Annona squamosa polysaccharide extracting solution as well as preparation method and application thereof

Abstract

The invention discloses an annona squamosa polysaccharide extracting solution as well as a preparation method and application thereof, and the preparation method comprises the following steps: S1, selecting mature annona squamosa from Hainan, then washing, peeling, deseeding and juicing to obtain annona squamosa pulp; s2, mixing the annona squamosa pulp with water, performing ultrasonic treatment, and sequentially performing first extraction and suction filtration to obtain primary filtrate and primary filter residues; s3, mixing the primary filter residues with water, and sequentially performing secondary extraction and suction filtration to obtain secondary filtrate and secondary filter residues; s4, mixing the secondary filter residue with water, and sequentially carrying out third extraction and suction filtration to obtain a third filtrate; and S5, mixing the primary filtrate, the secondary filtrate and the third filtrate, and concentrating to obtain the annona squamosa polysaccharide extracting solution. The annona squamosa polysaccharide extracting solution prepared by adopting the preparation method is rich in active ingredients such as amino acid, polypeptide and polysaccharide, has the effects of moisturizing, resisting oxidation and whitening, and can meet various skin care requirements without adding other functional ingredients.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, and in particular relates to an extract of custard apple polysaccharide, its preparation method and application. Background Technology

[0002] Annona squamosa is an evergreen fruit tree native to tropical and subtropical regions. Its fruit is loved by consumers for its sweet taste and rich nutrition. Besides being a high-quality edible fruit, the entire Annona squamosa plant contains abundant bioactive components, including polysaccharides, flavonoids, alkaloids, and terpenoids. These active components possess various biological functions, such as antioxidant, antibacterial, anti-inflammatory, and moisturizing effects, thus having broad research and application value in the food, pharmaceutical, and cosmetic fields.

[0003] Currently, there are still relatively few technologies for the extraction, separation, and application of custard apple polysaccharides, especially in the cosmetics field, where their potential as functional ingredients has not been fully explored. Traditional polysaccharide extraction methods typically include enzymatic hydrolysis and organic solvent extraction, but these methods may suffer from low extraction efficiency, significant loss of active ingredients, or complex operation, thus limiting the efficient utilization of custard apple polysaccharides.

[0004] Therefore, there is an urgent need for an extract of custard apple polysaccharides, its preparation method, and its application to address the shortcomings of existing technologies. Summary of the Invention

[0005] In view of the above problems, the purpose of this invention is to provide an Annona squamosa polysaccharide extract, its preparation method and application. The Annona squamosa polysaccharide extract, rich in polysaccharides, polypeptides and free amino acids, is obtained by using the ultrasonic-assisted hot water extraction process of this invention. This Annona squamosa polysaccharide extract is derived from natural plants, is rich in nutrients, has high safety, is not likely to cause allergic reactions, has good moisturizing effect, can scavenge free radicals, inhibit tyrosinase activity and reduce melanin production, thereby achieving moisturizing, antioxidant and whitening effects. It can be directly used as a functional additive in the cosmetics field.

[0006] To achieve the above objectives, the first aspect of the present invention provides a method for preparing an extract of custard apple polysaccharides, comprising the following steps: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice the custard apples to obtain custard apple pulp. S2. Mix the custard apple pulp with water and sonicate it at 180-220W for 15-25 minutes. Then perform the first extraction and filtration to obtain the first filtrate and the first filter residue. S3. Mix the primary filter residue with water and perform a second extraction and filtration sequentially to obtain a secondary filtrate and a secondary filter residue; S4. Mix the secondary filter residue with water and perform a third extraction and filtration sequentially to obtain a third filtrate. S5. Combine the first, second and third filtrates and concentrate them to obtain custard apple polysaccharide extract; The conditions for the first, second, and third extractions are independent: constant temperature extraction at 80~100℃ for 2.5~3.5h.

[0007] Compared with existing technologies, this application selects ripe custard apples from Hainan as raw materials, which are characterized by high polysaccharide content and stable quality. Simultaneously, an ultrasound-assisted hot water extraction process is employed, where ultrasound (180~220W for 15~25 min) effectively breaks down cell wall structures and significantly enhances solvent permeability, thereby increasing polysaccharide yield. Gentle hot water extraction (80~100℃ constant temperature extraction for 2.5~3.5 h) avoids the damage to active ingredients (such as amino acids and peptides) caused by high temperatures or organic solvents, thus effectively increasing the content of active ingredients (such as amino acids and peptides). Therefore, this application uses ripe custard apples from Hainan as raw materials. Through an ultrasonic-assisted hot water extraction process (180-220W ultrasound for 15-25 minutes combined with constant temperature extraction at 80-100℃ for 2.5-3.5 hours) coupled with a three-stage progressive filtration technique, the yield of custard apple polysaccharides is significantly improved, and the content of active ingredients (such as amino acids and peptides) is effectively increased. The resulting custard apple polysaccharide extract is highly safe, unlikely to cause allergic reactions, and has good moisturizing effects. It can scavenge free radicals, inhibit tyrosinase activity, and reduce melanin production, thereby achieving moisturizing, antioxidant, and whitening effects. Furthermore, this method is simple to operate and environmentally friendly and economical.

[0008] Further, in step S2 of the present invention, the volume ratio of custard apple pulp to water is 1:10~20. Preferably, in step S2 of the present invention, the volume ratio of custard apple pulp to water is 1:15.

[0009] Furthermore, step S2 of the present invention includes mixing custard apple pulp with water and then ultrasonically treating it at 200W for 20 minutes.

[0010] Furthermore, the conditions for the first extraction, second extraction, and third extraction of the present invention are all: constant temperature extraction at 100°C for 3 hours.

[0011] Accordingly, a second aspect of the present invention provides an Annona squamosa polysaccharide extract, prepared using the aforementioned method for preparing Annona squamosa polysaccharide extract. This Annona squamosa polysaccharide extract is derived from a natural plant, is rich in nutrients, has high safety, is unlikely to trigger allergic reactions, and possesses excellent moisturizing effects. It can scavenge free radicals, inhibit tyrosinase activity, and reduce melanin production, thereby achieving moisturizing, antioxidant, and whitening effects. It can be directly used as a functional additive in the cosmetics field.

[0012] Accordingly, a third aspect of the present invention provides the application of the aforementioned custard apple polysaccharide extract in cosmetics. This custard apple polysaccharide extract has multiple functions, including moisturizing, whitening, and antioxidant effects, and can be widely used as a functional additive in the cosmetics field, possessing high market application potential.

[0013] Specifically, the dosage forms of cosmetics are selected from pastes, ointments, sprays, gels, liniments, coatings, films, patches, plasters, or films.

[0014] Accordingly, a fourth aspect of the present invention provides a binary spray, comprising, by weight, 86-95 parts water, 0.03-0.06 parts chelating agent, 0.3-0.6 parts atomization improver, 0.3-0.6 parts preservative, 2-3 parts humectant, and 1-10 parts D phase, wherein the D phase is the custard apple polysaccharide extract prepared by the aforementioned method or the aforementioned custard apple polysaccharide extract.

[0015] Compared with existing technologies, this invention combines custard apple polysaccharide extract with moisturizers, atomization improvers, and chelating agents to obtain a binary spray with multiple functions. This not only fully preserves the moisturizing, antioxidant, and whitening active effects of custard apple polysaccharides, but its unique binary packaging design also effectively ensures the stability of the active ingredients while improving ease of use. Testing has shown that this binary spray exhibits excellent safety, stability, moisturizing properties, soothing effects, firming effects, whitening effects, and atomization performance.

[0016] Furthermore, the chelating agent of the present invention is selected from disodium ethylenediaminetetraacetate; the preservative is selected from p-hydroxyacetophenone.

[0017] Furthermore, the moisturizer of the present invention is selected from at least one of 1,2-hexanediol and 2,3-butanediol.

[0018] Further, the atomization improver of the present invention, by weight parts, comprises 40-50 parts of polydimethylsiloxane alcohol, 40-50 parts of water, 0.1-0.5 parts of tetrasodium diacetate of glutamate, 0.1-1 part of 1,3-propanediol, 2-5 parts of polysorbate-20, 0.1-1 part of capryloyl hydroxamic acid, and 0.1-1 part of glyceryl caprylate. Attached Figure Description

[0019] Figure 1 The graph shows the water loss rate of the custard apple polysaccharide extract from Example 10.

[0020] Figure 2 The moisture retention curve of the custard apple polysaccharide extract in Example 10 is shown.

[0021] Figure 3The graph shows the inhibition rate of tyrosinase by the polysaccharide extract of Annona squamosa in Example 10.

[0022] Figure 4 The graph shows the effect of the concentration of custard apple polysaccharide extract in Example 10 on the initial velocity of tyrosinase.

[0023] Figure 5 The Lineweaver-Bulk curve of the inhibitory effect of the custard apple polysaccharide extract of Example 10 on the inhibition of tyrosinase on L-tyrosine catalysis by tyrosinase.

[0024] Figure 6 This is a diagram illustrating the melanin-inhibiting effect of zebrafish in Example 11.

[0025] Figure 7 This is a bar chart comparing the melanin signal intensity in the head of zebrafish in Example 11 with that of the normal control group.

[0026] Figure 8 This is a diagram illustrating the melanin-inhibiting effect of zebrafish in Example 13.

[0027] Figure 9 This is a bar chart comparing the melanin signal intensity in the head of zebrafish in Example 13 with that of the normal control group. Detailed Implementation

[0028] To better illustrate the purpose, technical solution, and beneficial effects of this invention, the invention will be further described below in conjunction with specific embodiments and accompanying drawings. It should be noted that the methods described below are further explanations of this invention and should not be construed as limiting it.

[0029] Example 1 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp with water at a volume ratio of 1:10, sonicate at 200W for 15 minutes, and then extract at 80℃ for 2.5 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 80℃ for 2.5h. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 80℃ for 2.5h. After extraction, filter to obtain the third filtrate. S5. Combine the first, second, and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract.

[0030] Example 2 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp and water at a volume ratio of 1:10, then sonicate at 200W for 20 minutes, and then extract at 90℃ for 3 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 90℃ for 3 hours. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 90℃ for 3 hours. After extraction, filter to obtain the third filtrate. S5. Combine the first, second, and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract.

[0031] Example 3 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp and water at a volume ratio of 1:10, then sonicate at 200W for 25 minutes, and then extract at 100℃ for 3.5 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 100℃ for 3.5 hours. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 100℃ for 3.5h. After extraction, filter to obtain the third filtrate. S5. Combine the first, second, and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract.

[0032] Example 4 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp and water at a volume ratio of 1:15, then sonicate at 200W for 15 minutes, and then extract at 80℃ for 3 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at 80℃ for 3 hours. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at 80℃ for 3 hours. After extraction, filter to obtain the third filtrate. S5. Combine the first, second, and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract.

[0033] Example 5 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp and water at a volume ratio of 1:15, then sonicate at 200W for 15 minutes, and then extract at 90℃ for 3.5 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 90℃ for 3.5h. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 90℃ for 3.5h. After extraction, filter to obtain the third filtrate. S5. Combine the first, second, and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract.

[0034] Example 6 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp with water at a volume ratio of 1:15, sonicate at 200W for 20 minutes, and then extract at 100℃ for 2.5 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 100℃ for 2.5h. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 100℃ for 2.5h. After extraction, filter to obtain the third filtrate. S5. Combine the first, second, and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract.

[0035] Example 7 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp with water at a volume ratio of 1:20, sonicate at 200W for 20 minutes, and then extract at 80℃ for 3.5 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 80℃ for 3.5h. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 80℃ for 3.5h. After extraction, filter to obtain the third filtrate. S5. Combine the first, second, and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract.

[0036] Example 8 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp with water at a volume ratio of 1:20, sonicate at 200W for 25 minutes, and then extract at 90℃ for 2.5 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 90℃ for 2.5h. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 90℃ for 2.5h. After extraction, filter to obtain the third filtrate. S5. Combine the first, second, and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract.

[0037] Example 9 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp with water at a volume ratio of 1:20, sonicate at 200W for 15 minutes, and then extract at 100℃ for 3 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 100℃ for 3 hours. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at 100℃ for 3 hours. After extraction, filter to obtain the third filtrate. S5. Combine the first, second, and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract.

[0038] Example 10 This embodiment provides a method for preparing custard apple polysaccharide extract, the steps of which include: S1. Select ripe custard apples from Hainan, then wash, peel, remove seeds, and juice them to obtain custard apple pulp. S2. Mix custard apple pulp and water at a volume ratio of 1:15, then sonicate at 200W for 20 minutes, and then extract at 100℃ for 3 hours. After extraction, filter to obtain a first filtrate and a first filter residue. S3. Mix the primary filter residue with water at a mass ratio of 1:10 and extract at a constant temperature of 100℃ for 3 hours. After extraction, filter to obtain secondary filtrate and secondary filter residue. S4. Mix the secondary filter residue with water at a mass ratio of 1:10 and extract at 100℃ for 3 hours. After extraction, filter to obtain the third filtrate. S5. Combine the first, second and third filtrates and concentrate them to 20 mL to obtain custard apple polysaccharide extract. After freeze-drying the custard apple polysaccharide extract, the mass of the solid was weighed, and the concentration of the custard apple polysaccharide extract was obtained by dividing the mass of the solid by the volume (20 mL). The concentration of the custard apple polysaccharide extract was 15 mg / mL.

[0039] Comparative Example 1 Comparative Example 1 and Example 10 are basically the same, except that: Step S2 of Comparative Example 1 includes mixing custard apple pulp and water at a volume ratio of 1:15 and extracting at a constant temperature of 100°C for 3 hours. After extraction, the mixture is filtered to obtain a filtrate and a residue.

[0040] Comparative Example 2 Comparative Example 2 and Example 10 are basically the same, except that: Step S2 of Comparative Example 2 includes mixing custard apple pulp with 40% alcohol at a volume ratio of 1:15 and extracting at 100°C for 3 hours. After extraction, the mixture is filtered to obtain a filtrate and a residue.

[0041] Comparative Example 3 Comparative Example 3 and Example 10 are basically the same, except that: Step S2 of Comparative Example 3 includes mixing custard apple pulp with 40% alcohol at a volume ratio of 1:15, ultrasonically treating it at 200W for 20 minutes, and then extracting it at 100°C for 3 hours. After extraction, the mixture is filtered to obtain a filtrate and a residue.

[0042] The polysaccharide content of the custard apple polysaccharide extracts obtained in Examples 1-10 and Comparative Examples 1-3 was determined according to the following test methods, and the results are shown in Table 1.

[0043] Methods for determining polysaccharide content: Dissolve 10 mg of anhydrous glucose in 100 mL of distilled water to prepare a glucose standard solution. Accurately pipette 0.2, 0.4, 0.6, 0.8, and 1.0 mL of the glucose standard solution, add 1.0 mL of 6% phenol solution to each, and quickly add 5.0 mL of concentrated sulfuric acid. Mix thoroughly and heat in a boiling water bath for 15 minutes, then cool to room temperature. Measure the absorbance at 490 nm using a UV spectrophotometer. Use distilled water as a blank control. Plot a standard curve: Y = 0.01451X + 0.02548, R0 2 =0.9973.

[0044] Sample preparation: Take 0.2 mL of the diluted custard apple polysaccharide extract and measure the absorbance according to the above method. Repeat three times and take the average value. Calculate the total sugar yield using the following formula.

[0045] Z=(ρ×V×N×10 -3 ) / m×100%; where Z is the polysaccharide yield of the custard apple polysaccharide extract, %; ρ is the mass concentration of the diluted custard apple polysaccharide extract, mg / mL; V is the volume of the custard apple polysaccharide extract after dilution with water, mL; N is the dilution factor; and m is the mass of the custard apple pulp, g.

[0046] Table 1

[0047] Comparing Examples 1 to 10, it can be seen that Example 10 achieved the highest polysaccharide yield of 17.79% under the conditions of a material-to-liquid ratio of 1:15, ultrasonic time of 20 min, extraction temperature of 100℃, and extraction time of 3.0 h. Therefore, in the preparation method of the present invention, the preferred volume ratio of custard apple pulp to water is 1:15; the mixture of custard apple pulp and water is preferably ultrasonically treated at 200W for 20 min; and the conditions for the first, second, and third extractions are preferably all constant temperature extraction at 100℃ for 3 h.

[0048] The polypeptide content, amino acid content, and molecular weight of the custard apple polysaccharide extracts obtained in Example 10 and Comparative Examples 1-3 were tested using the following methods. The results are shown in Table 2.

[0049] Polypeptide content determination: Pipette 0, 0.2, 0.4, 0.6, 0.8, 1.0 mL and 4 mL of biuret reagent from bovine serum albumin solution (10 mg / mL) into test tubes, respectively, and bring the volume to 5 mL with distilled water. Incubate at room temperature for 30 minutes. Plot a standard curve with protein concentration on the x-axis and OD540 on the y-axis: Y = 0.04869X + 0.05924, R 2 =0.9989. Sample determination: Add an equal volume of 10% trichloroacetic acid (TCA) solution to the sample, mix well, let stand for 30 minutes, centrifuge at 4000 rpm for 10 minutes, collect the supernatant and filter through a 0.45 μm filter membrane. Take 1 mL of the filtrate, add 4 mL of biuret reagent, and determine OD540 according to the above method. Calculate the polypeptide content (D) according to the following formula.

[0050] D = (C × V) / m; where D is the polypeptide content in the custard apple polysaccharide extract, mg / g; C is the polypeptide concentration in the custard apple polysaccharide extract, mg / mL; V is the volume of the custard apple polysaccharide extract, 20 mL; and m is the mass of the custard apple pulp, g.

[0051] Amino acid content determination: The experimental method follows GB / T2409-1998. Plot the standard curve: Y = 1.832X - 0.5057, R0 2 =0.9914. Calculate the amino acid content using the following formula.

[0052] A = (C × V × N) / m; where A is the amino acid content in the custard apple polysaccharide extract, mg / g; C is the mass concentration of the diluted custard apple polysaccharide extract calculated by substituting the absorbance value into the standard curve, mg / mL; V is the volume of the custard apple polysaccharide extract, 20 mL; N is the dilution factor; and m is the mass of custard apple pulp, g.

[0053] Molecular weight test: The Ubbelohde viscometer has a capillary inner diameter of 0.59 mm and a viscometer constant of 0.01170 mm. 2 / S 2 The constant temperature water bath was 30℃. Sample concentrations: 0, 0.015, 0.012, 0.009, 0.006, 0.003, 0.00150 mg / mL. Eluting times were recorded: three times for each concentration, and the average of the three times was taken as the elution time (T0, T). The relative viscosity (n) was calculated. r =T / T0), specific viscosity (n)sp =n r -1) nsp / c, lnnr / c. Plot the fitting curves of nsp / c versus c and lnnr / c versus c. Calculate the molecular weight using [η] = KMa to obtain the molecular weight range M. Note: K = 0.01089, a = 0.7, [η] = intercept. The fitting curve equations are shown in Table 3.

[0054] Table 2

[0055] Table 3

[0056] A comparison of Example 10 and Comparative Examples 1-3 shows that, compared to traditional extraction methods (hot water extraction, alcohol hot extraction, and alcohol ultrasound-assisted hot extraction), the ultrasound-assisted hot water extraction method used in this application not only significantly improves the yield of custard apple polysaccharides but also effectively increases the content of active ingredients such as amino acids and peptides. These ingredients can exert key bioactive functions such as moisturizing, anti-oxidation, and whitening. Furthermore, this method avoids the use of organic solvents (such as alcohol), meets the requirements of green extraction processes, and is suitable for the preparation of natural functional cosmetic raw materials.

[0057] The antioxidant, moisturizing, soothing, and whitening properties of the custard apple polysaccharide extract obtained in Example 10 were tested using the following methods.

[0058] 1. Antioxidant performance testing: 1.1 DPPH free radical scavenging ability test: Extracts of Annona squamosa polysaccharides at different concentrations (0.3, 0.6, 1.2, 1.5, 1.8 mg / mL) were prepared. A 1 mmol / L DPPH solution was prepared with anhydrous ethanol as the stock solution. The stock solution was diluted 4 times to achieve an absorbance value in the range of 0.8-1 (stored in the dark) to prepare the DPPH working solution. 2 mL of the DPPH working solution was added to each of the Annona squamosa polysaccharide extracts at different concentrations (0.3, 0.6, 1.2, 1.5, 1.8 mg / mL), followed by distilled water to a final volume of 4 mL. After mixing, the solution was allowed to stand for 30 min, and the OD value (A1) was measured at 517 nm. 2 mL of anhydrous ethanol was mixed with 2 mL of the DPPH working solution, and the OD value (A0) was measured. 2 mL of the sample solution was mixed with 2 mL of anhydrous ethanol to obtain the A2 value. Simultaneously, 1 mg / mL of Vitamin C was used as a positive control, with the same volume as the Annona squamosa polysaccharide extract. The DPPH free radical scavenging rate was calculated using the following formula: ; Where A0 is the absorbance value after the reaction with anhydrous ethanol instead of the sample solution; A1 is the absorbance value after the reaction with the sample solution; and A2 is the absorbance value after the reaction with anhydrous ethanol instead of the DPPH solution.

[0059] The results of DPPH free radical scavenging ability are shown in Table 4.

[0060] Table 4

[0061] 1.2 Hydroxyl radical scavenging ability test: Different concentrations (1.5, 3.0, 4.5, 6.0, 7.5 mg / mL) of Annona squamosa polysaccharide extract were taken, and 2.0 mL of 9.0 mmol / L ferrous sulfate solution, 2.0 mL of 9.0 mmol / L salicylic acid solution, and 2.0 mL of 8.8 mmol / L hydrogen peroxide solution were added. Distilled water was added to a final volume of 8 mL, and the mixture was thoroughly shaken and incubated in a water bath at 37.0℃ for 25.0 min. After cooling, the absorbance was measured at a wavelength of 510 nm. Each group was measured in triplicate, and the average value was taken. Distilled water was used to zero the instrument. A 1 mg / mL concentration of vitamin C was used as a positive control group.

[0062] The formula for calculating hydroxyl radical scavenging rate is as follows: Clearance rate (%) = [A0 - (A1 - A2) / A0] × 100% A0: Absorbance after the reaction is performed with distilled water instead of the sample solution; A1: Absorption of light after the sample solution reacts; A2: Absorbance after the reaction with distilled water instead of hydrogen peroxide solution.

[0063] The results of the hydroxyl radical scavenging ability test are shown in Table 5.

[0064] Table 5

[0065] 1.3 FRAP Total Antioxidant Capacity Test FRAP working solution: It is prepared by mixing 25 mL of 300 mmol / L pH 3.6 acetate buffer (5.1 g sodium acetate and 20 mL glacial acetic acid diluted with water to 250 mL), 2.5 mL of 10 mmol / L TPTZ solution, and 2.5 mL of 20 mmol / L FeCl3 solution.

[0066] Construction of the standard curve: Prepare 0.3 mL of FeSO4 standard solution at six concentration gradients (0.1, 0.2, 0.4, 0.6, 0.8, and 1.0 mmol / L), add 3 mL of FRAP working solution and 0.3 mL of ultrapure water. After mixing, react for 5 min, and measure the absorbance at 593 nm to construct the standard curve.

[0067] Sample determination: 1 mg / mL VC was used as the positive control group. 0.3 mL of 4.5 mg / mL custard apple polysaccharide extract was taken, 3 mL of FRAP working solution was added, and then 0.3 mL of ultrapure water was added. The mixture was mixed and reacted accurately for 5 min. The absorbance was measured at 593 nm and zeroed with ultrapure water.

[0068] The formula for calculating FRAP value (mmol / L) for liquid samples is: FRAP value = X × D; where X is the concentration of the sample after substituting the absorbance value into the standard curve, in mmol / L; and D is the sample dilution factor. The results of the total antioxidant capacity test of FRAP are shown in Table 6.

[0069] Table 6

[0070] As shown in Tables 4 to 6, the custard apple polysaccharide extract prepared by the method of the present invention has excellent antioxidant properties.

[0071] 2. Moisturizing and soothing performance test: 2.1 Water Loss Rate Test Sodium hyaluronate was prepared as a 1.0 mg / mL solution as a positive control. Three 4×7 cm glass plates were used as the experimental group, positive control group, and negative control group, respectively. Filter paper of the same size was laid flat and fixed on each plate, and the initial weights were recorded. 0.3 mL of 4.5 mg / mL custard apple polysaccharide extract was applied to the glass plate of the experimental group, 0.3 mL of sodium hyaluronate was applied to the glass plate of the positive control group, and 0.3 mL of distilled water was applied to the glass plate of the negative control group. The weights were recorded. Then, the three glass plates were placed in a 36℃ electric drying oven, and their weights were recorded every 5 minutes. The difference in mass between the plates after sample addition and after removal from the electric constant temperature incubator was calculated to determine the amount of water loss. The water loss rate was used to determine the moisturizing effect. The water loss rate test results of the custard apple polysaccharide extract are shown below. Figure 1 As shown.

[0072] 2.2 Hyaluronidase Activity Test The experiment used the EIson-Morgan method. 0.1 mL of 2.5 mmol / L CaCl2 solution was added to 0.5 mL of hyaluronidase (500 U / mL), and the mixture was incubated at 37℃ for 20 min. Then, 0.5 mL of sample solution was added, and the mixture was incubated at 37℃ for 20 min. Next, 2 mL of 0.4 mg / mL sodium hyaluronate was added, and the mixture was incubated at 37℃ for 40 min. Then, 1 mL of acetylacetone solution (1.5 mL of acetylacetone dissolved in 50 mL of 1 mol / L sodium carbonate solution, freshly prepared) was added, and the mixture was incubated at 90℃ for 1 h. After cooling with water, 10 mL of 95% ethanol was slowly added, followed by 1.0 mL of Ehrlich reagent (prepared by dissolving 0.8 g DMAB in 15 mL of concentrated hydrochloric acid and 15 mL of 95% ethanol). The mixture was thoroughly mixed, and after standing at room temperature for 1 h, the absorbance was measured at 547 nm.

[0073] Calculate the hyaluronidase inhibition rate using the following formula: Hyaluronidase inhibition rate = [(DC) - (AB)] / DC × 100%; In the formula for hyaluronidase inhibition rate: A represents the OD values ​​of 1.5, 3.0, 4.5, 6.0, and 7.5 mg / mL custard apple polysaccharide extract and 1 mg / mL tannic acid solution, respectively; B represents the OD value of the blank sample (using acetate buffer solution instead of enzyme solution); C represents the OD value of the control blank solution (using acetate buffer solution instead of sample and enzyme solution); and D represents the OD value of the control solution (using acetate buffer solution instead of sample).

[0074] The results of the hyaluronidase activity test are shown in Table 7: Table 7

[0075] 2.3 Determination of Moisture Retention Rate Weigh 0.3 mg of custard apple polysaccharide extract and place it in a centrifuge tube. Expose the tube to a desiccator with RH=5% (blue silica gel) and measure the mass change of the sample over a certain period of time. The positive control is glycerol and the negative control is water. Calculate the moisture retention rate according to the formula.

[0076] Moisturizing rate (%) = (H1-H2) / H1×100%; where H2 is the mass (mg) of the custard apple polysaccharide extract after water loss, and H1 is the mass (mg) of the custard apple polysaccharide extract before it was added.

[0077] The moisturizing rate curve of custard apple polysaccharide extract is as follows: Figure 2 As shown.

[0078] pass Figures 1-2As shown in Table 7, the custard apple polysaccharide extract prepared by the method of the present invention has excellent moisturizing and soothing properties.

[0079] 3. Whitening performance test: 3.1 Tyrosinase inhibition test: Preparation of tyrosinase solution: Accurately weigh tyrosinase and dissolve it in PBS to prepare an enzyme solution of approximately 200 U / mg. Store at -20℃ and thaw at 4℃ before use. Tyrosine solution: Accurately weigh L-tyrosine, dissolve it completely in a small amount of 0.1 mol / mL hydrochloric acid solution, then add PBS to prepare a 1.5 mmol / mL tyrosine solution. Store at 4℃ protected from light. Samples: Anthocyanin extracts and arbutin solutions at concentrations of 5, 10, 15, and 20 mg / mL. According to the reaction groups in Table 8, transfer PBS solution, sample, and tyrosinase solution into 5 mL capped plastic centrifuge tubes (EP tubes), mix well, and incubate in a water bath at 37℃ for 10 min. Quickly add 0.5 mL of tyrosine solution, mix well, and incubate in a water bath at 37℃ for another 4 min. Quickly transfer the mixture to a cuvette, and measure the absorbance at 475 nm at the 5th minute (PBS solution serves as a blank control) to obtain C1, C2, T1, and T2. Perform the experiment in triplicate. According to the formula: Inhibition rate (%) = [1 - (T1 - T2) / (C1 - C2)] × 100%; The inhibition rate curve of sucrase polysaccharide extract on tyrosinase is shown in the figure below. Figure 3 As shown.

[0080] Table 8

[0081] 3.2 Tyrosinase kinetic analysis: Kinetic analysis of tyrosinase inhibition: The mechanisms of action between inhibitors and enzymes are classified into reversible and irreversible inhibition. Reversible inhibition is further divided into competitive, non-competitive, anti-competitive, and linear mixed types, which can be determined using a Lineweaver-Burk double reciprocal plot. By fixing the tyrosinase concentration at 200 U / mL and using L-tyrosine as the substrate, the concentrations of custard apple polysaccharide extract (5, 10, 15, and 20 mg / mL) and L-tyrosine solution (0.5, 1, 1.5, and 2 mg / mL) were varied.

[0082] Preparation of tyrosinase solution: Accurately weigh tyrosinase and dissolve it in PBS to prepare approximately 200 U / mg solution. -1 The enzyme solution should be frozen at -20°C and thawed at 4°C before use.

[0083] Tyrosine solution: Accurately weigh L-tyrosine, first using a small amount of 0.1 mol•L... -1Dissolve completely in hydrochloric acid solution, then add PBS solution to prepare L-tyrosine solutions with concentration gradients of 0.5, 1, 1.5, and 2 mg / mL. Store at 4°C protected from light.

[0084] Samples: Anthopogon japonicus polysaccharide extracts at concentrations of 5, 10, 15, and 20 mg / mL.

[0085] Transfer 1.5 mL of PBS solution, 0.5 mL of solutions with different sample concentrations, and 0.5 mL of solutions with different L-tyrosine concentrations to 5 mL capped plastic centrifuge tubes (EP tubes), mix thoroughly, and incubate in a water bath at 37℃ for 10 min. Quickly add 0.5 mL of tyrosine solution, mix thoroughly, and incubate in a water bath at 37℃ for another 4 min. Quickly transfer the mixture to a cuvette, and measure the absorbance at 475 nm at the 5th minute (PBS solution serves as a blank control). Obtain the absorbance value A after subtracting the blank. Perform the experiment in triplicate. The result is obtained using the formula V0 = (A × 1000) / (…). ×l×t)( The absorbance coefficient of dopachrome is 3600 (L / (mol·cm)); l is the optical path length of the cuvette (1 cm); t is the reaction time (1 min); 1000 is for unit conversion; V0 is in units of (mmol / L / min). The Michaelis-Menten equation was fitted after calculating the initial velocity. Based on the results, the initial velocity and the concentration of Annona squamosa polysaccharide extract were plotted using the Lineweaver-Burk double reciprocal equation method, and a table was created to analyze the kinetics of the inhibitory effect of Annona squamosa polysaccharide extract on tyrosinase. Specifically, the effect of Annona squamosa polysaccharide extract concentration on the initial velocity of tyrosinase is as follows: Figure 4 As shown; the Lineweaver-Bulk curve of the inhibitory effect of custard apple polysaccharide extract on tyrosinase-catalyzed L-tyrosine is shown in the figure. Figure 5 As shown in Table 9, the kinetic parameters of the inhibition of tyrosinase by custard apple polysaccharide extract on tyrosinase monophenolase are shown in Table 9.

[0086] Table 9

[0087] Depend on Figure 3 It can be seen that when the concentration of custard apple polysaccharide extract is between 5 and 20 mg / mL, the inhibition rate of tyrosinase increases with the increase of the extract concentration, showing a dose-dependent inhibition.

[0088] Depend on Figure 4 It can be seen that as the concentration of custard apple polysaccharide extract increases, the initial velocity decreases and the inhibitory effect on monophenolase increases. This indicates that custard apple polysaccharide extract and substrate tyrosine compete for the inhibition of tyrosinase, and its inhibitory effect on tyrosinase monophenolase is a reversible effect.

[0089] according to Figure 5 As shown in the Lineweaver-Burk double reciprocal plot and Table 9, the slope and intercept of the line change with the increase of the concentration of the custard apple polysaccharide extract on tyrosinase monophenolase. The Vmax decreases and Km increases in the Michaelis-Menten kinetic equation, indicating that the inhibitory effect of the custard apple polysaccharide extract on tyrosinase is a mixed inhibition type, that is, it can bind not only to the free enzyme (E) but also to the enzyme-substrate complex (ES).

[0090] In summary, the custard apple polysaccharide extract prepared by the method of the present invention has excellent whitening properties.

[0091] The application of the custard apple polysaccharide extract obtained in Example 10 in a binary spray will be described in detail below through specific examples 11 to 15. The atomization improvers in Examples 11 to 15, by weight, comprise 45 parts polydimethylsiloxane alcohol, 50 parts water, 0.3 parts tetrasodium glutamate diacetate, 0.3 parts 1,3-propanediol, 3 parts polysorbate-20, 0.5 parts capryloyl hydroxamic acid, and 0.6 parts glyceryl caprylate.

[0092] Example 11 This embodiment provides a binary spray, comprising, by weight, 1 part custard apple polysaccharide extract, 2 parts 2,3-butanediol, 0.5 parts 1,2-hexanediol, 0.5 parts p-hydroxyacetophenone, 0.5 parts atomization improver, 0.05 parts disodium ethylenediaminetetraacetate, and 95.45 parts water.

[0093] This embodiment provides a method for preparing a binary spray, the steps of which include: (1) Mix the prescribed amounts of p-hydroxyacetophenone, 1,2-hexanediol and 2,3-butanediol, then heat to 65°C and stir until dissolved to obtain the C phase. (2) Add the prescribed amount of water and disodium ethylenediaminetetraacetate to the emulsifying pot, heat to 82°C, and stir until dissolved to obtain phase A; (3) Cool phase A to 45°C, then add the amount of atomization improver specified in the formula and stir until homogeneous; (4) Add the C phase to the mixture obtained after step (3) and stir until homogeneous; (5) Cool the mixture obtained in step (4) to below 45°C and then add the formula amount of custard apple polysaccharide extract, stir evenly to obtain the finished product liquid; (6) The obtained finished liquid is filled into a spray container and nitrogen is introduced to remove oxygen. Then the container is sealed and packaged to obtain a binary spray agent.

[0094] Example 12 This embodiment provides a binary spray, which, by weight, comprises 3 parts of custard apple polysaccharide extract, 2 parts of 2,3-butanediol, 0.5 parts of 1,2-hexanediol, 0.5 parts of p-hydroxyacetophenone, 0.5 parts of atomization improver, 0.05 parts of disodium ethylenediaminetetraacetate, and 93.45 parts of water.

[0095] The preparation method of the binary spray in this embodiment is exactly the same as that in Example 11.

[0096] Example 13 This embodiment provides a binary spray, comprising, by weight, 5 parts of custard apple polysaccharide extract, 2 parts of 2,3-butanediol, 0.5 parts of 1,2-hexanediol, 0.5 parts of p-hydroxyacetophenone, 0.5 parts of atomization improver, 0.05 parts of disodium ethylenediaminetetraacetate, and 91.45 parts of water.

[0097] The preparation method of the binary spray in this embodiment is exactly the same as that in Example 11.

[0098] Example 14 This embodiment provides a binary spray, comprising, by weight, 10 parts of custard apple polysaccharide extract, 2 parts of 2,3-butanediol, 0.5 parts of 1,2-hexanediol, 0.5 parts of p-hydroxyacetophenone, 0.5 parts of atomization improver, 0.05 parts of disodium ethylenediaminetetraacetate, and 86.45 parts of water.

[0099] The preparation method of the binary spray in this embodiment is exactly the same as that in Example 11.

[0100] Example 15 This embodiment provides a binary spray, comprising, by weight, 5 parts of custard apple polysaccharide extract, 2 parts of 2,3-butanediol, 0.5 parts of 1,2-hexanediol, 0.5 parts of p-hydroxyacetophenone, 0.1 parts of atomization improver, 0.05 parts of disodium ethylenediaminetetraacetate, and 91.85 parts of water.

[0101] The preparation method of the binary spray in this embodiment is exactly the same as that in Example 11.

[0102] The binary sprays of Examples 11 to 15 were subjected to patch testing, stability testing, moisturizing efficacy testing, soothing efficacy testing, firming efficacy testing, whitening efficacy testing, and atomized particle size testing according to the following testing methods.

[0103] Patch test: Test substances: binary sprays from Examples 11 to 15; Participants: 30 in total (male and female); Test method: Using qualified patch testing equipment, the test substance (approximately 0.020 mL to 0.025 mL) was placed in the patch testing device using the closed patch test method. The device was then applied to the back of the subject with hypoallergenic adhesive tape. The test substance was removed after 24 hours. Skin reactions were observed at 0.5, 24, and 48 hours after removal. The results were recorded according to the skin reaction grading standards in the "Cosmetic Safety Technical Specifications" (2015 edition).

[0104] The results of the patch test are shown in Table 10.

[0105] Stability test: Test substances: binary sprays from Examples 11 to 15; Test method: The test material was stored upright and upside down at -5℃, 25℃ and 50℃ respectively. It was taken out on the 7th, 15th, 30th, 60th and 90th days respectively to observe and record its internal pressure, weight loss, spray rate, spray effect and compatibility with packaging materials.

[0106] The stability test results are shown in Table 10.

[0107] Moisturizing efficacy test: Test substances: binary sprays from Examples 11 to 15; Testing basis: Laboratory method (LC-WI-HZ-180 Moisturizing efficacy test experimental guide) Test Method: This method is an in vitro method. Relevant literature shows that the trend of in vivo methods is somewhat correlated with the results of in vitro methods. Through experimental design, the test sample is compared with the test results of a positive control sample with moisturizing ability. If the moisture change rate Δm curve of the test sample is smaller than (better than) the moisture change rate Δm curve of the positive control, then the test sample can be considered to have a certain moisturizing ability. Test environment temperature: 15℃~30℃, ambient humidity: 20%~75%; Test steps: Weigh the sample using an electronic balance → Pre-treat the test sample; Select a reagent with known hygroscopic ability as a positive control (the positive control reagent in this test is glycerol) → Place the sample and positive control in a constant temperature and humidity chamber under certain temperature and humidity conditions → Weigh the sample periodically using an electronic balance → Record the data, calculate the moisture change rate Δm, and create a curve.

[0108] The results of the moisturizing efficacy test are shown in Table 11.

[0109] Soothing efficacy test: Test substances: binary sprays from Examples 11 to 15; Detection basis: Laboratory method (LC-WI-HZ-185 Hyaluronidase Inhibition Test Operating Instructions) Test Method: This method is an in vitro method, applicable to cosmetics claiming soothing effects. Relevant literature shows a certain correlation between in vivo and in vitro methods. Through experimental design, the inhibition rate of the test sample on hyaluronidase is determined by measuring the N-acetylglucosamine content in the reaction system. If the inhibition rate of the test sample is better than (higher than) that of the blank control group and the statistical difference P value < 0.05, the test sample can be considered to have a certain soothing effect. Test environment temperature: 15℃~30℃, ambient humidity: 20%~75%; Test steps: Weigh the sample using an electronic balance → Pre-treat the test sample, prepare sample tubes with different dilutions, and simultaneously prepare parallel blank groups and positive control groups → Add the reactants and mix well, then place in a constant temperature water bath at 37°C for 40 minutes → Add pH adjuster, then in a boiling water bath for 15 minutes, followed by an ice bath for 10 minutes, and finally cool to room temperature → Measure the OD value at a wavelength of 530nm using an enzyme-linked immunosorbent assay (ELISA) analyzer, and calculate the hyaluronidase activity inhibition rate and statistical difference P-value.

[0110] The results of the soothing efficacy test are shown in Table 11.

[0111] Firming efficacy test: Test substances: binary sprays from Examples 11 to 15; Test basis: Laboratory method (LC-WI-HZ-176 Elastase Inhibition Assay Operating Instructions) Test Method: This method is an in vitro method, applicable to cosmetics claiming firming effects. Relevant literature shows a certain correlation between the trends of in vivo methods and the results of in vitro methods. Through experimental design, the results of elastase inhibition rate tests were compared between the test sample and the blank control group. If the inhibition rate of the test sample is better than (higher than) that of the blank control group and the statistical difference P value < 0.05, the test sample can be considered to have a certain firming effect. Test environment temperature: 15℃~30℃, ambient humidity: 20%~75%; Test steps: Weigh the sample using an electronic balance → Pre-treat the test sample to prepare a mixed solution → Incubate at 25°C for 15 minutes in an incubator; then add a certain amount of elastase using a pipette and continue incubation for 15 minutes → Measure the elastase inhibition rate (OD value) at a wavelength of 410 nm using an enzyme-linked immunosorbent assay (ELISA) analyzer → Record the data and calculate the elastase inhibition rate and statistical difference (P-value).

[0112] The results of the firming efficacy test are shown in Table 11.

[0113] Whitening efficacy test: Test substances: binary sprays from Examples 11 to 15; Testing basis: Standard Operating Procedures for Evaluating the Whitening Efficacy of Zebrafish Test Method: Zebrafish are transparent throughout their bodies during early development. Melanin begins to grow from the retinal epithelium at 24 hours of embryonic development. Pigment cells originate from a group of cells differentiated from the dorsal ectoderm—neural crest cells—which then proliferate, migrate, and differentiate into melanocytes. Intervention during melanin formation can inhibit melanin production. The skin whiteness of the zebrafish is used to evaluate whether the sample has a whitening effect. Test steps: 1. Randomly select zebrafish into a 6-well plate, with 15 fish per well.

[0114] 2. The sample was administered in water, and a normal control group was set up at the same time. The volume of each well was 3 mL.

[0115] 3. Incubate at 28℃ in the dark for 45 hours.

[0116] 4. Ten zebrafish were randomly selected from each experimental group and photographed under a dissecting microscope. Advanced image processing software was used to analyze and collect data to analyze the melanin signal intensity (S) in the zebrafish head. The whitening effect of the sample is calculated using a formula to determine whether it has a whitening effect.

[0117]

[0118] Judgment criteria: Statistical analysis p<0.05, efficacy value ≥20%, judged as effective.

[0119] The whitening efficacy test results are shown in Table 12; among them, the zebrafish in Example 11 showed the following effect on melanin inhibition: Figure 6 As shown; the histogram of melanin signal intensity in the head of zebrafish in Example 11 is compared with that of the normal control group. Figure 7 As shown, ***p<0.001; among which, the zebrafish in Example 13 showed the following melanin-inhibiting effect. Figure 8 As shown; the histogram of melanin signal intensity in the head of zebrafish in Example 13 is compared with that of the normal control group. Figure 9 As shown, ***p<0.001; Atomized particle size test: Test substances: binary sprays from Examples 11 to 15; Test method: Place the test sample upright at a distance of 30cm from the test light source of the P180-C laser particle size analyzer, press and spray out, and record the atomized particle size D50 value respectively.

[0120] The atomized particle size test results are shown in Table 13.

[0121] Table 10

[0122] Table 11

[0123] Table 12

[0124] Table 13

[0125] In summary, the custard apple polysaccharide extract of the present invention exhibits multiple advantages in cosmetic applications: First, it has low allergenicity, good stability, and excellent spray performance, making it convenient for practical application; second, it is rich in active ingredients such as amino acids, peptides, and polysaccharides, which can independently exert multiple skin care effects, including deep moisturizing, soothing and repairing, firming the skin, and inhibiting melanin production to achieve a whitening effect, without the need to add other functional ingredients to meet various skin care needs.

[0126] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A preparation method of annona polysaccharide extract solution, characterized by the steps of The preparation method comprises the following steps: S1, selecting ripe annona squamosa from Hainan, washing, peeling, removing seeds, and juicing to obtain annona squamosa fruit pulp; S2, mixing the annona squamosa fruit pulp with water, and then ultrasonically treating at 180-220 W for 15-25 min, and then sequentially performing first extraction and suction filtration to obtain first filtrate and first filter residue; S3, mixing the first filter residue with water, and then sequentially performing second extraction and suction filtration to obtain second filtrate and second filter residue; S4, mixing the second filter residue with water, and then sequentially performing third extraction and suction filtration to obtain third filtrate; S5, mixing the first filtrate, the second filtrate, and the third filtrate, and then concentrating to obtain annona squamosa polysaccharide extract. The conditions of the first extraction, the second extraction, and the third extraction are each independently constant temperature extraction at 80-100 ℃ for 2.5-3.5 h.

2. The preparation method of the sugar apple polysaccharide extract according to claim 1, characterized in that, The volume ratio of the annona squamosa fruit pulp to water in step S2 is 1:10-20.

3. The preparation method of the sugar apple polysaccharide extract according to claim 1, characterized in that, Step S2 comprises ultrasonically treating the mixture of the annona squamosa fruit pulp and water at 200 W for 20 min.

4. The preparation method of the annona polysaccharide extract according to claim 1, characterized in that, The conditions of the first extraction, the second extraction, and the third extraction are each constant temperature extraction at 100 ℃ for 3 h.

5. A cherimoya polysaccharide extract, characterized in that, The annona squamosa polysaccharide extract is prepared by the preparation method of any one of claims 1-4.

6. The annona squamosa polysaccharide extract prepared by the preparation method of any one of claims 1-4 or the annona squamosa polysaccharide extract of claim 5 is applied in cosmetics.

7. Use according to claim 6, wherein The dosage form of the cosmetic is selected from the group consisting of paste, ointment, spray, gel, liniment, coating agent, film coating agent, patch, plaster, and film.

8. A binary spray, characterized in that, The cosmetic comprises, in mass fraction, 86-95 parts of water, 0.03-0.06 parts of chelating agent, 0.3-0.6 parts of atomization improver, 0.3-0.6 parts of preservative, 2-3 parts of humectant, and 1-10 parts of D phase substance, wherein the D phase substance is the annona squamosa polysaccharide extract prepared by the preparation method of any one of claims 1-4 or the annona squamosa polysaccharide extract of claim 5.

9. The binary aerosol of claim 8 wherein, The chelating agent is selected from disodium ethylenediaminetetraacetate; and the preservative is selected from p-hydroxyacetophenone.

10. The binary aerosol of claim 8 wherein, The humectant is selected from at least one of 1,2-hexanediol and 2,3-butanediol.

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