Freeze-drying extraction method and application of moringa oleifera seed polysaccharide

Abstract

The application discloses a freeze-drying extraction method of moringa oleifera seed polysaccharide, which comprises the following steps: (1) defatting the shelled and crushed moringa oleifera seed kernel to obtain defatted moringa oleifera seed meal; (2) preparing a natural deep eutectic solvent by using betaine and 1,3-butanediol; (3) moringa oleifera seed polysaccharide extraction: preparing a DESs solution, extracting polysaccharide from the moringa oleifera seed meal to obtain a moringa oleifera seed meal mixed homogenate, performing fractional filtration on the moringa oleifera seed meal mixed homogenate by using a filtering device to obtain a clear moringa oleifera seed polysaccharide extraction liquid, and obtaining the moringa oleifera seed polysaccharide by performing twice pressure reduction, concentration, drying and washing filtration; and (4) freeze-drying of the moringa oleifera seed polysaccharide. The application combines the natural deep eutectic solvent extraction technology with the freeze-drying process, innovatively optimizes the extraction process of the moringa oleifera seed meal raw material, and cooperates with the corresponding filtering device to improve the extraction and separation efficiency of the moringa oleifera seed polysaccharide, promotes the deep development of the moringa oleifera seed industry, and also provides a new idea for cosmetic raw materials.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of cosmetics, in particular to a filtering device for freeze-drying moringa seed polysaccharide extraction, an extraction method and application. BACKGROUND

[0002] Moringa originated in India, and moringa seeds, as the fruit of moringa, contain rich plant oils and various active substances, and have good edible and medicinal values. At present, the extraction methods of moringa seed oil commonly used are solvent extraction, enzymatic extraction, ultrasonic-assisted extraction, etc., which have low extraction rates and the risk of solvent residue.

[0003] Moringa seed meal, as a by-product after oil extraction of moringa seed oil, has been shown to contain rich proteins, polysaccharides and polyphenols. Research data has proved that moringa seed polysaccharides have antioxidant, antibacterial, hypoglycemic and immune-regulating effects, and as natural plant-derived functional ingredients, they have potential development prospects in the field of cosmetics. The current extraction method of plant polysaccharides is mostly water extraction, which is relatively single, has a long extraction period and low efficiency, resulting in an increase in extraction cost and being not conducive to industrial scale production.

[0004] At present, moringa seed meal is mostly used as animal feed, which has relatively low economic value and is not fully utilized. SUMMARY

[0005] The present application solves the technical problem of providing a filtering device for freeze-drying moringa seed polysaccharide extraction, an extraction method and application.

[0006] The present application extracts freeze-dried moringa seed polysaccharides from moringa seed meal by using natural eutectic solvent, and applies them to cosmetic formulations. Through in vitro efficacy tests, cell tests and human efficacy tests, the safety and effectiveness of moringa seed polysaccharides are verified, and the by-products of moringa seed oil extraction are successfully utilized and the extraction process is optimized.

[0007] The filtering device for freeze-drying moringa seed polysaccharide extraction has 5 layers of filtering structures fixed in it from top to bottom: a first filter screen, a second filter screen, a macroporous resin, an activated carbon filter layer and a filter paper and ceramic filter sheet layer.

[0008] The filtering device for freeze-drying moringa seed polysaccharide extraction, wherein the first filter screen is 80 mesh, the second filter screen is 120 mesh, the macroporous resin is D101 macroporous resin, the thickness of the activated carbon filter layer is 5 mm, and the filter paper and ceramic filter sheet layer is a 10 ppi ceramic filter sheet on which a single layer of qualitative filter paper is provided.

[0009] The filtering device for freeze-drying moringa seed polysaccharide extraction, wherein the 5-layer filtering structure is sequentially fixed on the filtering layer frame from top to bottom, a liquid collecting hopper is arranged at the lower part of the filtering layer frame and is communicated with the filter paper and the ceramic filter sheet layer, the filtering layer frame and the liquid collecting hopper are fixed in the interior of the filtering cylinder wall, a liquid collecting funnel, a liquid valve and a pressure adjusting pipeline are arranged at the top, bottom and side of the filtering cylinder wall respectively; the liquid collecting funnel is communicated with the interior space of the filtering layer frame, the liquid valve is communicated with the bottom of the liquid collecting hopper, the upper part and the lower part of the pressure adjusting pipeline are respectively communicated with the upper space of the first filter screen and the upper space of the liquid collecting hopper, and a pressure adjusting valve is arranged on the pressure adjusting pipeline.

[0010] The material of the liquid collecting funnel, the filtering cylinder wall, the filtering layer frame, the first filter screen, the second filter screen, the liquid collecting hopper, the liquid valve, the pressure adjusting pipeline and the pressure adjusting valve is 304 stainless steel.

[0011] A freeze-drying moringa seed polysaccharide extraction method, comprising the following steps:

[0012] (1) The shelled and crushed moringa seed kernels are subjected to defatting treatment by using a supercritical carbon dioxide fluid extraction method to obtain defatted moringa seed meal;

[0013] (2) A natural low eutectic solvent, namely DESs, is prepared by using betaine and 1,3-butanediol;

[0014] (3) Moringa seed polysaccharide extraction: The DESs solution is prepared, the polysaccharide extraction is carried out on the moringa seed meal, the moringa seed meal mixed homogenate is obtained, the moringa seed meal mixed homogenate is subjected to fractional filtration by using the filtering device, the clear moringa seed polysaccharide extraction liquid is obtained, and the moringa seed polysaccharide is obtained after two times of pressure reduction, concentration, drying and washing filtration;

[0015] (4) Freeze-drying of the moringa seed polysaccharide to obtain the final product, moringa seed polysaccharide freeze-dried powder.

[0016] The freeze-drying moringa seed polysaccharide extraction method, characterized in that the supercritical carbon dioxide fluid extraction method comprises the following steps:

[0017] The moringa seed is crushed and loaded into an extraction kettle, the carbon dioxide switch is opened, the pressure and temperature of the extraction kettle, the first separation kettle and the second separation kettle are adjusted, when the extraction pressure reaches 35 MPa and the extraction temperature reaches 40 DEG C, the pressure of the first separation kettle is 12 MPa and the separation temperature is 45 DEG C, the pressure of the second separation kettle is 5 MPa and the separation temperature is 35 DEG C, the valve is adjusted to be stable, the carbon dioxide fluid is fully contacted with the moringa seed material in the extraction kettle, the carbon dioxide fluid carrying the moringa seed oil is introduced into the first separation kettle through a pipeline to carry out the first separation of the moringa seed oil, then a small amount of the moringa seed oil which is not separated is introduced into the second separation kettle with the carbon dioxide fluid to carry out the second separation, and the separated carbon dioxide is recycled through a loop, when the preset pressure and temperature of the extraction kettle, the first separation kettle and the second separation kettle are stable for a period of time, the moringa seed oil is collected from the first separation kettle and the second separation kettle every 30 min, the overall extraction time is 3 h, the moringa seed oil is obtained by being combined after the extraction is completed, and the defatted moringa seed meal in the extraction kettle is collected.

[0018] The extraction method of the freeze-dried moringa seed polysaccharide provided by the application, wherein step (2) specifically comprises the following steps: placing betaine and 1,3-butanediol in a beaker in a proportion of 1:1 in terms of molar ratio, heating and stirring under the condition of 80 DEG C until a transparent and uniform liquid, natural deep eutectic solvent, namely DESs, is formed, and the DESs is cooled to room temperature for storage and standby.

[0019] The extraction method of the freeze-dried moringa seed polysaccharide provided by the application, wherein step (3) specifically comprises the following steps: uniformly mixing the DESs with distilled water to prepare a DESs solution with a water content of 85%, and extracting polysaccharides from the moringa seed meal under the condition of a material-liquid ratio of 1:5, an extraction temperature of 70 DEG C and an extraction time of 120 min to obtain a moringa seed meal mixed homogenate.

[0020] The moringa seed meal mixed homogenate is subjected to fractional filtration through a filtering device to obtain a clear moringa seed polysaccharide extraction liquid, the moringa seed polysaccharide extraction liquid is concentrated and dried under reduced pressure to 1 / 3 of the original volume under the condition of a pressure of-0.1 MPa and a temperature of 50-60 DEG C, then anhydrous ethanol is added in a proportion of 1:3 in terms of the volume ratio of the moringa seed polysaccharide extraction liquid to anhydrous ethanol, alcohol precipitation is allowed to stand for 12 h, supernatant is removed by high-speed centrifugation at 10,000 r / min for 10 min, the moringa seed polysaccharide is obtained by dissolving in water, single-layer qualitative filter paper filtration, concentration under reduced pressure, pressure of-0.1 MPa and temperature of 50-60 DEG C, addition of anhydrous ethanol in a proportion of 1:3 in terms of the volume ratio of the moringa seed polysaccharide extraction liquid to anhydrous ethanol, alcohol precipitation standing for 12 h and centrifugation.

[0021] The extraction method of the freeze-dried moringa seed polysaccharide, wherein step (4) specifically comprises the following steps: the obtained moringa seed polysaccharide is pre-frozen at-80 DEG C for 12h, and freeze-drying treatment is carried out under the condition of-80 DEG C and 0.37mbar for 3-4h, and finally the freeze-dried moringa seed polysaccharide powder is obtained.

[0022] The application of the freeze-dried moringa seed polysaccharide in cosmetics.

[0023] The application, wherein the cosmetics are one of the following:

[0024] (1) Moringa seed polysaccharide essence water is prepared from the following raw materials by mass:

[0025] 1,3-butanediol 2-3%, acryloyldimethyl ammonium taurate / VP copolymer 0.1-0.5%, carbomer 0.05-0.1%, betaine 0.2-0.5%, p-hydroxyacetophenone 0.3-0.5%, moringa seed polysaccharide 0.1-0.15%, 1,2-hexanediol 0.3-0.5%, triethanolamine 0.1-0.3%, and the balance is water;

[0026] (3) Moringa seed polysaccharide essence milk is prepared from the following raw materials by mass:

[0027] Acrylic acid (ester) / C10-30 alkanol acrylate cross-linked polymer 0.1-0.15%, xanthan gum 0.05-0.1%, 1,3 butanediol 3-5%, dipropylene glycol 3-5%, glycerol 2-3%, sodium hyaluronate 0.01-0.02%, freeze-dried moringa seed polysaccharide powder 0.1-0.12%, dimethicone 2-5%, squalane 1-2%, 9% KOH 0.3-0.5%, phenoxyethanol 0.5-0.8%, and the balance is water;

[0028] (2) Moringa seed polysaccharide essence cream is prepared from the following raw materials by mass:

[0029] Sodium hyaluronate 0.01-0.05%, freeze-dried moringa seed polysaccharide powder 0.05-0.1%, xanthan gum 0.1-0.3%, EDTA-Na 0.05-0.1%, 1,3-butanediol 2-3%, glycerol 2-5%, cetyl stearyl alcohol 0.5-1%, squalane 2-5%, squalane 1-3%, caprylic acid / capric acid triglyceride 2-3%, p-hydroxyacetophenone 0.5-0.8%, 1,2-hexanediol 0.3-0.6%, sebamed 305 (polyacrylamide, C13-14 isoparaffin, laureth-7) 0.8-1%, and the balance is water.

[0030] The application is different from the prior art, the technical problems and the processing method (beneficial effects):

[0031] First, the utilization rate of moringa seed is relatively low, and the moringa seed meal after oil extraction is directly used as animal feed, and the protein, polysaccharide, polyphenol and other active substances in the moringa seed are not fully utilized, and the income of farmers is relatively low. The present application uses natural eutectic solvent to extract moringa seed polysaccharide from moringa seed meal, improves the application efficiency of moringa seed, and improves the deep processing degree of related industry. The extraction solvent used is a raw material included in the "List of Raw Materials for Cosmetics", which maximizes the safety of the raw material, avoids contact with toxic and harmful reagents, and ensures the safety of consumers.

[0032] Second, a corresponding filtering device is designed for the process of extracting moringa seed polysaccharide with natural eutectic solvent. The moringa seed meal after extraction has high viscosity, which greatly reduces the filtration efficiency of the moringa seed polysaccharide extract, wastes manpower and material resources, and further increases the production cost. The filtering device designed in the present application is classified and filtered, which improves the filtering effect to a certain extent and saves time and cost.

[0033] Third, the moringa polysaccharide is freeze-dried to remove water in the moringa seed polysaccharide in a low-temperature environment, and to extend the storage time of the moringa seed polysaccharide and save storage area without preservatives. Fourth, the present application verifies the feasibility and stability of the application of moringa seed polysaccharide in cosmetic formulations based on the extraction of moringa seed polysaccharide, which provides support for the application of natural eutectic solvent extracted moringa seed polysaccharide in cosmetics.

[0034] Natural eutectic solvent, as a substitute for traditional solvent, is a eutectic mixture composed of (HBA) hydrogen bond acceptor or (HBD) hydrogen bond donor in natural substances, which is a new type of solvent. Compared with traditional extraction solvents, natural eutectic solvent has high solubility and extraction effect, relatively low melting point, easy to obtain materials, simple synthesis method, low toxicity, high environmental protection, good recyclability and good biodegradability. In summary, the natural eutectic solvent composed of natural compounds such as sugar, organic acid and polyol in the "List of Raw Materials for Cosmetics" is more suitable for the extraction of cosmetic raw materials with high safety and efficacy requirements, which effectively avoids the contact with toxic or non-biodegradable organic solvents.

[0035] This invention utilizes a natural eutectic solvent to extract moringa seed polysaccharides from the residue remaining after supercritical fluid extraction of oil. Betaine is used as the hydrogen bond acceptor, and propylene glycol as the hydrogen bond donor, in a 1:1 molar ratio. The extracted moringa seed polysaccharides are then concentrated under reduced pressure, precipitated with alcohol, washed repeatedly with anhydrous ethanol, and freeze-dried at 0.37 bar and -80°C. The moringa seed polysaccharides are then applied in cosmetic formulations at specific ratios, and their moisturizing effects and safety are verified through weighing moisturizing tests, cell experiments, and human efficacy trials, resulting in products with moisturizing properties and providing support for their application in cosmetics. Moringa seed polysaccharides, as a safe, effective, and green-process extracted plant-derived raw material, can be widely used in cosmetic products to achieve good moisturizing effects. Extracting plant polysaccharides from the byproducts of moringa seed oil is of profound significance for developing moringa seed resources, increasing the economic added value of related industries, promoting farmers' income, and expanding the application range of moringa seed raw materials.

[0036] Currently, moringa seed meal is mostly processed as animal feed, resulting in relatively low economic value and insufficient resource utilization. This invention combines natural eutectic solvent extraction technology with freeze-drying to innovate and optimize the extraction process of moringa seed meal raw materials. Furthermore, it incorporates a corresponding filtration device to improve the extraction and separation efficiency of moringa seed polysaccharides. This not only promotes the in-depth development of the moringa seed industry but also provides new ideas for cosmetic raw materials.

[0037] The following description, in conjunction with the accompanying drawings, further illustrates the extraction apparatus, extraction method, and applications of the freeze-dried Moringa seed polysaccharide of the present invention. Attached Figure Description

[0038] Figure 1 This is a longitudinal cross-sectional structural diagram of the filtration device for extracting polysaccharides from freeze-dried Moringa seeds according to the present invention.

[0039] Figure 2 This is a flowchart of the method for extracting freeze-dried Moringa seed polysaccharides according to the present invention. Detailed Implementation

[0040] 1. Filtration device for extracting polysaccharides from freeze-dried Moringa seeds

[0041] like Figure 1 As shown, the filtration device for extracting polysaccharides from freeze-dried Moringa seeds has five layers of filtration structure fixed inside from top to bottom: a first filter screen 4, a second filter screen 5, macroporous resin 6, an activated carbon filter layer 7, and a filter paper and ceramic filter sheet layer 8.

[0042] The first filter screen 4 is 80 mesh, the second filter screen 5 is 120 mesh, the macroporous resin 6 is D101 macroporous resin, the activated carbon filter layer 7 is 5 mm thick, and the filter paper and ceramic filter sheet layer 8 is a 10 ppi ceramic filter sheet with a single layer of qualitative filter paper placed on it.

[0043] A 5-layer filter structure is fixed on the filter layer frame 3 from top to bottom. A liquid collection hopper 9 is provided at the bottom of the filter layer frame 3 and is connected to the filter paper and ceramic filter sheet layer 8. The filter layer frame 3 and the liquid collection hopper 9 are fixed inside the filter cylinder wall 2. A liquid collection funnel 1, a liquid valve 10 and a pressure regulating pipe 11 are respectively provided at the top, bottom and side of the filter cylinder wall 2. The liquid collection funnel 1 is connected to the internal space of the filter layer frame 3, the liquid valve 10 is connected to the bottom of the liquid collection hopper 9, and the upper and lower parts of the pressure regulating pipe 11 are connected to the upper space of the first filter screen 4 and the upper space of the liquid collection hopper 9, respectively. A pressure regulating valve 12 is provided on the pressure regulating pipe 11.

[0044] The liquid collecting funnel 1, filter cylinder wall 2, filter layer rack 3, first filter screen 4, second filter screen 5, liquid collecting hopper 9, liquid valve 10, pressure regulating pipe 11 and pressure regulating valve 12 are all made of 304 stainless steel.

[0045] II. Methods for Extracting Polysaccharides from Freeze-dried Moringa Seeds

[0046] like Figure 2 As shown, the method for extracting polysaccharides from freeze-dried Moringa seeds includes the following steps:

[0047] 1. Defatted Moringa seeds

[0048] The shelled and crushed Moringa seeds were defatted using supercritical carbon dioxide fluid extraction to obtain defatted Moringa seed meal.

[0049] The supercritical carbon dioxide fluid extraction method includes the following steps:

[0050] The moringa seed is crushed and loaded into the extraction kettle of a HA221-50-10-C supercritical carbon dioxide extraction device (Nantong Huaan Supercritical Extraction Co., Ltd., one extraction kettle and two separation kettles, i.e., a first separation kettle and a second separation kettle, of the extraction device are used in the present application), a carbon dioxide switch is opened, the pressure and temperature of the extraction kettle, the first separation kettle and the second separation kettle are adjusted, when the extraction pressure is 35 MPa and the extraction temperature is 40℃, the pressure of the first separation kettle is 12 MPa and the separation temperature is 45℃, the pressure of the second separation kettle is 5 MPa and the separation temperature is 35℃, the valve is adjusted to be stable, the carbon dioxide fluid is fully contacted with the moringa seed material in the extraction kettle, the carbon dioxide fluid carrying the moringa seed oil is introduced into the first separation kettle through a pipeline to perform the first separation of the moringa seed oil, and then a small amount of the moringa seed oil not separated is introduced into the second separation kettle with the carbon dioxide fluid to perform the second separation, and the separated carbon dioxide is recycled through a loop; when the preset pressure and temperature of the extraction kettle, the first separation kettle and the second separation kettle are stable for a period of time, the moringa seed oil is collected from the first separation kettle and the second separation kettle every 30 min; the overall extraction time is 3 h, the moringa seed oil is obtained after the extraction is completed, and the defatted moringa seed meal in the extraction kettle is collected.

[0051] 2. Preparation of natural deep eutectic solvent

[0052] The betaine and 1,3-butanediol are placed in a beaker in a proportion of 1:1 by mole ratio, heated and stirred at 80℃ until a transparent and uniform liquid is formed, and then cooled to room temperature for storage (DESs).

[0053] 3. Extraction of moringa seed polysaccharide

[0054] The DESs are mixed with distilled water to prepare a DESs solution with a water content of 85%, and the polysaccharide is extracted from the moringa seed meal according to the conditions of the material-liquid ratio (g:mL) of the moringa seed meal:DESs solution in Table 1. The mixed homogenate of the moringa seed meal is subjected to fractional filtration through the extraction and filtration device of the present application to obtain a clear moringa seed polysaccharide extract. After vacuum concentration and drying (-0.1 MPa; 50-60℃), the volume is concentrated to 1 / 3 of the original volume, anhydrous ethanol is added in a proportion of 1:3 (mL:mL) of the moringa seed polysaccharide extract:anhydrous ethanol, and alcohol precipitation is allowed to stand for 12 h. The supernatant is removed by high-speed centrifugation at 10,000 r / min for 10 min, dissolved with water, filtered through a single-layer qualitative filter paper (model: 101; filter speed: fast), vacuum concentrated (-0.1 MPa; 50-60℃) to 1 / 3 of the original volume, anhydrous ethanol is added in a proportion of 1:3 (mL:mL) of the moringa seed polysaccharide extract:anhydrous ethanol, and alcohol precipitation is allowed to stand for 12 h, and the moringa seed polysaccharide is obtained by centrifugation. The moringa seed polysaccharide is repeatedly washed with 95% anhydrous ethanol until the washing liquid is clear, and then filtered to obtain the moringa seed polysaccharide.

[0055] Table 1 Extraction conditions of moringa seed polysaccharides

[0056] Feed liquid ratio (g:mL) Extraction temperature (°C) Extraction time (min) Polysaccharide yield (%) 1:5 70 120 24.5％ 1:20 70 120 3.6％ 1:30 70 120 3.2％ 1:50 70 120 2.3％

[0057] Note: The conditions in Table 1 were optimized, and the final results showed that the highest yield of polysaccharides was obtained when the solid-liquid ratio (g:mL) of moringa seed meal to DESs solution was 1:5.

[0058] 4. Freeze-drying of moringa seed polysaccharides

[0059] The obtained moringa seed polysaccharides were pre-frozen at -80°C for 12h, and freeze-dried under the conditions of -80°C and 0.37mbar for 3-4h to obtain moringa seed polysaccharide freeze-dried powder.

[0060] III. Application of moringa seed polysaccharide freeze-dried powder

[0061] 1. Moringa seed polysaccharide essence water

[0062] (1) Formulation preparation

[0063] The raw materials are calculated based on the total mass of 100%, 1,3-butanediol 2-3%, acryloyldimethyl ammonium taurate / VP copolymer 0.1-0.5%, carbomer 0.05-0.1%, betaine 0.2-0.5%, p-hydroxyacetophenone 0.3-0.5%, moringa seed polysaccharide 0.1-0.15%, 1,2-hexanediol 0.3-0.5%, triethanolamine 0.1-0.3%, and the balance is water.

[0064] The preparation method of the moringa seed polysaccharide essence water comprises the following steps: sequentially adding the A-phase raw materials, stirring and heating to 80-85°C, and maintaining for 20min; cooling after the materials are completely dispersed and uniform; cooling to 75°C, adding the B-phase raw materials, and stirring uniformly; cooling to below 45°C, sequentially adding the C and D-phase raw materials, and stirring for 5-10min until dispersed uniformly.

[0065] Table 2 Formulation table of moringa seed polysaccharide essence water

[0066]

[0067] (2) Cytotoxicity test

[0068] The cytotoxicity of the moringa seed polysaccharide essence water was evaluated by testing its effect on the survival rate of HaCaT cells in the stratum corneum of human skin.

[0069] Table 3 Analysis of cytotoxicity test results

[0070] Concentration (% W / V) Cell survival rate (%) Standard deviation (%) Negative control 100.00 0.01 0.05 99.93 0.02 0.10 99.69 0.02 0.25 99.53 0.01 0.50 99.25 0.04 1.00 91.84 0.02 2.00 86.96 0.04

[0071] Moringa oleifera seed polysaccharide essence water has no obvious cytotoxicity to HaCaT cells at a concentration of 2% or below for 24 h.

[0072] (3) Human patch test

[0073] Patch test method: qualified patch test materials are selected, and the tested substance about 0.020 g-0.025 g (solid) or 0.020 mL-0.025 mL (liquid) is placed in the patch tester by the closed patch test method. The outer low sensitization adhesive tape is applied to the forearm of the subject, and the tested substance is removed after 24 h. The skin reaction is observed at 0.5, 24, and 48 h after removal, respectively, and the results are recorded according to the skin reaction grading standard in the current effective technical specification.

[0074] Table 4 Results of human skin patch test

[0075]

[0076] Test results: the human skin patch test results show that no skin adverse reactions occur in the tested parts of 30 subjects after 0.5 h, 24 h, and 48 h of removal of the tested substance patch tester, indicating that the Moringa oleifera seed polysaccharide essence water has no irritation to the human body surface.

[0077] (4) Moisturizing effect efficacy evaluation (physical weighing test)

[0078] Evaluation method: under constant temperature and humidity conditions, the mass difference before and after the sample is placed is weighed, the loss of sample amount is calculated, and the moisturizing effect is calculated.

[0079] Table 5 Physical moisturizing weighing test

[0080] Test time point (h) 16 24 40 48 64 72 Moisture retention rate (%) 98.76 98.16 96.15 95.34 93.43 92.16

[0081] Test results: the moisturizing rate of Moringa oleifera seed polysaccharide essence water is more than 90% at 16 h, 24 h, 40 h, 48 h, 64 h, and 72 h detection time, indicating that it has moisturizing efficacy under the physical weighing test conditions.

[0082] (5) Human efficacy test

[0083] The data information of the skin stratum corneum moisture content of 31 subjects is collected by the skin stratum corneum moisture content test probe, and the moisturizing effect of Moringa oleifera seed polysaccharide essence water is analyzed.

[0084] Table 6 Statistics table of cheek skin stratum corneum moisture content

[0085]

[0086] Note: improvement rate (%) after using the product = (test value after using the product N days-basic value) / basic value x 100%

[0087] Test results: After using the Moringa seed polysaccharide essence water for 20 minutes, the water content in the stratum corneum of the skin increased by 16.95% (p<0.05) compared with before use, indicating that the Moringa seed polysaccharide essence water has a moisturizing effect.

[0088] (6) Consumer use test

[0089] Test method: The subjects should visit the laboratory after using the Moringa seed polysaccharide essence water for 14 days (D14) and 28 days (D28), clean the test site with a cleaning product, and sit in a constant temperature and humidity chamber after drying. During the sitting period, fill in the questionnaire according to the subjective feeling after using the sample.

[0090] Table 7 Results of part of the sensory evaluation of the questionnaire survey of the subjects (n=30)

[0091]

[0092] Scoring criteria: 5 - very agree; 4 - agree; 3 - general; 2 - disagree; 1 - very disagree. Scores > 3 are considered positive options.

[0093] Consumer sensory evaluation test results: After using the Moringa seed polysaccharide essence water, 93.10% of the subjects thought that the product smell was appropriate; 96.67% of the subjects thought that the product was absorbed quickly when used; 96.67% of the subjects thought that the product had a good skin feel; 93.33% of the subjects thought that the product was mild and non-irritating when used; 96.67% of the subjects thought that the product did not stick to the skin after use.

[0094] Table 8 Results of part of the efficacy evaluation of the questionnaire survey of the subjects (n=30)

[0095]

[0096] Scoring criteria: 5 - very agree / satisfied; 4 - agree / satisfied; 3 - general; 2 - disagree / dissatisfied; 1 - very disagree / very dissatisfied. Scores > 3 are considered positive options.

[0097] Consumer efficacy evaluation test results: After using the Moringa oleifera seed polysaccharide essence water for 14 days (D14) and 28 days (D28), the subjects answered the questionnaire for efficacy evaluation, and 63.33%, 79.31% of the subjects respectively believed that

skin sensitivity and redness were improved

skin barrier ability was improved

skin elasticity was improved

skin dry lines and fine lines were improved

skin color was improved

skin water retention was improved

skin felt moisturized

overall, the product was satisfactory

[0098] 2. Moringa oleifera seed polysaccharide essence cream

[0099] (1) Formulation preparation

[0100] The raw materials are calculated based on a total mass of 100%, and the acrylate / C10-30 alkyl acrylate crosspolymer is 0.1-0.15%, the xanthan gum is 0.05-0.1%, the 1,3-butanediol is 3-5%, the dipropylene glycol is 3-5%, the glycerin is 2-3%, the sodium hyaluronate is 0.01-0.02%, the Moringa oleifera seed polysaccharide lyophilized powder is 0.1-0.12%, the dimethicone is 2-5%, the squalane is 1-2%, the 9% KOH is 0.3-0.5%, the phenoxyethanol is 0.5-0.8%, and the balance is water.

[0101] The preparation method of the Moringa oleifera seed polysaccharide essence cream includes the following steps: uniformly dispersing A phase in water; pre-dispersing B phase in water, adding A phase, and uniformly stirring at room temperature; heating A+B phase and C phase to 75℃ respectively; adding C phase to A+B phase while stirring, homogenizing with a homogenizer at 4500 cp for 10 seconds; reducing to room temperature, adding KOH, and adjusting the pH to about 6.5; adding phenoxyethanol, and discharging.

[0102] Table 9. Moringa oleifera seed polysaccharide essence cream formula

[0103]

[0104] (2) Cytotoxicity test

[0105] The cytotoxicity of the Moringa oleifera seed polysaccharide essence cream was evaluated by testing its effect on the survival rate of HaCaT cells in the human skin stratum corneum.

[0106] Table 10. Analysis of cytotoxicity test results

[0107] Concentration (% W / V) Cell survival rate (%) Standard deviation (%) Negative control 100.00 0.02 0.05 97.36 0.01 0.10 91.23 0.04 0.25 90.44 0.01 0.50 89.41 0.03 1.00 85.34 0.02

[0108] Moringa oleifera seed polysaccharide essence cream has no obvious cytotoxicity to HaCaT cells at a concentration of 1.00% for 24 h.

[0109] (3) Human patch test

[0110] Patch test method: qualified patch test materials are selected, and the tested substance about 0.020 g-0.025 g (solid) or 0.020 mL-0.025 mL (liquid) is placed in the patch test device by closed patch test method. The outer low sensitization adhesive tape is applied to the forearm of the subject, and the tested substance is removed after 24 hours. The skin reaction is observed at 0.5, 24 and 48 hours after removal, respectively, and the results are recorded according to the skin reaction grading standard in the current effective technical specification.

[0111] Table 11 Results of human skin patch test

[0112]

[0113] Test results: the human skin patch test results show that no skin adverse reactions occur at the tested sites of 30 subjects after 0.5 h, 24 h and 48 h of removal of the patch test device of the tested substance, indicating that the Moringa oleifera seed polysaccharide essence cream has no irritation to the human body surface.

[0114] (4) Moisturizing effect efficacy evaluation (physical weighing test)

[0115] Evaluation method: under constant temperature and humidity conditions, the mass difference before and after the sample is placed is weighed, the loss of sample amount is calculated, and the moisturizing effect is calculated.

[0116] Table 12 Physical moisturizing weighing test

[0117] Test time point (h) 16 24 40 48 64 72 Moisture retention rate (%) 98.15 97.56 96.25 95.19 93.54 92.25

[0118] Test results: the Moringa oleifera seed polysaccharide essence cream has a moisturizing rate of more than 90% at 16 h, 24 h, 40 h, 48 h, 64 h and 72 h detection time, indicating that it has moisturizing efficacy under the test conditions of the physical weighing method.

[0119] (5) Human efficacy test

[0120] The data information of the skin stratum corneum water content of 31 subjects is collected by the skin stratum corneum water content test probe, and the moisturizing effect of the Moringa oleifera seed polysaccharide essence cream is analyzed.

[0121] Table 13 Statistics table of cheek skin stratum corneum water content

[0122]

[0123] Note: Improvement rate (%) after using the product = (test value after using the product for N days - base value) / base value x 100%

[0124] Test results: After using the Moringa seed polysaccharide essence cream for 20 minutes, the water content in the stratum corneum of the skin increased by 18.47% (p < 0.05) compared with before use, indicating that the Moringa seed polysaccharide essence cream has a moisturizing effect.

[0125] (6) Consumer use test

[0126] Test method: The subjects should visit the laboratory after using the Moringa seed polysaccharide essence cream for 14 days (D14) and 28 days (D28), clean the test site with a cleaning product, and sit in a constant temperature and humidity chamber after drying. During the sitting period, fill in the questionnaire according to the subjective feeling after using the sample.

[0127] Table 14 Statistical table of part of the results of the sensory evaluation of the questionnaire survey of the subjects (n = 30)

[0128]

[0129] Scoring criteria: 5 - very agree; 4 - agree; 3 - general; 2 - disagree; 1 - very disagree. Scores > 3 are considered positive options.

[0130] Consumer sensory evaluation test results: After using the Moringa seed polysaccharide essence cream, 96.67% of the subjects thought that the product smell was appropriate; 93.55% of the subjects thought that the product was quickly absorbed when used; 93.33% of the subjects thought that the product had a good skin feel; 96.67% of the subjects thought that the product was mild and non-irritating when used; 93.33% of the subjects thought that the product did not stick to the skin after use.

[0131] Table 15 Statistical results of the efficacy evaluation part of the questionnaire survey of the subjects (n = 30)

[0132]

[0133] Scoring criteria: 5 - very agree / satisfied; 4 - agree / satisfied; 3 - general; 2 - disagree / dissatisfied; 1 - very disagree / very dissatisfied. Scores > 3 are considered positive options.

[0134] The results of the efficacy evaluation test by consumers: After using the Moringa oleifera seed polysaccharide essence cream for 14 days (D14) and 28 days (D28), respectively, 66.67%, 83.33% of the subjects believed that the skin sensitivity and redness were improved; 73.33%, 93.10% of the subjects believed that the skin barrier ability was improved; 73.33%, 90.00% of the subjects believed that the skin elasticity was improved; 73.33%, 86.67% of the subjects believed that the skin dry and fine lines were improved; 76.67%, 86.67% of the subjects believed that the skin color was improved; 83.33%, 96.67% of the subjects believed that the skin water retention was improved; 80.00%, 93.33% of the subjects believed that the skin felt moisturized; 100.00%, 100.00% of the subjects believed that the overall use of the product was satisfactory.

[0135] 3. Moringa oleifera seed polysaccharide essence cream

[0136] (1) Formulation preparation

[0137] The raw materials are 0.01-0.05% sodium hyaluronate, 0.05-0.1% Moringa oleifera seed polysaccharide freeze-dried powder, 0.1-0.3% xanthan gum, 0.05-0.1% EDTA-Na, 2-3% 1,3-butanediol, 2-5% glycerol, 0.5-1% cetylstearyl alcohol, 2-5% squalane, 1-3% squalane, 2-3% caprylic / capric triglyceride, 0.5-0.8% p-hydroxyacetophenone, 0.3-0.6% 1,2-hexanediol, 0.8-1% sebamed 305 (polyacrylamide, C13-14 isoparaffin, laureth-7), and the balance is water, based on a total mass of 100%.

[0138] The preparation method of the Moringa oleifera seed polysaccharide essence cream comprises the following steps: uniformly mixing A phase, stirring or low-speed homogenizing to disperse uniformly, then heating to 70-80℃; mixing B phase and heating to 70-80℃, then adding B phase to A phase and homogenizing uniformly, stirring and then cooling to 40-45℃; pre-dissolving all except sebamed 305 (polyacrylamide, C13-14 isoparaffin, laureth-7) in C phase, cooling to room temperature, then adding sebamed 305 (polyacrylamide, C13-14 isoparaffin, laureth-7), mixing, adding the mixed C phase to A and B phases, homogenizing uniformly, and cooling to room temperature to discharge.

[0139] Table 16 Moringa oleifera seed polysaccharide essence cream formula

[0140]

[0141] (2) Cytotoxicity test

[0142] The cytotoxicity of the sample was evaluated by testing the effect of the moringa seed polysaccharide essence cream on the survival rate of HaCaT cells in the stratum corneum of human skin.

[0143] Table 17 Analysis of the results of the cytotoxicity test

[0144] Concentration (% W / V) Cell survival rate (%) Standard deviation (%) Negative control 100.00 0.02 0.05 96.51 0.02 0.10 90.25 0.04 0.25 89.15 0.02 0.50 85.21 0.03

[0145] The moringa seed polysaccharide essence cream had no significant cytotoxicity to HaCaT cells at a concentration of 0.5% for 24 h.

[0146] (3) Human patch test

[0147] Patch test method: qualified patch test materials were selected, and the test substance about 0.020g-0.025g (solid) or 0.020mL-0.025mL (liquid) was placed in the patch tester by the closed patch test method. The low sensitization adhesive tape was applied to the forearm of the test subject. After 24 hours, the test substance was removed, and the skin reaction was observed at 0.5, 24, and 48 hours after removal, respectively. The results were recorded according to the grading standard of skin reaction in the current effective technical specification.

[0148] Table 18 Results of human skin patch test

[0149]

[0150] Test results: The human skin patch test results showed that no skin adverse reactions occurred in the test sites of 29 subjects at 0.5h, 24h, and 48h after removal of the test substance patch tester, indicating that the moringa seed polysaccharide essence cream had no irritation to the human body surface.

[0151] (4) Evaluation of moisturizing effect (physical weighing test)

[0152] Evaluation method: under constant temperature and humidity conditions, the mass difference before and after the sample was weighed, the loss of sample amount was calculated, and the moisturizing effect was calculated.

[0153] Table 19 Physical moisturizing weighing test

[0154] Test time point (h) 16 24 40 48 64 72 Moisture retention rate (%) 98.75 97.63 96.27 95.55 93.12 91.56

[0155] Test results: The moringa seed polysaccharide essence cream had a moisturizing rate of more than 90% at 16h, 24h, 40h, 48h, 64h, and 72h detection times, indicating that it had moisturizing efficacy under the physical weighing test conditions.

[0156] (5) Human efficacy test

[0157] The moisture content of the stratum corneum of 31 subjects was collected by a skin corneum moisture content test probe, and the moisturizing effect of the moringa seed polysaccharide essence cream was analyzed.

[0158] Table 20 Statistical table of the moisture content of the stratum corneum of the cheek skin

[0159]

[0160] Note: Improvement rate (%) after using the product = (test value after using the product N days - basic value) / basic value x 100%

[0161] Test results: After using the moringa seed polysaccharide essence cream for 20 minutes, the moisture content of the stratum corneum of the skin increased by 32.12% (p<0.05), indicating that the moringa seed polysaccharide essence cream had a moisturizing effect.

[0162] (6) Consumer use test

[0163] Test method: The subjects should visit the laboratory after using the moringa seed polysaccharide essence cream for 14 days (D14) and 28 days (D28), clean the test site with a cleaning product, and sit in a constant temperature and humidity room after drying. During the sitting period, fill in the questionnaire according to the subjective feeling after using the sample.

[0164] Table 21 Statistical table of part of the results of the sensory evaluation of the questionnaire survey of the subjects (n=30)

[0165]

[0166] Scoring criteria: 5 - strongly agree; 4 - agree; 3 - general; 2 - disagree; 1 - strongly disagree. Scores > 3 are considered positive options.

[0167] Consumer sensory evaluation test results: After using the moringa seed polysaccharide essence cream, 93.33% of the subjects thought that the product smell was appropriate; 96.67% of the subjects thought that the product was quickly absorbed when used; 96.67% of the subjects thought that the product had a good skin feel; 93.33% of the subjects thought that the product was mild and non-irritating when used; 90.00% of the subjects thought that the product did not stick to the skin after use.

[0168] Table 22 Statistical results of the efficacy evaluation part of the questionnaire survey of the subjects (n=30)

[0169]

[0170] Scoring criteria: 5 - strongly agree / satisfied; 4 - agree / satisfied; 3 - general; 2 - disagree / dissatisfied; 1 - strongly disagree / very dissatisfied. Scores > 3 are considered positive options.

[0171] The results of the efficacy evaluation test: after using the Moringa oleifera seed polysaccharide essence cream for 14 days (D14) and 28 days (D28), respectively, 65.52% and 80.00% of the subjects believed that the skin sensitivity and redness were improved; 73.33% and 90.00% of the subjects believed that the skin barrier ability was improved; 73.33% and 96.67% of the subjects believed that the skin elasticity was improved; 76.67% and 83.33% of the subjects believed that the skin fine lines and wrinkles were improved; 73.33% and 90.00% of the subjects believed that the skin dullness was improved; 86.67% and 100.00% of the subjects believed that the skin water retention was improved; 83.33% and 96.67% of the subjects believed that the skin felt moisturized; and 100.00% and 100.00% of the subjects believed that the overall satisfaction with the product was high.

[0172] The above-described embodiments are merely preferred embodiments of the present application and are not intended to limit the scope of the present application. Various modifications and improvements to the technical solutions of the present application made by those of ordinary skill in the art without departing from the design spirit of the present application shall fall within the scope of protection of the present application.

Claims

1. A method of extracting freeze-dried polysaccharides from moringa oleifera seeds, characterized by: It comprises the following steps: (1) defatting treatment of shelled and crushed Moringa seed kernels by supercritical carbon dioxide fluid extraction method to obtain defatted Moringa seed meal; The supercritical carbon dioxide fluid extraction method comprises the following steps: The HA221-50-10-C type supercritical carbon dioxide extraction device is used to crush Moringa seed and load it into the extraction kettle, open the carbon dioxide switch, adjust the pressure and temperature of the extraction kettle, the first separation kettle and the second separation kettle, when the extraction pressure is 35 MPa and the extraction temperature is 40℃, the pressure of the first separation kettle is 12 MPa and the separation temperature is 45℃, the pressure of the second separation kettle is 5 MPa and the separation temperature is 35℃, adjust the valve to make it stable; the carbon dioxide fluid fully contacts with the Moringa seed material in the extraction kettle, the carbon dioxide fluid carrying Moringa seed oil is led into the first separation kettle through the pipeline to separate the Moringa seed oil for the first time, and then a small amount of Moringa seed oil which is not separated is led into the second separation kettle with the carbon dioxide fluid to separate the Moringa seed oil for the second time, and the separated carbon dioxide is recycled through the loop; when the preset pressure and temperature of the extraction kettle, the first separation kettle and the second separation kettle are stable for a period of time, the Moringa seed oil is collected from the first separation kettle and the second separation kettle every 30 min; the overall extraction time is 3 h, and the Moringa seed oil is obtained after the extraction is completed, and the defatted Moringa seed meal in the extraction kettle is collected; (2) natural deep eutectic solvent (DESs) is prepared by using betaine and 1,3-butanediol with a molar ratio of 1:1; (3) Moringa seed polysaccharide extraction: the DESs is uniformly mixed with distilled water to prepare a DESs solution with a water content of 85%, and the Moringa seed meal is extracted according to the solid-liquid ratio of 1:5 to obtain Moringa seed meal mixed homogenate; The Moringa seed meal mixed homogenate is filtered by a filtering device to obtain a clear Moringa seed polysaccharide extraction solution, which is concentrated and dried under reduced pressure to 1 / 3 of the original volume at a pressure of-0.1 MPa and a temperature of 50-60℃, then anhydrous ethanol is added according to the volume ratio of 1:3, alcohol precipitation is static for 12 h, 10000 r / min, 10 min high-speed centrifugation is used to remove the supernatant, water is added for dissolution, single-layer qualitative filter paper is used for filtration, and the solution is concentrated under reduced pressure at a pressure of-0.1 MPa and a temperature of 50-60℃ to 1 / 3 of the original volume, then anhydrous ethanol is added according to the volume ratio of 1:3, alcohol precipitation is static for 12 h, and Moringa seed polysaccharide is obtained by centrifugation, which is repeatedly washed with 95% anhydrous ethanol until the washing solution is clear, and then filtered to obtain Moringa seed polysaccharide; The inside of the filtering device is sequentially fixed with 5 layers of filtering structures from top to bottom: a first filtering screen (4), a second filtering screen (5), a macroporous resin (6), an activated carbon filtering layer (7), and a filter paper and ceramic filter sheet layer (8); the first filtering screen (4) is 80 mesh, the second filtering screen (5) is 120 mesh, the macroporous resin (6) is D101 macroporous resin, the activated carbon filtering layer (7) has a thickness of 5 mm, and the filter paper and ceramic filter sheet layer (8) is a 10 ppi ceramic filter sheet, on which a single layer of qualitative filter paper is arranged; (4) Freeze-drying the moringa seed polysaccharide to obtain a final product of moringa seed polysaccharide freeze-dried powder.

2. The process for extraction of freeze-dried polysaccharides from moringa seeds as claimed in claim 1, wherein: The 5 layers of filtering structures are sequentially fixed on a filtering layer frame (3) from top to bottom, and a liquid collecting hopper (9) is arranged at the lower part of the filtering layer frame (3) and is in communication with the filter paper and ceramic filter sheet layer (8); the filtering layer frame (3) and the liquid collecting hopper (9) are fixed in the inside of the filtering cylinder wall (2), and a liquid collecting funnel (1), a liquid valve (10), and a pressure adjusting pipeline (11) are arranged at the top, bottom, and side of the filtering cylinder wall (2), respectively; the liquid collecting funnel (1) is in communication with the inside space of the filtering layer frame (3), the liquid valve (10) is in communication with the bottom of the liquid collecting hopper (9), and the upper part and lower part of the pressure adjusting pipeline (11) are in communication with the upper space of the first filtering screen (4) and the upper space of the liquid collecting hopper (9), respectively; a pressure adjusting valve (12) is arranged on the pressure adjusting pipeline (11). The material of the liquid collecting funnel (1), the filtering cylinder wall (2), the filtering layer frame (3), the first filtering screen (4), the second filtering screen (5), the liquid collecting hopper (9), the liquid valve (10), the pressure adjusting pipeline (11), and the pressure adjusting valve (12) is 304 stainless steel.

3. The process for extraction of lyophilized moringa seed polysaccharides as claimed in claim 2, wherein: Step (2) specifically comprises the following steps: placing betaine and 1,3-butanediol in a beaker in a proportion of 1:1 by mole ratio, heating and stirring at 80 DEG C until a transparent and uniform liquid, a natural deep eutectic solvent, i.e., DESs, is formed, and cooling to room temperature for storage.

4. The process for extraction of lyophilized moringa seed polysaccharides as claimed in claim 3, wherein: Step (4) specifically comprises the following steps: placing the obtained moringa seed polysaccharide under the condition of-80 DEG C for pre-freezing for 12 hours, and performing freeze-drying treatment under the condition of-80 DEG C and 0.37 mbar for 3-4 hours to finally obtain moringa seed polysaccharide freeze-dried powder.

5. Application of the moringa seed polysaccharide freeze-dried powder prepared by the method of any one of claims 1-4 in cosmetics.

6. Use according to claim 5, characterized in that: The cosmetic is one of the following: (1) Moringa seed polysaccharide essence water, which is prepared from the following raw materials by mass fraction: 1,3-butanediol 2-3%, acryloyldimethyltaurate ammonium / VP copolymer 0.1-0.5%, carbomer 0.05-0.1%, betaine 0.2-0.5%, p-hydroxyacetophenone 0.3-0.5%, moringa oleifera seed polysaccharide lyophilized powder 0.1-0.15%, 1,2-hexanediol 0.3-0.5%, triethanolamine 0.1-0.3%, and the balance being water; (3) Moringa oleifera seed polysaccharide essence milk is made from the following raw materials in parts by mass: acrylic acid (ester) / C10-30 alkanol acrylate cross-linked polymer 0.1-0.15%, xanthan gum 0.05-0.1%, 1,3-butanediol 3-5%, dipropylene glycol 3-5%, glycerol 2-3%, sodium hyaluronate 0.01-0.02%, moringa oleifera seed polysaccharide lyophilized powder 0.1-0.12%, dimethicone 2-5%, squalane 1-2%, 9% KOH 0.3-0.5%, phenoxyethanol 0.5-0.8%, and the balance being water; (4) Moringa oleifera seed polysaccharide essence cream is made from the following raw materials in parts by mass: sodium hyaluronate 0.01-0.05%, moringa oleifera seed polysaccharide lyophilized powder 0.05-0.1%, xanthan gum 0.1-0.3%, EDTA-Na 0.05-0.1%, 1,3-butanediol 2-3%, glycerol 2-5%, cetylstearyl alcohol 0.5-1%, squalane 2-5%, squalane 1-3%, caprylic / capric triglyceride 2-3%, p-hydroxyacetophenone 0.5-0.8%, 1,2-hexanediol 0.3-0.6%, sebamed 305 (polyacrylamide, C13-14 isoparaffin, laureth-7) 0.8-1%, and the balance being water.

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