CN115444086A - Health-care beverage with whitening and skin-brightening functions and preparation method and application thereof - Google Patents
Health-care beverage with whitening and skin-brightening functions and preparation method and application thereof Download PDFInfo
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- CN115444086A CN115444086A CN202211167291.4A CN202211167291A CN115444086A CN 115444086 A CN115444086 A CN 115444086A CN 202211167291 A CN202211167291 A CN 202211167291A CN 115444086 A CN115444086 A CN 115444086A
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- skin
- whitening
- extract
- parts
- health beverage
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The invention discloses a health-care beverage with whitening and skin brightening functions and a preparation method thereof, and relates to the technical field of health-care beverages. The composition is characterized by comprising the following raw materials in parts by weight: 1-1.5 parts of angelica extract solution, 0.05-0.08 part of sodium hyaluronate, 3-5 parts of grape concentrated juice, 2-4 parts of pomegranate concentrated juice, 3-5 parts of pear concentrated juice, 0.08-0.12 part of yeast extract, 0.07-0.1 part of cubilose acid, 0.03-0.05 part of acerola cherry powder, 0.01-0.12 part of tea tree pollen, 0.04-0.06 part of jasmine pollen and 25-35 parts of coix seed extract. The beverage has the advantages of soft taste, strong fragrance, bright color, uniform texture and good stability. Can remove free radicals, inhibit melanin synthesis, and whiten and lighten skin.
Description
Technical Field
The invention relates to the field of health-care beverages, in particular to a health-care beverage with whitening and skin brightening functions and a preparation method and application thereof.
Background
The color of the skin is generally determined by the amount and type of melanin present in the skin, and generally, the more the amount of melanin contained in the skin, the darker the skin color, and the less the amount of melanin, the whiter the skin color. Since melanin is mainly produced by melanocytes located in the basal layer of skin, blocking the formation of melanin in the basal layer of skin is an important way to lighten the skin color of a human body.
In the prior art, in order to make the skin brighter, people usually choose some cosmetics with whitening and skin brightening functions to be externally applied (smeared or externally applied) on the skin surface, but the active ingredients of the cosmetics are difficult to permeate into the basal layer or the dermis layer of the skin in the actual use process, so that the expected whitening effect can be obtained by using the cosmetics for a long time.
Therefore, the oral preparation can be used for purposefully supplementing functional substances with the functions of whitening and brightening the skin in an oral administration mode so as to inhibit the formation of melanin, and is a new method for brightening the skin chromaticity.
For example, an antioxidant whitening type protein beverage with the application number of CN202010123820.5 and a preparation method thereof belong to the technical field of beverage preparation. According to the technical scheme, the ginseng which is a main whitening extraction product is extracted, and the ginseng polyphenol which is effectively extracted can effectively inhibit the generation of tyrosinase, effectively reduce the production of melanin and overcome the defect that the traditional whitening product cannot radically improve the white skin.
Disclosure of Invention
The invention provides a health-care beverage with whitening and skin-brightening functions, a preparation method thereof and application thereof to overcome the defect that effective components in an external cosmetic with whitening and skin-brightening functions in the prior art do not permeate into a basal layer or a dermis layer of skin to cause poor whitening effect.
In order to realize the purpose of the invention, the invention is realized by the following technical scheme:
in a first aspect, the invention firstly provides a health beverage with whitening and skin brightening functions, which is prepared from the following raw materials in parts by weight:
1-1.5 parts of angelica dahurica extract solution, 0.05-0.08 part of sodium hyaluronate, 0.08-0.12 part of yeast extract, 0.07-0.1 part of cubilose acid, 0.03-0.05 part of acerola cherry powder, 25-35 parts of coix seed extract and 8.05-13.18 parts of mouthfeel regulator.
The invention adopts the extracts of the traditional Chinese medicines of the dahurian angelica root and the coix seed as basic formulas, and the two traditional Chinese medicines are both medicinal and edible products and have the function of whitening skin. Radix angelicae dahuricae, recorded in the Shennong herbal Jing, can grow and moisten skin and can be used as face cream, was the first whitening product since ancient times, and according to statistics, radix angelicae dahuricae is used most frequently in the beauty and skin care formula in ancient books, and has a positive whitening and skin care effect. Coix seeds, which are dry mature seeds of the Gramineae plant Coix seeds, contain various nutrient substances and have the functions of promoting diuresis, excreting dampness and whitening skin.
The sodium hyaluronate can improve the physiological condition of skin, provide excellent external environment for the synthesis of dermal collagen and elastic fiber, strengthen the supply of nutrient substances and achieve the effects of protecting skin and beautifying.
The acerola cherry is rich in a large amount of vitamin C which is more than thirty times that in the lemon, can accelerate the metabolism and regeneration of skin cells, invigorate qi and blood, and avoid the generation of dark yellow skin and color spots.
The yeast extract can inhibit the transfer of melanin to surface cells, promote the metabolism of melanocytes containing melanin to accelerate cutin exfoliation, and has been proved to have whitening effect.
The cubilose acid can rapidly mobilize the cell activity, improve the nutrient absorption rate of the skin by times, stimulate collagen and elastin in the skin to accelerate synthesis, repair broken elastic fiber nets, and have the functions of regulating the immunity and resisting aging.
Cell experiments prove that the whitening beverage prepared by the invention has certain free radical scavenging capacity and tyrosinase activity inhibiting capacity and has an inhibiting effect on melanin synthesis.
Preferably, the mouthfeel regulator consists of the following raw materials in parts by weight: 3-5 parts of grape concentrated juice, 2-4 parts of pomegranate concentrated juice, 3-5 parts of pear concentrated juice, 0.01-0.12 part of tea tree pollen and 0.04-0.06 part of jasmine pollen.
The grape, pomegranate and pear concentrated juice, jasmine flower and tea tree pollen in the taste regulator are mainly used for regulating the taste and the color, so that the beverage is integrally coordinated in flavor, proper in sugar/acid ratio, soft in taste, rich in fragrance, bright in color, uniform in texture and good in stability.
Preferably, the preparation method of the angelica dahurica extract solution comprises the following steps: reflux-extracting radix Angelicae Dahuricae decoction pieces with distilled water to obtain decoction, filtering the decoction, freeze-drying to obtain radix Angelicae Dahuricae extract powder, and re-dissolving the radix Angelicae Dahuricae extract powder in water to obtain radix Angelicae Dahuricae extract.
Preferably, the concentration of the angelica dahurica extract is 1.8% -2.2%.
Preferably, the preparation method of the coix seed extract comprises the following steps: reflux-extracting Coicis semen Chinese medicinal decoction pieces with distilled water to obtain decoction, filtering the decoction, and concentrating to obtain Coicis semen extract.
Preferably, the relative density of the coix seed extract is between 1.03 and 1.06.
In a second aspect, the present invention secondly provides a method for preparing the above health beverage with skin whitening and brightening functions,
the method comprises the following steps:
(1) Adding sodium hyaluronate into the angelica dahurica extract solution, mixing and stirring uniformly;
(2) Sequentially adding a taste regulator, yeast extract, cubilose acid and acerola cherry powder, and stirring and mixing uniformly;
(3) Then adding the coix seed extract water solution, and stirring and mixing uniformly;
(4) Sterilizing in water bath to obtain the health beverage.
In a third aspect, the invention also provides application of the health beverage with the whitening and skin-brightening functions in inhibiting melanin synthesis.
In a fourth aspect, the invention also provides application of the health-care beverage with the functions of whitening and brightening the skin in removing free radicals.
In a fifth aspect, the invention also provides an application of the health beverage with the whitening and skin-brightening functions in inhibiting tyrosinase activity.
Therefore, the invention has the following beneficial effects:
(1) The health-care beverage can effectively eliminate the free radical force in the skin and inhibit the activity of tyrosinase in an oral mode, thereby having an inhibiting effect on melanin synthesis;
(2) The health-care beverage can accelerate the metabolism and regeneration of skin cells, and has the functions of regulating the immunity and resisting aging;
(3) The health-care beverage has the advantages of overall harmonious flavor, proper sugar/acid ratio, soft taste, rich fragrance, bright color, uniform texture and better stability.
Drawings
Fig. 1 is a rutin standard curve in fig. 1.
FIG. 2 is a gallic acid standard curve.
FIG. 3 is a graph of DPPH radical clearance of vitamin C.
FIG. 4 is a graph of DPPH radical clearance of beverages.
FIG. 5 is a graph of the ABTS radical clearance for Vc.
FIG. 6 is a graph of ABTS free radical clearance of a beverage.
FIG. 7 is a graph showing the effect of Vc on the proliferation of B16 mouse melanoma cells.
FIG. 8 is a graph of the effect of beverages on the proliferation of melanoma cells in B16 mice.
FIG. 9 is a graph of the effect of beverages on tyrosinase activity in B16 mouse melanoma cells.
FIG. 10 is a graph showing the effect of beverages on melanin synthesis by the melanoma cells of B16 mice.
Detailed Description
The invention is further described with reference to the drawings and the detailed description. Those skilled in the art will be able to implement the invention based on these teachings. Furthermore, the embodiments of the present invention described in the following description are generally only a part of the embodiments of the present invention, and not all of the embodiments. Therefore, all other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present invention without any creative effort shall fall within the protection scope of the present invention.
The raw materials and manufacturers used in the invention are as follows:
sodium hyaluronate: shanghai Guangsho Biotech, inc.;
concentrated grape juice: yue Peng tai (tianjin) food ltd;
pomegranate concentrated juice: licheng food science and technology (shanghai) limited;
concentrating pear juice: xiamen Ziguo food Co Ltd;
yeast extract: shanghai Guanshuo Biotechnology Co., ltd (agent, manufacturer is PFI in Japan, chinese name: fuerma in Japan);
bird's nest acid: wuxi Aikopie Biotechnology, inc.;
acerola cherry powder: glard technologies (Shanghai) Inc.;
tea tree pollen: shanghai mountain fungus science and technology Limited;
jasmine pollen: shanxi New Tian Yuan Biotech Co., ltd.
Example 1
A health beverage with whitening and skin brightening functions is prepared from the following raw materials in parts by weight: 1.2g of angelica extract solution, 0.07g of sodium hyaluronate, 4g of grape concentrated juice, 3g of pomegranate concentrated juice, 4g of pear concentrated juice, 0.1g of yeast extract, 0.1g of cubilose acid, 0.05g of acerola cherry powder, 0.01g of tea tree pollen, 0.05g of jasmine pollen and 30g of coix seed extract.
The whitening and skin-brightening health-care beverage is prepared by the following steps:
(1) Weighing 80g of angelica dahurica traditional Chinese medicine decoction pieces, adding 800mL of distilled water for decoction, heating and refluxing for 2 times, extracting for 1 hour each time, filtering the two water decoctions by eight layers of gauze, combining the filtrates, and freeze-drying to obtain 12.8g of angelica dahurica extract powder. Dissolving radix Angelicae Dahuricae extract powder 1g in water 50g to obtain radix Angelicae Dahuricae extract solution with concentration of 1.96%.
(2) Preparation of coix seed extract: 500g of coix seed traditional Chinese medicine decoction pieces are added with 4L of distilled water for decoction, the decoction is heated and refluxed for extraction for 3 times, each time is 75min, the decoction of the three times is filtered by eight layers of gauze, the filtrate is combined and concentrated to about 800mL, and the relative density is 1.04.
(3) Firstly, adding sodium hyaluronate into an aqueous solution of an angelica dahurica extract, mixing and stirring uniformly, then sequentially adding concentrated grape juice, concentrated pomegranate juice, concentrated pear juice, yeast extract, cubilose acid, acerola cherry powder, tea tree pollen and jasmine pollen, stirring and mixing uniformly, then adding an aqueous solution of a coix seed extract, stirring and mixing uniformly, filling, and sterilizing for 30min at 90 ℃ in a water bath.
Example 2
A health beverage with whitening and skin brightening functions is composed of the following raw materials in parts by weight: 1g of angelica dahurica extract solution, 0.05g of sodium hyaluronate, 3g of grape concentrated juice, 4g of pomegranate concentrated juice, 3g of pear concentrated juice, 0.12g of yeast extract, 0.07g of cubilose acid, 0.03g of acerola cherry powder, 0.05g of tea tree pollen, 0.04g of jasmine pollen and 25g of coix seed extract.
The whitening and skin-brightening health-care beverage is prepared by the following steps:
(1) Weighing 80g of angelica dahurica traditional Chinese medicine decoction pieces, adding 800mL of distilled water for decoction, heating and refluxing for 2 times, extracting for 1 hour each time, filtering the two water decoctions by eight layers of gauze, combining the filtrates, and freeze-drying to obtain 12.8g of angelica dahurica extract powder. Taking 1g of angelica dahurica extract powder, adding 50g of water for dissolving to obtain an angelica dahurica extract solution with the concentration of 1.8% for later use.
(2) Preparation of coix seed extract: 500g of coix seed traditional Chinese medicine decoction pieces are added with 4L of distilled water for decoction, the decoction is heated and refluxed for extraction for 3 times, each time is 75min, the decoction of the three times is filtered by eight layers of gauze, the filtrate is combined and concentrated to about 800mL, and the relative density is 1.06.
(3) Firstly, adding sodium hyaluronate into an aqueous solution of an angelica dahurica extract, mixing and stirring uniformly, then sequentially adding concentrated grape juice, concentrated pomegranate juice, concentrated pear juice, yeast extract, cubilose acid, acerola cherry powder, tea tree pollen and jasmine pollen, stirring and mixing uniformly, then adding an aqueous solution of a coix seed extract, stirring and mixing uniformly, filling, and sterilizing for 30min at 90 ℃ in a water bath.
Example 3
A health beverage with whitening and skin brightening functions is prepared from the following raw materials in parts by weight: 1.5g of angelica extract solution, 0.08g of sodium hyaluronate, 5g of grape concentrated juice, 2g of pomegranate concentrated juice, 53g of pear concentrated juice, 0.08g of yeast extract, 0.07g of cubilose acid, 0.04g of acerola cherry powder, 0.12g of tea tree pollen, 0.06g of jasmine pollen and 35g of coix seed extract.
The whitening and skin-brightening health-care beverage is prepared by the following steps:
(1) Weighing 80g of angelica dahurica traditional Chinese medicine decoction pieces, adding 800mL of distilled water for decoction, heating and refluxing for 2 times, extracting for 1 hour each time, filtering the two water decoctions by eight layers of gauze, combining the filtrates, and freeze-drying to obtain 12.8g of angelica dahurica extract powder. Dissolving radix Angelicae Dahuricae extract powder 1g in water 50g to obtain radix Angelicae Dahuricae extract solution with concentration of 2.2%.
(2) Preparation of coix seed extract: 500g of coix seed traditional Chinese medicine decoction pieces are added with 4L of distilled water for decoction, the decoction is heated and refluxed for extraction for 3 times, each time is 75min, the decoction of the three times is filtered by eight layers of gauze, the filtrate is combined and concentrated to about 800mL, and the relative density is 1.03.
(3) Firstly, adding sodium hyaluronate into an aqueous solution of an angelica dahurica extract, mixing and stirring uniformly, then sequentially adding concentrated grape juice, concentrated pomegranate juice, concentrated pear juice, yeast extract, cubilose acid, acerola cherry powder, tea tree pollen and jasmine pollen, stirring and mixing uniformly, then adding an aqueous solution of a coix seed extract, stirring and mixing uniformly, filling, and sterilizing for 30min at 90 ℃ in a water bath.
Example 4
A health beverage with whitening and skin brightening functions is composed of the following raw materials in parts by weight: 9.8g of angelica extract solution, 0.42g of sodium hyaluronate, 35g of grape concentrated juice, 21g of pomegranate concentrated juice, 28g of pear concentrated juice, 0.56g of yeast extract, 0.56g of cubilose acid, 0.35g of acerola cherry powder, 0.08g of tea tree pollen, 0.42g of jasmine pollen and 210g of coix seed extract aqueous solution.
The whitening and skin-brightening health-care beverage is prepared by the following steps:
(1) Weighing 80g of angelica dahurica traditional Chinese medicine decoction pieces, adding 800mL of distilled water for decoction, heating and refluxing for 2 times, extracting for 1 hour each time, filtering the two water decoctions by eight layers of gauze, combining the filtrates, and freeze-drying to obtain 12.8g of angelica dahurica extract powder. Dissolving radix Angelicae Dahuricae extract powder 1g in water 50g to obtain radix Angelicae Dahuricae extract solution with concentration of 1.96%.
(2) Preparation of coix seed extract: decocting Coicis semen decoction pieces 4kg with 32L distilled water under reflux for 75min 3 times, filtering the three decoctions with eight layers of gauze, mixing filtrates, and concentrating to about 5L with relative density of 1.05.
(3) Firstly, adding sodium hyaluronate into an aqueous solution of an angelica dahurica extract, mixing and stirring uniformly, then sequentially adding concentrated grape juice, concentrated pomegranate juice, concentrated pear juice, yeast extract, cubilose acid, acerola cherry powder, tea tree pollen and jasmine pollen, stirring and mixing uniformly, then adding an aqueous solution of a coix seed extract, stirring and mixing uniformly, filling, and sterilizing for 30min at 90 ℃ in a water bath.
[ functional ingredient analysis of beverages ]
1 Total Flavonoids content determination
Reagent: rutin control (Shanghai Yuan leaf Biotech Co., ltd.), 60% ethanol, 5% NaNO2 solution, and 10% Al (NO 3) 3 Solution, 4% NaOH solution,The whitening beverage prepared in the first embodiment.
The experimental process comprises the following steps:
precisely weighing 10.07mg rutin reference substance, dissolving with anhydrous ethanol, and diluting with 60% ethanol to constant volume in 50mL volumetric flask to obtain 0.2mg/mL rutin reference substance mother liquor. Respectively sucking 0, 1mL, 2mL, 3mL, 4mL, 5mL, and 6mL of the mother solution into a 25mL volumetric flask, adding water to 6mL, adding 4mL of 5% NaNO2 solution, shaking, standing for 6min, and adding 1mL10% Al (NO) 3 ) 3 Shaking the solution, standing for 6min, adding 10mL of 4% NaOH solution, adding water to constant volume, shaking, and standing for 15min. The absorbance was measured at 510nm in an ultraviolet spectrophotometer and 3 times in parallel. A standard curve was plotted with the absorbance (A) as the abscissa and the concentration (C) as the ordinate.
0.5mL of the whitening beverage prepared in example I was taken, and the pretreatment was performed in the same manner. The absorbance was measured at 510nm in an ultraviolet spectrophotometer and 3 times in parallel.
(3) Calculation of flavone content in beverage
The standard curve equation of rutin is y =0.00734x +0.00484 2 =0.99507 as shown in fig. 1.
The content of total flavonoids in the measured sample is obtained by a standard curve, and then the content of total flavonoids in the whitening beverage prepared in the first example is calculated to be 0.059mg/mL according to the formula W = c × n (W is the content of flavonoids (mg/mL), c is the mass concentration of flavonoids (mg/mL), and n is the dilution factor).
2 Total phenol content determination
(1) Reagent: 10% Folin-Ciocalteu solution, gallic acid control (Shanghai-derived leaf Biotech Co., ltd.), 10% sodium carbonate solution, and the whitening beverage prepared in example one.
(2) The experimental process comprises the following steps:
weighing 21.39mg of gallic acid reference substance, dissolving with ultrapure water, and diluting to 100mL to obtain 213.9mg/L gallic acid mother liquor. Respectively sucking 1mL, 2mL, 3.5mL, 5mL, 6mL and 7.5mL of mother liquor, adding the mother liquor into 6 25mL volumetric flasks, and fixing the volume to the scale to obtain series of standard solutions with mass concentrations of 8.556mg/L, 17.112mg/L, 29.946mg/L, 42.78mg/L, 51.336mg/L and 64.17mg/L respectively. Respectively sucking 1mL of the 6 standard solutions into a 25mL centrifuge tube, adding 5mL of 10% Folin-Ciocalteu solution, mixing, adding 4mL of 7.5% sodium carbonate solution within 3-8min, mixing, and standing at room temperature in the dark for 1h. The absorbance (A) at 765nm was measured with a UV-visible spectrophotometer, 3 times in parallel, and a standard curve was drawn.
The whitening beverage prepared in the first example was diluted by 100 times, 1mL of each of the diluted beverages was transferred to a 25mL centrifuge tube, 1mL of the diluted sample solution was accurately aspirated, the diluted sample solution was transferred to a 10mL centrifuge tube, 5mL of a 10% folin-Ciocalteu solution was added thereto, the mixture was mixed, 4mL of a 7.5% sodium carbonate solution was added thereto after 3 minutes, the mixture was mixed, and the mixture was left to stand at room temperature for 1 hour in the dark. The absorbance (A) at 765nm was measured with a UV-visible spectrophotometer and 3 replicates were measured.
(3) Calculation of the Total phenol content of the beverage
The standard curve equation of gallic acid is y =0.01064x +0.01795 2 =0.99744 as shown in fig. 2.
The total phenol content in the measured sample is obtained by a standard curve, and then the total phenol content in the whitening beverage prepared in the first example is calculated to be 1.50mg/mL according to the formula W = c × n (W is the polyphenol content (mg/mL), c is the polyphenol mass concentration (mg/mL), and n is the dilution factor).
[ evaluation of functionality of beverage ]
1. DPPH radical scavenging
(1) Reagent: DPPH reagent (Shanghai chemical industry development Co., ltd., taishiai), vitamin C (Shanghai leaf Biotechnology Co., ltd.), and ethanol as analytical pure, and whitening beverage prepared in example two
(2) The experimental process comprises the following steps:
and (3) preparation of a DPPH reagent: 2.56mg of DPPH powder is weighed out finely, and 5mL of 95% ethanol is added to prepare a saturated DPPH reagent.
Preparing a vitamin C solution: precisely weighing Vc1.95mg, placing the Vc1.95mg in a 10mL brown volumetric flask, adding distilled water to dissolve the Vc to a constant volume, and shaking up to obtain the Vc original solution with the concentration of 0.195mg/mL. Then, the concentrations were 19.5. Mu.g/mL, 13.0. Mu.g/mL, 10.8. Mu.g/mL, 9.75. Mu.g/mL, 8.86. Mu.g/mL, and 6.5. Mu.g/mL, respectively.
Preparing a sample solution: the whitening beverage prepared in example two was diluted with distilled water and then prepared to a concentration of 0.6mL/mL,0.5mL/mL,0.4mL/mL,0.35mL/mL,0.30mL/mL,0.25mL/mL, respectively, and centrifuged at 10000rpm for 10min.
DPPH clearance calculation: adding a sample into a 96-well plate, placing the 96-well plate in the shade, reacting for 30min, and measuring the OD value by using a microplate reader, wherein the measuring wavelength is 517nm. The DPPH clearance calculation formula is as follows.
(3) Results of the experiment
The clearance curves of the beverage and the positive control Vc versus DPPH free radical are shown in fig. 3, fig. 4, table 1.
TABLE 1DPPH radical scavenging Rate
As can be seen from the table, the whitening beverage prepared in example two had a certain radical scavenging ability, although the radical scavenging ability was not as good as that of vitamin C.
(4) Discussion of the experiments
The beverage contains components such as saccharides and proteins, a large amount of flocculent precipitates are generated when a DPPH solvent is added, the flocculent precipitates wrap DPPH reaction products, and detection results are influenced if the flocculent precipitates are directly centrifuged, so that when a DPPH solution is prepared, a saturated solution is prepared and the addition amount is reduced, which is different from the common concentration.
2ABTS free radical scavenging
(1) Reagent: ABTS kit (Solarbio BC 4775) and whitening beverage prepared in example two
(2) Procedure of experiment
According to the detection instruction, reaction solutions with different concentrations are prepared according to the steps, a sample is added, the reaction solutions are fully and uniformly mixed and then are kept stand for 6min in a dark place at room temperature, an enzyme-linked immunosorbent assay (OD) value is measured by using an enzyme-linked immunosorbent assay, and the measurement wavelength is selected to be 405nm. The DPPH clearance calculation formula is as follows.
A0 represents the absorbance value of 190. Mu.L of the developing solution + 10. Mu.L of the aqueous solution; a1 represents the absorbance value of 190. Mu.L of the developing solution + 10. Mu.L of the test sample; a2 represents the absorbance value of 190. Mu.L of the developing solution solvent + 10. Mu.L of the test sample.
(3) Results of the experiment
The clearance curves of the beverages and the positive control Vc against ABTS free radicals are shown in fig. 5, fig. 6, table 2.
TABLE 2ABTS radical clearance
As can be seen from fig. 5, fig. 6 and table 2, the clearance curves of the beverage and the positive control Vc for ABTS radicals are less linear, and the effect of the clearance of radicals tends to saturate with increasing concentration. Predicting SC from a graph 50 It was 0.076mL/mL.
3 tyrosinase activity inhibition
(1) Reagent: tyrosinase (Sigma), L-tyrosine (Sigma), arbutin (Shanghai-derived leaf Biotech Co., ltd.), and the whitening beverage prepared in example two
(2) Procedure of experiment
Preparation of tyrosinase: accurately weighing 0.17mg of tyrosinase, placing the tyrosinase in a brown volume of 5mL, adding PBS buffer solution, and fixing the volume to a scale mark, wherein the final concentration is 170 mu g/mL.
Preparing tyrosine: tyrosine 50.30mg is precisely weighed and placed in a 100mL brown volumetric flask, PBS buffer solution is added, and the volume is determined to the scale mark.
Preparing an arbutin positive control solution: accurately weighing 10.45mg of arbutin, placing the arbutin in a 5mL brown volumetric flask, and adding PBS to dissolve the arbutin to obtain an arbutin control solution with the concentration of 2.09 mg/mL.
Adding an L-tyrosine solution, a sample solution to be detected, a phosphate buffer solution and a tyrosinase solution into a 96-well plate, placing the 96-well plate in an incubator at 37 ℃ for culturing for 30min, then measuring an OD value by using an enzyme-labeling instrument, and selecting a measuring wavelength of 475nm. The tyrosinase inhibition rate was calculated as follows.
A0 represents the absorbance value of 120. Mu.L tyrosine solution + 80. Mu.L PBS; a1 represents the absorbance value of 120. Mu.L of tyrosine solution + 40. Mu.L of tyrosine + 40. Mu.L of sample; a1 represents the absorbance value of 120. Mu.L tyrosine solution + 40. Mu.LPBS
(3) Results of the experiment
The results of the inhibition of tyrosinase by the beverages are shown in Table 3.
TABLE 3 tyrosinase inhibition
| Sample (I) | Tyrosinase inhibition% |
| Arbutin (2.09 mg/mL) | 65.3 |
| Beverage (0.5 mL/mL) | 56.7 |
The results show that the whitening beverage prepared in the second embodiment has a certain inhibition effect on tyrosinase activity.
4 mouse B16 melanoma cells
(1) Reagent
Fetal bovine serum (Zhejiang Hangzhou Biotechnology, inc.); PBS buffer (pH = 7.4) (Hyclone); 0.25% trypsin (0.02% EDTA) (Gibco); cyan-streptomycin solution (Gibco); 1640 culture medium (monatin biotechnology limited); CCK-8 (Dojindo chemical research institute); dimethyl sulfoxide (Sigma) and whitening beverage prepared in example II
(2) Procedure of experiment
(1) Mouse B16 melanoma cell culture and sample handling
Culturing B16 melanoma cells in RPMI-1640 medium containing 10% fetal bovine serum, standing at 37 deg.C, 5% 2 Culturing in saturated humidity incubator, collecting cells in logarithmic growth phase, and adjusting cell suspension density to 1 × 10 5 Inoculating to 96-well plate (100 uL per well) (melanin synthesis amount experiment is inoculated to 12-well plate, 1mL per well, each concentration is set to 3 times), culturing for 12h for cell adherence, adding culture solution containing different sample concentrations and V into each well C At 4 replicates per concentration setting, fresh medium was added to the control and incubated for 24h.
(2) MTT method for measuring proliferation rate of B16 mouse melanoma cells
After 24h sample treatment, 10uL of CCK8 reagent was added to each well, taking care not to generate bubbles in the wells, which would affect the OD. The culture plate is incubated in an incubator for 1-4h, and the absorbance of each well at 450nm is measured by a microplate reader.
(3) Determination of tyrosinase Activity
The activity of tyrosinase in B16 cells was determined by L-DOPA oxidation. According to 1 × 10 4 Cell density per mL 96-well plate inoculation, 100. Mu.L of culture medium per well, 37 ℃,5% CO 2 Incubator, after 24h incubation, add different samples in groups of 4 parallel each, 37 ℃,5% CO 2 Culturing in an incubator for 24h, discarding the supernatant, washing with PBS for 3 times, adding 50 μ L of 1% octyl phenol polyoxyethylene ether (Triton X-100) solution to each well, rapidly freezing at-81 deg.C for 30min, thawing at room temperature to completely break the cells, pre-heating at 37 deg.C for 5min, adding 10 μ L of 1mg/ml L-DOPA solution, reacting at 37 deg.C for 2h, and measuring the absorbance at 490nm with a microplate reader.
(4) Determination of amount of melanin synthesis
After 24 hours of sample treatment, the supernatant was aspirated and discarded, washed 2 times with PBS solution, the cells were collected in a centrifuge tube, centrifuged at 1500rpm for 10min, the supernatant was discarded and 1mL of PBS was added for resuspension, then 500uL of ethyl ether with ethanol at a volume ratio of 1: 1 was added, placed at room temperature for 30min, centrifuged at 3000rpm for 5min, the supernatant was discarded, 1mL of 1.0mol/L NaOH solution containing 10% DMSO was added, and subjected to 80 ℃ water bath for 45min, and the absorbance at 405nm was measured.
(3) Results of the experiment
The effect of vitamin C and beverages on the proliferation rate of melanoma cells in B16 mice is shown in fig. 7 and fig. 8.
The experimental result shows that the whitening beverage prepared in the second example has the strongest inhibition effect on the proliferation of melanoma cells of a B16 mouse when the concentration of the whitening beverage is 0.125 mL/mL.
The effect of the drink on the tyrosinase activity of the B16 mouse melanoma cells is shown in FIG. 9.
The effect of the drink on melanin synthesis by the melanoma cells of B16 mice is shown in FIG. 10.
The experimental result shows that the whitening beverage prepared in the second embodiment has a strong inhibition effect on tyrosinase of melanoma cells of a B16 mouse and has a certain melanin synthesis inhibition effect.
[ microbiological assay for beverages ]
Detection time: 2021.09.27;2021.10.03;2021.10.06
Temperature: 47 deg.C
The microorganisms were detected according to the national standards for food safety, and the results are shown in Table 4.
TABLE 4 detection results of microorganisms
| Basis of detection | Detecting items | Detection value | Unit of detection value | Detection conclusion |
| GB 4789.2-2016 | Total number of colonies × 5 | ＜1 | CFU/mL | Conform to |
| GB 4789.3-2016 (second method) | Coliform group is 5 | ＜1 | CFU/mL | Meet with |
| GB 4789.15-2016 (first method) | Mould fungus | ＜1 | CFU/mL | Conform to |
| GB 4789.15-2016 (first method) | Yeast | ＜1 | CFU/mL | Conform to |
| GB 4789.4-2016 | Salmonella X5 | Not detected out | /25mL | Conform to |
| GB 4789.10-2016 (second method) | Staphylococcus aureus X5 | ＜1 | CFU/mL | Conform to |
Claims (10)
1. A health beverage with whitening and skin-brightening functions is characterized in that,
the composition comprises the following raw materials in parts by weight: 1-1.5 parts of angelica extract solution, 0.05-0.08 part of sodium hyaluronate, 0.08-0.12 part of yeast extract, 0.07-0.1 part of cubilose acid, 0.03-0.05 part of acerola cherry powder, 25-35 parts of coix seed extract and 8.005-13.18 parts of mouthfeel regulator.
2. The health beverage with whitening and skin-brightening functions as claimed in claim 1,
the mouthfeel regulator is composed of the following raw materials in parts by weight: 3-5 parts of grape concentrated juice, 2-4 parts of pomegranate concentrated juice, 3-5 parts of pear concentrated juice, 0.01-0.12 part of tea tree pollen and 0.04-0.06 part of jasmine pollen.
3. The health beverage with whitening and skin-brightening functions as claimed in claim 1,
the preparation method of the angelica dahurica extract solution comprises the following steps: reflux-extracting radix Angelicae Dahuricae Chinese medicinal decoction pieces with distilled water to obtain decoction, filtering the decoction, freeze-drying to obtain radix Angelicae Dahuricae extract powder, and re-dissolving the radix Angelicae Dahuricae extract powder in water to obtain radix Angelicae Dahuricae extract.
4. The health beverage with skin whitening and brightening functions as claimed in claim 1 or 3,
the concentration of the angelica dahurica extract is 1.8% -2.2%.
5. The health beverage with whitening and skin-brightening functions as claimed in claim 1,
the preparation method of the coix seed extract comprises the following steps: reflux-extracting Coicis semen Chinese medicinal decoction pieces with distilled water to obtain decoction, filtering the decoction, and concentrating to obtain Coicis semen extract.
6. The health beverage with whitening and skin-brightening functions as claimed in claim 1 or 5,
the relative density of the coix seed extract is between 1.03 and 1.06.
7. A method for preparing the health beverage with whitening and skin-brightening functions of any one of claims 1 to 6,
the method comprises the following steps:
(1) Adding sodium hyaluronate into the angelica dahurica extract solution, mixing and stirring uniformly;
(2) Sequentially adding a taste regulator, yeast extract, cubilose acid and acerola cherry powder, and stirring and mixing uniformly;
(3) Then adding the coix seed extract water solution, and stirring and mixing uniformly;
(4) Sterilizing in water bath to obtain the health beverage.
8. Use of the health beverage with skin whitening and brightening functions as claimed in any one of claims 1 to 6 in inhibiting melanin synthesis.
9. Use of the health beverage with whitening and skin-brightening functions as claimed in any one of claims 1 to 6 in scavenging free radicals.
10. Use of the health beverage with whitening and skin-brightening functions as claimed in any one of claims 1 to 6 in inhibiting tyrosinase activity.
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