protein that maintains the elasticity of connective tissue, allowing many tissues in the body to recover their shape after stretching or contraction. With the increasing age and the external injuries received, the content of elastin in skin will gradually decrease. When elastin is lost, the skin will lose elasticity and cause laxity and wrinkles, even if there is a certain proportion of collagen.
In addition, when elastin and/or collagen are lost, the thickness of the skin is indirectly reduced, so that the epidermis layer is closer to blood vessels in the dermis layer, and the whole skin is in a reddish state, thereby affecting the appearance of the skin. Skin in this state is also more sensitive than skin in a healthy state, and is more likely to develop other skin symptoms such as inflammation.
Disclosure of Invention
In view of the above, there is a need to develop or research a composition that can reduce the number or area of red spots on the skin to maintain or improve the appearance of the skin.
Accordingly, an object of the present invention is to provide a use of an extract from seed coats of Fagopyrum tataricum for preparing a composition for reducing the number or area of red spots on skin, wherein the extract is obtained by extracting seed coats of Fagopyrum tataricum with water.
According to some embodiments of the invention, the composition is administered to a subject daily for at least 2 weeks, wherein the composition contains 5 grams to 10 grams of tartary buckwheat seed coat extract.
According to some embodiments of the invention, the composition is further prepared as a pharmaceutical composition, a food composition, or a nutraceutical composition.
According to some embodiments of the present invention, the extract of tartary buckwheat seed coat is obtained by the following steps: pulverizing and sieving testa Fagopyri Esculenti to form testa Fagopyri Esculenti powder; and extracting the tartary buckwheat seed coat powder with water to form a tartary buckwheat seed coat extract, wherein the solid-liquid weight ratio of the tartary buckwheat seed coat powder to the water is 10: 1 to 12: 1.
according to some embodiments of the present invention, the step of extracting the tartary buckwheat seed hull powder with water to form the tartary buckwheat seed hull extract comprises extracting the tartary buckwheat seed hull powder with water at 80 ℃ to 90 ℃ for 0.5 to 3.0 hours to form a first extract; cooling the first extract to room temperature; centrifuging the cooled first extraction liquid, and taking the supernatant as a first extraction supernatant; filtering the first extraction supernatant: and concentrating the filtered first extraction supernatant under reduced pressure to form a tartary buckwheat seed coat extract.
In some embodiments, the temperature range for concentration under reduced pressure is 45-70 ℃.
The invention is described in detail below with reference to the drawings and specific examples, but the invention is not limited thereto.
Drawings
FIG. 1 shows that the extract of seed coat of Fagopyrum tataricum reduces the amount of the final product of collagen saccharification.
FIG. 2 shows that extract of seed coat of Fagopyrum tataricum reduces the amount of Desmosine (Desmosine) in blood of a subject. P value < 0.01; p value < 0.001.
FIG. 3 shows the improvement rate of skin texture after four consecutive weeks of application of the extract from the seed coat of Fagopyrum tataricum to the facial skin of the subject.
Fig. 4 to 6 are photographs of the facial skin of the subject taken at week 0, week 2 and week 4, respectively (texture portions are marked by software).
FIG. 7 shows the change in the relative amount of red spots on the skin of a subject after four consecutive weeks of application of the extract from the seed coat of Fagopyrum tataricum to the facial skin of the subject. P value < 0.05.
Fig. 8 to 10 are images of the facial skin of the subject taken by the detector at week 0, week 2 and week 4, respectively.
Detailed Description
Some embodiments of the invention will be described below. The invention may be embodied in many different forms without departing from the spirit thereof, and the scope of protection should not be limited to the details given herein.
The invention uses Excel software to perform statistical analysis. Data are expressed as mean ± Standard Deviation (SD) and differences between groups were analyzed by student's t-test (student's t-test).
Examples of the buckwheat species that are commonly used and edible in the genus Fagopyrum include common buckwheat (Fagopyrum esculentum, also referred to as "buckwheat") and tartary buckwheat (Fagopyrum mtaraticum, also referred to as "tartary buckwheat"). Common buckwheat is commonly grown in continents, while tartary buckwheat, which is derived from eastern parts of the Tibet or northwestern parts of Yunnan, is grown only in Asia, Europe, and North America. Compared with rice, buckwheat has a shorter growth period, can grow in poor acid soil, does not need excessive nutrients and nitrogen, but needs sufficient moisture, and can flower in a cooler climate. Therefore, the fertilizer can be used as grain, can be used as reseeding after water disasters, and can also be used as green manure, feed or covering plants for preventing water and soil loss.
In general, plant seeds are generally composed of three parts, namely, a seed coat, an embryo and an endosperm, and seeds of the genus Fagopyrum are also the same. The invention finds that the extract of the seed coat of the tartary buckwheat in the genus of buckwheat has the effects of reducing the loss speed of elastin and collagen in the body and reducing the skin redness, and can be used for preparing a composition for reducing the loss of elastin and collagen in the body and reducing the number or area of red spots on the skin. The extract of the seed coat of tartary buckwheat has the above effects, so that the extract can be used for improving skin problems and further improving the appearance of the whole skin.
The term "extract of seed coat of Fagopyrum tataricum" as used herein is obtained by extracting seed coat of Fagopyrum tataricum with a solvent (e.g. water) for a specific time and temperature.
The tartary buckwheat seed coat extract can be used for preparing a composition for reducing in-vivo elastin loss or a composition for inhibiting collagen glycosylation, and the composition can be a pharmaceutical composition, a food composition and a health-care food composition. Alternatively, the pharmaceutical composition of the present invention may be contained in a food or a cosmetic. Or the tartary buckwheat seed coat extract can be further prepared into a pharmaceutical composition, a health-care product composition, a food composition or a health-care food composition.
The pharmaceutical composition may be manufactured using techniques well known to those skilled in the art into a dosage form suitable for parenteral (parenterally) or topical (topocalily) administration, including, but not limited to: injections (injection) [ e.g., sterile aqueous solution (sterile aqueous solution) or dispersion (dispersion) ], sterile powders (sterile powder), external preparations (external preparation), and the like.
The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier (pharmaceutically acceptable carrier) which is widely used in pharmaceutical manufacturing technology. For example, the pharmaceutically acceptable carrier may comprise one or more agents selected from the group consisting of: solvents (solvents), buffers (buffers), emulsifiers (emulsifiers), suspending agents (suspending agents), disintegrating agents (disintegrants), disintegrating agents (disintegrating agents), dispersing agents (dispersing agents), binding agents (binding agents), excipients (excipients), stabilizers (stabilizing agents), chelating agents (chelating agents), diluents (diluents), gelling agents (gelling agents), preservatives (preserving), wetting agents (wetting agents), lubricants (lubricants), absorption delaying agents (absorbing agents), liposomes (lipids), and the like. The choice and amount of such agents will be readily selected by the skilled artisan.
In some embodiments, the pharmaceutically acceptable carrier comprises a solvent selected from the group consisting of: water, normal saline (normal saline), Phosphate Buffered Saline (PBS), aqueous alcohol-containing solutions (aqueous solution linking alcohol), and combinations thereof.
In some embodiments, the pharmaceutical composition may be administered by a parenteral route (parenteral routes) selected from the group consisting of: subcutaneous injection (subecanal injection), intradermal injection (intraepithelial injection), and intralesional injection (intralesion).
In some embodiments, the pharmaceutical composition can be manufactured into an external preparation (external preparation) suitable for topical application to the skin using techniques well known to those skilled in the art, including, but not limited to: creams (lotion), gels (gel), ointments (ingredient), creams (cream), patches (patch), liniments (liniment), powders (powder), aerosols (aerosol), sprays (spray), emulsions (lotion), serum (serum), pastes (paste), foams (foam), drops (drop), suspensions (suspension), ointments (salve) and bandages (bandage).
In some embodiments, the external preparation is prepared by mixing the pharmaceutical composition with a base (base) as is well known to those skilled in the art.
In some embodiments, the substrate may include one or more additives (additives) selected from the group consisting of: water, alcohols, glycols, hydrocarbons such as petroleum jelly and white petrolatum]Wax (wax) [ such as Paraffin and yellow wax (yellow wax)]Preserving agents (preserving agents), antioxidants (antioxidants), surfactants (surfactants), absorption enhancers (absorption enhancers), stabilisers (stabilizing agents), gelling agents (gelling agents) [ such as
974P(
974P), microcrystalline cellulose (microcrystalline cellulose) and carboxymethyl cellulose (carboxymethyl cellulose)]Active agents (actives), moisturizers (humectants), odor absorbers (odor absorbers), perfumes (fragrans), pH adjusting agents (pH adjusting agents), chelating agents (chelating agents), emulsifiers (emulsifiers), occlusive agents (occlusive agents), softeners (emulsifiers), thickeners (thickeners), solubilizing agents (solubilizing agents), penetration enhancers (penetration enhancers), anti-irritants (anti-irritants), colorants (colorants), and propellants (propellants). The selection and amounts of such additives are within the skill and routine skill of those skilled in the art.
In some embodiments, the care product may include an acceptable adjuvant (acceptable adjuvant) that is widely used in the art of care product manufacture. For example, the acceptable adjuvant may comprise one or more agents selected from the group consisting of: solvents, gelling agents, active agents, preservatives, antioxidants, screening agents, chelating agents, surfactants, colouring agents, thickening agents, fillers, fragrances and odour absorbers. The selection and the amount of the reagents can be properly adjusted according to actual requirements.
In some embodiments, the treatment composition may be manufactured in a form suitable for skin care (skincare) or makeup (makeup) using techniques well known to those skilled in the art, including, but not limited to: aqueous solutions (aqueous solutions), aqueous-alcoholic solutions (aqueous-alcoholic solutions) or oily solutions (oil solutions), emulsions in the form of oil-in-water type, water-in-oil type or compound type, gels, ointments, creams, masks (masks), patches, wipes, powders, aerosols, sprays, lotions, serums, pastes, foams, dispersions, drops, mousses (mousses), sunblocks, lotions (toiletries), foundations (foundations), make-up removal products (make-up removal products), soaps (soaps) and other body cleansing products (body cleansing products), and the like.
In some embodiments, the treatment may also be combined with one or more known active topical agents (external use agents) selected from the following: whitening agents (whitening agents) [ such as vitamin a acid (tretinoin), catechin (catechin), kojic acid, arbutin and vitamin C ], moisturizing agents, bactericides (bacteriodes), ultraviolet absorbers (ultravirosomes), plant extracts (plant extracts) [ such as aloe vera extract (aloe extract) ], skin nutrients (skin nutrients), anesthetics (anesthesics), antiacne agents (anti-acne agents), antipruritics (antipruritics), analgesics (analgesis), anti-dermatitis agents (anti-hyperkeratosis agents), anti-xerosis agents (anti-dry skin agents), antiperspirants (antiperspirancy agents), anti-aging agents (anti-aging agents), anti-aging agents (anti-inflammation agents), anti-aging agents (anti-acne agents), anti-xerosis agents (anti-seborrheic agents), anti-perspiration agents (anti-aging agents), anti-aging agents (anti-corticosteroid), anti-seborrheic agents (anti-seborrheic agents), and anti-corticosteroid (anti-seborrheic agents). The selection and amounts of such agents for external use are within the skill and routine skill of those skilled in the art.
In some embodiments, the pharmaceutical composition may be used as a food additive (food additive) to be added during the preparation of raw materials or during the manufacture of food by conventional methods, to be formulated with any edible material into a food product for ingestion by humans and non-human animals.
In some embodiments, the types of food products include, but are not limited to: beverages (leafages), fermented foods (fermented foods), bakery products (bakery products), health foods (health foods) and dietary supplements (dietary supplements).
Hereinafter, a method for extracting the seed coat extract of Fagopyrum tataricum and a method for detecting the seed coat extract of Fagopyrum tataricum will be described in detail.
[ example 1] method for preparing extract of seed coat of Fagopyrum tataricum
In this example, the extract of seed coat of Fagopyrum tataricum was prepared as follows:
1. pulverizing seed coat of Fagopyrum tataricum (producing area: Taiwan area), sieving with 10 mesh sieve, and removing oversize granules to obtain Fagopyrum tataricum seed coat powder. In other embodiments, the mesh size of the screen may be 8 mesh or 12 mesh.
2. Mixing water and tartary buckwheat seed husk powder in a weight ratio of 11: 1 and soaking at 85 + -5 deg.C for about 1.0 hr to extract Fagopyrum tataricum seed coat powder to form a first extract containing solids. In other embodiments, the weight ratio of the solvent to the tartary buckwheat seed husk powder can also be 10: 1 to 12: 1, the temperature for soaking can be 80-90 ℃, and the extraction time can be 0.5-3.0 hours. If the solvent is too little or the extraction time is too short, the extraction efficiency will be obviously reduced; if the extraction time is too long, the active ingredients in the extract may be degraded.
3. The first extract was cooled to room temperature (25 ℃ -30 ℃).
4. The first extract with the solid after cooling was centrifuged in a centrifuge (brand/model: Thermo Scientific Heraeus Fresco 17) for about 10 minutes, and the supernatant was taken as the first extract supernatant.
5. The first extraction supernatant was filtered through a 400 mesh sieve to remove fine solids.
6. Concentrating the filtered first extractive supernatant with a concentrator (brand/model: BUCHI-Rotavapor R-100) at 60 + -5 deg.C under reduced pressure until the Brix value (Degrees Brix) of the solution is 2.0 + -0.5, and stopping concentrating to obtain Fagopyrum tataricum seed coat extract. In other embodiments, concentration under reduced pressure may be carried out at 45 ℃ to 70 ℃.
[ example 2] detection of the amount of collagen glycation
As previously mentioned, one of the factors of collagen chain cleavage is the glycosylation of collagen. When proteins undergo glycosylation reactions, many kinds of glycosylation end products (AGEs) are produced, which are non-reducible substances and alter and affect the normal function of the proteins. Therefore, if the glycosylation of collagen can be inhibited, the breakage of collagen chains can be prevented, and the loss rate of collagen can be effectively reduced. To confirm whether the extract of tartary buckwheat seed coat has the effect of inhibiting collagen glycosylation, the collagen glycosylation amount is tested as follows.
Preparing a medicine:
1. using the extract of seed coat of Fagopyrum tataricum obtained in the previous example 1, Phosphate-Buffered Saline (PBS, NaH) was used as Phosphate-Buffered Saline with a concentration of 200mM 2 PO 3 (Honeywell, model #04269), Na 2 HPO 4 (Sigma, model # V900061) was mixed with water at a pH of 7.4) to prepare solutions of the extract of the seed coat of Fagopyrum tataricum at concentrations of 0.1mg/mL, 0.5mg/mL, 1.0mg/mL, and 5mg/mL, respectively, as a solvent.
2. A collagen solution having a concentration of 60mg/mL was prepared using the aforementioned phosphate buffered saline at a concentration of 200mM and collagen powder (collagen available from Rousselot, model # P2000 HD). Sodium azide (Sodium azide) was added to the collagen solution in an amount of 0.06 wt%. (sodium azide from Sigma, model # S2002)
3. A fructose solution was prepared at a concentration of 1.5M using the aforementioned phosphate buffered saline at a concentration of 200mM with fructose (fructose purchased from Sigma, model # F0127).
Collagen glycosylation test:
1. 0.2mL of the seed coat extract solution of tartary buckwheat of the 4 kinds of concentrations is mixed with 0.2mL of the collagen solution and 0.2mL of the fructose solution respectively to prepare 4 groups of sample solutions.
2. 0.2mL of deionized water was mixed with 0.2mL of the collagen solution and 0.2mL of the fructose solution to prepare a blank solution as a control group.
3. The fluorescence intensity (excitation wavelength 360nm, emission wavelength 460nm) of the 4 sets of sample solutions and the blank solution was measured using a spectrofluorometer (BioTekFLx 800, manufactured and sold by Ltd.) before the collagen glycosylation reaction was performed.
4. The 4 groups of sample solutions and the blank solution were left at 50 ℃ for 24 hours to allow each solution to undergo a collagen glycosylation reaction.
5. The fluorescence intensities (excitation wavelength 360nm, emission wavelength 460nm) of the 4 groups of sample solutions after the reaction and the blank solution were measured by the spectrofluorometer.
6. The relative production rate of the final product of protein glycosylation is calculated according to the following formula:
[ (intensity of fluorescence of sample) After the reaction Fluorescence intensity of the sample Before reaction ) /(control group fluorescence intensity) After the reaction Control group fluorescence intensity Before reaction )]×100％
The relative production rates of the end products of protein glycosylation for each sample relative to the control group are shown in figure 1.
As shown in FIG. 1, each concentration of the extract of Fagopyrum tataricum seed coat (0.1mg/mL, 0.5mg/mL, 1.0mg/mL, and 5mg/mL) has the effect of inhibiting the production of the collagen-containing end product, and has a correlation with the concentration of the extract of Fagopyrum tataricum seed coat. In the sample solutions of the tartary buckwheat seed coat extracts of 0.1mg/mL, 0.5mg/mL, 1.0mg/mL and 5mg/mL, the relative production rates of the protein glycosylation final products are respectively 92%, 48%, 31% and 19% of the control group. The tartary buckwheat seed coat extract can really inhibit the glycosylation reaction of the collagen, reduce the generation of glycosylation final products, further reduce the loss of the collagen in vivo and achieve the effect of slowing down the skin aging.
[ example 3] measurement of desmosine content in vivo
Elastin has a specific component, Desmosine (Desmosine), which is a cross-linker formed by three aldysine side chains and one lysine side chain, and can covalently cross-link more than 2 polypeptide chains in a collagen molecule. The special structure and properties of desmosine impart elasticity and connectivity to elastin, allowing elastin to act as a "latch" between collagen in the skin.
When the elastin is decomposed, the catenin contained in the elastin is released into the blood, so that the concentration of the catenin in the blood is increased; conversely, if the amount of elastin decomposed is decreased, the concentration of desmosine in the blood will decrease. Therefore, the concentration of desmosine in the blood can be used as an index of the amount of elastin decomposed. In order to confirm whether the extract from the seed coat of Fagopyrum tataricum can inhibit the decomposition of elastin, the content of desmosine in vivo was measured as follows.
The extract of the seed coat of tartary buckwheat prepared in the example 1 is mixed with water to prepare a beverage of the extract of the seed coat of tartary buckwheat. And 8 subjects (general women 30-55 years old) were asked to drink the prepared tartary buckwheat seed coat extract drink at a daily intake dose of 5g of tartary buckwheat seed coat extract for 4 weeks. And the subjects were bled before the start of the experiment (week 0, not drunk), at weeks 2 and 4, and the Desmosine ELISA Kit (CUSABIO, CSB-E12871h) was used to measure the Desmosine concentration in the blood (the drugs used in the following steps, unless otherwise specified, were provided by the Desmosine ELISA Kit).
Measurement of desmosine concentration in blood:
1. a serum sample is separated from the collected whole blood sample.
2. Each serum sample, a set of serially diluted Desmosine standards (used to make a calibration line) with known concentrations, and a blank solution were pipetted 100. mu.l into a 96-well plate dedicated to the Desmosine ELISA Kit.
3. The aforementioned 96-well plate was placed in a 37 ℃ oven for 2 hours of reaction.
4. Remove the liquid from the 96-well plate.
5. Pipette 100 μ l of Biotin-antibody (1 ×) into each well of a 96-well plate.
6. The 96-well plate was then placed back in the 37 ℃ oven for 1 hour.
7. Remove the liquid from the 96-well plate.
8. The 96-well plates were washed 3 times with Wash buffer (phosphate buffered saline at 200mM, pH 7.4) each time as follows:
(1) 200. mu.l of Wash buffer was added to each well.
(2) And standing for 2 minutes.
(3) Remove the liquid from the 96-well plate.
9. Pipette 100 μ l of HRP-avidin (1X) into each well of a 96-well plate.
10. The 96-well plate was placed back in the 37 ℃ oven for 1 hour of reaction.
11. The 96-well plate was washed 3 times with Wash buffer, i.e., step 8 was repeated three times.
12. Mu.l of TMB (3,3 ', 5, 5' -Tetramethylbenzidine) Substrate was pipetted into each well of a 96-well plate.
13. The 96-well plate was placed back in the 37 ℃ oven for 15 minutes.
14. 50 μ l of Stop Solution was pipetted into each well of a 96 well plate.
15. The absorbance at 450nm and 540nm was measured by ELISA reader (BioTek, FLx 800).
16. A standard curve was established by Excel, and the absorbance at 450nm was corrected for the absorbance at 540nm (absorbance at 450nm and 540nm was measured for each well, and then the absorbance at 540nm was subtracted from the absorbance at 450 nm), and quantitative conversion of the sample was performed.
17. The samples were statistically analyzed using Student-t test in Excel.
The results of measurement of the relative concentration of desmosine in blood, with the concentration at week 0 taken as 100%, are shown in FIG. 2. Graph denotes p <0.01 relative to the reference group; denotes p <0.001 relative to the reference group.
As shown in fig. 2, after the subjects continuously drink the tartary buckwheat seed coat extract for 2 weeks, the desmosine content in the bodies of the subjects is remarkably reduced and is only about 78.1 percent before the subjects take the tartary buckwheat seed coat extract. After the tartary buckwheat seed coat extract is continuously drunk for 4 weeks, the chain element content in the body can be reduced to about 65.7 percent before the tartary buckwheat seed coat extract is ingested. It is shown that the damage amount of elastin in the body of the subject is greatly reduced after the subject ingests the tartary buckwheat seed coat extract, and the reduction effect can be more obvious along with the increase of the ingesting period. Therefore, the tartary buckwheat seed coat extract can effectively slow down the loss of elastin in the body.
Example 4 skin texture improvement test/relative amount of skin Red spots test
The extract of the seed coat of tartary buckwheat prepared in example 1 and water are prepared into a drink of the extract of the seed coat of tartary buckwheat so as to provide a proper effective dose for a subject. In this example, 8 subjects (women of 30-55 years old) were asked to drink the prepared fagopyrum tataricum seed coat extract drink at a daily intake dose of 5g of fagopyrum tataricum seed coat extract for 4 weeks. The subjects were tested for their cleansed facial skin before the start of the experiment (week 0, not drunk), week 2 and week 4 by a whole-face skin texture tester (7th Generation VISIA Complex Analysis System; Canfield, USA).
The results of the skin texture detection and the actual appearance change are shown in fig. 3 and fig. 4 to 6, respectively. Fig. 3 is a graph showing the skin roughness of the test subjects before taking the extract of the seed coat of Fagopyrum tataricum as 100%, and recording the relative skin roughness of the test subjects after taking the extract of the seed coat of Fagopyrum tataricum for 2 weeks and 4 weeks. Fig. 4-6 are photographs of the facial skin of a subject taken at week 0, week 2, and week 4, wherein the texture portion was marked by soft body manipulation.
As shown in fig. 3, the improvement of skin texture was observed when the subjects took the extract of tartary buckwheat seed coat for 2 weeks, and the relative roughness of skin texture was reduced to 87.6% after 4 weeks of continuous taking. It can be seen from the real photographs of fig. 4 to 6 that the fine lines on the skin are significantly reduced or lightened as the period of taking the extract from the seed coat of tartary buckwheat increases.
The results of the number and area of red spots on the skin of the subject and the actual appearance change are shown in fig. 7 and fig. 8 to 10, respectively. Fig. 7 is a value of 100% measured by an instrument according to the number and area of red spots before the test subject ingests the extract of the seed coat of tartary buckwheat, and a value measured according to the number/area of red spots after the test subject continuously ingests the extract of seed coat of tartary buckwheat for 2 weeks and 4 weeks is recorded. Fig. 8 to 10 are images of the facial skin of one subject taken by the detector at week 0, week 2 and week 4.
As shown in fig. 7, when the subjects took the extract of the seed coat of tartary buckwheat for 2 weeks, the reduction in the number/area of red spots began to be observed, and when the subjects took the extract for 4 weeks continuously, the number/area of red spots on the facial skin decreased to 95.1% of the original number/area. In the real-shot images of fig. 8 to 10, it can be found that the number/area of red spots on the skin is significantly reduced or faded as the period of taking the extract from the seed coat of tartary buckwheat increases.
From the results of the skin texture improvement test and the relative amount of skin red spots and other experiments, it can be known that the extract from the seed coat of Fagopyrum tataricum can inhibit the glycosylation of collagen and reduce the concentration of desmosine in blood, and further can inhibit the loss of collagen and elastin in the body, and thus can be used for slowing down the aging of skin and/or improving the appearance of skin.
The foregoing is by way of example only, and not limiting. It is intended that all equivalent modifications or variations without departing from the spirit and scope of the present invention shall be included in the appended claims.
Claims (6)
1. Use of an extract of testa Fagopyri Tatarici for preparing a composition for reducing the number or area of red spots on skin, wherein the extract is obtained by extracting a testa Fagopyri Tatarici with water.
2. The use of claim 1, wherein the composition is a pharmaceutical composition.
3. The use of claim 1, wherein the composition is further formulated as a cosmetic composition, a food composition, or a nutraceutical composition.
4. The use according to any one of claims 1 to 3, wherein the extract of the seed coat of Fagopyrum tataricum is obtained by a method comprising the steps of:
pulverizing and sieving a seed coat of Fagopyrum tataricum to form a seed coat powder of Fagopyrum tataricum; and
extracting the tartary buckwheat seed coat powder with water to form the tartary buckwheat seed coat extract, wherein the solid-liquid weight ratio of the tartary buckwheat seed coat powder to the water is 10: 1 to 12: 1.
5. the use of claim 4, wherein the step of extracting the testa Fagopyri Tatarici powder with water to form the testa Fagopyri Tatarici extract comprises:
extracting the seed coat powder of tartary buckwheat with water at 80-90 deg.C for 0.5-3.0 hr to form a first extractive solution;
cooling the first extract to room temperature;
centrifuging the cooled first extraction liquid, and taking the supernatant as a first extraction supernatant;
filtering the first extraction supernatant: and
concentrating the filtered first extraction supernatant under reduced pressure to form the tartary buckwheat seed coat extract.
6. Use according to claim 5, wherein the temperature range for concentration under reduced pressure is 45-70 ℃.
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|---|---|---|---|---|
| KR100711007B1 (en) * | 2005-07-28 | 2007-04-25 | 재단법인춘천바이오산업진흥원 | Functional Cosmetic Composition Comprising Extract of Fagopyrum esculentum |
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2020
- 2020-06-11 TW TW109119752A patent/TWI801738B/en active
- 2020-06-11 CN CN202210896717.3A patent/CN115040564A/en active Pending
- 2020-06-11 CN CN202010533517.2A patent/CN112057502A/en active Pending
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| CN105795480A (en) * | 2016-04-08 | 2016-07-27 | 吉林鑫水科技开发有限公司 | Extraction and analysis method and application of flavone in buckwheat |
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| Publication number | Publication date |
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| TW202045134A (en) | 2020-12-16 |
| TWI801738B (en) | 2023-05-11 |
| CN112057502A (en) | 2020-12-11 |
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