CN116617120A - Preparation method of sea grape extract, product and application thereof - Google Patents
Preparation method of sea grape extract, product and application thereof Download PDFInfo
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- CN116617120A CN116617120A CN202310858782.1A CN202310858782A CN116617120A CN 116617120 A CN116617120 A CN 116617120A CN 202310858782 A CN202310858782 A CN 202310858782A CN 116617120 A CN116617120 A CN 116617120A
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- sea grape
- grape extract
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- sea
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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
- A61K8/9706—Algae
- A61K8/9722—Chlorophycota or Chlorophyta [green algae], e.g. Chlorella
-
- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- 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
-
- 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
-
- 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
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Chemical & Material Sciences (AREA)
- Dermatology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Mycology (AREA)
- Botany (AREA)
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a preparation method of a sea grape extract, a product and application thereof, wherein the preparation method of the sea grape extract comprises the following steps: crushing and squeezing the sea grape raw material to obtain a squeezed product; carrying out ultrahigh pressure treatment on the pressed product to obtain a treated product; centrifuging the treated product, mixing the supernatant with a flocculant for flocculation treatment, centrifuging again, and carrying out fine filtration on the supernatant to obtain the sea grape extract. The preparation method adopts the ultra-high pressure to treat the squeezed product of the sea grape raw material, and combines the subsequent flocculant treatment, so that the obtained product has excellent moisturizing, antioxidation and whitening effects, and the whole process is simple and easy to operate.
Description
Technical Field
The invention belongs to the technical field of plant extraction, relates to a preparation method of a sea grape extract, a product and application thereof, and in particular relates to a preparation method of a sea grape extract with excellent skin moisturizing, whitening and antioxidation effects, a product and application thereof.
Background
Botrytis cinerea belonging to genus Botrytis of family Botryococcus of phylum Chlorophyta, the appearance of the grape is known as a string of sparkling and crystal clear grapes, and the grape is also called "sea grape". In recent years, various algae and extracts thereof are gradually paid attention to by researchers and consumers, and Botrytis cinerea contains rich polysaccharide, seaweed polyphenol, amino acid, plant tannin, high dietary fiber, amino acid, unsaturated fatty acid, inorganic salt and various trace elements, has resistance to fungi and bacteria, can prevent certain chronic diseases, and has better effects on resisting cancer, tumor, oxidization and the like. The sea grape extract also has good efficacy in the aspects of beauty and health care, has the name of a beauty magic artist, has higher economic value, and has wide development prospect in the cosmetic market.
The prior art sea grape extract is mostly extracted under high temperature or extracted by using organic solvent, for example, CN111419743a discloses a preparation method of sea grape extract, which comprises the following steps: washing the sea grape with deionized water, crushing, adding deionized water and antioxidant, heating, stirring and extracting; cooling, coarse filtering and fine filtering; suction filtration, sterilization and corrosion prevention of filtrate; precipitating with ethanol, and lyophilizing to obtain the extract. The organic solvent is used for extraction or extraction under high temperature condition, so that the residual organic solvent in the target product is easy to sensitize, the harm to human body is caused, and the stability of the effective active ingredients cannot be fully ensured.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a preparation method of a sea grape extract and a product and application thereof, in particular to a preparation method of a sea grape extract with excellent skin moisturizing, whitening and antioxidation effects and a product and application thereof.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
in a first aspect, the present invention provides a method for preparing a sea grape extract, the method comprising the steps of:
(1) Crushing and squeezing the sea grape raw material to obtain a squeezed product;
(2) Carrying out ultrahigh pressure treatment on the pressed product to obtain a treated product;
(3) Centrifuging the treated product, mixing the supernatant with a flocculant for flocculation treatment, centrifuging again, and carrying out fine filtration on the supernatant to obtain the sea grape extract.
The preparation method of the sea grape (also called as Botrytis cinerea) extract provided by the invention creatively adopts ultrahigh pressure to treat the squeezed product of the sea grape raw material, and combines with subsequent flocculant treatment, so that the obtained product has excellent moisturizing, antioxidation and whitening effects, the whole process is simple and easy to operate, and compared with other extraction processes (such as high-temperature extraction, organic solvent extraction and the like), the extract can be ensured to be permanently and stably exerted on the effects to the greatest extent, and is safer and milder and wider in applicability.
Preferably, the pressure of the ultra-high pressure treatment is 180-280MPa, for example 180MPa, 190MPa, 200MPa, 210MPa, 220MPa, 230MPa, 240MPa, 250MPa, 260MPa, 280MPa, etc.; the dwell time is 5-15min, such as 5min, 7min, 8min, 9min, 10min, 12min, 13min, 14min, 15min, etc. Other specific point values in the numerical ranges are selectable, and will not be described in detail herein.
When the specific technological parameters are selected by ultrahigh pressure treatment, the prepared sea grape extract has higher content of active ingredients, and the moisturizing, antioxidant and whitening effects can be ensured to a greater extent.
Preferably, the centrifugation is performed at 2000-4000rpm (e.g., 2000rpm, 2500rpm, 3000rpm, 3500rpm, 4000rpm, etc.) for 20-40min (e.g., 20min, 25min, 30min, 35min, 40min, etc.). Other specific point values in the numerical ranges are selectable, and will not be described in detail herein.
Preferably, the fine filtration is performed using a filter plate having a pore size of 0.5 to 3 μm (e.g., 0.5 μm, 1 μm, 1.5 μm, 2 μm, 2.5 μm, 3 μm, etc.). Other specific point values within the numerical range can be selected, and will not be described in detail herein.
Preferably, the flocculant comprises any one or a combination of at least two of a water-soluble polysaccharide, a cationic chitosan, guar gum, a protein colloid, gelatin, an animal glue, a cationic starch, or tannin.
Preferably, the flocculant comprises a combination of cationic chitosan and guar gum.
The flocculant was pretreated as follows prior to use: mixing and stirring cationic chitosan and guar gum in a small amount of acetic acid aqueous solution to form paste, and then mixing and swelling with water.
The flocculant with specific composition in the preparation method can remove inactive impurities, so that the composition mode of the treated product is obviously improved in the effects of moisture preservation, antioxidation and whitening.
Preferably, the mass ratio of the cationic chitosan to the guar gum is 1:1-1:3, for example, 1:1, 2:3, 1:2, 2:5, 1:3, etc., and other specific values within the numerical range are selectable, which will not be described in detail herein.
The invention also discovers that the flocculating agent formed by compounding the cationic chitosan and the guar gum in a mass ratio of 1:1-1:3 is preferably adopted, and the composition mode of the product after flocculation treatment is further improved in the effects of moisture preservation, antioxidation and whitening.
Preferably, the fine filtration is followed by a decolorizing treatment and/or a deodorizing treatment.
The decoloring and/or deodorizing treatment uses, for example, any one or a combination of at least two of activated carbon, diatomaceous earth, silica gel, or neutral alumina.
In a second aspect, the present invention provides a marine grape extract prepared according to the preparation method of the first aspect.
In a third aspect, the present invention provides the use of a marine grape extract according to the second aspect for the preparation of a product with moisturizing effect.
Preferably, the product comprises a cosmetic, the content of the sea grape extract in the cosmetic being 0.001-0.1%, for example 0.001%, 0.005%, 0.008%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1% etc. by solid content. Other specific point values within the numerical range can be selected, and will not be described in detail herein.
In a fourth aspect, the present invention provides the use of a marine grape extract according to the second aspect for the preparation of a product having antioxidant efficacy.
Preferably, the product comprises a cosmetic, the content of the sea grape extract in the cosmetic being 0.001-0.1%, for example 0.001%, 0.005%, 0.008%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1% etc. by solid content. Other specific point values within the numerical range can be selected, and will not be described in detail herein.
In a fifth aspect, the present invention provides the use of a marine grape extract according to the second aspect for the preparation of a product having skin whitening efficacy.
Preferably, the product comprises a cosmetic, the content of the sea grape extract in the cosmetic being 0.001-0.1%, for example 0.001%, 0.005%, 0.008%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1% etc. by solid content. Other specific point values within the numerical range can be selected, and will not be described in detail herein.
Compared with the prior art, the invention has the following beneficial effects:
the preparation method of the sea grape extract creatively adopts ultrahigh pressure to treat the squeezed product of the sea grape raw material, and combines the subsequent flocculant treatment, so that the obtained product has excellent moisturizing, antioxidation and whitening effects, the whole process is simple and easy to operate, and compared with other extraction processes (such as high-temperature extraction, organic solvent extraction and the like), the extract can be ensured to be permanently and stably exerted on the efficacy to the greatest extent, and the preparation method is safer and milder and has wider applicability.
Drawings
FIG. 1 is a graph showing the effect of a blank control group, a positive control group and a sea grape extract group on the formation of melanin on the body surface of a zebra fish embryo.
Detailed Description
The technical scheme of the invention is further described by the following specific embodiments. It will be apparent to those skilled in the art that the examples are merely to aid in understanding the invention and are not to be construed as a specific limitation thereof.
The marine grape raw materials according to the following examples were purchased from guangzhou-one-pill biotechnology limited;
the cationic chitosan is purchased from Guangzhou Taikang daily chemical industry Co., ltd;
guar gum is a product available from sorv under the model number JAGUARC-14S.
Example 1
The embodiment provides a sea grape extract and a preparation method thereof:
(1) Crushing the sea grape raw material to 60 meshes, and performing physical pressing under the pressure of 0.5MPa for 10min to obtain a pressed product;
(2) Carrying out ultrahigh pressure treatment on all the pressed products, wherein the pressure is 200MPa, and the dwell time is 10min, so as to obtain treated products;
(3) Centrifuging all the treated products at 3000rpm for 30min, mixing the supernatant with 1.5% flocculant (obtained by mixing cationic chitosan and guar gum with 1% acetic acid aqueous solution with a mass ratio of 1:2, stirring to paste, and swelling with 70 times of water for 24 h), performing flocculation treatment, centrifuging at 2000rpm for 40min, fine filtering the supernatant with a filter plate with a pore diameter of 1 μm, adding 1% active carbon into the filtrate, and decolorizing and deodorizing to obtain the sea grape extract.
Example 2
The embodiment provides a sea grape extract and a preparation method thereof:
(1) Crushing the sea grape raw material to 60 meshes, and performing physical pressing under the pressure of 0.5MPa for 10min to obtain a pressed product;
(2) Carrying out ultrahigh pressure treatment on all the pressed products, wherein the pressure is 250MPa, and the dwell time is 5min, so as to obtain treated products;
(3) Centrifuging all the treated products at 2500rpm for 35min, mixing the supernatant with 1.5% flocculant (obtained by mixing cationic chitosan and guar gum with a mass ratio of 1:3 with 3 times of 1% acetic acid aqueous solution, stirring to paste, and swelling with 70 times of water for 24 h), flocculating, centrifuging at 2500rpm for 35min, fine filtering the supernatant with a filter plate with a pore diameter of 1 μm, adding 1% active carbon into the filtrate, decolorizing and deodorizing to obtain the sea grape extract.
Example 3
The embodiment provides a sea grape extract and a preparation method thereof:
(1) Crushing the sea grape raw material to 60 meshes, and performing physical pressing under the pressure of 0.5MPa for 10min to obtain a pressed product;
(2) Carrying out ultrahigh pressure treatment on all the pressed products, wherein the pressure is 180MPa, and the dwell time is 12min, so as to obtain treated products;
(3) Centrifuging all the treated products at 3500rpm for 20min, mixing the supernatant with 1.5% flocculant (prepared by mixing cationic chitosan and guar gum with a mass ratio of 1:1 with 3 times of 1% acetic acid aqueous solution, stirring to paste, and swelling with 70 times of water for 24 h), flocculating, centrifuging at 3500rpm for 20min, fine filtering the supernatant with a filter plate with a pore diameter of 1 μm, adding 1% active carbon into the filtrate, decolorizing and deodorizing to obtain the sea grape extract.
Example 4
The present example provides a sea grape extract and a method for preparing the same, which are different from example 1 only in that all the pressed products are subjected to ultra-high pressure treatment in step (2), the pressure is 150MPa, and the dwell time is 15min, so as to obtain the treated products. Other conditions remained unchanged.
Example 5
The present example provides a sea grape extract and a method for preparing the same, which are different from example 1 only in that all the pressed products are subjected to ultra-high pressure treatment in step (2), the pressure is 350MPa, and the dwell time is 5min, so as to obtain the treated products. Other conditions remained unchanged.
Example 6
The present example provides a sea grape extract and a preparation method thereof, which are different from the example 1 only in that the flocculant in the step (3) is obtained by mixing and stirring a single cationic chitosan with 3 times of 1% aqueous acetic acid solution to form a paste, then mixing and swelling with 70 times of water for 24 hours, and the addition amount of the flocculant is kept unchanged. Other conditions remained unchanged.
Example 7
The present example provides a sea grape extract and a preparation method thereof, which are different from the example 1 only in that the flocculant in the step (3) is obtained by mixing and stirring a single guar gum with 3 times of 1% aqueous acetic acid solution to form a paste, then mixing and swelling with 70 times of water for 24 hours, and the addition amount of the flocculant is kept unchanged. Other conditions remained unchanged.
Comparative example 1
This comparative example provides a sea grape extract and a preparation method thereof, which differ from example 1 only in that no ultrahigh pressure treatment is performed, specifically as follows:
(1) Crushing the sea grape raw material to 60 meshes, and performing physical pressing under the pressure of 1MPa for 20min to obtain a pressed product;
(2) Centrifuging all the squeezed products at 3000rpm for 30min, mixing the supernatant with 1.5% flocculant (obtained by mixing cationic chitosan and guar gum with 1% acetic acid aqueous solution with a mass ratio of 1:2, stirring to paste, and swelling with 70 times of water for 24 h), flocculating, centrifuging at 2000rpm for 40min, fine filtering the supernatant with a filter plate with a pore diameter of 1 μm, adding 1% active carbon into the filtrate, decolorizing and deodorizing to obtain the sea grape extract.
Comparative example 2
This comparative example provides a sea grape extract and a method for preparing the same, which differ from example 1 only in that flocculation treatment is not performed, specifically as follows:
(1) Crushing the sea grape raw material to 60 meshes, and performing physical pressing under the pressure of 0.5MPa for 10min to obtain a pressed product;
(2) Carrying out ultrahigh pressure treatment on all the pressed products, wherein the pressure is 200MPa, and the dwell time is 10min, so as to obtain treated products;
(3) Centrifuging all the treated products at 3000rpm for 30min, collecting supernatant, fine filtering with a filter plate with aperture of 1 μm, adding 1% active carbon into the filtrate, decolorizing and deodorizing to obtain the sea grape extract.
Comparative example 3
The comparative example provides a sea grape extract and a preparation method thereof:
(1) Crushing the sea grape raw material to 60 meshes, and performing physical pressing under the pressure of 0.5MPa for 10min to obtain a pressed product;
(2) Heating and reflux extracting all the pressed products under normal pressure, wherein the extraction conditions are as follows: extracting for 30min at 80deg.C for 2 times to obtain treated product;
(3) Centrifuging all the treated products at 3000rpm for 30min, mixing the supernatant with 1.5% flocculant (obtained by mixing cationic chitosan and guar gum with 1% acetic acid aqueous solution with a mass ratio of 1:2, stirring to paste, and swelling with 70 times of water for 24 h), performing flocculation treatment, centrifuging at 2000rpm for 40min, fine filtering the supernatant with a filter plate with a pore diameter of 1 μm, adding 1% active carbon into the filtrate, and decolorizing and deodorizing to obtain the sea grape extract.
Comparative example 4
The comparative example provides a sea grape extract and a preparation method thereof:
(1) Crushing the sea grape raw material to 60 meshes, and performing physical pressing for 10 minutes under the pressure of 0.5MPa to obtain a pressed product;
(2) Extracting all the squeezed products by ultrasonic-enzyme assistance under the following extraction conditions: the addition amount of cellulase is 3%, the pH is 5.5, simultaneously, the ultrasonic treatment is carried out, the power is 400W, the ultrasonic time is 20min, the ultrasonic temperature is 45 ℃, and the enzyme deactivation treatment is carried out after the completion of the ultrasonic treatment, so as to obtain a treated product;
(3) Centrifuging all the treated products at 3000rpm for 30min, mixing the supernatant with 1.5% flocculant (obtained by mixing cationic chitosan and guar gum with 1% acetic acid aqueous solution with a mass ratio of 1:2, stirring to paste, and swelling with 70 times of water for 24 h), performing flocculation treatment, centrifuging at 2000rpm for 40min, fine filtering the supernatant with a filter plate with a pore diameter of 1 μm, adding 1% active carbon into the filtrate, and decolorizing and deodorizing to obtain the sea grape extract.
Test example 1
The moisturizing effect was evaluated on the sea grape extracts prepared in examples 1 to 7 and comparative examples 1 to 4:
(1) The test principle is as follows: the moisture retention rate of the humectant can be characterized by testing the moisture retention rate of the humectant by a laboratory method (weighing method) based on the different forces of the humectant on water molecules and the slight differences in the ability to absorb and retain moisture.
(2) Instrument and material:
instrument: a constant temperature and humidity box;
test material: sea grape extract (test), water (blank), 10% glycerol solution (standard reference).
(3) Testing environmental conditions: the test environment temperature is 20+/-1 ℃ and the humidity is 45+/-2 RH%.
(4) The testing method comprises the following steps: weighing the sample, placing the sample in a constant temperature and humidity box, weighing the mass of the sample at 2 hours, 4 hours, 8 hours and 24 hours, and recording data to calculate the moisture retention rate. And selecting a reagent known to have moisturizing ability as a standard reference.
(5) Calculating a test result:
the Mt (mass of the test), yt (mass of the blank control) and Ht (mass of the standard reference) values of each period are respectively measured according to the experimental design, and the formula is calculated according to the moisture retention rate:
moisture retention rate P Sample ＝(Mt/M0)×100％
Moisture retention rate P Empty space ＝(Yt/Y0)×100％
Moisture retention rate P Label (C) ＝(Ht/H0)×100％
Relative moisture retention% = [ P Sample -P Empty space ]/[P Label (C) -P Empty space ]×100％
The relative moisture retention of each test article was calculated for each time period.
(6) Evaluation criteria:
based on the calculated relative moisture retention of the samples after 24 hours of continued placement, the data can be divided into four classes: good, high, medium and low moisturizing effect.
The relative moisture retention percentage is lower than 0% -50% "and has low moisture retention performance;
the relative moisture retention percentage is 50% -100%, and the moisture retention performance is moderate;
the relative moisture retention rate is between 100 and 150 percent, and the moisture retention performance is high;
the relative moisture retention rate is more than 150 percent, and the moisture retention performance is good.
(7) The test results are shown in table 1:
TABLE 1
As can be seen from the data in table 1, compared with the comparative example, the sea grape extract prepared by the preparation method of the present invention has more excellent moisturizing effect, and the selection of the flocculant and the conditions of the ultra-high pressure treatment also affect the moisturizing effect of the product.
Test example 2
Evaluation of antioxidant effect on the sea grape extracts prepared in examples 1 to 7 and comparative examples 1 to 4:
(1) The test principle is as follows: free radical overproduction is a major cause of natural aging and photoaging of the skin. Therefore, the free radical is removed to delay skin aging and reduce wrinkles. 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH) is a stable long-life free radical, and when a free radical scavenger exists, the light absorption of the DPPH ethanol solution is weakened due to the single electron pairing with the free radical scavenger. The extent of discoloration of the DPPH ethanol solution is linearly related to the number of electrons it receives, and thus the ability of the test sample to scavenge free radicals, i.e., the magnitude of antioxidant activity, can be evaluated.
(2) Instrument and material:
the main instrument is as follows: spectronicGENESYS-5 spectrophotometer;
the main reagent comprises: DPPH, analytically pure; methanol, analytically pure; ascorbic acid, analytically pure.
Preparing a solution: first, DPPH 1mg is dissolved in about 20mL of methanol, and the solution is subjected to ultrasonic treatment for 5min and is fully shaken to ensure that the upper part and the lower part are uniform. Taking 1mL of the DPPH solution, and measuring A value at 519nm to ensure that A=1.2-1.3; and preparing various sample solutions: the sea grape extracts of examples 1 to 7 and comparative examples 1 to 4 were diluted with methanol to prepare solutions having an extract mass fraction of 1%.
(3) The testing method comprises the following steps:
the sample addition systems for each group are shown in the following table, the reaction solution was shaken and then placed in a water bath at 37℃for 15 minutes, and then taken out, and the absorbance value of the solution was measured by an ultraviolet-visible spectrophotometer.
| Reagent(s) | C1/mL | C2/mL | C3/mL |
| DPPH solution | 2 | 2 | 0 |
| Sample solution | 0 | 2 | 2 |
| Anhydrous methanol | 2 | 0 | 2 |
The calculation formula of the DPPH free radical clearance is shown as follows:
DPPH radical scavenging = 1- (A2-A3)/a1×100%
Wherein A1, A2 and A3 are absorbance values of C1, C2 and C3, respectively.
The test results are shown in table 2:
TABLE 2
As can be seen from the data in table 2, compared with the comparative example, the sea grape extract prepared by the preparation method of the present invention has more excellent antioxidant effect, and the selection of the flocculant and the conditions of the ultra-high pressure treatment also affect the antioxidant effect of the product.
Test example 3
Evaluation of whitening effect on the sea grape extracts prepared in examples 1 to 7 and comparative examples 1 to 4:
(1) The test is based on: the experimental design is carried out according to the standards of the group of the method for measuring the whitening efficacy of the cosmetic of the T/HPCIA 005-2021.
(2) Test principle: and (3) constructing a whitening model by using zebra fish embryos, and evaluating the whitening efficacy of the sample. The color spots, including freckle, chloasma, senile plaque, etc., are formed due to the increase of melanin of skin. Zebra fish has 87% of high homology with human genes and has related genes for controlling melanin formation. The zebra fish body surface has marked zebra stripes, which mainly comprise 3 kinds of melanocytes, yellow pigment cells and silver pigment cells. The zebra fish pigment cells originate from the nerve ridge, and at 24hpf, early pigments of zebra fish are formed and distributed in spots in the whole body skin and retina. Tyrosinase is necessary for the conversion of tyrosine to melanin. At 16.5hpf, the tyrosinase gene was transcribed in the developing retinal pigment epithelium, with pigmentation P visible at about 24 hpf. Therefore, the whitening effect of the sample was evaluated by detecting tyrosinase activity and melanin formation and distribution in zebra fish.
(3) Test materials:
test fish: zebra fish of AB line;
test instrument: a Z-A-D5 five-layer single-row independent culture unit (Shanghai sea Saint biological experiment device Co., ltd.) is a SZ680 continuous zoom stereoscopic microscope (Chongqing Orte optical instruments Co., ltd.) is a ZXSD-A1090 biochemical incubator (Shanghai intelligent city analytical instruments manufacturing Co., ltd.) is a SOP ten-thousandth electronic scale (Sidorischen instruments Co., ltd.), a KO5200DA type numerical control ultrasonic cleaner (Kunskawa ultrasonic instruments Co., ltd.) is a THZ-92B gas bath Wen Zhendang device (Shanghai Bosch medical biological instruments Co., ltd.).
Test article: the sea grape extracts of each group are directly dissolved by using fish embryo culture solution to prepare a solution with the concentration of 1.25 percent.
Positive control group: 6mg/mL arbutin solution;
blank control group: standard dilution water;
(4) The test method comprises the following steps:
(4.1) collection of experimental fish and fertilized eggs:
the sexually mature zebra fish is bred in the zebra fish breeding unit in a male-female separate tank. Water temperature: 26+/-2 ℃; ph=7.2; conductivity: 520 μs/cm; light/dark cycle: 14h to 10h. The day before the exposure experiment starts, the sexually mature zebra fish are paired in a ratio of 1:2, naturally mate and spawn, and embryos are collected.
(4.2) exposure experiments:
the AB-series zebra fish embryos with normal development of 6-9hpf are selected under a stereoscopic microscope and placed in a 6-hole cell culture plate, 30 embryos are placed in each hole, 3 compound holes are arranged in parallel, and 5mL of corresponding solution is added in each hole. The blank control group is added with standard dilution water, the positive control group is added with 6mg/mL arbutin solution, and the sample group is added with 1.25% of the sea grape extract solution of each group. The zebra fish embryos were then placed in an incubator for 72 hours of incubation, and the zebra fish embryos were observed under a stereoscopic microscope for body surface melanin deposition at 48 hours and 72 hours and photographed. The head and yolk back regions of each fish embryo are marked by analysis software, then the average measured intensity in the column of measurement is selected, and the average signal intensity in the test result is selected as the melanin signal intensity. The melanin inhibition rate was calculated.
Zebra fish embryo melanin inhibition rate = [ (normal zebra fish embryo melanin level-sample treatment group zebra fish embryo melanin level) ×100/(normal zebra fish embryo melanin level-positive control group zebra fish embryo melanin level) ]%. The results are shown in Table 3.
TABLE 3 Table 3
| Group of | 72h zebra fish embryo melanin inhibition rate (%) |
| Example 1 | 80.79％ |
| Example 2 | 77.11％ |
| Example 3 | 75.46％ |
| Example 4 | 68.29％ |
| Example 5 | 72.63％ |
| Example 6 | 73.19％ |
| Example 7 | 74.37％ |
| Comparative example 1 | 24.16％ |
| Comparative example 2 | 59.39％ |
| Comparative example 3 | 50.28％ |
| Comparative example 4 | 49.21％ |
The photographs of the blank control group, the positive control group and the sample group of the example 1 are shown in fig. 1, and in the blank control group, the zebra fish embryo appears melanin on the body surface at 48 hours and is more obvious at 72 hours; compared with a blank control group, the formation and distribution of the black pigment on the body surface of the zebra fish of the positive control group are obviously reduced, which proves that the positive control sample has good effect of inhibiting the black pigment formation; when the zebra fish embryo is treated by 1.25% of the sea grape extract for 72 hours, compared with a blank control group, the formation and distribution of the melanin on the surface of the zebra fish are obviously reduced, which indicates that the sea grape extract can inhibit the formation of the melanin on the surface of the zebra fish under the experimental condition. And in combination with the data in table 3, it is clear that the selection of the flocculant and the conditions of the ultra-high pressure treatment also affect the effect of the product in inhibiting the formation of melanin on the surface of the zebra fish.
The applicant states that the present invention is illustrated by the above examples as a preparation method of a sea grape extract and its products and applications, but the present invention is not limited to the above examples, i.e. it is not meant that the present invention must be practiced in dependence on the above examples. It should be apparent to those skilled in the art that any modification of the present invention, equivalent substitution of raw materials for the product of the present invention, addition of auxiliary components, selection of specific modes, etc., falls within the scope of the present invention and the scope of disclosure.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the scope of the technical concept of the present invention, and all the simple modifications belong to the protection scope of the present invention.
In addition, the specific features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described further.
Claims (10)
1. The preparation method of the sea grape extract is characterized by comprising the following steps of:
(1) Crushing and squeezing the sea grape raw material to obtain a squeezed product;
(2) Carrying out ultrahigh pressure treatment on the pressed product to obtain a treated product;
(3) Centrifuging the treated product, mixing the supernatant with a flocculant for flocculation treatment, centrifuging again, and carrying out fine filtration on the supernatant to obtain the sea grape extract.
2. The method for preparing a sea grape extract according to claim 1, wherein the pressure of the ultra-high pressure treatment is 180-280MPa and the dwell time is 5-15min.
3. The method for preparing a sea grape extract according to claim 1 or 2, characterized in that the centrifugation is carried out at 2000-4000rpm for 20-40min;
preferably, the fine filtration is performed using a filter plate having a pore size of 0.5 to 3 μm.
4. A method of preparing a marine grape extract according to any one of claims 1-3, wherein the flocculant comprises any one or a combination of at least two of a water-soluble polysaccharide, a cationic chitosan, guar gum, a protein colloid, gelatin, a cationic starch, or a tannin;
preferably, the flocculant comprises a combination of cationic chitosan and guar gum.
5. The method of claim 4, wherein the mass ratio of cationic chitosan to guar gum is 1:1-1:3.
6. The method for producing a sea grape extract according to any one of claims 1 to 5, wherein the fine filtration is followed by a decolorizing treatment and/or a deodorizing treatment.
7. A maritime grape extract produced by the process of any one of claims 1-6.
8. Use of the extract of sea grape according to claim 7 for the preparation of a product with moisturizing effect;
preferably, the product comprises a cosmetic, the content of the sea grape extract in the cosmetic being 0.001-0.1% by solid content.
9. Use of a marine grape extract according to claim 7 for the preparation of a product with antioxidant efficacy;
preferably, the product comprises a cosmetic, the content of the sea grape extract in the cosmetic being 0.001-0.1% by solid content.
10. Use of the extract of vitis vinifera according to claim 7 for preparing a product with skin whitening effect;
preferably, the product comprises a cosmetic, the content of the sea grape extract in the cosmetic being 0.001-0.1% by solid content.
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