Method for affinity co-extraction of plant whitening composition and product and application thereof

Abstract

The invention provides a method for affinity co-extraction of a plant whitening composition and a product and application thereof, and the method comprises the following steps: (1) mixing phyllanthus emblica and / or roselle with an extraction reagent, extracting, and carrying out solid-liquid separation to obtain a filtrate 1; mixing the filter residue with an extraction reagent, extracting, and carrying out solid-liquid separation to obtain filtrate 2; or mixing the phyllanthus emblica and the roselle with the extraction reagent, extracting, carrying out solid-liquid separation, collecting liquid and mixing to obtain filtrate 1; respectively mixing the filter residues of the emblic leafflower fruit and the roselle with an extraction reagent, extracting, carrying out solid-liquid separation, collecting liquid, and mixing to obtain filtrate 2; (2) mixing mulberry leaves with the filtrate 2, extracting, and carrying out solid-liquid separation to obtain a filtrate 3; mixing the filtrate 3 and the filtrate 1, adjusting the pH value, reacting, standing, carrying out solid-liquid separation, and collecting the precipitate; after the precipitate is dissolved in an alkaline aqueous solution, solid-liquid separation is conducted, liquid is collected, and the plant whitening composition is obtained and has the effects of whitening, saccharification resistance, oxidation resistance, water supplementing and moisturizing.

Description

Technical Field

[0001] This invention belongs to the field of plant extraction technology, specifically relating to a method for affinity co-extraction of plant whitening compositions, its products, and applications. Background Technology

[0002] Most Chinese people have skin types between II and IV, characterized by an abundance of melanosomes and a tendency to tan easily. Tyrosinase is a key enzyme in melanin synthesis; inhibiting tyrosinase activity can reduce melanin production and effectively whiten the skin. Furthermore, with age, skin is prone to dullness and yellowing, which is closely related to skin glycation. Skin glycation refers to the reaction between proteins and sugars, producing a series of substances called advanced glycation end products (AGEs). Existing literature has shown that reducing the production and accumulation of AGEs can prevent skin yellowing.

[0003] Hibiscus extract and Phyllanthus emblica extract are rich in polyphenols and polysaccharides, possessing whitening, antioxidant, and anti-inflammatory effects. DNJ (1-deoxynojirimycin) is a polyhydroxy alkaloid extracted from mulberry leaves. Current research has shown that DNJ has hypoglycemic and anti-glycation effects, effectively inhibiting the formation of AGEs. Furthermore, mulberry leaves are rich in flavonoids, chlorogenic acid, and other bioactive components, exhibiting antioxidant and anti-inflammatory effects, and are currently widely used in cosmetics.

[0004] Patent CN115778996A discloses a method for preparing amla extract. This method uses water as the sole extraction solvent, which cannot effectively dissolve the polyphenols in amla, directly affecting the whitening effect of subsequent cosmetics.

[0005] Patent CN119157790A discloses a method for preparing mulberry leaf extract using "compound bacterial fermentation + aqueous two-phase extraction + ultrafiltration". Compared with traditional water extraction / alcohol extraction, the active ingredients after fermentation have higher purity and are green and low in toxicity. However, the process is complex, costly, and has poor technical controllability, making it unsuitable for large-scale production.

[0006] Patent CN105125436A discloses a method for preparing a compound whitening composition. This method involves mixing traditional Chinese medicinal herbs such as roselle and astragalus, soaking them in a solvent for 2 hours, then extracting at 60℃ with stirring for 1-2 hours. This extraction is repeated 2-3 times. The mixture is then filtered, concentrated, purified using macroporous resin, mixed with pea extract, stirred, and allowed to stand for 24-48 hours before filtration to obtain the whitening composition. This method is relatively complex, consumes a large amount of ethanol, and the 24-48 hour standing time is significant, limiting production capacity. Furthermore, the properties of different plant active ingredients vary, and mutual reactions and dissolution inhibition may occur during co-extraction.

[0007] In summary, existing extraction and preparation processes for plant-based skin whitening ingredients suffer from various problems, including high complexity, high cost, and ineffective extraction of active ingredients. Therefore, developing a simple, low-cost, and highly efficient method for preparing plant-based skin whitening compositions is of great significance. Summary of the Invention

[0008] In view of the shortcomings of the prior art, the purpose of this invention is to provide a method for co-extracting plant whitening compositions, as well as the products and applications thereof. The compositions prepared by the method of this invention can significantly inhibit tyrosinase activity, inhibit the generation of advanced glycation end products (AGEs), scavenge free radicals, and increase skin moisture content, thus exerting excellent whitening, anti-glycation, antioxidant, and moisturizing effects.

[0009] To achieve this objective, the present invention adopts the following technical solution:

[0010] In a first aspect, the present invention provides a method for co-extracting a plant whitening composition, the method comprising:

[0011] (1) Mix Phyllanthus emblica and / or Hibiscus rosa-sinensis with the extraction reagent, extract, and separate the solid and liquid to obtain filtrate 1; mix the residue with the extraction reagent again, extract, and separate the solid and liquid to obtain filtrate 2;

[0012] Alternatively, amla and roselle are mixed with the extraction reagent separately, extracted, separated into solid and liquid, and the liquid is collected and mixed to obtain filtrate 1; the residues of amla and roselle are then mixed with the extraction reagent separately, extracted, separated into solid and liquid, and the liquid is collected and mixed to obtain filtrate 2.

[0013] (2) Mulberry leaves and filtrate 2 are mixed, extracted, and separated into solid and liquid to obtain filtrate 3; filtrate 3 and filtrate 1 are mixed, pH value is adjusted, reaction is carried out, and the mixture is allowed to stand, separated into solid and liquid, and the precipitate is collected; after the precipitate is dissolved in an alkaline aqueous solution, the solid and liquid are separated, and the liquid is collected to obtain the plant whitening composition.

[0014] This invention achieves the enrichment and separation of active ingredients from mulberry leaves through two extractions of Phyllanthus emblica and / or Roselle. The Phyllanthus emblica and / or Roselle extracts contain abundant polyphenols, anthocyanins, and vitamin C, and are acidic (pH 3-4.5). The second extract (filtrate 2) has slightly lower levels of polyphenols, anthocyanins, and vitamin C. Under acidic conditions, the natural alkaloid 1-deoxynojirimycin (DNJ) in mulberry leaves undergoes protonation. The amine group of DNJ combines with hydrogen ions in the solution to form a soluble salt, which facilitates the dissolution of DNJ. The first extract (filtrate 1) has higher levels of polyphenols, anthocyanins, and vitamin C. By adjusting the pH, the electrostatic attraction between the polyphenols and organic acids in the extract and DNJ is strongest, resulting in a complex precipitate. The complex precipitate dissolves in an alkaline aqueous solution, while insoluble proteins are removed, ultimately achieving the effective enrichment of active ingredients from Phyllanthus emblica and / or Roselle and mulberry leaves.

[0015] This invention is the first to utilize extracts of Phyllanthus emblica and / or hibiscus to enrich the effective components of mulberry leaves. Through multi-stage regulatory design, the natural active components in Phyllanthus emblica and / or hibiscus and mulberry leaves are fully extracted. The resulting plant composition can significantly inhibit tyrosinase activity, inhibit the formation of advanced glycation end products (AGEs), scavenge free radicals, and increase skin hydration, exhibiting excellent whitening, anti-glycation, antioxidant, and moisturizing effects.

[0016] Preferably, in step (1), the extraction reagent includes an organic solvent and water.

[0017] Preferably, the organic solvent includes any one or a combination of at least two of ethanol, methanol, or ethyl acetate.

[0018] Preferably, the volume percentage of the organic solvent in the extraction reagent is 30-50%, for example, it can be 30%, 31%, 32%, 35%, 37%, 40%, 43%, 45%, 48% or 50%, etc.

[0019] Other specific point values ​​within the range of the above values ​​can be selected, and will not be elaborated on here.

[0020] Preferably, in step (1), the ratio of the extracted material to the liquid is 1 g:(15-25) mL.

[0021] The specific point values ​​in (15-25) can be 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25, etc.

[0022] Other specific point values ​​within the range of the above values ​​can be selected, and will not be elaborated on here.

[0023] Preferably, in step (1), the extraction temperature is 50-60℃, for example, it can be 50℃, 51℃, 52℃, 53℃, 54℃, 55℃, 56℃, 57℃, 58℃, 59℃ or 60℃, etc.; the time is 0.5-1.5 h, for example, it can be 0.5 h, 0.6 h, 0.7 h, 0.8 h, 0.9 h, 1 h, 1.1 h, 1.2 h, 1.3 h, 1.4 h or 1.5 h, etc.

[0024] Preferably, in step (2), the extraction temperature is 55-65℃, for example, it can be 55℃, 56℃, 57℃, 58℃, 59℃, 60℃, 61℃, 62℃, 63℃, 64℃ or 65℃, etc.; the time is 2-3 h, for example, it can be 2 h, 2.1 h, 2.2 h, 2.3 h, 2.4 h, 2.5 h, 2.6 h, 2.7 h, 2.8 h, 2.9 h or 3 h, etc.

[0025] Preferably, in step (2), the pH value is adjusted to 2-4, for example, 2, 2.3, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.7 or 4, etc.

[0026] Preferably, in step (2), the reaction temperature is 25-40℃, for example, it can be 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, 32℃, 35℃, 38℃ or 40℃, etc.; the time is 0.3-0.8 h, for example, it can be 0.3 h, 0.35 h, 0.4 h, 0.45 h, 0.5 h, 0.55 h, 0.6 h, 0.65 h, 0.7 h, 0.75 h or 0.8 h, etc.

[0027] Other specific point values ​​within the range of the above values ​​can be selected, and will not be elaborated on here.

[0028] Preferably, in step (2), the reaction is carried out under stirring.

[0029] Preferably, in step (2), the settling time is 0.5-1.5 h, for example, it can be 0.5 h, 0.6 h, 0.7 h, 0.8 h, 0.9 h, 1 h, 1.1 h, 1.2 h, 1.3 h, 1.4 h or 1.5 h.

[0030] Preferably, in step (2), the pH value of the alkaline aqueous solution is 8-9, for example, it can be 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9 or 9.

[0031] Other specific point values ​​within the range of the above values ​​can be selected, and will not be elaborated on here.

[0032] Preferably, in step (2), the solute in the alkaline aqueous solution includes any one or a combination of at least two of arginine, triethanolamine, sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, or ammonia.

[0033] Preferably, in step (2), the collected liquid is further concentrated and dried.

[0034] Preferably, the mass ratio of Phyllanthus emblica to mulberry leaf is (20-30):(30-60).

[0035] The specific point values ​​in (20-30) can be 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30, etc.

[0036] The specific point values ​​in (30-60) can be 30, 31, 32, 35, 37, 40, 43, 45, 48, 50, 55 or 60, etc.

[0037] Other specific point values ​​within the range of the above values ​​can be selected, and will not be elaborated on here.

[0038] Preferably, the mass ratio of roselle to mulberry leaves is (15-30):(30-60).

[0039] The specific point values ​​in (15-30) can be 15, 16, 17, 18, 19, 20, 22, 25, 28 or 30, etc.

[0040] The specific point values ​​in (30-60) can be 30, 31, 32, 35, 37, 40, 43, 45, 48, 50, 55 or 60, etc.

[0041] Other specific point values ​​within the range of the above values ​​can be selected, and will not be elaborated on here.

[0042] In a second aspect, the present invention provides a plant-based whitening composition prepared by the method described in the first aspect.

[0043] Thirdly, the present invention provides the use of the plant-based whitening composition as described in the second aspect in the preparation of cosmetics, food or health products.

[0044] Compared with the prior art, the present invention has the following beneficial effects:

[0045] (1) This invention is the first to use extracts of Phyllanthus emblica and / or hibiscus to enrich the effective components of mulberry leaves. Through multi-stage regulation design, the natural active components in Phyllanthus emblica and / or hibiscus and mulberry leaves are fully extracted.

[0046] (2) The plant composition obtained can significantly inhibit tyrosinase activity, inhibit the generation of advanced glycation end products (AGEs), scavenge free radicals and increase skin moisture content, and exert excellent whitening, anti-glycation, antioxidant and moisturizing effects.

[0047] (3) Compared with extracting Phyllanthus emblica and / or roselle extract and mulberry leaf extract separately and then combining them, the composition obtained by the present invention has better anti-glycation, whitening, antioxidant and moisturizing effects.

[0048] (4) The preparation process used in this invention is simple and suitable for large-scale production. Detailed Implementation

[0049] To further illustrate the technical means and effects of the present invention, the following describes the technical solution of the present invention in conjunction with preferred embodiments of the present invention. However, the present invention is not limited to the scope of the embodiments.

[0050] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0051] Example 1

[0052] This embodiment provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0053] (1) Take 25 g of Phyllanthus emblica and 20 g of Hibiscus rosa-sinensis and mix them with 900 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter, and obtain filtrate 1. Mix the residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter, and obtain filtrate 2.

[0054] (2) Add 45 g of mulberry leaves to filtrate 2, extract at 60℃ for 2.5 h, filter to obtain filtrate 3. Mix filtrate 3 and filtrate 1, adjust the pH to 3, react at 30℃ for 0.5 h with stirring; then let stand for 1 h, filter, and collect the precipitate. Dissolve the precipitate in an aqueous solution of arginine with a pH of 8.5, filter, collect the liquid, concentrate and dry to obtain the plant whitening composition.

[0055] Example 2

[0056] This embodiment provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0057] (1) Take 30 g of Phyllanthus emblica and 30 g of Hibiscus rosa-sinensis and mix them with 900 mL of 50% (v / v) methanol aqueous solution, extract at 60℃ for 0.5 h, filter, and obtain filtrate 1. Mix the filter residue with 50% (v / v) methanol aqueous solution at a material-liquid ratio of 1 g: 15 mL, extract at 60℃ for 0.5 h, filter, and obtain filtrate 2.

[0058] (2) Add 60 g of mulberry leaves to filtrate 2, extract at 55℃ for 3 h, filter to obtain filtrate 3. Mix filtrate 3 and filtrate 1, adjust the pH to 2.5, react at 40℃ for 0.3 h with stirring; then let stand for 0.5 h, filter, and collect the precipitate. Dissolve the precipitate in an aqueous sodium hydroxide solution with a pH of 8, filter, collect the liquid, concentrate and dry to obtain the plant whitening composition.

[0059] Example 3

[0060] This embodiment provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0061] (1) Take 20 g of Phyllanthus emblica, 15 g of Hibiscus rosa-sinensis and 875 mL of 30% (v / v) ethyl acetate aqueous solution, extract at 50℃ for 1.5 h, filter to obtain filtrate 1. Mix the residue with 30% (v / v) ethyl acetate aqueous solution at a material-to-liquid ratio of 1 g: 25 mL, extract at 50℃ for 1.5 h, filter to obtain filtrate 2.

[0062] (2) Add 30 g of mulberry leaves to filtrate 2, extract at 65℃ for 2 h, filter to obtain filtrate 3. Mix filtrate 3 and filtrate 1, adjust the pH to 3.5, react at 25℃ for 0.8 h with stirring; then let stand for 1.5 h, filter, and collect the precipitate. Dissolve the precipitate in an aqueous solution of arginine with a pH of 9, filter, collect the liquid, concentrate and dry to obtain the plant whitening composition.

[0063] Example 4

[0064] This embodiment provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0065] (1) Take 25 g of Phyllanthus emblica and mix it with 500 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter, and collect the first filtrate of Phyllanthus emblica; take 20 g of Hibiscus rosa-sinensis and mix it with 400 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter, and collect the first filtrate of Hibiscus rosa-sinensis; mix the first filtrate of Phyllanthus emblica and the first filtrate of Hibiscus rosa-sinensis to obtain filtrate 1. Mix the Phyllanthus emblica residue with 40% (v / v) ethanol aqueous solution at a material-to-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter, and collect the second filtrate of Phyllanthus emblica; mix the Hibiscus rosa-sinensis residue with 40% (v / v) ethanol aqueous solution at a material-to-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter, and collect the second filtrate of Hibiscus rosa-sinensis; mix the second filtrate of Phyllanthus emblica and the second filtrate of Hibiscus rosa-sinensis to obtain filtrate 2.

[0066] (2) Add 45 g of mulberry leaves to filtrate 2, extract at 60℃ for 2.5 h, filter to obtain filtrate 3. Mix filtrate 3 and filtrate 1, adjust the pH to 3, react at 30℃ for 0.5 h with stirring; then let stand for 1 h, filter, and collect the precipitate. Dissolve the precipitate in an aqueous solution of arginine with a pH of 8.5, filter, collect the liquid, concentrate and dry to obtain the plant whitening composition.

[0067] Example 5

[0068] This embodiment provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0069] (1) Take 25 g of Phyllanthus emblica and mix it with 500 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter, and obtain filtrate 1. Mix the filter residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter, and obtain filtrate 2.

[0070] (2) Add 45 g of mulberry leaves to filtrate 2, extract at 60℃ for 2.5 h, filter to obtain filtrate 3. Mix filtrate 3 and filtrate 1, adjust the pH to 3, react at 30℃ for 0.5 h with stirring; then let stand for 1 h, filter, and collect the precipitate. Dissolve the precipitate in an aqueous solution of arginine with a pH of 8.5, filter, collect the liquid, concentrate and dry to obtain the plant whitening composition.

[0071] Example 6

[0072] This embodiment provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0073] (1) Take 20 g of roselle and mix it with 400 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter, and obtain filtrate 1. Mix the filter residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter, and obtain filtrate 2.

[0074] (2) Add 45 g of mulberry leaves to filtrate 2, extract at 60℃ for 2.5 h, filter to obtain filtrate 3. Mix filtrate 3 and filtrate 1, adjust the pH to 3, react at 30℃ for 0.5 h with stirring; then let stand for 1 h, filter, and collect the precipitate. Dissolve the precipitate in an aqueous solution of arginine with a pH of 8.5, filter, collect the liquid, concentrate and dry to obtain the plant whitening composition.

[0075] Comparative Example 1

[0076] This comparative example provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0077] (1) Take 25 g of Phyllanthus emblica and 20 g of Hibiscus rosa-sinensis and mix them with 900 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter, and obtain filtrate 1. Mix the residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter, and obtain filtrate 2.

[0078] (2) Add 45 g of mulberry leaves to filtrate 1, extract at 60℃ for 2.5 h, filter to obtain filtrate 3. Mix filtrate 3 and filtrate 2, adjust the pH to 3, react at 30℃ for 0.5 h with stirring; then let stand for 1 h, filter, and collect the precipitate. Dissolve the precipitate in an aqueous solution of arginine with a pH of 8.5, filter, collect the liquid, concentrate and dry to obtain the plant whitening composition.

[0079] Comparative Example 2

[0080] This comparative example provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0081] (1) Take 25 g of Phyllanthus emblica and mix it with 500 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter and collect the first filtrate of Phyllanthus emblica; mix the Phyllanthus emblica residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter and collect the second filtrate of Phyllanthus emblica; mix the first filtrate of Phyllanthus emblica and the second filtrate of Phyllanthus emblica to obtain Phyllanthus emblica extract.

[0082] (2) Take 20 g of roselle and mix it with 400 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter and collect the first filtrate of roselle; mix the roselle residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter and collect the second filtrate of roselle; mix the first filtrate and the second filtrate of roselle to obtain roselle extract.

[0083] (3) Mix 45 g of mulberry leaves with 900 mL of 40% (v / v) ethanol aqueous solution, extract at 60℃ for 2.5 h, filter to obtain filtrate, adjust the pH of the filtrate to 3, react at 30℃ for 0.5 h with stirring; then let stand for 1 h, filter, and collect the precipitate. Dissolve the precipitate in arginine aqueous solution with pH 8.5, filter, collect the liquid, and obtain mulberry leaf extract.

[0084] (4) Mix the amla extract, roselle extract and mulberry leaf extract, concentrate and dry to obtain the plant whitening composition.

[0085] Comparative Example 3

[0086] This comparative example provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0087] (1) Take 25 g of Phyllanthus emblica and mix it with 500 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter and collect the first filtrate of Phyllanthus emblica; mix the Phyllanthus emblica residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter and collect the second filtrate of Phyllanthus emblica; mix the first filtrate of Phyllanthus emblica and the second filtrate of Phyllanthus emblica to obtain Phyllanthus emblica extract.

[0088] (2) Mix 45 g of mulberry leaves with 900 mL of 40% (v / v) ethanol aqueous solution, extract at 60℃ for 2.5 h, filter to obtain filtrate, adjust the pH of the filtrate to 3, react at 30℃ for 0.5 h with stirring; then let stand for 1 h, filter, and collect the precipitate. Dissolve the precipitate in arginine aqueous solution with pH 8.5, filter, collect the liquid, and obtain mulberry leaf extract.

[0089] (3) Mix the amla extract and mulberry leaf extract, concentrate and dry to obtain the plant whitening composition.

[0090] Comparative Example 4

[0091] This comparative example provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0092] (1) Take 20 g of roselle and mix it with 400 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter and collect the first filtrate of roselle; mix the roselle residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter and collect the second filtrate of roselle; mix the first filtrate and the second filtrate of roselle to obtain roselle extract.

[0093] (2) Mix 45 g of mulberry leaves with 900 mL of 40% (v / v) ethanol aqueous solution, extract at 60℃ for 2.5 h, filter to obtain filtrate, adjust the pH of the filtrate to 3, react at 30℃ for 0.5 h with stirring; then let stand for 1 h, filter, and collect the precipitate. Dissolve the precipitate in arginine aqueous solution with pH 8.5, filter, collect the liquid, and obtain mulberry leaf extract.

[0094] (3) Mix roselle extract and mulberry leaf extract, concentrate and dry to obtain the plant whitening composition.

[0095] Comparative Example 5

[0096] This comparative example provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0097] (1) Take 25 g of Phyllanthus emblica and 20 g of Hibiscus rosa-sinensis and mix them with 900 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter, and obtain filtrate 1. Mix the residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter, and obtain filtrate 2.

[0098] (2) Add 45 g of mulberry leaves to filtrate 2, extract at 60℃ for 2.5 h, filter to obtain filtrate 3. Adjust the pH of filtrate 3 to 3, react at 30℃ for 0.5 h with stirring; then let stand for 1 h, filter, and collect the precipitate. Dissolve the precipitate in an aqueous solution of arginine with a pH of 8.5, filter, collect the liquid, add it to filtrate 1, mix, concentrate and dry to obtain the plant whitening composition.

[0099] Comparative Example 6

[0100] This comparative example provides a method for co-extracting a plant-based whitening composition with affinity, the method comprising:

[0101] (1) Take 25 g of Phyllanthus emblica and mix it with 500 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter and collect the first filtrate of Phyllanthus emblica; mix the Phyllanthus emblica residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter and collect the second filtrate of Phyllanthus emblica; mix the first filtrate of Phyllanthus emblica and the second filtrate of Phyllanthus emblica to obtain Phyllanthus emblica extract.

[0102] (2) Take 20 g of roselle and mix it with 400 mL of 40% (v / v) ethanol aqueous solution, extract at 55℃ for 1 h, filter and collect the first filtrate of roselle; mix the roselle residue with 40% (v / v) ethanol aqueous solution at a material-liquid ratio of 1 g: 20 mL, extract at 55℃ for 1 h, filter and collect the second filtrate of roselle; mix the first filtrate and the second filtrate of roselle to obtain roselle extract.

[0103] (3) Take 45 g of mulberry leaves and mix them with 900 mL of 40% (v / v) ethanol aqueous solution. Extract at 60℃ for 2.5 h, filter, collect the liquid, and obtain mulberry leaf extract.

[0104] (4) Mix the amla extract, roselle extract and mulberry leaf extract, concentrate and dry to obtain the plant whitening composition.

[0105] Test Example 1: Tyrosinase Inhibition Rate

[0106] The compositions obtained in Examples 1-6 and Comparative Examples 1-6 were prepared into 1 mg / mL solutions using disodium hydrogen phosphate-citric acid buffer at pH 6.8 as test samples. The inhibitory ability of the test samples on tyrosinase was determined according to T / SHRH 015-2018 "Cosmetics - Test Method for Inhibition of Tyrosinase Activity".

[0107] The test results are shown in Table 1. Comparative Examples 2-6 show that, compared to the method of extraction followed by combination, the composition obtained by enriching the active ingredients of mulberry leaves through two extractions of Phyllanthus emblica and / or Roselle, and then purifying them through mixed acid precipitation, significantly inhibits tyrosinase activity. Comparative Example 1 shows that the different contents of polyphenols, organic acids, etc., in the two extractions of Phyllanthus emblica and / or Roselle affect the solubility of DNJ in mulberry leaves, further affecting the tyrosinase inhibition rate of the final composition. Examples 5-6 show that the combination of Phyllanthus emblica and Roselle has a better enrichment effect on the active ingredients of mulberry leaves compared to Phyllanthus emblica or Roselle alone, resulting in a better tyrosinase inhibition rate in the final composition. Example 4 shows that the co-extraction of Phyllanthus emblica and Roselle results in a better tyrosinase inhibition rate in the final composition compared to extraction followed by combination of the two.

[0108] Table 1

[0109]

[0110] Test Example 2: AGEs Inhibition Rate

[0111] (1) Zebrafish preparation: Wild-type AB strain zebrafish were selected 5 days after fertilization, divided into groups of 20 fish and placed in 1.5 mL centrifuge tubes. A glycosylation model was established by zebrafish absorbing high-sugar solution through the skin to rapidly enhance the glycosylation reaction, and the effect of the sample on inhibiting AGEs synthesis was tested.

[0112] (2) Sample preparation: Using water as a solvent, the whitening compositions prepared in Examples 1-6 and Comparative Examples 1-6 were prepared into solutions of 100 mg / mL as test samples.

[0113] (3) Set up sample group, normal control group and model control group. 20 μL of the test sample with 100 mg / mL, 20 μL of 200 mg / mL glucose solution and 160 μL of zebrafish culture water were added to the zebrafish in the sample group. 200 μL of zebrafish culture water was added to the zebrafish in the normal control group. 200 μL of 20 mg / mL glucose solution was added only to the model control group.

[0114] (4) After incubating each group of zebrafish in the dark on a shaker at 60℃ for 18 h, the zebrafish were ground and crushed, and then incubated in the dark on a shaker at 60℃ for another 22 h. After centrifugation, 100 μL of supernatant was taken from each tube and transferred to a 96-well microplate. The microplate was used to detect and analyze the data under 370 nm excitation light and 440 nm absorption light. The fluorescence value (S) of zebrafish AGEs was analyzed, and the AGEs inhibition rate was calculated. The AGEs inhibition rate (%) = ((S model control group - S sample group) / (S model control group - S normal control group)) × 100%.

[0115] The test results are shown in Table 2. Comparative Examples 2-6 show that, compared to the method of extraction followed by combination, the composition prepared by enriching the active ingredients of mulberry leaves with two extracts of Phyllanthus emblica and / or Roselle, combined with a mixed acid precipitation purification method, significantly inhibits the formation of advanced glycation end products (AGEs). Comparative Example 1 shows that the different contents of polyphenols, organic acids, etc., in the two extracts of Phyllanthus emblica and / or Roselle affect the solubility of DNJ in mulberry leaves, further affecting the AGEs inhibition rate of the final composition. Examples 5-6 show that the combination of Phyllanthus emblica and Roselle has a better enrichment effect on the active ingredients of mulberry leaves than Phyllanthus emblica or Roselle alone, resulting in a better AGEs inhibition rate in the final composition. Example 4 shows that the co-extraction of Phyllanthus emblica and Roselle results in a better AGEs inhibition rate in the final composition compared to extraction followed by combination of individual extracts.

[0116] Table 2

[0117]

[0118] Test Example 3 DPPH Scavenging Rate

[0119] The compositions obtained in Examples 1-6 and Comparative Examples 1-6 were prepared into 0.1 mg / mL solutions using disodium hydrogen phosphate-citric acid buffer at pH 6.8 as test samples. The scavenging ability of the test samples against DPPH free radicals was determined according to T / SHRH 006-2018 "Cosmetics - Test Method for Free Radical (DPPH) Scavenging".

[0120] The test results are shown in Table 3. Comparative Examples 2-6 show that, compared to the method of extraction followed by combination, the composition obtained by enriching the active ingredients of mulberry leaves through two extractions of Phyllanthus emblica and / or Roselle, combined with a mixed acid precipitation optimization method, exhibits excellent free radical scavenging effects. Comparative Example 1 shows that the different contents of polyphenols, organic acids, etc., in the two extractions of Phyllanthus emblica and / or Roselle further affect the free radical scavenging effect of the final composition. Examples 5-6 show that the combination of Phyllanthus emblica and Roselle has a better enrichment effect on the active ingredients of mulberry leaves compared to Phyllanthus emblica or Roselle alone, resulting in a better free radical scavenging effect in the final composition. Example 4 shows that when extracting mulberry leaves using a combination of Phyllanthus emblica and Roselle, the co-extraction of Phyllanthus emblica and Roselle results in a better free radical scavenging effect in the final composition compared to extraction followed by combination of the two.

[0121] Table 3

[0122]

[0123] Test Example 4: Skin Safety and Moisture Content Test

[0124] (1) Sample preparation: The compositions provided in Examples 1-6 and Comparative Examples 1-6 were prepared into face creams (the composition content was 0.5%) as test samples, and the face cream base without any composition was used as a blank control sample.

[0125] (2) Volunteer selection: 130 volunteers aged 20-45 with healthy skin were selected as trial subjects and randomly divided into 13 groups of 10 each.

[0126] (3) Safety test: Using the closed patch test method, 0.025g of the test sample was placed in the patch tester (with deionized water as a reference), and low-sensitivity adhesive tape was applied to the flexor side of the subject's forearm. The test sample was removed after 24 hours, and the skin reaction was observed at 0.5 h, 24 h, and 48 h after removal. The results were recorded according to the skin reaction grading standard in the "Cosmetic Safety Technical Specifications" (2015 edition). Only samples that pass the safety test can be used for human trials.

[0127] (4) Moisture content test: After cleansing and applying toner in the morning and evening, the subjects took an appropriate amount of the test sample or control sample and applied it evenly to their face, gently massaging until absorbed, twice a day, once in the morning and once in the evening, for 28 consecutive days. Each group of subjects selected the same test sample or blank control sample.

[0128] The relative moisture content of the skin surface was measured using a skin moisture content meter. The relative moisture content of the skin surface was recorded before and 28 days after the sample was used. The rate of change of the relative moisture content of the skin surface relative to the sample was calculated as follows: Moisture content growth rate (%) = ((skin moisture content after use - skin moisture content before use) / skin moisture content before use) × 100%.

[0129] The safety test results are shown in Table 4. All subjects showed negative skin reactions, proving that the cosmetics prepared using the composition of this invention are safe and non-irritating. The skin moisture content test results are shown in Table 5. As can be seen from Comparative Examples 2-6, compared to the method of single extraction followed by combination, this invention enriches the active ingredients of mulberry leaves through two extractions of Phyllanthus emblica and / or Roselle, resulting in a composition that significantly increases skin moisture content and has excellent moisturizing effects. As can be seen from Comparative Example 1, the different contents of polyphenols, organic acids, etc., in the two extracts of Phyllanthus emblica and / or Roselle further affect the moisturizing effect of the final composition.

[0130] Table 4

[0131]

[0132] Table 5

[0133]

[0134] This invention illustrates a method for co-extracting plant whitening compositions, its products, and applications through the above embodiments. However, this invention is not limited to the above embodiments, meaning that it does not necessarily rely on the above embodiments for implementation. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials in the products of this invention, additions of auxiliary components, and selection of specific methods all fall within the protection and disclosure scope of this invention.

[0135] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0136] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A method of affinity co-extraction of a plant whitening composition, characterized in that, The method comprises: (1) mixing Phyllanthus emblica and / or Roselle with an extraction reagent, extracting, solid-liquid separation, to obtain filtrate 1; mixing the residue with an extraction reagent, extracting, solid-liquid separation, to obtain filtrate 2; Or, mixing Phyllanthus emblica and Roselle with an extraction reagent respectively, extracting, solid-liquid separation, collecting the liquid and mixing, to obtain filtrate 1; mixing the residue of Phyllanthus emblica and Roselle with an extraction reagent respectively, extracting, solid-liquid separation, collecting the liquid and mixing, to obtain filtrate 2; (2) mixing mulberry leaves and filtrate 2, extracting, solid-liquid separation, to obtain filtrate 3; mixing filtrate 3 and filtrate 1, adjusting pH value, reacting, standing, solid-liquid separation, collecting the precipitate; dissolving the precipitate in an alkaline aqueous solution, solid-liquid separation, collecting the liquid, to obtain the plant whitening composition.

2. The method of claim 1, wherein, In step (1), the extraction reagent comprises an organic solvent and water; Preferably, the organic solvent comprises any one or a combination of at least two of ethanol, methanol or ethyl acetate; Preferably, the volume percentage content of the organic solvent in the extraction reagent is 30-50%.

3. The method according to claim 1 or 2, characterized in that, In step (1), the solid-liquid ratio of the extraction is 1 g:(15-25) mL; Preferably, the extraction temperature is 50-60℃, and the time is 0.5-1.5 h.

4. The method according to any one of claims 1-3, characterized in that, In step (2), the extraction temperature is 55-65℃, and the time is 2-3 h.

5. The method according to any one of claims 1-4, characterized in that, In step (2), the pH value is adjusted to 2-4; Preferably, the reaction temperature is 25-40℃, and the time is 0.3-0.8 h; Preferably, the reaction is carried out under stirring.

6. The method according to any one of claims 1-5, characterized in that, In step (2), the standing time is 0.5-1.5 h.

7. The method according to any one of claims 1 to 6, characterized in that, In step (2), the pH value of the alkaline aqueous solution is 8-9; Preferably, after collecting the liquid, a step of concentrating and drying is further included.

8. The method according to any one of claims 1-7, characterized in that, The mass ratio of Phyllanthus emblica and mulberry leaves is (20-30):(30-60); Preferably, the mass ratio of Roselle and mulberry leaves is (15-30):(30-60).

9. The plant whitening composition prepared by the method according to any one of claims 1-8.

10. The use of the plant whitening composition according to claim 9 in the preparation of cosmetics, food or health products.

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