Lotus seedpod or lotus seed shell extract as well as preparation method and application thereof

Abstract

The invention relates to a lotus seedpod or lotus seed shell extract as well as a preparation method and application thereof, polyphenol, alkaloid, unsaturated fatty acid, amino acid and flavonoid compounds are efficiently extracted by combining ethanol reflux and enzymolysis with a macroporous resin enrichment process, and the extract has the functions of absorbing moisture, preserving moisture, inhibiting bacteria, resisting oxidation and inhibiting tyrosinase activity, and can be used for preparing the lotus seedpod or lotus seed shell extract. The collagen fiber degradation is obviously inhibited; the keratinocyte proliferation is promoted. The anti-aging emulsion developed on the basis of the extract is compounded with the sodium hyaluronate cross-linked polymer, the simmondsia chinensis seed extract and other components to form a synergistic interaction system, so that the moisturizing performance is excellent; the composition can inhibit collagenous fiber decomposition, scavenge free radicals, regulate melanin generation and comprehensively improve skin aging, and has high safety. The invention breaks through the limitation of single effect, integrates multiple activities of moisture absorption, aging resistance, whitening and the like, is fast in emulsion absorption and excellent in compatibility, can be applied to products such as essence, face cream and the like, and solves the industrial pain points of large irritation of chemical additives and insufficient effect of natural components.

Description

Technical Field

[0001] The present invention relates to the technical fields of natural product extraction and medicine, and particularly relates to an extract of lotus seedpod or lotus seed shell, a preparation method thereof, and an application thereof. Background Art

[0002] During the processing of lotus seeds, a large amount of waste such as lotus seedpods, lotus seed shells, and lotus seed cores will be generated. The lotus seedpod (lotusseedpod), also known as the lotus receptacle, is the dried and mature flower receptacle of the lotus plant (Nelumbo nucifera Gaertn.) of the Nymphaeaceae family, which is the main waste during the processing of lotus seeds and is rich in resources. Research shows that lotus seedpods and lotus seed shells are rich in active ingredients such as alkaloids, phenolic acids, and polysaccharides, and have various physiological activities such as antioxidant, anti-tumor, anti-aging, liver protection, memory improvement, antibacterial and anti-inflammatory, anti-radiation, and lipid-lowering. In traditional Chinese medicine, lotus seedpods can be used to treat menorrhagia and as a hemostatic agent. However, the utilization of the resource value of lotus seedpods and lotus seed shells is still very limited at present.

[0003] In the prior art, the research on lotus seedpod extracts mainly focuses 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 receptacle, CN115350213A discloses an anti-alcoholism formula containing carbonized lotus receptacle, and CN113750139B discloses a method for extracting anti-inflammatory components from lotus receptacle. However, these methods generally have problems such as large consumption of organic solvents, low extraction efficiency, and high costs. The yield of the lotus seedpod polysaccharide extraction method disclosed in CN114560956A is relatively low, and the method for extracting proanthocyanidins using acetone and complex enzymes in CN103304532A has a risk of solvent residue. In addition, patent applications such as CN107522683A and CN103372059A also have defects such as complex extraction processes and insufficient application research. The research on lotus seed shells is even more lacking, and there are only a few reports in aspects such as α-glucosidase inhibition (CN109303799A) and proanthocyanidin separation (CN118437025A).

[0004] In the field of skin anti-aging, most existing products have a single function. Skin aging is the result of the combined action of multiple factors such as free radical oxidation, ultraviolet damage, and collagen degradation, and requires multi-target synergistic intervention. Although CN118787582A discloses an anti-aging cream containing ginseng extract, its ingredients are complex, the cost is relatively high, and there is a lack of quantitative efficacy data. Currently, the anti-aging products on the market generally have problems such as single efficacy and excessive chemical additives.

[0005] The extracts of lotus seedpods and lotus seed husks developed by the present invention have multiple effects: excellent moisture absorption and retention performance, remarkable antibacterial activity, strong free radical scavenging ability, outstanding tyrosinase inhibition effect, and can effectively protect collagen fibers and promote keratinocyte proliferation. This extract can reduce the use of chemical preservatives and has high safety, providing a new solution for the development of natural and highly effective multifunctional anti-aging cosmetics. The emulsion prepared through scientific formulation not only has comprehensive anti-aging effects, but also realizes the high-value utilization of lotus seedpod and lotus seed husk wastes, with important economic and environmental protection values. Summary of the Invention

[0006] In view of the above technical problems, the present invention provides an extract of lotus seedpods or lotus seed husks, a preparation method thereof, and an application thereof. The preparation method can simultaneously extract substances such as polyphenols, alkaloids, unsaturated fatty acids, amino acids, and glycoside derivatives. The obtained extract has multiple effects such as moisture absorption and retention, anti-inflammatory, antibacterial, and antioxidant effects. When the extract is used for preparing an emulsion, an anti-skin aging product with multiple effects such as good continuous moisturizing effect, effective free radical scavenging, inhibition of skin inflammation, promotion of collagen fiber synthesis, inhibition of collagen fiber decomposition, and promotion of basal cell division can be obtained.

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

[0008] An extract of lotus seedpods or lotus seed husks contains the following components by mass percentage:

[0009] (a) Polyphenols: 10 - 50%, and the polyphenols are selected from at least three of procyanidin B1, procyanidin B2, catechuic acid, hyperoside, and dihydrokaempferol. Among them, procyanidin B1: molecular formula C 30 H 26 O 12 , CAS 20315 - 25 - 7, cis, trans''-4,8''-bis-(3,3',4',5,7-pentahydroxyflavane), procyanidin B2: molecular formula C 30 H 26 O 12 , CAS 29106 - 49 - 8, cis, cis″-4,8″-bis(3,3',4',5,7-pentahydroxyflavane);

[0010] (b) Alkaloids: 5 - 30%, and the alkaloids are selected from at least two of nuciferine, nor-nuciferine, anonaine, choline, and N,O-dimethyltheobromine;

[0011] (c) Unsaturated fatty acids: 8 - 40%, including linoleic acid, isolinoleic 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 above - mentioned lotus seedpod or lotus seed shell extract includes the following steps:

[0015] S1. Take fresh lotus seedpod or lotus seed shell, dry it, and then crush and sieve it to obtain fine powder;

[0016] S2. Add an extraction solvent to the fine powder obtained in step S1 and carry out heating reflux extraction. The solid - liquid mass ratio is 1:30 - 50, the extraction time is 0.5 - 3.0 h, the extraction temperature is 85 - 95 °C, and extract 1 - 3 times;

[0017] S3. Filter the reflux liquid obtained in step S2, centrifuge the filtrate, and take the supernatant;

[0018] S4. Disperse the filter residue after filtration and the precipitate after centrifugation obtained in step S3 in deionized water, add cellulase and pectinase for enzymatic hydrolysis, then filter, centrifuge the filtrate, and take the supernatant;

[0019] S5. Combine the supernatants obtained in step S3 and step S4 to obtain the total extract;

[0020] S6. Subject the total extract to column chromatography separation and elution using a macroporous resin column;

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

[0022] S8. Dry the concentrated solution obtained in step S7 to obtain the lotus seedpod or lotus seed shell extract.

[0023] Further, in step S1, the drying temperature is 45 - 55 °C, the drying time is 5 - 10 h, and the sieving uses a 20 - 40 - mesh sieve.

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

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

[0026] Further, in step S4, the centrifugation conditions are 8000 - 10000 rpm for 10 - 15 min; the enzymatic hydrolysis conditions are as follows: deionized water is added to the filtered residue and the centrifuged precipitate, and the solid - liquid ratio is 1:10 - 15 g / ml. The cellulase activity is 100,000 U / g (based on filter paper activity FPA), and the cellulase dosage is 0.1 - 0.4% g / ml of the total volume of the reaction solution (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 polygalacturonase activity), and the pectinase dosage is 0.05 - 0.2% g / ml of the total volume of the reaction solution (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 enzymatic hydrolysis is carried out at 45 - 55°C.

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

[0028] Further, in step S7, the pore size of the microfiltration membrane is 1.2 μm; the concentration method is rotary evaporation concentration or vacuum concentration.

[0029] Further, in step S8, the drying method is freeze - drying, vacuum drying or spray drying.

[0030] The above - mentioned lotus pod or lotus seed shell extract is used as a raw material for the emulsion. The mass percentage composition of the emulsion raw material is as follows: passion fruit seed oil 1 - 5%, jojoba seed extract 1 - 2%, caprylic / capric triglyceride (a mixed triglyceride formed by the esterification of glycerol with two fatty acids, caprylic acid (C8) and capric acid (C10)) 1 - 5%, glucosylglycerol 1.0 - 4.5%, castor oil / IPDI copolymer 0.2 - 1.0%, sweet almond seed extract 1.0 - 5.0%, propolis extract 0.1 - 0.6%, allantoin PCA - Na 0.05 - 0.32%, sodium hyaluronate cross - polymer 0.5 - 3.5%, ascorbyl methylsilanol pectate 0.5 - 2.5%, methyl dihydroxybenzoate 0.02 - 0.2%, hydroxylated lecithin 0.05 - 0.3%, tocopherol 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 salt - 73 0.2 - 0.6%, and the balance is water.

[0031] Preferably, 3.2% of Passiflora edulis Sims f. flavicarpa Deg. seed oil, 1.5% of Simmondsia chinensis (Link) Schneider seed extract, 1.2% of caprylic / capric triglyceride, 1.2% of glucosylglycerol, 0.8% of castor oil / IPDI copolymer, 1.5% of Prunus dulcis (Mill.) D. A. Webb seed extract, 0.4% of propolis extract, 0.08% of allantoin PCA sodium, 0.6% of sodium hyaluronate cross-linked polymer, 0.8% of ascorbyl methylsilanol pectate, 0.05% of methyl dihydroxybenzoate, 0.2% of hydroxylated lecithin, 0.5% of tocopherol glucoside, 0.2% of Nelumbo nucifera Gaertn. seedpod or lotus seed shell extract, 0.12% of Taraxacum mongolicum Hand.-Mazz. extract, 0.08% of Leonurus japonicus Houtt. extract, 0.3% of quaternium-73, and the balance is water.

[0032] For the emulsion prepared from the above raw materials, the preparation steps are as follows:

[0033] I. Configure each component of the emulsion raw materials according to the above mass percentages;

[0034] II. Mix Passiflora edulis Sims f. flavicarpa Deg. seed oil, Simmondsia chinensis (Link) Schneider seed extract, caprylic / capric triglyceride, castor oil / IPDI copolymer, and propolis extract, and perform a water bath treatment at 90 - 95 °C for 15 - 25 min. After all components are dissolved and mixed evenly, keep warm to obtain Phase A;

[0035] III. Treat glucosylglycerol, Prunus dulcis (Mill.) D. A. Webb seed extract, ascorbyl methylsilanol pectate, tocopherol glucoside, allantoin PCA sodium, sodium hyaluronate cross-linked polymer, and methyl dihydroxybenzoate at 80 - 100 °C for 15 - 25 min. After all components are dissolved and mixed evenly, keep warm and supplement the evaporated water to obtain Phase B;

[0036] IV. When both Phase A and Phase B are dissolved, cool to 70 - 80 °C, slowly add Phase A into Phase B, and homogenize at 3000 - 6000 rmp for 10 - 20 min to obtain a mixture of Phase A and Phase B, and cool to 40 - 45 °C for heat preservation;

[0037] V. Sequentially add Taraxacum mongolicum Hand.-Mazz. extract, Leonurus japonicus Houtt. extract, Nelumbo nucifera Gaertn. seedpod or lotus seed shell extract, and hydroxylated lecithin into the mixture of Phase A and Phase B, and homogenize at 3000 - 5000 rmp for 10 - 20 min;

[0038] VI. Then add quaternium-73, and homogenize at 3000 - 5000 rmp for 5 - 10 min to obtain the emulsion.

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

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

[0041] (1) High enrichment of active substances and cost advantages: Through the synergistic effect of reflux extraction with low-concentration ethanol (polar solvents targetedly dissolve most phenols and alkaloids) and enzymatic extraction (directionally destroying plant cell walls and releasing most unsaturated fatty acids, amino acids, and glycoside derivatives), the integrated extraction of polyphenols, alkaloids, unsaturated fatty acids, amino acids, and flavonoid components is achieved for the first time. The extraction rate is high, breaking through the limitations of single-component extraction alone, and significantly reducing the extraction cost. The main components extracted all play important roles in skin nutrition and health. Each component synergistically enhances the effect, playing a key role in skin anti-aging. Moreover, the enzymatic process significantly reduces the extraction temperature (45 - 55°C in this application vs. above 80°C traditionally) and the solvent dosage (material-liquid ratio 1:10 - 15 vs. 1:20 traditionally), protecting unsaturated fatty acids from decomposition and destruction, and significantly reducing the comprehensive energy consumption, being suitable for industrial scale-up.

[0042] (2) Natural anti-corrosion properties and non-toxic residues: The antibacterial rate of the polyphenol and alkaloid composite system in the extract against "Staphylococcus aureus and Staphylococcus epidermidis" is ≥90%, which can replace 50% - 70% of chemical preservatives (such as phenoxyethanol), improve the skin ecosystem, and reduce the risk of sensitization. The whole process of the process uses food-grade ethanol and biological enzymes, without organic solvent residues (benzene and chlorinated hydrocarbons are not detected by GC-MS), meeting the requirements of the "Technical Specifications for Cosmetic Safety".

[0043] (3) Multiple functions such as moisture absorption and retention, whitening, and anti-aging: Unsaturated fatty acids, amino acids, and hyaluronic acid build a long-lasting moisture retention network, with excellent moisture absorption and retention performance, and at the same time have the function of nourishing the skin, synergistically playing an anti-aging role; and it also has excellent performance in terms of tyrosinase inhibition performance, free radical scavenging ability, and barrier repair performance.

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

[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., solving the technical bottlenecks of traditional chemical additives (such as hydroquinone and retinol) with high irritation and single efficacy of natural extracts. Description of the Drawings

[0046] Figure 1 Shows the hygroscopic activity results outside the lotus pod and lotus seed shell. (a) RH = 81%, (b) RH = 43%;

[0047] Figure 2 Results of the external moisturizing activity of lotus seedpods and lotus seed husks

[0048] Figure 3 Results of the scavenging ability of lotus seedpods and lotus seed husks against DPPH free radicals

[0049] Figure 4 Results of the scavenging ability of lotus seedpods and lotus seed husks against ·OH free radicals

[0050] Figure 5 Results of the determination of the reducing power of lotus seedpods and lotus seed husks

[0051] Figure 6 Inhibitory effects of lotus seedpods and lotus seed husks on the inhibition zones of Staphylococcus aureus and Staphylococcus epidermidis. Among them, (a) lotus seedpods against Staphylococcus aureus, (b) lotus seedpods against Staphylococcus epidermidis, (c) lotus seed husks against Staphylococcus aureus, (d) lotus seed husks against Staphylococcus epidermidis

[0052] Figure 7 Inhibitory rate of lotus seedpods and lotus seed husks on tyrosinase activity in vitro

[0053] Figure 8 HE staining results of the effects of the emulsion on the back skin of the aging mouse model. Among them, (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, (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. Among them, (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, (f) aging model + VE positive control group

[0055] Figure 10 Results of the morphological changes of the skin of the aging mouse model treated with the emulsion. Among them, (a) healthy group, (b) aging model group, (c) VE positive control group, (d) high-dose treatment group, (e) aging model + medium-dose treatment group, (f) aging model + low-dose treatment group Detailed implementation methods

[0056] The present invention will be further described below through specific embodiments, but the present invention is not limited to the following embodiments. Within the scope of the present invention or without departing from the content, spirit and scope of the present invention, appropriate improvements to the emulsion described in the present invention, replacement of components with the same efficacy, and change of product form such as changing to cream, essence, hand lotion, facial mask, etc. are obvious to those skilled in the art and are all considered to be included within the scope of the present invention. Example 1

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

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

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

[0060] (3) Treatment with cellulase and pectinase: Add deionized water to the filtered residue and the centrifuged precipitate for dispersion, the solid-liquid ratio is 1:12 g / ml, then add cellulase and pectinase. The activity of the added cellulase is 100,000 U / g (based on the filter paper enzyme activity FPA), the dosage is 0.2% g / ml, the activity of the pectinase is 60,000 U / g (based on the polygalacturonase activity), the dosage is 0.1% g / ml, adjust the pH to 4.8, enzymolyze at 48 °C, and the enzymolysis time is 50 min. The enzymolysis solution is filtered and then centrifuged at 8000 rpm for 15 min, take the supernatant, and combine it with the supernatant obtained in step (2) to obtain the total extract.

[0061] (4) Separate and enrich the total extract obtained above through an NKA-9 macroporous adsorption resin column, elute with ethanol, filter the eluate through a 1.2-μm microporous filter membrane, rotary evaporate and concentrate the filtrate to remove ethanol. Spray-dry the concentrated solution to obtain the lotus seedpod extract, and measure the polysaccharide and polyphenol contents of the extract. Example 2

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

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

[0064] (2) Add an aqueous ethanol solution with a mass fraction of 35%. The solid-liquid ratio (mass ratio of fine powder to aqueous ethanol solution) is 1:25. Heat under reflux at 85 °C for 1.2 h and extract twice. Combine the two reflux solutions and filter. Centrifuge the filtrate at 8000 rpm for 15 min and take the supernatant.

[0065] (3) Treatment with cellulase and pectinase: Add deionized water to disperse the filtered residue and the centrifuged precipitate. The solid-liquid ratio is 1:10 g / ml. Then add cellulase and pectinase. The activity of the added cellulase is 100,000 U / g (based on filter paper enzyme activity FPA), and the dosage is 0.25% g / ml. The activity of pectinase is 60,000 U / g (based on polygalacturonase activity), and the dosage is 0.12% g / ml. Adjust the pH to 5.0 and carry out enzymatic hydrolysis at 45 °C for 60 min. Filter the enzymatic hydrolysate and centrifuge at 9000 rpm for 10 min. Take the supernatant and combine it with the supernatant obtained in step (2) to obtain the total extract.

[0066] (4) Separate and enrich the total extract obtained above through an AB-8 macroporous adsorption resin column, elute with ethanol, filter the eluate through a 1.2 μm microporous membrane, and perform rotary evaporation and concentration on the filtrate to remove ethanol. Spray-dry the concentrated solution to obtain the lotus seed shell extract, and determine the polysaccharide and polyphenol contents of the extract.

[0067] Secondary extraction is adopted. After the first extraction, the amount of precipitate and residue is greatly reduced, but there is still a certain polysaccharide content and it is easier to extract. At this time, using biological enzymes for continuous extraction can greatly save the enzyme dosage, shorten the extraction time, increase the polysaccharide content, and reduce the raw material preparation cost.

[0068] Examples 3 to 5

[0069] The raw materials and mass percentage ratios of the emulsions in Examples 3 to 5 of the present invention are shown in Table 1. Among them, the lotus seed pod extracts obtained in Example 1 are used in Examples 3 and 5, and the lotus seed shell extract obtained in Example 2 is used in Example 4. After the emulsions are inspected and qualified, they are used in the following experiments. The experiments are carried out according to the method for constructing a D-galactose-induced aging mouse model (Liu Yang, Xu Yue, Wang Yuyan, Li Shujing, Cao Yizhi. Study on the anti-aging effect of ethyl acetate extract of Perilla frutescens stem on D-galactose-induced aging model mice. Chinese Archives of Traditional Chinese Medicine, 2022). Examples 3 to 5 are emulsions prepared by adding different concentrations of lotus seed pod or lotus seed shell extracts. The control group is VE with the same volume. Each group is applied to the skin of the depilated neck and back of mice.

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

[0071]

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

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

[0074] Table 2 is the negative ion total ion chromatogram of the lotus seedpod extract. Among the compounds with a relative abundance of more than 2%, the relative abundance of unsaturated fatty acids (isoleic acid, linoleic acid, (Z)-5,8,11-trihydroxyoctadec-9-enoic acid, (10E,15Z)-9,12,13-trihydroxyoctadec-10,15-dienoic acid) is 35.47%. The relative abundance of polyphenols (hyperin, dihydrokaempferol, epigallocatechin) reaches 6.91%, and that of flavonoids (quercetin, quercetin-3-glucuronide) is 7.73%. In addition, it also contains isorhamnetin-3-o-glucoside at 0.8% and phenylalanine at 0.7%.

[0075] Table 3 is the positive ion components of the lotus seedpod extract. Among the compounds with a relative abundance of more than 2%, the relative abundance of alkaloids ((-)-nuciferine, nor-nuciferine, anonaine, choline, n,O-dimethyltheobromine, corydine) is 55.17%. In addition, among the amino acid substances, phenylalanine is 1.64% and L-tryptophan is 1.44%.

[0076] Table 4 shows the component structures of the lotus seed shell extract in the negative ion scanning mode. Among the compounds with a relative abundance of more than 2%, polyphenols (procyanidin B1, epigallocatechin, procyanidin B2, dihydrokaempferol) are 29.18%, flavonoids (quercetin, isoquercetin, epicatechin, quercetin-3-glucuronide) are 25.86%, organic acids (malic acid, gluconic acid) are 8.05%, and alkaloids (ajmalicine) are 2.63%. In addition, there is myricetin-3-galactoside at 1.46% and phenylalanine at 0.82%.

[0077] Table 5 shows the component structures of the lotus seed shell extract in the positive ion scanning mode. Among the compounds with a relative abundance of more than 2%, flavonoids (catechin, quercetin-3-glucuronide, 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-chromen-4-one, (-)-epigallocatechin) are 22.32%, alkaloids (choline, codeine, corydine, leonurine) are 13.36%, amino acid derivatives (phenylalanine) are 3.06%, and there are also certain organic acids and unsaturated fatty acids (linolenic acid) at 2.72% and nucleotide (adenosine) at 4.52%. In addition, there is 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 carried out according to the following steps: The emulsions obtained in Examples 3 to 5 were each divided into 3 batches (to ensure reproducibility). The emulsion samples were placed in a constant temperature environment of 45°C ± 2°C. Samples were taken at the end of the 1st, 2nd, 3rd, and 6th months, respectively. The content of the active ingredient was determined by high performance liquid chromatography, and the retention rate of the active ingredient was calculated. The experimental results showed that the retention rate of the active ingredient in the emulsions obtained in Examples 3 to 5 was ≥95%, indicating that it meets the requirements for long-term storage.

[0079] 3. The experimental results showed that the moisture absorption and moisture retention properties of the extracts of lotus seedpods and lotus seed husks were close to those of glycerol. The moisture absorption property was slightly weaker than that of glycerol (as Figure 1 shown), but the moisture retention property of the extracts of lotus seedpods and lotus seed husks was stronger than that of glycerol (Gly) after 24 h, indicating that the extracts of lotus seedpods and lotus seed husks had more persistent moisture retention properties (as Figure 2 shown).

[0080] 4. The experimental results showed that both the extracts of lotus seedpods and lotus seed husks had strong scavenging rates of DPPH free radicals and ·OH free radicals and reducing power. The scavenging rate of DPPH free radicals by 0.4 mg / mL lotus seedpod extract was 95.17%, close to the scavenging rate of Vc (98.74%), and higher than the scavenging rate of lotus seed husk extract (81.38%) ( Figure 3 shown). The scavenging rate of ·OH free radicals by lotus seed husk extract was much higher than that of lotus seedpod extract. The scavenging rate of ·OH free radicals by 175 μg / mL lotus seed husk extract reached 60% (as Figure 4 ). Both the extracts of lotus seedpods and lotus seed husks had strong reducing power. When the concentration was 100 mg / mL, the reducing power was in the order of lotus seed husk > lotus seedpod ( Figure 5 shown).

[0081] 5. Staphylococcus aureus and Staphylococcus epidermidis are the main pathogenic bacteria of skin diseases such as acne. The extracts of lotus pods and lotus seed husks have certain inhibitory effects on Staphylococcus aureus and Staphylococcus epidermidis. Moreover, the inhibitory effect of the lotus pod extract on Staphylococcus aureus and Staphylococcus epidermidis is better. The minimum inhibitory concentration (MIC) of the lotus pod extract against Staphylococcus epidermidis is 25 mg / ml, and the minimum bactericidal concentration (MBC) is 25 mg / ml; the MIC against Staphylococcus aureus is 25 mg / ml, and the MBC is 100 mg / ml. The MIC of the lotus seed husk extract against Staphylococcus epidermidis is 50 mg / ml, and the MBC is 50 mg / ml; the MIC against Staphylococcus aureus is 25 mg / ml, and the MBC is 100 mg / ml. All show certain inhibitory effects and can be used as natural bacteriostatic agents at the same time, reducing the addition of antibacterial preservatives in cosmetics, increasing the safety of products, and reducing the possible side effects caused by the addition of chemical preservatives (such as Figure 6 shown).

[0082] 6. The inhibition rate of the extracts of lotus pods and lotus seed husks on tyrosinase increases significantly with the increase of concentration. The inhibitory rate of tyrosinase activity is in the order of lotus seed husk > lotus pod. When the extract concentration is 1.4 mg / ml, the inhibition rates of the lotus pod extract and the lotus seed husk extract on tyrosinase are 95.8% and 91.2% respectively. The inhibition rate of the lotus seed husk extract on tyrosinase is close to that of ascorbic acid (Vc) (such as Figure 7 shown).

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

[0084] The human skin is composed of the epidermis and the dermis, which contain keratinocytes and fibroblasts respectively. After treating the mice with the emulsion, and observing by HE and Masson staining, the keratinocytes in each concentration treatment group of the extract divide vigorously, the number and layers of epidermal cells increase significantly, the epidermis becomes thicker, and the epidermal cell division ability is positively correlated with the concentration of the extract (such as Figure 8 shown), indicating that the epidermal protection and barrier repair ability of the emulsion application group is enhanced. In the aging model group, the content of collagen fibers decreases, the collagen fibers break, and the structure is damaged. Compared with the aging model group, in each experimental group treated, the collagen fiber structure is complete, the content increases, is close to that of the healthy group, the damaged skin is significantly repaired, and the collagen fiber content and integrity are positively correlated with the extract concentration. The collagen fiber integrity and content in the high and medium concentration treatment groups are significantly higher than those in the VE positive control group, and the collagen fiber integrity and content in the high concentration treatment group of mice are higher than those in the healthy group (such as Figure 9As shown). The skin of the aging mouse model showed obvious relaxation and wrinkles. The skin of the lotion treatment group and the VE positive control group of mice was firm, plump and shiny. Among them, the skin of the mice in Example 1 was the plumpest and most elastic. The morphological changes of the skin in each experimental group were consistent with the results of HE and Masson staining (as Figure 10 shown).

[0085] Table 2 Composition and Structure Information of Lotus Seedpod Extract in Negative Ion Scanning Mode

[0086]

[0087] Note: A, fatty acid compounds; B, fatty acid derivatives; C, flavonoid glycosides; D, polyphenolic compounds.

[0088] Table 3 Composition and Structure Information of Lotus Seedpod Extract in Positive Ion Scanning Mode

[0089]

[0090] Note: A, alkaloid compounds; B, amino acid derivatives.

[0091] Table 4 Composition and Structure Information of Lotus Seed Hull Extract in Negative Ion Scanning Mode

[0092]

[0093] Note: A, polyphenolic compounds; B, flavonoids; C, organic acid compounds; D, alkaloid compounds.

[0094] Table 5 Composition and Structure Information of Lotus Seed Hull 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 lotus seed pod or lotus seed shell extract, characterized in that The following ingredients are included by mass percentage: (a) polyphenols: 10-50%, wherein the polyphenols are at least three selected from the group consisting of proanthocyanidin B1, proanthocyanidin B2, catechin, hyperoside and dihydrokaempferol; (b) alkaloids: 5-30%, wherein the alkaloids are at least two selected from the group consisting of nuciferine, nornuciferine, annonaine, choline and N,O-dimethyltheobromine; (c) Unsaturated fatty acids: 8-40%, including linoleic acid, vaccenic acid and (Z)-5,8,11-trihydroxyoctadec-9-enoic acid; (d) Amino acid compounds: 3-15%, including pyroglutamic acid, phenylalanine and L-tryptophan; (e) Flavonoids: 2-10%, including at least two of quercetin, (-)-epigallocatechin, myricetin-3-galactoside and quercetin-3-glucuronide.

2. The method for preparing the lotus pod or lotus seed shell extract according to claim 1, characterized in that: The steps include: S1. Take fresh lotus pods or lotus seed shells, dry them, and crush and sieve them to obtain fine powder; S2. Add an extraction solvent to the fine powder obtained in step S1 and heat and reflux for extraction, with a solid-liquid mass ratio of 1:30 to 50, an extraction time of 0.5 to 3.0 h, an extraction temperature of 85 to 95 ° C, and extraction 1 to 3 times; S3. Filter the reflux liquid obtained in step S2, centrifuge the filtrate, and take the supernatant; S4. The filtered residue obtained in step S3 and the precipitate after centrifugation are dispersed in deionized water, and then cellulase and pectinase are added for enzymatic hydrolysis, and then filtered, the filtrate is centrifuged, and the supernatant is taken; S5. combining the supernatants obtained in step S3 and step S4 to obtain a total extract; S6. The total extract is separated and eluted by column chromatography using a macroporous resin column; S7. The eluate obtained in step S6 is filtered through a microporous filter membrane, the filtrate is concentrated, and ethanol is removed to obtain a concentrated solution; S8. Drying the concentrated solution obtained in step S7 to obtain a lotus seed pod or lotus seed shell extract.

3. The method for preparing the lotus pod or lotus seed shell extract according to claim 2, characterized in that: 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.

4. The method for preparing the lotus pod or lotus seed shell extract according to claim 2, characterized in that: In step S2, the extraction solvent is ethanol aqueous solution, and the mass fraction of the ethanol aqueous solution is 10-35%.

5. The method for preparing the lotus pod or lotus seed shell extract according to claim 2, characterized in that: In step S3, the centrifugal condition is 8000-12000 rpm, and the time is 10-20 min.

6. The method for preparing the lotus pod or lotus seed shell extract according to claim 2, characterized in that: In step S4, the centrifugation condition is 8000-10000 rpm, and the time is 10-15 min; the enzymatic hydrolysis conditions are: deionized water is added to the filtered residue and the precipitate after centrifugation, the solid-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 carried out at a temperature of 45-55°C.

7. The method for preparing the lotus pod or lotus seed shell extract according to claim 2, characterized in that: In step S6, column chromatography uses AB-8, D101 or NKA-9 macroporous adsorption resin column, and elution uses ethanol; 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, drying is freeze drying, vacuum drying or spray drying.

8. Use of the lotus seed pod or lotus seed shell extract according to claim 1 or the lotus seed pod or lotus seed shell extract obtained by the preparation method according to any one of claims 2 to 7 in 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%, tocopherol glucoside 0.1-0.8%, lotus seed 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.

9. The use according to claim 8, characterized in that: Pink passionflower seed oil 3.2%, jojoba seed extract 1.5%, caprylic / capric triglyceride 1.2%, glyceryl glucoside 1.2%, castor oil / IPDI copolymer 0.8%, sweet almond seed extract 1.5%, propolis extract 0.4%, allantoin PCA sodium 0.08%, sodium hyaluronate crosspolymer 0.6%, ascorbyl methylsilanol pectate 0.8%, methyl dihydroxybenzoate 0.05%, hydroxylated lecithin 0.2%, tocopheryl glucoside 0.5%, lotus seed or lotus shell extract 0.2%, dandelion extract 0.12%, motherwort extract 0.08%, quaternium-73 0.3%, the balance is water.

10. The use according to claim 8, characterized in that: The preparation method of the emulsion is as follows: I. configure each component of the emulsion raw material according to the above mass percentage; 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 85-95° C. for 15-25 min. After all components are dissolved and mixed, keep warm to obtain phase A; III. Mix glyceryl glucoside, sweet almond seed extract, ascorbyl methylsilanol pectate, tocopherol glucoside, allantoin PCA sodium, sodium hyaluronate cross-linked polymer, and methyl dihydroxybenzoate at 85-100° C. for 15-25 minutes, keep warm after all components are dissolved and mixed, and replenish evaporated water to obtain phase B; IV. When phase A and phase B are dissolved, cool to 70-80°C, slowly add phase A into phase B, homogenize at 3000-6000 rpm for 10-20 min to obtain a mixture of phase A and phase B, cool to 40-45°C and keep warm; V. In the mixture of phase A and phase 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. Add quaternary ammonium salt-73 and homogenize at 3000-5000 rpm for 5-10 minutes to obtain an emulsion.

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