Lotus pod or lotus seed shell extract, preparation method and application thereof

Abstract

The present invention relates to a lotus pod or lotus seed shell extract, and its preparation method and application. Through the synergistic combination of ethanol reflux and enzymatic hydrolysis and macroporous resin enrichment process, polyphenols, alkaloids, unsaturated fatty acids, amino acids and flavonoids are efficiently extracted. The extract has the functions of moisture absorption and moisturizing, antibacterial, antioxidant and tyrosinase inhibition, and significantly inhibits collagen fiber degradation and promotes keratinocyte proliferation. The anti-aging emulsion developed based on the extract is compounded with sodium hyaluronate cross-linked polymer, jojoba seed extract and other ingredients to form a synergistic enhancement system: it has excellent moisturizing properties; it can inhibit collagen fiber decomposition, scavenge free radicals and regulate melanin production, comprehensively improve skin aging, and has high safety. The present invention breaks through the limitation of a single efficacy and integrates multiple activities such as moisture absorption, anti-aging and whitening. The emulsion has fast absorption and excellent compatibility. It can be applied to products such as essences and creams, solving the industry pain points of high irritation of chemical additives and insufficient efficacy of natural ingredients.

Description

Technical Field

[0001] The present invention relates to the field of natural product extraction and pharmaceutical technology, and in particular to a lotus pod or lotus seed shell extract, a preparation method and application thereof. Background Art

[0002] Lotus seed processing generates a significant amount of waste, including lotus seed pods, lotus seed shells, and lotus seed cores. Lotus seed pods, also known as lotus pods, are the dried, mature receptacles of the lotus (Nelumbo nucifera Gaertn.) plant, a member of the Nymphaeaceae family. They are a major waste product during lotus seed processing and are abundant resources. Research has shown that lotus seed pods and lotus seed shells are rich in active ingredients such as alkaloids, phenolic acids, and polysaccharides, exhibiting a variety of physiological activities, including antioxidant, anti-tumor, anti-aging, liver-protective, memory-enhancing, antibacterial, anti-inflammatory, radiation-resistant, and lipid-lowering properties. In traditional Chinese medicine, lotus seed pods are used to treat menorrhagia and as a hemostatic agent. However, the current utilization of the lotus seed pods and lotus seed shells as resources remains very limited.

[0003] Prior art research on lotus pod extracts has primarily focused on the extraction of single components, such as flavonoids, anthocyanins, polysaccharides, polyphenols, and proanthocyanidins. For example, CN114028494A discloses a traditional Chinese medicine composition containing lotus pods, CN115350213A discloses a hangover-relief formula containing lotus pod charcoal, and CN113750139B discloses a method for extracting anti-inflammatory components from lotus pods. However, these methods generally suffer from issues such as high organic solvent usage, low extraction efficiency, and high costs. The lotus pod polysaccharide extraction method disclosed in CN114560956A has a low yield, and the method in CN103304532A, which uses acetone and a complex enzyme to extract proanthocyanidins, carries the risk of residual solvents. Furthermore, patent applications such as CN107522683A and CN103372059A also suffer from complex extraction processes and insufficient applied research. Research on lotus seed shell is even more lacking, with only a few reports on α-glucosidase inhibition (CN109303799A) and proanthocyanidin separation (CN118437025A).

[0004] In the field of skin anti-aging, most existing products have a single effect. Skin aging is the result of multiple factors, including free radical oxidation, UV damage, and collagen degradation, requiring multi-target synergistic intervention. Although CN118787582A discloses an anti-aging cream containing ginseng extract, its ingredients are complex, the cost is high, and there is a lack of quantitative efficacy data. Anti-aging products currently on the market generally suffer from single efficacy and excessive chemical additives.

[0005] The lotus pod and seed shell extract developed by this invention possesses multiple benefits: excellent moisture absorption and moisturizing properties, significant antibacterial activity, powerful free radical scavenging ability, prominent tyrosinase inhibition, effective collagen fiber protection, and keratinocyte proliferation promotion. This extract reduces the use of chemical preservatives and offers high safety, providing a new solution for the development of natural, highly effective, and multifunctional anti-aging cosmetics. The emulsion prepared through scientific formulation not only exhibits comprehensive anti-aging benefits but also achieves high-value utilization of lotus pod and seed shell waste, possessing significant economic and environmental value. Summary of the Invention

[0006] To address the above technical issues, the present invention provides a lotus seed pod or lotus seed shell extract, its preparation method, and its use. The preparation method can simultaneously extract polyphenols, alkaloids, unsaturated fatty acids, amino acids, and glycoside derivatives. The resulting extract has multiple benefits, including moisturizing, hygroscopicity, anti-inflammatory, antibacterial, and antioxidant properties. The extract can be used in lotions to produce anti-aging products with multiple benefits, including long-lasting moisturizing, effective free radical scavenging, inhibition of skin inflammation, promotion of collagen fiber synthesis, inhibition of collagen fiber decomposition, and promotion of basal cell division.

[0007] In order to achieve the above object, the technical solution of the present invention is as follows:

[0008] A lotus seed pod or lotus seed shell extract, comprising the following components by mass percentage:

[0009] (a) Polyphenols: 10-50%, wherein the polyphenols are selected from at least three of procyanidin B1, procyanidin B2, catechin, hyperoside and dihydrokaempferol, wherein procyanidin B1: molecular formula C 30 H 26 O 12 , CAS 20315-25-7, cis, trans''-4,8''-di-(3,3',4',5,7-pentahydroxyflavan), proanthocyanidin B2: molecular formula C 30 H 26 O 12 , CAS29106-49-8, cis, cis″-4,8″-bis(3,3′,4′,5,7-pentahydroxyflavan);

[0010] (b) alkaloids: 5-30%, wherein the alkaloids are at least two selected from the group consisting of nuciferine, nornuciferine, annonine, choline, and N,O-dimethyltheobromine;

[0011] (c) Unsaturated fatty acids: 8-40%, including linoleic acid, vaccenic acid and (Z)-5,8,11-trihydroxyoctadec-9-enoic acid;

[0012] (d) Amino acids: 3-15%, including pyroglutamic acid, phenylalanine and L-tryptophan;

[0013] (e) Flavonoids: 2-10%, including at least two of quercetin, (-)-epigallocatechin, myricetin-3-galactoside, and quercetin-3-glucuronide.

[0014] The preparation method of the lotus seed pod or lotus seed shell extract comprises the following steps:

[0015] S1. Take fresh lotus pods or lotus seed shells, dry them, crush them and sieve them to obtain fine powder;

[0016] S2. The extraction solvent is added to the fine powder obtained in step S1 and the mixture is heated under reflux for extraction. The solid-liquid mass ratio is 1:30-50, the extraction time is 0.5-3.0h, the extraction temperature is 85-95°C, and the extraction is performed 1-3 times.

[0017] S3. The reflux obtained in step S2 was filtered, the filtrate was centrifuged, and the supernatant was taken;

[0018] S4. The filtered residue and the precipitate obtained in step S3 were dispersed in deionized water, and then cellulase and pectinase were added for enzymatic hydrolysis, and then filtered, the filtrate was centrifuged, and the supernatant was taken;

[0019] S5. The supernatants obtained in step S3 and step S4 are combined to obtain a total extract;

[0020] S6. The total extract was separated and eluted by column chromatography using a macroporous resin column;

[0021] S7. The eluate obtained in step S6 is filtered through a microporous membrane, and the filtrate is concentrated to remove ethanol to obtain a concentrate;

[0022] S8. Drying the concentrated solution obtained in step S7 to obtain a lotus seed pod or lotus seed shell extract.

[0023] Furthermore, in step S1, the drying temperature is 45-55° C., the drying time is 5-10 hours, and the sieving is performed using a 20-40 mesh screen.

[0024] Furthermore, in step S2, the extraction solvent is an ethanol aqueous solution, and the mass fraction of the ethanol aqueous solution is 10 to 35%.

[0025] Furthermore, in step S3, the centrifugation condition is 8000-12000 rpm, and the time is 10-20 min.

[0026] Furthermore, in step S4, the centrifugation conditions are 8,000-10,000 rpm for 10-15 minutes. The enzymatic hydrolysis conditions are as follows: deionized water is added to the filtered residue and the centrifuged precipitate at a solid-to-liquid ratio of 1:10-15 g / ml. The added cellulase activity is 100,000 U / g (based on the filter paper enzyme activity (FPA)), and the cellulase dosage is 0.1-0.4% g / ml of the total reaction solution volume (i.e., 0.1 g to 0.4 g of cellulase is added per 100 mL of solution). The pectinase activity is 60,000 U / g (based on the polygalacturonase activity), and the pectinase dosage is 0.05-0.2% g / ml of the total reaction solution volume (i.e., 0.05 g to 0.2 g of pectinase is added per 100 mL of solution). The pH of the enzymatic hydrolysis solution is adjusted to 4.5-5.0, and the enzymatic hydrolysis is performed at 45-55°C.

[0027] Furthermore, in step S6, the macroporous adsorption resin is AB-8, D101 or NKA-9 macroporous adsorption resin column; and ethanol is used for elution.

[0028] Furthermore, in step S7, the pore size of the microporous filter membrane is 1.2 μm; and the concentration method is rotary evaporation concentration or vacuum concentration.

[0029] Furthermore, in step S8, the drying is freeze drying, vacuum drying or spray drying.

[0030] The lotus seed pod or lotus seed shell extract is used as the emulsion raw material, and the mass percentage composition of the emulsion raw material is: pink passionflower seed oil 1-5%, jojoba seed extract 1-2%, caprylic / capric triglyceride (a mixed triglyceride formed by esterifying glycerol with caprylic (C8) and capric (C10) fatty acids) 1-5%, glyceryl glucoside 1.0-4.5%, castor oil / IPDI Copolymer 0.2-1.0%, Prunus Amygdalus Dulcis Seed Extract 1.0-5.0%, Propolis Extract 0.1-0.6%, Allantoin PCA Sodium 0.05-0.32%, Sodium Hyaluronate Crosspolymer 0.5-3.5%, Methyl Ascorbyl Silane Pectate 0.5-2.5%, Methyl Dihydroxybenzoate 0.02-0.2%, Hydroxylated Lecithin 0.05-0.3%, Tocopheryl Glucoside 0.1-0.8%, Lotus Pod or Lotus Seed Shell Extract 0.10-0.3%, Dandelion Rhizome / Root Extract 0.05-0.25%, Motherwort Extract 0.05-0.15%, Quaternary Ammonium-73 0.2-0.6%, and the balance is water.

[0031] Preferably, the ingredients include 3.2% pink passionflower seed oil, 1.5% jojoba seed extract, 1.2% caprylic / capric triglyceride, 1.2% glyceryl glucoside, 0.8% castor oil / IPDI copolymer, 1.5% sweet almond seed extract, 0.4% propolis extract, 0.08% allantoin PCA sodium, 0.6% sodium hyaluronate crosspolymer, 0.8% ascorbyl methylsilanol pectate, 0.05% methyl dihydroxybenzoate, 0.2% hydroxylated lecithin, 0.5% tocopheryl glucoside, 0.2% lotus seed pod or lotus seed shell extract, 0.12% dandelion extract, 0.08% motherwort extract, 0.3% quaternium-73, and the balance is water.

[0032] The emulsion composed of the above raw materials is prepared in the following steps:

[0033] I. configure the various components of the emulsion raw material according to the above mass percentages;

[0034] II. Mix pink passionflower seed oil, jojoba seed extract, caprylic / capric triglyceride, castor oil / IPDI copolymer, and propolis extract and treat in a water bath at 90-95°C for 15-25 minutes. Mix until all components are dissolved and then heat to obtain phase A.

[0035] III. Treat glyceryl glucoside, prunus amygdalus dulcis seed extract, ascorbyl methylsilanol pectate, tocopheryl glucoside, sodium allantoin PCA, sodium hyaluronate crosspolymer, and methyl dihydroxybenzoate at 80-100°C for 15-25 minutes, dissolve and mix until all components are uniformly dissolved, then incubate and replenish evaporated water to obtain phase B.

[0036] IV. When phases A and B are dissolved, cool to 70-80°C, slowly add phase A to phase B, homogenize at 3000-6000 rpm for 10-20 minutes to obtain a mixture of phases A and B, and cool to 40-45°C and keep warm;

[0037] V. In the mixture of phases A and B, dandelion extract, motherwort extract, lotus or lotus seed shell extract and hydroxylated lecithin were sequentially added and homogenized at 3000 to 5000 rpm for 10 to 20 min;

[0038] VI. Then add quaternary ammonium salt-73 and homogenize at 3000-5000 rpm for 5-10 minutes to obtain an emulsion.

[0039] The present invention obtains lotus pod or lotus seed shell extract through a two-step extraction method (ethanol aqueous solution heating reflux extraction and cellulase-pectinase synergistic enzymatic extraction) combined with a macroporous resin enrichment process. The extract is rich in polyphenols, alkaloids and unsaturated fatty acids, and thus has multiple functions such as strong moisturizing, anti-inflammatory, antibacterial and antioxidant effects.

[0040] The beneficial effects of the present invention are:

[0041] (1) High active substance enrichment and cost advantage: Through the synergistic effect of low-concentration ethanol reflux extraction (polar solvent targeted dissolution of most phenols and alkaloids) and enzymatic extraction (directional destruction of plant cell walls to release most unsaturated fatty acids, amino acids, and glycoside derivatives), the integrated extraction of polyphenols, alkaloids, unsaturated fatty acids, amino acids, and flavonoids was achieved for the first time, with a high extraction rate, breaking through the limitations of single component extraction and significantly reducing the extraction cost. The main extracted components all play an important role in skin nutrition and health, and the synergistic effect of each component plays a key role in skin anti-aging. In addition, the enzymatic hydrolysis process significantly reduces the extraction temperature (45~55℃ in this application vs. traditional 80℃ or above) and solvent usage (material-liquid ratio 1:10~15 vs. traditional 1:20), protecting unsaturated fatty acids from decomposition and destruction, significantly reducing overall energy consumption, and is suitable for industrial scale-up.

[0042] (2) Natural preservative properties and no toxic residue: The polyphenol and alkaloid complex system in the extract has an antibacterial rate of ≥90% against Staphylococcus aureus and Staphylococcus epidermidis, which can replace 50% to 70% of chemical preservatives (such as phenoxyethanol), improve skin ecology, and reduce the risk of allergies. The entire process uses food-grade ethanol and biological enzymes, and there is no organic solvent residue (GC-MS test did not detect benzene and chlorinated hydrocarbons), which meets the requirements of the "Cosmetic Safety Technical Specifications".

[0043] (3) Multiple functions such as moisture absorption, moisturizing, whitening and anti-aging: Unsaturated fatty acids, amino acids and hyaluronic acid build a long-lasting moisturizing network with excellent moisture absorption and moisturizing properties. At the same time, it has the function of nourishing the skin and synergistically exerting anti-aging effects. It also has excellent performance in tyrosinase inhibition, free radical scavenging ability and barrier repair performance.

[0044] (4) Good formula compatibility and stability: The extract has good compatibility with common emulsifiers. The prepared emulsion has an "active ingredient retention rate ≥ 95%" in the 45°C / 6-month accelerated test, and the pH stability (4.5~6.5) meets the long-term storage requirements.

[0045] (5) Good application prospects: As a core functional ingredient, it can be developed into products such as "moisturizing essence, whitening mask, anti-aging cream", etc., to solve the technical bottleneck of traditional chemical additives (such as hydroquinone, retinol) being highly irritating and natural extracts having a single efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 The results of hygroscopic activity outside the shell of lotus pods and lotus seeds are (a) RH=81%, (b) RH=43%;

[0047] Figure 2 It is the result of moisturizing activity outside the shell of lotus pods and lotus seeds;

[0048] Figure 3 This is the result of the scavenging ability of lotus pods and lotus seed shells on DPPH free radicals;

[0049] Figure 4 This is the result of the scavenging ability of lotus pods and lotus seed shells on OH free radicals;

[0050] Figure 5 The results are the results of the determination of the reducing power of lotus pods and lotus seed shells;

[0051] Figure 6 The inhibition zones of inhibition of lotus seed pods and lotus seed shells on Staphylococcus aureus and Staphylococcus epidermidis are shown in Figure 2, where (a) lotus seed pods inhibit Staphylococcus aureus, (b) lotus seed pods inhibit Staphylococcus epidermidis, (c) lotus seed shells inhibit Staphylococcus aureus, and (d) lotus seed shells inhibit Staphylococcus epidermidis.

[0052] Figure 7 is the inhibition rate of tyrosinase activity outside the shell of lotus pods and seeds;

[0053] Figure 8 HE staining results of the effects of the emulsion on the back skin of the aging mouse model, including (a) healthy group, (b) aging model group, (c) aging model + high concentration treatment group, (d) aging model + medium concentration treatment group, (e) aging model + low concentration treatment group, and (f) aging model + VE positive control group;

[0054] Figure 9 Masson staining results of the effects of the emulsion on the back skin of the aging mouse model, including (a) healthy group, (b) aging model group, (c) aging model + high concentration treatment group, (d) aging model + low concentration treatment group, (e) aging model + medium concentration treatment group, and (f) aging model + VE positive control group;

[0055] Figure 10 Results of skin morphological changes in aging mouse models treated with emulsion, including (a) healthy group, (b) aging model group, (c) VE positive control group, (d) high-dose treatment group, (e) aging model + medium-dose treatment group, and (f) aging model + low-dose treatment group. DETAILED DESCRIPTION

[0056] The present invention is further described below through specific embodiments, but the present invention is not limited to the following examples. It would be apparent to those skilled in the art that appropriate modifications, substitutions of components with equivalent efficacy, or changes in product form, such as to a cream, serum, hand lotion, facial mask, etc., within the scope of the present invention or without departing from the content, spirit, and scope of the present invention, are all considered to be within the scope of the present invention. Example 1

[0057] The preparation method of lotus seed extract is as follows:

[0058] (1) Raw material processing: Take fresh and clean lotus pods, dry them at 50℃ for 10 hours, crush them with a grinder, sieve them through a 40-mesh sieve, and take the fine powder for extraction.

[0059] (2) Add 25% ethanol aqueous solution with a material-liquid ratio (mass ratio of fine powder to ethanol aqueous solution) of 1:30, heat and reflux at 90°C for 1.5 hours, extract twice, combine the two reflux liquids and filter, centrifuge the filtrate at 8000 rpm for 15 minutes, and take the supernatant.

[0060] (3) Cellulase and pectinase treatment: Deionized water was added to the filtered residue and the precipitate after centrifugation for dispersion, with a solid-liquid ratio of 1:12 g / ml. Cellulase and pectinase were then added, with the added cellulase activity being 100,000 U / g (based on the filter paper enzyme activity FPA) and the dosage being 0.2% g / ml. The added pectinase activity was 60,000 U / g (based on the polygalacturonase activity) and the dosage being 0.1% g / ml. The pH was adjusted to 4.8, and enzymatic hydrolysis was performed at 48°C for 50 min. The hydrolyzed solution was filtered and centrifuged at 8000 rpm for 15 min. The supernatant was collected and combined with the supernatant obtained in step (2) to obtain the total extract.

[0061] (4) The total extract obtained above was separated and enriched by NKA-9 macroporous adsorption resin column, eluted with ethanol, and the eluate was filtered through a 1.2 μm microporous filter membrane. The filtrate was concentrated by rotary evaporation to remove ethanol. The concentrate was spray-dried to obtain the lotus seed extract, and the polysaccharide and polyphenol content of the extract was determined. Example 2

[0062] The preparation method of lotus seed shell extract is as follows:

[0063] (1) Raw material processing: Take fresh and clean lotus seed shells, dry them at 55℃ for 8 hours, crush them with a grinder, sieve them through a 30-mesh sieve, and take the fine powder for extraction.

[0064] (2) Add 35% ethanol aqueous solution with a material-liquid ratio (mass ratio of fine powder to ethanol aqueous solution) of 1:25, heat and reflux at 85°C for 1.2 hours, extract twice, combine the two reflux liquids and filter, centrifuge the filtrate at 8000 rpm for 15 minutes, and take the supernatant.

[0065] (3) Cellulase and pectinase treatment: Deionized water was added to the filtered residue and the precipitate after centrifugation for dispersion, with a solid-liquid ratio of 1:10 g / ml. Cellulase and pectinase were then added, with the added cellulase activity being 100,000 U / g (based on the filter paper enzyme activity FPA) and the dosage being 0.25% g / ml. The added pectinase activity was 60,000 U / g (based on the polygalacturonase activity) and the dosage being 0.12% g / ml. The pH was adjusted to 5.0, and enzymatic hydrolysis was performed at 45°C for 60 min. The hydrolyzed solution was filtered and centrifuged at 9000 rpm for 10 min. The supernatant was collected and combined with the supernatant obtained in step (2) to obtain the total extract.

[0066] (4) The total extract obtained above was separated and enriched by passing it through an AB-8 macroporous adsorption resin column, eluted with ethanol, and the eluate was filtered through a 1.2 μm microporous filter membrane. The filtrate was concentrated by rotary evaporation to remove the ethanol. The concentrate was spray-dried to obtain the lotus seed shell extract, and the polysaccharide and polyphenol contents of the extract were determined.

[0067] Secondary extraction is used because the amount of precipitation and residue after the first extraction is greatly reduced, but there is still a certain amount of polysaccharide content and it is easier to extract. At this time, using biological enzymes to continue extraction can greatly save the amount of enzyme used, shorten the extraction time, increase the polysaccharide content, and reduce the cost of raw material preparation.

[0068] Examples 3 to 5

[0069] The raw materials and mass percentages of the emulsions in Examples 3-5 of the present invention are shown in Table 1. Examples 3 and 5 utilize the lotus pod extract obtained in Example 1, while Example 4 utilizes the lotus seed shell extract obtained in Example 2. After passing inspection, the emulsions were used in the following experiments. The experiments were conducted according to the method for constructing a D-galactose aging mouse model (Liu Yang, Xu Yue, Wang Yuyan, Li Shujing, Cao Yizhi. Anti-aging Effects of Perilla Stem Ethyl Acetate Extract on D-galactose Aging Mice. Chinese Journal of Traditional Chinese Medicine, 2022). Examples 3-5 were prepared by adding varying concentrations of either lotus pod or lotus seed shell extract. The control group received an equal volume of VE, and each group was smeared on the depilated skin on the back of the neck of the mice.

[0070] Table 1 Raw materials and proportions of Examples 3 to 5

[0071]

[0072] The efficacy characteristics of lotus seed pod or lotus seed shell extract are as follows:

[0073] 1. The extract components were analyzed by ultra-high pressure liquid chromatography-mass spectrometry (UPLC-MS). The results are shown in Tables 2 to 5:

[0074] Table 2 shows the total negative ion profile of the lotus seed pod extract. Among compounds with a relative abundance of 2% or more, the relative abundance of unsaturated fatty acids (vaccenic acid, linoleic acid, (Z)-5,8,11-trihydroxyoctadecan-9-enoic acid, and (10E,15Z)-9,12,13-trihydroxyoctadecan-10,15-dienoic acid) is 35.47%. Polyphenols (hyperoside, dihydrokaempferol, and epigallocatechin) are 6.91% in relative abundance, while flavonoids (quercetin and quercetin-3-glucuronide) account for 7.73%. Furthermore, the extract contains 0.8% isorhamnetin-3-o-glucoside and 0.7% phenylalanine.

[0075] Table 3 shows the positive ion components of the lotus seed extract. Among the compounds with a relative abundance of 2% or more, the relative abundance of alkaloids ((-)-nuciferine, nornuciferine, annonine, choline, n,O-dimethyltheobromine, and collagenine) was 55.17%. Furthermore, among the amino acids, phenylalanine accounted for 1.64% and L-tryptophan 1.44%.

[0076] Table 4. Component structure of lotus seed shell extract in negative ion scanning mode. Among the compounds with a relative abundance greater than 2%, polyphenols (procyanidin B1, epigallocatechin, procyanidin B2, and dihydrokaempferol) account for 29.18%, flavonoids (quercetin, isoquercetin, epicatechin, and quercetin-3-glucuronide) for 25.86%, organic acids (malic acid and gluconic acid) for 8.05%, and alkaloids (siraitine) for 2.63%. Additionally, myricetin-3-galactoside accounts for 1.46% and phenylalanine for 0.82%.

[0077] Table 5 shows the structural components of the lotus seed shell extract in positive ion scanning mode. Among the compounds with a relative abundance greater than 2%, the following compounds were present: flavonoids (catechin, quercetin-3-glucuronide, 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-chromen-4-one, and (-)-epigallocatechin) at 22.32%, alkaloids (choline, codeine, collagenine, and leonurine) at 13.36%, an amino acid derivative (phenylalanine) at 3.06%, organic acids and unsaturated fatty acids (linolenic acid) at 2.72%, and nucleotides (adenosine) at 4.52%. Furthermore, isorhamnetin-3-o-glucoside at 0.8% and phenylalanine at 0.7%.

[0078] 2. The emulsions obtained in Examples 3 to 5 were subjected to an accelerated test at 45°C for 6 months. The accelerated test was conducted as follows: The emulsions obtained in Examples 3 to 5 were divided into three batches (to ensure reproducibility). The emulsion samples were placed in a constant temperature environment at 45°C ± 2°C. Samples were taken at the end of the first, second, third, and sixth months, and the active ingredient content was determined by high-performance liquid chromatography. The active ingredient retention rate was calculated. The experimental results showed that the active ingredient retention rate of the emulsions obtained in Examples 3 to 5 was ≥95%, indicating that the long-term storage requirement was met.

[0079] 3. The experimental results show that the moisture absorption and moisturizing properties of lotus seed pod and lotus seed shell extracts are close to those of glycerin, and the moisture absorption properties are slightly weaker than glycerin (such as Figure 1 However, the moisturizing properties of lotus seed pod and lotus seed shell extracts were stronger than those of glycerol (Gly) after 24 hours, indicating that lotus seed pod and lotus seed shell extracts have more lasting moisturizing properties (as shown in Figure 2). Figure 2 shown).

[0080] 4. The experimental results show that the extracts of lotus seed pods and lotus seed shells have strong DPPH free radical and ·OH free radical scavenging rates and reducing power. The scavenging rate of 0.4mg / mL lotus seed pod extract on DPPH free radicals is 95.17%, which is close to the scavenging rate of Vc (98.74%) and higher than the scavenging rate of lotus seed shell extract (81.38%) ( Figure 3 The OH free radical scavenging rate of lotus seed shell extract is much higher than that of lotus pod extract. The OH free radical scavenging rate of 175ug / ml lotus seed shell extract reaches 60% (as shown in Figure 2). Figure 4 ). Both lotus pod and lotus seed shell extracts have strong reducing power. When the concentration is 100mg / ml, the reducing power is lotus seed shell> lotus pod ( Figure 5 shown).

[0081] 5. Staphylococcus aureus and Staphylococcus epidermidis are the main pathogens of skin diseases such as acne. Lotus seed pod and lotus seed shell extracts have a certain inhibitory effect on Staphylococcus aureus and Staphylococcus epidermidis, and lotus seed pod extract has a better inhibitory effect on Staphylococcus aureus and Staphylococcus epidermidis. The minimum inhibitory concentration (MIC) of lotus seed pod extract on Staphylococcus epidermidis is 25 mg / ml, and the minimum bactericidal concentration (MBC) is 25 mg / ml; the MIC for Staphylococcus aureus is 25 mg / ml, and the MBC is 100 mg / ml. The MIC of lotus seed shell extract against Staphylococcus epidermidis is 50mg / ml, and the MBC is 50mg / ml; against Staphylococcus aureus, the MIC is 25mg / ml, and the MBC is 100mg / ml, both showing a certain inhibitory effect. It can also be used as a natural antibacterial agent to reduce the addition of antibacterial preservatives in cosmetics, increase product safety, and reduce the possible side effects of adding chemical preservatives (such as Figure 6 shown).

[0082] 6. The inhibition rate of lotus seed pod and lotus seed shell extracts on tyrosinase increased significantly with increasing concentration. The inhibition rate of tyrosinase activity showed that lotus seed shell > lotus seed pod. When the extract concentration was 1.4mg / ml, the inhibition rates of lotus seed pod and lotus seed shell extracts on tyrosinase were 95.8% and 91.2% respectively. The inhibition rate of lotus seed shell extract on tyrosinase was close to that of ascorbic acid (Vc) (such as Figure 7 shown).

[0083] 7. Experimental results of the lotion on skin aging model mice

[0084] Human skin is composed of the epidermis and dermis, which contain keratinocytes and fibroblasts respectively. After the mice were treated with the emulsion, HE and Masson staining showed that the keratinocytes in the extract treatment groups at all concentrations divided vigorously, the number and number of epidermal cell layers increased significantly, the epidermal layer became thicker, and the epidermal cell division ability was positively correlated with the concentration of the extract (such as Figure 8 As shown), it shows that the epidermal protection and barrier repair ability of the lotion application group was enhanced. The collagen fiber content in the aging model group decreased, the collagen fibers were broken, and the structure was destroyed. Compared with the aging model group, the collagen fiber structure of each treated experimental group was intact, the content increased, and was close to that of the healthy group. The damaged skin was significantly repaired, and the collagen fiber content and integrity were positively correlated with the extract concentration. The collagen fiber integrity and content of the high and medium concentration treatment groups were significantly higher than those of the VE positive control group, among which the collagen fiber integrity and content of the mice in the high concentration treatment group were higher than those in the healthy group (as shown Figure 9The skin of the aging mouse model showed obvious sagging and wrinkles, while the skin of the mice in the emulsion treatment group and the VE positive control group was firm, plump, and shiny. The skin of the mice in Example 1 was the most plump and elastic. The skin morphological changes of each experimental group were consistent with the results of HE and Masson staining (as shown in Figure 2). Figure 10 shown).

[0085] Table 2 Component structure information of lotus seed extract in negative ion scanning mode

[0086]

[0087] Note: A, fatty acid compounds, B, fatty acid derivatives, C, flavonoid glycoside compounds, D, polyphenol compounds.

[0088] Table 3 Component structure information of lotus seed extract in positive ion scanning mode

[0089]

[0090] Note: A. Alkaloid compounds, B. Amino acid derivatives.

[0091] Table 4 Component structure information of lotus seed shell extract in negative ion scanning mode

[0092]

[0093] Note: A. Polyphenol compounds, B. Flavonoid compounds, C. Organic acid compounds, D. Alkaloid compounds.

[0094] Table 5 Component structure information of lotus seed shell extract in positive ion scanning mode

[0095]

[0096] Note: A, flavonoids, B, alkaloids, C, nucleotides, D, organic acids, E, amino acids, F, fatty acids.

Claims

1. A method for preparing a lotus seed pod or lotus seed shell extract, characterized in that: The steps include: S1. Take fresh lotus pods or lotus seed shells, dry them, crush them and sieve them to obtain fine powder; S2. Add an extraction solvent to the fine powder obtained in step S1 and perform heating and reflux extraction. The extraction solvent is a 10-35% ethanol aqueous solution with a solid-liquid mass ratio of 1:30-50. The extraction time is 0.5-3.0h, the extraction temperature is 85-95°C, and the extraction is repeated 1-3 times. S3. The reflux obtained in step S2 was filtered, the filtrate was centrifuged, and the supernatant was taken; S4. The filtered residue obtained in step S3 and the precipitate after centrifugation were dispersed in deionized water, and then cellulase and pectinase were added for enzymatic hydrolysis, and then filtered, the filtrate was centrifuged, and the supernatant was taken; S5. The supernatants obtained in step S3 and step S4 are combined to obtain a total extract; S6. The total extract was separated and eluted by column chromatography using a macroporous resin column, column chromatography using AB-8, D101 or NKA-9 macroporous adsorption resin column, eluted with ethanol; S7. The eluate obtained in step S6 is filtered through a microporous membrane, and the filtrate is concentrated to remove ethanol to obtain a concentrate; S8. Drying the concentrated solution obtained in step S7 to obtain a lotus seed pod or lotus seed shell extract.

2. The method for preparing the lotus seed pod or lotus seed shell extract according to claim 1, wherein: In step S1, the drying temperature is 45-55° C., the drying time is 5-10 hours, and a 20-40 mesh screen is used for sieving.

3. The method for preparing the lotus seed pod or lotus seed shell extract according to claim 1, wherein: In step S3, the centrifugation condition is 8000-12000 rpm, and the time is 10-20 min.

4. The method for preparing the lotus seed pod or lotus seed shell extract according to claim 1, wherein: In step S4, the centrifugation conditions are 8000-10000 rpm and the time is 10-15 min. The enzymatic hydrolysis conditions are as follows: deionized water is added to the filtered residue and the precipitate after centrifugation, the material-liquid ratio is 1:10-15 g / ml, the added cellulase activity is 100,000 U / g, the cellulase dosage is 0.1-0.4% g / ml of the total volume of the reaction solution, the pectinase activity is 60,000 U / g, the pectinase dosage is 0.05-0.2% g / ml of the total volume of the reaction solution, the solution pH is adjusted to 4.5-5.0, and the enzymatic hydrolysis is performed at a temperature of 45-55°C.

5. The method for preparing the lotus seed pod or lotus seed shell extract according to claim 1, wherein: In step S7, the pore size of the microporous filter membrane is 1.2 μm; the concentration method is rotary evaporation concentration or vacuum concentration; in step S8, the drying is freeze drying, vacuum drying or spray drying.

6. The lotus seed pod or lotus seed shell extract obtained by the preparation method of the lotus seed pod or lotus seed shell extract according to any one of claims 1 to 5.

7. The use of the lotus seed pod or lotus seed shell extract according to claim 6 in preparing an emulsion, characterized in that: The raw materials of the emulsion are composed of 1-5% pink passionflower seed oil, 1-2% jojoba seed extract, 1-5% caprylic / capric triglyceride, 1.0-4.5% glyceryl glucoside, 0.2-1.0% castor oil / IPDI copolymer, 1.0-5.0% sweet almond seed extract, 0.1-0.6% propolis extract, and allantoin PCA. Sodium 0.05-0.32%, sodium hyaluronate crosspolymer 0.5-3.5%, ascorbyl methylsilanol pectate 0.5-2.5%, methyl dihydroxybenzoate 0.02-0.2%, hydroxylated lecithin 0.05-0.3%, tocopheryl glucoside 0.1-0.8%, lotus seed pod or lotus seed shell extract 0.10-0.3%, dandelion rhizome / root extract 0.05-0.25%, motherwort extract 0.05-0.15%, quaternary ammonium salt-73 0.2-0.6%, and the balance is water.

8. The use according to claim 7, characterized in that Pink Passionflower Seed Oil 3.2%, Simmondsia Chinensis (Jojoba) Seed Extract 1.5%, Caprylic / Capric Triglyceride 1.2%, Glyceryl Glucoside 1.2%, Castor Oil / IPDI Copolymer 0.8%, Prunus Amygdalus Dulcis (Sweet Almond) Seed Extract 1.5%, Propolis Extract 0.4%, Allantoin Sodium PCA 0.08%, Sodium Hyaluronate Crosspolymer 0.6%, Ascorbyl Methylsilanol Pectate 0.8%, Methyl Dihydroxybenzoate 0.05%, Hydroxylated Lecithin 0.2%, Tocopheryl Glucoside 0.5%, Nelumbo Nucifera (Lotus) Seed Shell Extract 0.2%, Dandelion Rhizome / Root Extract 0.12%, Leonurus Herba Extract 0.08%, Quaternium-73 0.3%, Balance: Water.

9. The use according to claim 7, characterized in that The preparation method of the emulsion is as follows: I. configure the various components of the emulsion raw material according to the above mass percentages; II. Mix pink passionflower seed oil, jojoba seed extract, caprylic / capric triglyceride, castor oil / IPDI copolymer, and propolis extract, and treat in a water bath at 85 to 95°C for 15 to 25 minutes. Mix until all components are dissolved and then heat to obtain phase A. III. Mix glyceryl glucoside, prunus amygdalus dulcis seed extract, ascorbyl methylsilanol pectate, tocopheryl glucoside, sodium allantoin PCA, sodium hyaluronate crosspolymer, and methyl dihydroxybenzoate at 85-100°C for 15-25 minutes. After all components are dissolved and mixed, incubate at this temperature to replenish evaporated water, thereby obtaining phase B. IV. When phases A and B are dissolved, cool to 70-80°C, slowly add phase A to phase B, homogenize at 3000-6000 rpm for 10-20 minutes to obtain a mixture of phases A and B, and cool to 40-45°C and keep warm; V. In the mixture of phases A and B, dandelion extract, motherwort extract, lotus or lotus seed shell extract and hydroxylated lecithin were sequentially added and homogenized at 3000 to 5000 rpm for 10 to 20 min; VI. Then add quaternary ammonium salt-73 and homogenize at 3000-5000 rpm for 5-10 minutes to obtain an emulsion.

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