Chamomile extract based on supramolecular solvent as well as preparation method and application of chamomile extract

Abstract

The invention relates to the technical field of natural product extraction, in particular to a chamomile extract based on a supramolecular solvent and a preparation method and application of the chamomile extract. The preparation method comprises the following steps: drying and crushing chamomile to prepare chamomile powder; adding a supramolecular solvent into the chamomile powder, mixing, and carrying out ultrasonic treatment on the obtained mixture at a first temperature to prepare a chamomile extracting solution; the supramolecular solvent comprises a supramolecular receptor, a supramolecular substrate and water; the supramolecular receptor comprises organic acid; the supramolecular substrate comprises at least one of betaine and choline chloride; the first temperature is 40-80 DEG C; and carrying out solid-liquid separation on the chamomile extracting solution, and collecting liquid to prepare the chamomile extract. The chamomile extract has better anti-inflammatory and soothing effects.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of natural product extraction, in particular to chamomile extract based on supramolecular solvent and preparation method and application thereof. BACKGROUND

[0002] As a traditional medicinal and edible plant, chamomile is rich in active ingredients such as flavonoids, volatile oils, and phenolic acids, and has the effects of anti-inflammatory, fungal inhibition, and spasmolysis, and is widely used in the fields of medicine, food, and cosmetics. At present, the extraction methods of chamomile active ingredients mainly include organic solvent extraction, steam distillation, ultrasonic-assisted extraction, etc. Among them, the organic solvent extraction method is widely used due to its simple operation and low cost, but this method has the following defects: the use of organic solvents such as methanol and ethanol can cause environmental pollution and has residual risks; the active ingredients are easily oxidized and degraded during the extraction process, resulting in low extraction rate; solvent recovery is difficult, which does not meet the concept of green development.

[0003] As a new type of green extraction agent, supramolecular solvent can form a uniform system through hydrogen bonds, van der Waals forces and other non-covalent bonds, and has the advantages of low toxicity, environmental protection, recyclability, and good compatibility with active ingredients, and has been gradually applied in the field of natural plant extraction. However, there is no chamomile extraction method based on supramolecular solvent so far.

[0004] In addition, the traditional extraction method is usually optimized from the perspective of improving the extraction amount and extraction efficiency, and lacks the combination of efficacy mechanism research targeting the characteristics of chamomile. SUMMARY

[0005] Therefore, one or more embodiments of the present application provide chamomile extract based on supramolecular solvent and preparation method and application thereof, which has better anti-inflammatory and soothing efficacy.

[0006] The technical solution of the present application includes the following contents:

[0007] In a first aspect, the present application provides a preparation method of chamomile extract based on supramolecular solvent, comprising the following steps:

[0008] The chamomile is dried and crushed to obtain chamomile powder;

[0009] The supramolecular solvent is added to the chamomile powder, and the obtained mixture is ultrasonicated at a first temperature to obtain a chamomile extract solution; the composition of the supramolecular solvent includes a supramolecular acceptor and a supramolecular substrate; the supramolecular acceptor includes an organic acid; the supramolecular substrate includes at least one of betaine and choline chloride; the first temperature is 40-80℃;

[0010] The chamomile extract solution is subjected to solid-liquid separation, and the liquid is collected to prepare the chamomile extract.

[0011] Further, after the liquid is collected, the obtained liquid is concentrated to prepare a concentrated liquid.

[0012] The concentrated liquid is subjected to freeze-drying to prepare the chamomile extract.

[0013] Further, the preparation method of the supramolecular solvent comprises the following steps:

[0014] The supramolecular acceptor and the supramolecular substrate are mixed, and the obtained mixture is stirred at 50-80°C for 2-12h, and then the water is added and mixed to prepare the supramolecular solvent.

[0015] Optionally, the volume ratio of the water in the supramolecular solvent is 20%-40%.

[0016] Further, the molar ratio of the supramolecular acceptor to the supramolecular substrate is 1:(0.5-5); and / or,

[0017] The mass-volume ratio of the chamomile powder to the supramolecular solvent is 1g:10-50mL.

[0018] In some embodiments, the organic acid comprises at least one of succinic acid, mandelic acid, salicylic acid, and malic acid.

[0019] In some embodiments, the step of subjecting the chamomile extract solution to solid-liquid separation comprises centrifuging the chamomile extract solution, and then filtering the obtained supernatant; optionally, the centrifugation speed is 8000-10000rpm.

[0020] In some embodiments, the preparation method satisfies at least one of the following conditions:

[0021] (1) the ultrasonic time is 20-60min, and the first temperature is 40-60°C;

[0022] (2) the freeze-drying temperature is -70~-60°C.

[0023] In a second aspect, the present application provides a chamomile extract based on a supramolecular solvent, which is prepared by the preparation method described above.

[0024] Further, the chamomile extract based on the supramolecular solvent has the effect of targetedly inhibiting the secretion and release of the key inflammatory mediator IL-33; and / or,

[0025] The chamomile extract based on the supramolecular solvent comprises at least one of the following active ingredients: apigenin, luteolin, quercetin, rutin and naringenin.

[0026] In a third aspect, the application provides use of the chamomile extract based on the supramolecular solvent described above in the preparation of a cosmetic product with anti-inflammatory soothing effect.

[0027] Compared with the prior art, the preparation method of the application has at least the following beneficial effects:

[0028] The supramolecular solvent used in the preparation method of the application is a cosmetic-grade raw material and can be recycled, and the extraction process does not emit volatile organic pollutants, fully meeting the safety standards for cosmetics. The prepared extract can be directly used for the research and development of anti-inflammatory soothing masks, moisturizing lotions and other skin care products, has a wide range of applications and broad industrialization prospects.

[0029] The chamomile extract of the application has good inflammation inhibition and soothing effect compared with the existing chamomile extract, solving the problems of low industrialization conversion efficiency and unclear efficacy caused by the traditional natural product research of “paying more attention to extraction and less attention to mechanism”. BRIEF DESCRIPTION OF DRAWINGS

[0030] In order to more clearly illustrate the specific embodiments of the application or the technical solutions in the prior art, the drawings needed in the description of the specific embodiments or the prior art will be briefly introduced. Obviously, the drawings in the following description are some embodiments of the application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0031] Figure 1 Figure is a graph showing the influence of different extraction conditions on the extraction rate of chamomile polyphenol extract.

[0032] Figure 2 Figure is a graph showing the DPPH free radical scavenging rate determination results of betaine-amygdalic acid-chamomile extract (BMMS).

[0033] Figure 3 Figure is a graph showing the ABTS free radical scavenging rate determination results of betaine-amygdalic acid-chamomile extract (BMMS).

[0034] Figure 4 Figure is a graph showing the influence of different concentrations of BMMS extract on the activity of macrophage RAW264.7.

[0035] Figure 5 Figure is a graph showing the down-regulation rate test results of BMMS extract on inflammatory factor IL-6.

[0036] Figure 6Figure for the down-regulation rate test results of the BMMS extract on the inflammatory factor IL-33.

[0037] Figure 7 Figure for the Venn diagram of the active ingredient target points of the chamomile in Example 1 and the sensitive skin related target points, showing the intersection of the two groups of target points.

[0038] Figure 8 Figure for the protein-protein interaction (PPI) network diagram of the intersection target points of the chamomile active ingredients and sensitive skin.

[0039] Figure 9 Figure for the core target point ranking diagram obtained after topological analysis of the PPI network by Cytoscape software.

[0040] Figure 10 Figure for the KEGG pathway enrichment analysis results of the chamomile active ingredient target points.

[0041] Figure 11 Figure for the "component-target" interaction network diagram of the chamomile active ingredients and the core target points of sensitive skin.

[0042] Figure 12 Figure for the node degree ranking diagram of the 9 active ingredients in the chamomile in the "component-target" network. DETAILED DESCRIPTION

[0043] The present application will be further described in conjunction with the embodiments and examples. It should be understood that these examples are only used to illustrate the present application and not intended to limit the scope of the present application. Furthermore, it should be understood that those skilled in the art can make various modifications or changes to the present application after reading the content taught by the present application, and these equivalent forms also fall within the protection scope of the claims of the present application.

[0044] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present application belongs. The terminology used in the description of the present application herein only for the purpose of describing specific embodiments of the present application and is not intended to limit the present application.

[0045] In a first aspect, the present application provides a preparation method of chamomile extract based on supramolecular solvent, comprising the following steps:

[0046] The chamomile is dried and pulverized to obtain chamomile powder;

[0047] adding a supramolecular solvent mixture to the chamomile powder, and obtaining a chamomile extract by ultrasonic treatment of the mixture at a first temperature; the supramolecular solvent comprises a supramolecular acceptor, a supramolecular substrate, and water; the supramolecular acceptor comprises an organic acid; the supramolecular substrate comprises at least one of betaine and choline chloride; and the first temperature is 40-80℃.

[0048] performing solid-liquid separation on the chamomile extract, and obtaining a concentrated solution by concentrating the obtained liquid;

[0049] performing freeze-drying on the concentrated solution to obtain the chamomile extract.

[0050] In some preferred embodiments, the first temperature is 50℃.

[0051] Further, the molar ratio of the supramolecular acceptor to the supramolecular substrate is 1:(0.5-5), such as 1:0.5, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, 1:5, etc.

[0052] In some preferred embodiments, the molar ratio of the supramolecular acceptor to the supramolecular substrate is 1:2.

[0053] Further, the supramolecular acceptor and the supramolecular substrate are mixed to form a deep eutectic solvent (DES) after stirring at 50-80℃ (such as 50℃, 60℃, 70℃, 80℃, etc.) for 2-12h (such as 2h, 4h, 6h, 8h, 10h, 12h, etc.), and then the water is added to obtain the supramolecular solvent.

[0054] Optionally, the volume ratio of the water in the supramolecular solvent is 20-40%, such as 20%, 25%, 30%, 35%, 40%, etc.

[0055] Further, the mass-volume ratio of the chamomile powder to the supramolecular solvent is 1g:10-50mL, such as 1g:10mL, 1g:20mL, 1g:30mL, 1g:40mL, 1g:50mL, etc.

[0056] In some embodiments, the organic acid comprises at least one of succinic acid, mandelic acid, salicylic acid, and malic acid.

[0057] In some embodiments, the step of performing solid-liquid separation on the chamomile extract comprises centrifuging the chamomile extract, and then filtering the obtained supernatant; optionally, the centrifugation speed is 8000-10000rpm.

[0058] In some embodiments, the time of ultrasonic is 20-60 min, and the first temperature is 40-60℃.

[0059] In some embodiments, the solid content of the concentrated solution is 10-15%.

[0060] In some embodiments, the temperature of freeze-drying is -70-60℃.

[0061] In some preferred embodiments, the supramolecular receptor is mandelic acid, the supramolecular substrate is betaine, and the volume ratio is 1:2.

[0062] In some preferred embodiments, the mass-volume ratio of the chamomile powder and the supramolecular solvent is 1 g:40 mL.

[0063] In some preferred embodiments, the power of ultrasonic is 200 W, and the time is 30 min.

[0064] In a second aspect, the present application provides a chamomile extract based on a supramolecular solvent, which is prepared by the above-mentioned preparation method.

[0065] Further, the chamomile extract based on a supramolecular solvent has the effect of targeting inhibiting the secretion and release of the key inflammatory mediator IL-33, can block the activation and cascade amplification of the local pro-inflammatory signal pathway of the skin, and thus effectively down-regulate the intensity of the inflammatory response, indirectly improve the sensitive state of the skin from the pathological mechanism level, and exhibit excellent anti-inflammatory and soothing biological activity.

[0066] Further, the chamomile extract based on a supramolecular solvent includes at least one of the following active ingredients: apigenin, luteolin, quercetin, rutin, and naringenin. The above-mentioned ingredients have good anti-inflammatory, antioxidant, and calming soothing effects in synergistic action, and the corresponding core target points include TNF, AKT1, EGFR, STAT3, CASP3, etc., and the key pathways include the NF-κB signal pathway, the JAK-STAT signal pathway, and the PI3K-Akt signal pathway.

[0067] Further, the advantage active ingredients are apigenin, luteolin, and quercetin.

[0068] In some embodiments, the chamomile extract based on a supramolecular solvent is subjected to network pharmacology analysis:

[0069] The potential action target points of the active ingredients are predicted by the SwissTarget Prediction platform, the target efficacy related disease target points are searched by the GeneCards database, the core target points are obtained by taking the intersection of the two, and the network pharmacology analysis is performed.

[0070] The core target protein-protein interaction (PPI) network was constructed using the String database, and the KEGG pathway analysis was performed using the Metascape database to construct the "component-target-pathway" network.

[0071] Based on the above analysis results, it is found that the chamomile extract based on the supramolecular solvent has good anti-inflammatory, antioxidant and soothing effects.

[0072] In some embodiments, the chamomile extract is combined with a rose extract to prepare the cosmetic product, and the combination of the chamomile extract and the rose fermentation liquid can further enhance the anti-inflammatory effect, thereby providing valuable application potential and scientific basis for the research and development of sensitive skin anti-inflammatory soothing skin care products.

[0073] In some embodiments, the cosmetic product has at least one of anti-inflammatory, antioxidant and soothing effects.

[0074] The cosmetic product can be in the form of, but is not limited to, a water-based product, an oil-based product, a cream-based product, a surfactant-based product, a gel-based product, a paste-based product, etc. The following are some specific embodiments.

[0075] In the following specific embodiments, the experimental parameters not specified can be preferably referred to the guidelines provided in the present application, and can also be referred to the experimental manuals in the art or other known experimental methods in the art, or the experimental conditions recommended by the manufacturers.

[0076] In the following specific embodiments, the raw materials and reagents involved can be commercially available or can be prepared by those skilled in the art according to known methods.

[0077] I. Preparation of chamomile extract based on supramolecular solvent

[0078] Embodiment 1

[0079] The present embodiment provides a method for preparing a chamomile extract based on a supramolecular solvent, comprising the following steps:

[0080] S1, raw material pretreatment: dry the chamomile flower head, crush it, pass it through a 50-mesh sieve, and obtain chamomile powder;

[0081] S2, extraction: the betaine and the almond acid are mixed in a molar ratio of 1:2, heated and stirred at 80°C for 8 hours to form a deep eutectic solvent (DES), and then mixed with 30% deionized water to prepare a supramolecular solvent; 400 mL of the supramolecular solvent is added to 10 g of chamomile powder (mass-volume ratio 1 g:40 mL), and the obtained mixture is placed in an ultrasonic extractor for extraction, with the extraction temperature set at 50°C, the extraction power set at 200W, and the extraction time set at 30 min, to prepare a chamomile extract;

[0082] S3, separation and concentration: the chamomile extract is centrifuged at 10000 r / min for 15 min, and then the supernatant is vacuum filtered at 0.05 MPa for 4 min; the obtained liquid is concentrated by rotary evaporation to obtain a concentrated liquid, and the solid content of the concentrated liquid is 5%.

[0083] S4, vacuum drying: the concentrated liquid is freeze-dried at -60°C to -70°C to obtain a chamomile extract.

[0084] Example 2

[0085] The example provides a preparation method of a chamomile extract based on a supramolecular solvent, which adopts a method basically same as that of Example 1, except that the preparation method of the supramolecular solvent is as follows: the betaine and the succinic acid are mixed in a molar ratio of 1:2, heated and stirred at 80°C for 8 hours to form a deep eutectic solvent (DES), and then mixed with 30% deionized water.

[0086] Example 3

[0087] The example provides a preparation method of a chamomile extract based on a supramolecular solvent, which adopts a method basically same as that of Example 1, except that the preparation method of the supramolecular solvent is as follows: the betaine and the malic acid are mixed in a molar ratio of 1:2, heated and stirred at 80°C for 8 hours to form a deep eutectic solvent (DES), and then mixed with 30% deionized water.

[0088] Example 4

[0089] The example provides a preparation method of a chamomile extract based on a supramolecular solvent, which adopts a method basically same as that of Example 1, except that the preparation method of the supramolecular solvent is as follows: the betaine and the salicylic acid are mixed in a molar ratio of 1:2, heated and stirred at 80°C for 8 hours to form a deep eutectic solvent (DES), and then mixed with 30% deionized water.

[0090] Example 5

[0091] The embodiment provides a preparation method of chamomile extract based on a supramolecular solvent, and the method is basically same as that in the embodiment 1, and the difference is that the preparation method of the supramolecular solvent is as follows: choline chloride and almond acid are mixed according to a molar ratio of 1:2, heated and stirred at 80 DEG C for 8 hours to form a deep eutectic solvent (DES), and then 30% deionized water is added and mixed.

[0092] Embodiment 6

[0093] The embodiment provides a preparation method of chamomile extract based on a supramolecular solvent, and the method is basically same as that in the embodiment 1, and the difference is that the preparation method of the supramolecular solvent is as follows: choline chloride and almond acid are mixed according to a molar ratio of 1:2, heated and stirred at 80 DEG C for 8 hours to form a deep eutectic solvent (DES), and then 30% deionized water is added and mixed.

[0094] Embodiment 7

[0095] The embodiment provides a preparation method of chamomile extract based on a supramolecular solvent, and the method is basically same as that in the embodiment 1, and the difference is that the preparation method of the supramolecular solvent is as follows: choline chloride and almond acid are mixed according to a molar ratio of 1:2, heated and stirred at 80 DEG C for 8 hours to form a deep eutectic solvent (DES), and then 30% deionized water is added and mixed.

[0096] Embodiment 8

[0097] The embodiment provides a preparation method of chamomile extract based on a supramolecular solvent, and the method is basically same as that in the embodiment 1, and the difference is that the preparation method of the supramolecular solvent is as follows: choline chloride and almond acid are mixed according to a molar ratio of 1:2, heated and stirred at 80 DEG C for 8 hours to form a deep eutectic solvent (DES), and then 30% deionized water is added and mixed.

[0098] Embodiment 9

[0099] The embodiment provides a preparation method of chamomile extract based on a supramolecular solvent, and the method is basically same as that in the embodiment 1, and the difference is that the preparation method of the supramolecular solvent is as follows: choline chloride and almond acid are mixed according to a molar ratio of 1:2, heated and stirred at 80 DEG C for 8 hours to form a deep eutectic solvent (DES), and then 30% deionized water is added and mixed.

[0100] Embodiment 10

[0101] The embodiment provides a preparation method of chamomile extract based on a supramolecular solvent, and the method is basically same as that in the embodiment 1, and the difference is that the preparation method of the supramolecular solvent is as follows: choline chloride and almond acid are mixed according to a molar ratio of 1:2, heated and stirred at 80 DEG C for 8 hours to form a deep eutectic solvent (DES), and then 30% deionized water is added and mixed.

[0102] Example 11

[0103] The present example provides a preparation method of chamomile extract based on supramolecular solvent, which uses substantially the same method as Example 1, except that in the step of placing the obtained mixture in the ultrasonic extraction instrument for extraction, the extraction temperature is set to 40℃, the extraction power is 200W, and the extraction time is 60min.

[0104] Example 12

[0105] The present example provides a preparation method of chamomile extract based on supramolecular solvent, which uses substantially the same method as Example 1, except that in the step of placing the obtained mixture in the ultrasonic extraction instrument for extraction, the extraction temperature is set to 80℃, the extraction power is 200W, and the extraction time is 20min.

[0106] Comparative Example 1

[0107] The present comparative example provides a preparation method of chamomile extract based on supramolecular solvent, which uses substantially the same method as Example 1, except that in the step of placing the obtained mixture in the ultrasonic extraction instrument for extraction, the extraction temperature is set to 40℃, the extraction power is 200W, and the extraction time is 60min.

[0108] Comparative Example 2

[0109] The present comparative example provides a preparation method of chamomile extract based on supramolecular solvent, which uses substantially the same method as Example 1, except that in the step of placing the obtained mixture in the ultrasonic extraction instrument for extraction, the extraction temperature is set to 40℃, the extraction power is 200W, and the extraction time is 60min.

[0110] Comparative Example 3

[0111] The present comparative example provides a preparation method of chamomile extract based on supramolecular solvent, which uses substantially the same method as Example 1, except that in the step of placing the obtained mixture in the ultrasonic extraction instrument for extraction, the extraction temperature is set to 40℃, the extraction power is 200W, and the extraction time is 60min.

[0112] Comparative Example 4

[0113] The present comparative example provides a preparation method of chamomile extract based on supramolecular solvent, which uses substantially the same method as Example 1, except that in the step of placing the obtained mixture in the ultrasonic extraction instrument for extraction, the extraction temperature is set to 40℃, the extraction power is 200W, and the extraction time is 60min.

[0114] The polyphenol extraction rate and hyaluronidase are used as screening indicators.

[0115] The determination and calculation method of the extraction rate is as follows:

[0116] Gallic acid standard solution preparation: 10 mg of gallic acid analytical standard (AR≥98%) was accurately weighed into a 50 mL volumetric flask, and then distilled water was added to make up the volume. The concentration of the gallic acid standard solution was 0.2 mg / mL, and it was stored in a 4°C refrigerator in the dark. Preparation of 10% (V / V) Folin-Ciocalteu reagent: 5 mL of Folin-Ciocalteu reagent was accurately measured and added to a 100 mL volumetric flask, which was then shaken to obtain a 10% Folin-Ciocalteu reagent solution. Preparation of Na2CO3 solution (7.5%): 7.5 g of Na2CO3 solid was accurately weighed, dissolved in distilled water, and then transferred to a 100 mL volumetric flask. Distilled water was added to make up the volume, and the solution was shaken to obtain a 7.5% Na2CO3 solution. The composition of the above gallic acid standard solution is shown in Table 1.

[0117] Table 1 Composition of gallic acid standard solution

[0118]

[0119] The absorbance value was measured at 765 nm, and three parallel samples were taken for each group.

[0120] The test solution was measured by the above method. The absorbance value at 760 nm was measured in triplicate for each group. The total phenol content in the crude extract was calculated from the standard curve. The absorbance value was substituted into the regression equation to calculate the total phenol content in the sample, and the result was expressed as the number of grams of gallic acid equivalent per gram of sample, with the unit mg / g.

[0121] The gallic acid standard curve was obtained: Y=0.009510X+0.154 R 2 =0.998; Y is the absorbance, and X is the yield of polyphenol extraction.

[0122] Table 2 Comparison of extraction yields of chamomile extract with different extraction solvents

[0123]

[0124] The results are shown in Table 2. The extraction rate of betaine-amygdalic acid is the highest, followed by ethanol. Most polyphenols are moderately or weakly polar molecules. If the polarity is too high, it is difficult to effectively dissolve moderately polar polyphenols, resulting in the lowest extraction rate, for example, the extraction rate of water is only 8.54. The betaine-organic acid system forms a deep eutectic solvent (DES), which breaks the plant cell wall through hydrogen bonding and van der Waals forces, releasing polyphenols. The type of acid affects the physicochemical properties (polarity, viscosity, pH) of the DES, which in turn affects the extraction efficiency. The extraction rate of amygdalic acid is higher because it contains a benzene ring structure and has strong hydrophobicity. It can produce π-π stacking with the aromatic ring of polyphenols, enhancing solubility. In summary, the extraction scheme of Example 1 improves the extraction efficiency of target components while meeting the technical requirements of green and environmentally friendly technology. The combination of hydrogen bond donors (HBD) and hydrogen bond acceptors (HBA) will result in different types of interaction forces between deep eutectic solvents (DESs) and target components during the extraction process, which in turn affects the solubility of target components and the solubility of solvents. According to the principle of "like dissolves like", the intermolecular forces formed in Example 1 are more suitable for the solubility characteristics of polyphenolic compounds, so they exhibit better extraction performance compared to traditional ethanol solvents. Although the DES system may limit the mass transfer efficiency due to its high viscosity, amygdalic acid can effectively reduce the viscosity of the system and improve the solvent permeability due to its strong hydrophobicity. At the same time, compared to the drawbacks of ethanol such as toxicity and high volatility, the environmental compatibility and safety of DESs are more prominent, making them more suitable for application in research fields such as cosmetics, food, medicine, and other fields with high requirements for raw material safety.

[0125] Example 13

[0126] This example is a response surface test for process optimization of the preparation method of chamomile extract based on supramolecular solvents.

[0127] Using the test results in Table 2, the Design-Expert 8.0.6.1 software was used to optimize the yield of chamomile polyphenols; the three conditions that have a greater impact on the yield of chamomile polyphenols were used as influencing factors, namely the time of ultrasonic extraction (A), the ultrasonic temperature (B) and the water content (C), as shown in Table 3. This experiment was a 3-factor 3-level response surface experiment; the Box-Behnken test was designed to perform response surface analysis, and the results are shown in Table 4 and Figure 1 .

[0128] Table 3 Influence factors of response surface test for extraction process optimization

[0129]

[0130] Table 4 Orthogonal test design for extraction process optimization

[0131]

[0132] The Design-Expert 8.0.6.1 software was used to perform multiple regression fitting on the test data, and a quadratic multinomial regression equation between the chamomile polyphenol extract yield (Y) and each factor of the test was obtained: chamomile polyphenol extraction rate = -88.12 + 1.89A + 3.09B + 0.59C + 0.0080AB + 0.0055AC + 0.0061BC - 0.028A 2 -0.043B 2 -0.019C 2 .

[0133] It can be known from the response surface analysis result that the optimal extraction process conditions of the chamomile extract polyphenol of the supramolecular solvent are: ultrasonic extraction temperature 41.76℃, ultrasonic extraction time 39.58min, and water content 33.70%. Under the above optimal extraction process, the theoretical yield of chamomile polyphenol is 25.13mg / g. Considering that the actual operation of the process needs to be simple and convenient, various possible process factors are comprehensively modified to: the process principle should be: ultrasonic extraction temperature 42℃, ultrasonic extraction time 40min, and water content 33%. The yield of chamomile polyphenol is 26.41mg / g.

[0134] Antioxidant property determination:

[0135] 1.1 DPPH free radical scavenging rate determination.

[0136] a. Prepare DPPH solution (2.3mg: 50mL anhydrous ethanol) and store it in the dark. Its absorbance value at 517nm should be between 1.2-1.3. Wait for use.

[0137] b. Take a centrifuge tube, add 2mL of betaine-amygdalic acid-chamomile extract solution, and then add 2mL of DPPH solution, marked as A1.

[0138] c. Take a centrifuge tube, add 2mL of betaine-amygdalic acid-chamomile extract solution, and then add 2mL of deionized water solution, marked as A2.

[0139] d. Take a centrifuge tube, add 2mL of deionized water, and then add 2mL of DPPH solution, marked as A3.

[0140] e. Calculate the DPPH free radical scavenging capacity of the betaine-amygdalic acid-chamomile extract solution according to the following formula (I):

[0141] (I);

[0142] In formula (I), A1 is the absorbance after mixing DPPH and sample; A2 is the absorbance after mixing sample solution and deionized water; A3 is the absorbance after mixing DPPH and deionized water; and A0 is the absorbance of deionized water (one group of detection can be achieved).

[0143] The results are shown in Table 1. Figure 2 As shown in Table 1, the DPPH free radical scavenging rate (vertical coordinate, %) of betaine-amyrin-echinacea extract (BMMS) (horizontal coordinate, mg / mL) at different concentrations was determined with vitamin C (VC) as a positive control. In the concentration range of 0.1-1.0 mg / mL, the DPPH free radical scavenging rate of the BMMS extract showed a gradient rising trend with the increase of concentration, and the scavenging rate reached 97.95% at a concentration of 1.0 mg / mL. Compared with the positive control VC, the DPPH free radical scavenging ability of the BMMS extract was basically the same, indicating that it had strong free radical scavenging ability and had the potential to be developed as a natural antioxidant.

[0144] 1.2 ABTS free radical scavenging rate determination.

[0145] a. 5 mL of 7.4 mmol / L ABTS stock solution was mixed with 88 μL of 2.6 mmol / L K2S2O8, and was left to stand for 12-16 hours to prepare a working solution of ABTS. 0.4 mL of ABTS solution was taken and diluted with PBS solution. The absorbance value at 734 nm was required at room temperature.

[0146] b. A test tube was taken, 0.5 mL of BMMS extract was added, and then 2 mL of ABTS solution was added, and A1 was recorded.

[0147] c. A test tube was taken, 0.5 mL of BMMS extract was added, and then 2 mL of PBS solution was added, and A2 was recorded.

[0148] d. A test tube was taken, 0.5 mL of PBS solution was added, and then 2 mL of ABTS solution was added, and A3 was recorded.

[0149] e. The ABTS free radical scavenging ability of the BMMS solution was calculated according to formula (II):

[0150] (II) ;

[0151] In formula (II), A1 is the absorbance of the sample solution and ABTS; A2 is the absorbance of the sample solution and distilled water; A3 is the absorbance of ABTS without the sample solution; and A0 is the absorbance of distilled water.

[0152] The results are shown in Table 1. Figure 3The ABTS free radical scavenging rate (vertical coordinate, %) of different concentrations of betaine-amygdalic acid-chamomile extract (BMMS) (horizontal coordinate, mg / mL) was investigated with vitamin C (VC) as a positive control. In the concentration range of 0.1-1.0 mg / mL, the ABTS free radical scavenging rate of the BMMS extract gradually increased with the increase of the concentration, and when the concentration was 1.0 mg / mL, the scavenging rate was as high as 99.86%. Compared with the positive control VC, the ABTS free radical scavenging ability of the BMMS extract was similar to that of VC, which confirmed that the extract had good antioxidant activity.

[0153] 1.3 Macrophage RAW264.7 viability assay.

[0154] a. RAW264.7 macrophages were plated in DMEM medium in a 96-well plate and incubated in a carbon dioxide cell incubator for 24 h.

[0155] b. The supernatant of the 96-well plate was discarded, and the BMMS sample was diluted with DMEM. The diluted sample was added to the 96-well plate, which was incubated in a carbon dioxide cell incubator for 24 h.

[0156] c. The supernatant of the 96-well plate was discarded, and the viability of the RAW264.7 macrophages was detected using a CCK-8 detection kit. (RAW264.7 macrophages + diluted BMMS sample + CCK-8 was recorded as the OD value of the experimental well; RAW264.7 macrophages + CCK-8 was recorded as the OD value of the blank well; RAW264.7 macrophages + DMEM medium + CCK-8 was recorded as the OD value of the control well).

[0157] d. The BMMS solution was calculated according to the following formula:

[0158] Cell survival rate = (experimental well OD value - blank well OD value) / (control well OD value - blank well OD value) x 100%;

[0159] The results are shown in Table 1. Figure 4As shown, the BMMS extract was divided into different concentrations (abscissa, mg / mL) for the macrophage RAW264.7 survival rate test (ordinate, %). The cell viability was not significantly different from the no BMMS extract concentration in the range of 50-100 μg / mL, the cell viability remained above 100% and had a promoting effect on cell proliferation, when the BMMS extract concentration reached 200 μg / mL, there was a significant difference compared with the no BMMS extract concentration, the cell viability reached 87.88%; when the BMMS extract concentration reached 400 μg / mL, there was a significant difference compared with the no BMMS extract concentration, the cell viability reached 70.50%, showing inhibition to the cells; when the BMMS extract concentration reached 800 μg / mL, there was a very significant difference compared with the no BMMS extract concentration, the cell viability reached 57.00%, showing a significant inhibition to the cells; in summary, 200 μg / mL was selected as the maximum concentration of the BMMS extract for a series of cell experiments.

[0160] 1.4 Inflammatory factor (IL)-6 down-regulation rate test.

[0161] Inflammatory factor IL-6 test: adjust the concentration of RAW264.7 cell suspension and add to a 96-well cell plate, culture overnight at 37°C, 5% CO2 for detection. Dilute the test sample to a non-cytotoxic concentration with LPS-containing cell culture medium, then add it, use LPS-containing medium as negative control (NC), and use LPS-free cell culture medium as blank control (BC). Dilute the positive control with LPS-containing cell culture medium as positive control (PC, dexamethasone sodium phosphate). After adding the sample, continue to culture the cells for 20-24 h, then take the supernatant after the action is over, and detect it according to the IL-6 Elisa kit operation procedure.

[0162] Result calculation and determination:

[0163] Calculate the inflammatory factor IL-6 down-regulation rate according to formula (III):

[0164] Down-regulation rate = (1-T / NC) x 100% (III);

[0165] In formula (III), T - average value of inflammatory factor IL-6 in the experimental group; NC - average value of inflammatory factor IL-6 in the negative control group.

[0166] Statistically analyze the data and significance, calculate the P value, P<0.05 indicates significant difference, otherwise no statistical difference.

[0167] When the inflammatory factor IL-6 content of the negative control group is much higher than that of the blank control group, and they have a significant difference (P<0.05), it means that the test is valid.

[0168] When the sample group has lower IL-6 content than the negative control group, and the difference between them is significant (P<0.05), it is determined that the sample has soothing effect.

[0169] The results are shown in Table 1. Figure 5 As shown in Table 1, dexamethasone sodium phosphate (DSP) was used as a positive control, and its inhibition rate of IL-6, a key inflammatory mediator, was 91.34%. The test results showed that the inhibition of IL-6 release by BMMS showed a significant dose-dependent effect: as the concentration gradient of BMMS increased, the anti-inflammatory efficacy was continuously enhanced, and when the concentration reached 200 μg / mL, the inhibition rate of IL-6 was as high as 9.39%, which was highly close to the anti-inflammatory activity of the positive control DSP; when BMMS was compounded with rose fermentation broth (RFL), the synergistic anti-inflammatory effect was significantly enhanced, and the inhibition rate of IL-6 was further increased to 84.50%, and the anti-inflammatory activity was more excellent. The results fully confirmed that BMMS can effectively reduce inflammatory cell infiltration and tissue damage by targeting the expression and release of IL-6, a key effector factor of the inflammatory pathway, and blocking the initiation and amplification of the skin inflammatory cascade from the upstream, thereby improving the sensitive state of the skin from the pathological mechanism, and showing excellent anti-inflammatory soothing biological activity; and the compounding system of BMMS and rose fermentation broth can further enhance the anti-inflammatory efficacy, providing valuable application potential and scientific basis for the research and development of sensitive skin anti-inflammatory soothing skin care products.

[0170] 1.5 Inflammation factor (IL)-33 down-regulation rate test.

[0171] IL-33 (Interleukin-33) is a potent pro-inflammatory factor and an important immune-regulating cytokine of type 2 immune response. It not only plays a key pro-inflammatory role in allergic diseases (such as asthma, atopic dermatitis), autoimmune diseases (such as rheumatoid arthritis), and chronic inflammation, but also has been widely concerned for its association with the pathophysiological process of pruritus. Studies have confirmed that the specific receptor (IL-33R) of IL-33 is functionally expressed in the dorsal root ganglion (DRG) sensory neurons. IL-33 can activate sensory neurons and participate in the transmission and amplification of itch signals by directly binding to the receptor and starting the downstream signaling pathway. Based on this, the IL-33 inflammation factor test was performed on the sample of the embodiment to evaluate its inhibitory effect on IL-33-mediated excessive inflammation (such as allergic diseases and chronic inflammation) and soothing effect.

[0172] IL-33 content determination experiments were performed using RAW 264.7 cells. The cell suspension was adjusted to an appropriate concentration and inoculated in a 96-well plate, and incubated at 37°C, 5% CO2 overnight. The sample group (containing LPS and chamomile extract), negative control (containing LPS), positive control (containing tranilast and LPS), and blank control (without LPS) were set up. After 24 hours of treatment, the culture medium was slowly washed with PBS, and an appropriate amount of PBS was added for repeated freeze-thawing. Then the supernatant was aspirated, vortexed and centrifuged, and the IL-33 content was determined by double antibody sandwich ELISA: standard / sample, biotinylated detection antibody, HRP-labeled streptavidin and TMB substrate were added in turn, and the absorbance at 450 nm was measured after color development. The content was calculated according to the formula IL-33 inhibition rate = 1-(sample-blank) / (stimulation-blank) x 100%, and all experiments were performed in triplicate.

[0173] The results are shown in Figure 6 The model verification results showed that the IL-33 secretion level in the cell supernatant was stably maintained at 117.20 pg / mL after LPS repeated freeze-thawing stimulation, which clearly confirmed the success of the IL-33-mediated inflammatory cell model; the inhibition rate of the positive control drug tranilast (TLT) on IL-33 was 92.45%, which further verified that the model had good drug response sensitivity and experimental reliability, laying a solid foundation for subsequent activity evaluation. In vitro experimental data showed that within the safe concentration range, the inhibition of IL-33 release by BMMS showed a significant dose-dependent effect, with the 200 μg / mL concentration group having the optimal inhibitory activity, with an IL-33 inhibition rate of 82.89%; when BMMS was combined with rose fermentation broth (RFL), the synergistic anti-inflammatory effect was significantly enhanced, and the IL-33 inhibition rate was further increased to 85.00%, which was closer to the inhibitory efficacy of the positive control drug. The above results fully confirmed that BMMS can target the secretion and release of the key inflammatory mediator IL-33, block the activation and cascade amplification of the local skin pro-inflammatory signaling pathway, and effectively down-regulate the intensity of the inflammatory response, indirectly improving the sensitive state of the skin from the pathological mechanism level, and exhibiting excellent anti-inflammatory soothing biological activity; and the combination of BMMS with rose fermentation broth can further enhance the anti-inflammatory efficacy, providing valuable application potential and scientific basis for the research and development of sensitive skin anti-inflammatory soothing skin care products.

[0174] II. Application of chamomile extract.

[0175] In order to further study the mechanism of chamomile in relieving skin sensitivity, network pharmacology analysis was performed.

[0176] 1. Network pharmacology method of chamomile active extract

[0177] (1) Screening of chamomile extract components and prediction of active targets.

[0178] Using database and literature retrieval, 9 relevant potential active ingredients were screened out as shown in Table 5 below.

[0179] Table 5 Active ingredients of chamomile extract

[0180]

[0181] After obtaining the structure information of the active ingredients of chamomile, the structure diagram of the compounds was converted into a format supported by the SwissTargetPrediction target prediction platform (http: / / swisstargetprediction.ch / ), and then uploaded to the platform to carry out prediction analysis of the potential action targets of the active ingredients. In the operation of the database, the species parameter was set as "human" to ensure the relevance of the predicted targets to the physiological and pathological processes of the human body and to avoid interference of non-target species targets in subsequent analysis. The obtained targets were integrated, and duplicate target entries were removed by de-duplication processing, and finally the core target set of chamomile active ingredients was obtained, laying a foundation for subsequent target-related analysis.

[0182] (2) Prediction of active ingredients of chamomile and sensitive skin related targets.

[0183] In order to screen the target genes related to sensitive skin disease, the human gene comprehensive database GeneCards (https: / / www.genecards.org / ) was used to carry out retrieval and screening of the disease related targets. During the retrieval process, "Sensitive skin" was used as the core keyword, and the retrieval species was set as "human" to ensure the relevance of the targets to the pathological process of the human body, and finally a database of sensitive skin disease related targets was constructed, laying a foundation for subsequent intersection analysis of disease targets and active ingredient targets.

[0184] In order to screen the common action targets of chamomile active ingredients and sensitive skin disease, the Venn diagram analysis was carried out using the online analysis platform Venny (http: / / www.liuxiaoyuyuan.cn / ), which can directly show the overlapping relationship between different data sets. Through the visualization result, the intersection targets of chamomile active ingredients and sensitive skin disease were confirmed, which can be preliminarily identified as the potential core action targets of chamomile in the intervention of sensitive skin, providing key research objects for subsequent functional enrichment analysis and mechanism analysis.

[0185] (3) PPI construction of chamomile active ingredient-sensitive skin targets, core ingredient target screening, signal pathway analysis.

[0186] To analyze the interaction between active ingredients of chamomile and the intersection target points of sensitive skin diseases, the protein-protein interaction (PPI) network analysis of the intersection target points was carried out by setting the species as "human" in the String database (https: / / cn.string-db.org / ). The results were uploaded to Cytoscape 3.9.1 software for data visualization and in-depth analysis.

[0187] To systematically analyze the biological function and signal pathway association of potential target points, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed on the potential target points obtained by screening using the Metascape database (https: / / metascape.org / gp / index.html# / main / step1).

[0188] To intuitively present the action association between active ingredients of chamomile and core target points of sensitive skin, the core target points of sensitive skin obtained by previous screening were integrated with the information of active ingredients of chamomile, and then imported into Cytoscape 3.9.1 software. The "potential component-disease core target point" visualization network diagram for sensitive skin and skin inflammation was generated.

[0189] 2. Network pharmacology results analysis of chamomile extract

[0190] (1) Prediction of active target points of chamomile extract components and sensitive skin disease target points

[0191] Through the SwissTarget Prediction target prediction platform, 270 potential action target points of active ingredients of chamomile were obtained after screening and integration. At the same time, 1814 target points related to sensitive skin (Sensitive skin) were retrieved and screened from the GeneCards database. The two groups of target points were compared by Venn diagram analysis, and the comparison results are shown in Figure 7 , it was found that there were 130 common target points between the target points of active ingredients of chamomile and the target points related to sensitive skin. Then the intersection target points were imported into the STRING database, and the species was limited to "Homo sapiens", and the free protein nodes (no interacting proteins) were removed. The protein interaction network data after screening was exported, and Cytoscape 3.9.1 software was used for network topology analysis and visualization processing, and the processing results are shown in Figure 8 , the importance of target points in the network was sorted by software calculation of node degree value, intermediate centrality and other topological parameters, Figure 9The core target point ranking chart obtained by topological analysis of PPI network through Cytoscape software. The top-ranking core target points are AKT1, TNF, EGFR, BCL-2, and STAT3. These target points can be preliminarily identified as the key role target points of active ingredients of chamomile in regulating sensitive skin.

[0192] (2) Core ingredient target point screening and signal pathway analysis of active ingredients of chamomile-skin target points.

[0193] Sensitive skin is a complex pathological state involving multiple biological processes such as inflammatory response, cell signal transduction, and metabolic regulation. Its clinical manifestations have both obvious subjective feelings and objective signs. Subjectively, the core performances are skin tightness, stinging, transient burning, and persistent itching. Objectively, it is accompanied by typical symptoms such as capillary dilation, flushing, erythema, desquamation, and dryness. The occurrence and development of this pathological state are regulated by internal and external factors. It is not only easily aggravated by external stimuli such as temperature fluctuations, ultraviolet radiation, and environmental dust, but also can be induced by the interaction of internal factors such as abnormal activation of inflammatory response, accumulation of psychological stress, and imbalance of skin microbial flora. The synergistic superposition of the above factors often leads sensitive skin to fall into a vicious cycle of repeated aggravation. To clarify the potential mechanism of chamomile in soothing sensitive skin, KEGG pathway enrichment analysis was performed, and the analysis results are as follows Figure 10 It can be seen that chamomile may play a role by regulating key signal pathways through multiple targets. A total of 185 significantly enriched signal pathways (P<0.05) were identified, including PI3K-AKT signaling pathway, NF-κB signaling pathway, JAK-STAT signaling pathway, and other core pathways closely related to inflammation regulation and cell function, providing important molecular biological basis for further elucidating the soothing mechanism of chamomile.

[0194] With the help of Cytoscape3.9.1 software, a component-target interaction network related to sensitive skin was constructed, which vividly displayed the complex interaction between each component and its corresponding target, thus forming a complete network structure. By constructing the compound-target interaction network, the interaction between 9 active ingredients and the core target points related to sensitive skin was analyzed, and the analysis results are shown in Figure 11 In this visual network, blue nodes represent active compounds, green nodes mark key target proteins, and connecting lines directly show the specific binding relationship between compounds and target points. Network topological analysis reveals that these phytochemicals may regulate sensitive skin response through multi-target synergistic action. The degree of each compound node in the network was calculated by Cytoscape3.9.1 software. The higher the node degree, the more target points the compound can act on, and the more critical the regulatory role in the network. Figure 12The "component-target" interaction network diagram of active ingredients of chamomile and core target points of sensitive skin. According to the degree ranking results, the order of the 9 active ingredients is 7-methoxycoumarin, apigenin, luteolin, quercetin, naringenin, caffeic acid, apigenin-7-o-glucoside, rutin and coumarin. Among them, 7-methoxycoumarin, apigenin, luteolin and quercetin have the highest node degree and can act on the most sensitive skin related target points, so they can be identified as the core active ingredients for regulating sensitive skin.

[0195] Based on the above result analysis, it can be concluded that chamomile may play a role in relieving skin sensitivity, which is due to polyphenolic compounds.

[0196] Each technical feature of the above-described embodiments and examples can be combined in any suitable manner. In order to make the description concise, not all possible combinations of the technical features in the above-described embodiments and examples are described, but it should be understood that any combination of these technical features that does not contradict should be considered within the scope of the present disclosure.

[0197] The above-described embodiments only express several embodiments of the present application, but should not be construed as limiting the scope of the patent application. It should be noted that for those skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of the present application. In addition, it should be understood that after reading the above teachings of the present application, those skilled in the art can make various changes or modifications to the present application, and the equivalent forms also fall within the scope of the present application. It should also be understood that those skilled in the art can obtain technical solutions based on the technical solutions provided in the present application through logical analysis, reasoning or limited experiments, which are within the scope of the appended claims of the present application. Therefore, the scope of the patent of the present application should be subject to the appended claims, and the description can be used to explain the content of the claims.

Claims

1. A process for the preparation of an extract of chamomile based on supramolecular solvents, characterized in that, The method comprises the following steps: drying and pulverizing the chamomile to obtain chamomile powder; adding a supramolecular solvent to the chamomile powder, and ultrasonically treating the mixture at a first temperature to obtain a chamomile extract; the supramolecular solvent comprises a supramolecular acceptor, a supramolecular substrate and water; the supramolecular acceptor comprises an organic acid; the supramolecular substrate comprises at least one of betaine and choline chloride; and the first temperature is 40-80℃; performing solid-liquid separation on the chamomile extract to collect a liquid to obtain the chamomile extract.

2. The production method according to claim 1, characterized by, After the liquid is collected, the liquid is concentrated to obtain a concentrated liquid; the concentrated liquid is freeze-dried to obtain the chamomile extract.

3. The production method according to claim 1 or 2, characterized by, The method for preparing the supramolecular solvent comprises the following steps: mixing the supramolecular acceptor and the supramolecular substrate, and then adding water to the mixture after stirring at 50-80℃ for 2-12h to obtain the supramolecular solvent; the volume fraction of the water in the supramolecular solvent is 20-40%.

4. The production method according to claim 1, characterized by, The mass-volume ratio of the chamomile powder and the supramolecular solvent is 1g:10-50mL; and / or, the molar ratio of the supramolecular acceptor and the supramolecular substrate is 1:(0.5-5).

5. The method of any one of claims 1-2, 4, wherein, The organic acid comprises at least one of succinic acid, mandelic acid, salicylic acid and malic acid.

6. The method of any one of claims 1-2, 4, wherein, The step of performing solid-liquid separation on the chamomile extract comprises centrifuging the chamomile extract, and then filtering the supernatant; optionally, the centrifugal speed is 8000-10000rpm.

7. The method of any one of claims 1-2, 4, wherein, At least one of the following conditions is met: (1) the ultrasonic treatment time is 20-60min, and the first temperature is 40-60℃; (2) the freeze-drying temperature is -70℃ to -60℃.

8. An extract of chamomile based on supramolecular solvents, characterized in that, The chamomile extract prepared by the method of any one of claims 1-7 has the effect of targetedly inhibiting the secretion and release of the key inflammatory mediator IL-33; and / or, 9. The supramolecular solvent-based chamomile extract according to claim 8, characterized in that, The chamomile extract based on the supramolecular solvent comprises at least one of the following active ingredients: apigenin, luteolin, quercetin, rutin and naringenin.

10. The chamomile extract based on the supramolecular solvent of claim 8 or 9 for use in the preparation of a cosmetic product having anti-inflammatory soothing efficacy. ​

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