Magnolia liliiflora composite extract and application thereof
The purple magnolia composite extract prepared through the dual extraction process solves the problem of insufficient utilization of the active ingredients of purple magnolia in the prior art, realizes multi-efficacy synergy, and significantly improves the safety and skin care effect of the extract.
Patent Information
- Application Number
- CN202510029778.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-08
- Publication Date
- 2025-05-06
AI Technical Summary
The prior art is difficult to fully utilize the active ingredients in purple magnolia, resulting in the inability to fully exert its anti-inflammatory, antibacterial, anti-allergic effects, and it is difficult to achieve the synergistic effect of multiple functions in a single extraction method.
The double extraction process was used to prepare the purple magnolia composite extract. The first purple magnolia extract was obtained by pH adjustment and ultrasonic extraction, and the second purple magnolia extract was obtained by reflux extraction and separation and purification of macroporous resin. The mass ratio of the two was 1:9-9:1, which significantly improved the safety and efficacy of the extract.
It has achieved more effective utilization of purple magnolia raw materials, significantly improved the safety of the end product and anti-inflammatory, oil control, delayed skin aging, and inhibited scars, so as to achieve skin care effects in multiple aspects and dimensions.
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Figure CN119925225A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of plant extracts and relates to a magnolia odorata composite extract and application thereof. Background Art
[0002] In the field of sensitive skin, skin inflammation has always been a focus of attention. As a complex physiological reaction, skin inflammation is mainly induced by a variety of internal and external factors such as microbial infection, allergic reaction and autoimmune disorder. Once skin inflammation enters a long-term or chronic adverse state, it will lead to impaired skin barrier function, damaged stratum corneum structural integrity, and increased skin sensitivity and fragility. In this state, the skin's defense ability decreases, thereby inducing problems such as acne, redness, inflammatory redness and skin aging. In addition, inflammation inhibits the skin's repair function, affects the wound healing process, and leads to scar formation.
[0003] Purple magnolia (Magnolia liliflora) belongs to the genus Magnolia of the Magnoliaceae family. It contains volatile oils, alkaloids, flavonoids, lignans and other ingredients, and has anti-inflammatory, antibacterial and anti-allergic effects. At present, there are few studies on the efficacy of purple magnolia buds in the direction of cosmetics. The active ingredients in purple magnolia have different solubility and activity, but a single extraction method is difficult to fully obtain its active ingredients, resulting in a single activity and failure to fully exert its multi-functional effects. Summary of the invention
[0004] In view of the deficiencies in the prior art, the object of the present invention is to provide a purple magnolia compound extract and its application, specifically to provide a purple magnolia compound extract and its application with anti-inflammatory and soothing, oil control, delaying skin aging, and inhibiting skin scar effects.
[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:
[0006] In a first aspect, the present invention provides a magnolia violet composite extract, wherein the magnolia violet composite extract consists of a first magnolia violet extract and a second magnolia violet extract;
[0007] The first purple magnolia extract is prepared by a preparation method comprising the following steps:
[0008] (S1) mixing the purple magnolia raw material with a 10-30% (e.g., 10%, 12%, 15%, 18%, 20%, 22%, 25%, 27%, 28%, 30%, etc.) ethanol aqueous solution containing an alkali at pH = 9-11 (e.g., pH = 9, pH = 9.5, pH = 10, pH = 10.5, pH = 11, etc.) and then soaking the mixture to obtain a soaking product;
[0009] (S2) soaking the product and performing ultrasonic extraction to obtain an extract;
[0010] (S3) filtering the extract, concentrating and drying the filtrate to obtain the first purple magnolia extract;
[0011] The second purple magnolia extract is prepared by a preparation method comprising the following steps:
[0012] (P1) mixing the purple magnolia raw material with 85-95% (e.g., 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 95%, etc.) ethanol aqueous solution and performing reflux extraction to obtain an extract;
[0013] (P2) filtering the extract, and subjecting the filtrate to separation and purification on a macroporous resin to obtain an eluent;
[0014] (P3) The eluate is concentrated and dried to obtain the second Magnolia grandiflora extract.
[0015] The purple magnolia extract complex developed by the present invention is composed of a first purple magnolia extract and a second purple magnolia extract, which not only realizes more effective utilization of purple magnolia raw materials, but also significantly improves the safety of the final product and its efficacy in anti-inflammatory and soothing, oil control, delaying anti-aging and inhibiting scars. Specifically, it is embodied in inhibiting the expression of inflammatory genes, lipase activity, elastase and scar gene expression, achieving skin care effects in multiple aspects and dimensions.
[0016] Preferably, the mass ratio of the first Magnolia violeta extract to the second Magnolia violeta extract is 1:9-9:1, such as 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, etc. More preferably, it is 1:5-4:3.
[0017] When the aforementioned first magnolia extract and the second magnolia extract are combined in a mass ratio of 1:9-9:1, more preferably 1:5-4:3, the product has better effects in anti-inflammatory and soothing, oil control, delaying skin aging, and inhibiting scars.
[0018] Preferably, the purple magnolia raw material is purple magnolia buds.
[0019] Preferably, the liquid ratio of the purple magnolia raw material to the 10-30% ethanol aqueous solution containing alkali in step (S1) is 1:(10-20) g / mL, for example 1:10 g / mL, 1:12 g / mL, 1:15 g / mL, 1:18 g / mL, 1:20 g / mL, etc.
[0020] Preferably, the solid-liquid ratio of the purple magnolia raw material to the 85-95% ethanol aqueous solution in step (P1) is 1:(7-10) g / mL, such as 1:7 g / mL, 1:8 g / mL, 1:9 g / mL, 1:10 g / mL, etc.
[0021] Preferably, the base in the ethanol aqueous solution in step (S1) comprises any one of ammonia water, sodium hydroxide, potassium hydroxide, sodium carbonate or arginine, or a combination of at least two thereof.
[0022] Preferably, the soaking in step (S1) is carried out at 30-45°C (e.g., 30°C, 35°C, 38°C, 40°C, 45°C, etc.) for 1-3h (e.g., 1h, 1.5h, 2h, 2.5h, 3h, etc.).
[0023] Preferably, the ultrasonic extraction in step (S2) is carried out at 40-50°C (for example, 40°C, 42°C, 44°C, 45°C, 46°C, 48°C, 50°C, etc.) for 1-4h (for example, 1h, 2h, 3h, 4h, etc.), and the ultrasonic power is 80-200W (for example, 80W, 100W, 120W, 150W, 180W, 200W, etc.).
[0024] Preferably, the reflux extraction in step (P1) is performed 1-3 times, for example, 1 time, 2 times or 3 times; each time is 1-3 hours, for example, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, etc.
[0025] Preferably, the macroporous resin in step (P2) comprises AB-8 type or D101 type.
[0026] Preferably, the separation and purification method in step (P2) is as follows: The separation and purification method in step (P2) is as follows: The filtrate is eluted with the activated macroporous resin column in sequence with water, 65-75% (for example, 65%, 67%, 68%, 70%, 73%, 75%, etc.) ethanol aqueous solution, and 90-95% (for example, 90%, 91%, 92%, 93%, 94%, 95%, etc.) ethanol aqueous solution, and the eluate of 90-95% ethanol aqueous solution is collected.
[0027] When preparing the second purple magnolia extract, the degree of elution and purification thereof is adopted as above so that the obtained product can have better effects in anti-inflammatory and soothing, oil control, delaying skin aging and inhibiting scars.
[0028] In a second aspect, the present invention provides a use of the purple magnolia composite extract according to the first aspect in preparing a product having any one or at least two of the following effects;
[0029] 1) Oil control; 2) Delay skin aging; 3) Inhibit scars.
[0030] In a third aspect, the present invention provides a use of the purple magnolia composite extract according to the first aspect in the preparation of an SDF-1 (Stromal-cell-derived factor-1) gene expression inhibitor, a CRLF2 (Cytokine Receptor-LikeFactor 2) gene expression inhibitor, or a CCR1 (CC Chemokine Receptor Type 1) gene expression inhibitor.
[0031] The SDF-1 gene encodes matrix-derived factor-1, which causes damaged keratinocytes to promote scar formation. Inhibiting it can promote tissue regeneration and reduce scarring of injured skin.
[0032] The CRLF2 gene encodes the thymic stromal lymphopoietin receptor, which binds to TSLP and causes immune cells to secrete pro-inflammatory factors, thereby triggering allergic inflammation. Regulating it can alleviate allergic inflammation and maintain skin health.
[0033] The protein encoded by the CCR1 gene promotes the recruitment and migration of inflammatory cells by recognizing CC chemokines such as MCP-3 and RANTES. By regulating the expression of CCR1, the infiltration of inflammatory cells is reduced and the spread of inflammatory response is inhibited.
[0034] The purple magnolia compound extract developed by the present invention can simultaneously block the SDF-1 gene, the CRLF2 gene, or the CCR1 gene, that is, simultaneously reduce allergic inflammation, reduce inflammatory cell infiltration and scar formation, thereby reducing skin inflammation and repairing tissue damage.
[0035] Compared with the prior art, the present invention has the following beneficial effects:
[0036] The purple magnolia composite extract developed by the present invention is composed of a first purple magnolia extract prepared by the above-mentioned specific preparation process and a second purple magnolia extract prepared by the above-mentioned specific preparation process. It not only realizes more effective utilization of purple magnolia raw materials, but also significantly improves the safety of the final product. The two extracts have different core functional components and can cooperate with each other in anti-inflammatory and soothing, oil control, delaying skin aging, and inhibiting scars. Compared with the first purple magnolia extract alone or the second purple magnolia extract alone, the composite extract has significantly improved effects in the above aspects. BRIEF DESCRIPTION OF THE DRAWINGS
[0037] Figure 1 This is the result diagram of the effect of Magnolia grandiflora compound extract on the expression of Crlf2 gene in HaCaT cells;
[0038] Figure 2 This is the result diagram of the effect of Magnolia grandiflora compound extract on the CCR1 gene expression of HaCaT cells;
[0039] Figure 3 This is a graph showing the effect of the compound extract of Magnolia grandiflora on the expression of SDF-1 gene in HaCaT cells. DETAILED DESCRIPTION
[0040] The technical solution of the present invention is further described below by specific implementation methods. It should be understood by those skilled in the art that the embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.
[0041] Preparation Example 1-1
[0042] This preparation example provides a magnolia extract, and the preparation method is as follows:
[0043] (S1) mixing a 60-mesh crushed product of purple magnolia buds with a 30% ethanol aqueous solution adjusted to pH=10 with a NaOH solution at a solid-liquid ratio of 1:15 g / mL, and soaking the mixture at 40° C. for 1 hour to obtain an soaking product;
[0044] (S2) the soaked product was subjected to ultrasonic extraction at 50° C. for 2 h with a power of 120 W to obtain an extract;
[0045] (S3) filtering the extract, concentrating and drying the filtrate to obtain the first purple magnolia extract.
[0046] Preparation Example 1-2
[0047] This preparation example provides a magnolia extract, and the preparation method is as follows:
[0048] (S1) mixing 80-mesh crushed material of purple magnolia buds with a 10% ethanol aqueous solution adjusted to pH 9 with a sodium carbonate solution at a solid-liquid ratio of 1:10 g / mL, and soaking the mixture at 30° C. for 2 h to obtain an soaking product;
[0049] (S2) the soaked product was subjected to ultrasonic extraction at 40° C. for 3 h with a power of 80 W to obtain an extract;
[0050] (S3) filtering the extract, concentrating and drying the filtrate to obtain the first purple magnolia extract.
[0051] Preparation Example 1-3
[0052] This preparation example provides a magnolia extract, and the preparation method is as follows:
[0053] (S1) mixing a 50-mesh crushed product of purple magnolia buds with a 20% ethanol aqueous solution adjusted to pH 9 with an arginine solution at a solid-liquid ratio of 1:20 g / mL, and soaking the mixture at 45° C. for 1 hour to obtain an soaking product;
[0054] (S2) the soaked product was subjected to ultrasonic extraction at 50° C. for 1 h with a power of 150 W to obtain an extract;
[0055] (S3) filtering the extract, concentrating and drying the filtrate to obtain the first purple magnolia extract.
[0056] Preparation Example 1-4
[0057] This preparation example provides a magnolia extract, and the preparation method is different from that of Preparation Example 1-1 in that the pH value of the 30% ethanol aqueous solution is not adjusted, and the 30% ethanol aqueous solution is directly mixed with the crushed material. Other conditions remain unchanged.
[0058] Preparation Example 1-5
[0059] This preparation example provides a magnolia extract, and the preparation method is different from that of Preparation Example 1-1 in that an equal amount of 30% ethanol aqueous solution adjusted to pH=10 with NaOH solution is replaced with 30% ethanol aqueous solution adjusted to pH=3 with citric acid solution. Other conditions remain unchanged.
[0060] Preparation Example 1-6
[0061] This preparation example provides a magnolia extract, and the preparation method is different from that of Preparation Example 1-1 only in that an equal volume of 30% ethanol aqueous solution is replaced with water in step (S1), and other conditions remain unchanged.
[0062] The comparison of the preparation process parameters of Preparation Example 1-1 to Preparation Example 1-6 is shown in Table 1:
[0063] Table 1
[0064]
[0065] Preparation Example 2-1
[0066] This preparation example provides a magnolia extract, and the preparation method is as follows:
[0067] (P1) Mixing 60-mesh crushed material of purple magnolia buds with 95% ethanol aqueous solution at a solid-liquid ratio of 1:8 g / mL, and performing reflux extraction twice, each time for 2 hours, to obtain an extract;
[0068] (P2) After filtering the extract, eluting with AB-8 macroporous resin in sequence with water, 70% ethanol aqueous solution, and 95% ethanol aqueous solution, 10 BV each time, and collecting the eluate with 95% ethanol aqueous solution;
[0069] (P3) The eluate is concentrated and dried to obtain a second magnolia extract.
[0070] Preparation Example 2-2
[0071] This preparation example provides a magnolia extract, and the preparation method is as follows:
[0072] (P1) Mixing 60-mesh crushed material of purple magnolia buds with 85% ethanol aqueous solution at a solid-liquid ratio of 1:10 g / mL, and performing reflux extraction 3 times, each time for 1 hour, to obtain an extract;
[0073] (P2) After filtering the extract, the extract was eluted with water, 65% ethanol aqueous solution, and 90% ethanol aqueous solution using D101 macroporous resin, with 6 BV of elution each time, and the eluate of 90% ethanol aqueous solution was collected;
[0074] (P3) The eluate is concentrated and dried to obtain a second magnolia extract.
[0075] Preparation Example 2-3
[0076] This preparation example provides a magnolia extract, and the preparation method is as follows:
[0077] (P1) Mixing a 50-mesh crushed product of purple magnolia buds with a 95% ethanol aqueous solution at a solid-liquid ratio of 1:7 g / mL, and performing reflux extraction twice, each time for 2 hours, to obtain an extract;
[0078] (P2) After filtering the extract, the extract was eluted with water, 75% ethanol aqueous solution, and 95% ethanol aqueous solution using D101 macroporous resin, with 8 BV of elution each time, and the eluate of 95% ethanol aqueous solution was collected;
[0079] (P3) The eluate is concentrated and dried to obtain a second magnolia extract.
[0080] Preparation Example 2-4
[0081] This preparation example provides a magnolia extract, and the preparation method is different from that of Preparation Example 2-1 only in the elution procedure: (P2) eluting with water, 30% ethanol aqueous solution, and 95% ethanol aqueous solution in sequence, eluting 10 BV each time, and collecting the eluate of 95% ethanol aqueous solution. Other conditions remain unchanged.
[0082] Preparation Example 2-5
[0083] This preparation example provides a magnolia extract, and the preparation method is different from that of Preparation Example 2-1 only in the elution procedure: (P2) eluting with water, 50% ethanol aqueous solution, and 95% ethanol aqueous solution in sequence, eluting 10 BV each time, and collecting the eluate of 95% ethanol aqueous solution. Other conditions remain unchanged.
[0084] The comparison of the preparation process parameters of Preparation Example 2-1 to Preparation Example 2-5 is shown in Table 2:
[0085] Table 2
[0086]
[0087] Example 1
[0088] This embodiment provides a composite extract of Magnolia purpurogena, which is obtained by mixing 10 parts of the Magnolia purpurogena extract of Preparation Example 1-1 and 40 parts of the Magnolia purpurogena extract of Preparation Example 2-1.
[0089] Example 2
[0090] This embodiment provides a composite extract of Magnolia purpurogena, which is obtained by mixing 30 parts of the Magnolia purpurogena extract of Preparation Example 1-2 and 70 parts of the Magnolia purpurogena extract of Preparation Example 2-2.
[0091] Example 3
[0092] This embodiment provides a composite extract of Magnolia purpurogena, which is obtained by mixing 50 parts of the Magnolia purpurogena extract of Preparation Examples 1-3 and 50 parts of the Magnolia purpurogena extract of Preparation Examples 2-3.
[0093] Example 4
[0094] This embodiment provides a composite extract of Magnolia purpurogena, which is obtained by mixing 10 parts of the Magnolia purpurogena extract of Preparation Example 1-1 and 90 parts of the Magnolia purpurogena extract of Preparation Example 2-1.
[0095] Example 5
[0096] This embodiment provides a composite extract of Magnolia purpurogena, which is obtained by mixing 80 parts of the Magnolia purpurogena extract of Preparation Example 1-1 and 20 parts of the Magnolia purpurogena extract of Preparation Example 2-1.
[0097] Embodiment 6-7
[0098] This example provides two magnolia odorata composite extracts, which differ from Example 1 only in that the magnolia odorata extract of Preparation Example 2-1 is replaced by the magnolia odorata extracts of Preparation Examples 2-4 to 2-5 in equal amounts, respectively.
[0099] Comparative Examples 1-3
[0100] This comparative example provides three kinds of magnolia odorata composite extracts, which differ from Example 1 only in that the magnolia odorata extract of Preparation Example 1-1 is replaced by the magnolia odorata extracts of Preparation Examples 1-4 to 1-6 in equal amounts, respectively.
[0101] Comparative Examples 4-5
[0102] The first purple magnolia extract prepared in Preparation Example 1-1 and the second purple magnolia extract prepared in Preparation Example 2-1 were separately prepared.
[0103] Test Example 1
[0104] Experiment on the effect of lipase inhibition:
[0105] The PBS dilutions (4 mg / mL) prepared in Examples 1-7 and Comparative Examples 1-5 and the pancreatic lipase working solution were added to a 96-well plate, pre-incubated at 37°C for 10 min, and then 4-nitrophenyl palmitate was added. After reacting for 30 min, the absorbance at 405 nm was measured. The blank control group was added with PBS solution without sample. The oil control effect of each group of samples on the skin was evaluated by the inhibitory effect of each group of samples on the enzyme. The results are shown in Table 3.
[0106] Test Example 2
[0107] Experiment on the effect on DPPH free radical scavenging activity:
[0108] The 96-well plate was divided into a sample group, a positive control group, and a blank control group, with 2 replicate wells in each group; the diluent (4 mg / mL) prepared in Example 1-7 and Comparative Example 1-5 was added to the DPPH solution in an equal volume of 1:1, with 150 mL added to each well, and the vitamin C solution was used as a positive control sample; the plate was placed in a dark environment, and the reaction was carried out at room temperature in a dark environment for 30 minutes, and the OD value was measured at 517 nm. The DPPH scavenging rate of the purple magnolia complex test according to Example 1-7 and Comparative Example 1-5 was determined, and the IC of DPPH free radical scavenging of each group of samples was calculated. 50 The results are shown in Table 3.
[0109] Test Example 3
[0110] Effects on collagenase activity:
[0111] This method uses N-[3-(2-furanyl) acryloyl]-L-leucine-glycine-L-proline-L-alanine (also known as FALGPA) as a substrate, selects type I collagenase from Bacillus histolyticus as a reaction enzyme, and evaluates the anti-aging effect of each group of samples on the skin by examining the inhibitory ability of Examples 1-7 and Comparative Examples 1-5 at 4 mg / mL on the enzyme. The results are shown in Table 3.
[0112] Table 3
[0113]
[0114]
[0115] As shown in Table 3, the purple magnolia extract complex involved in the present invention has excellent effects of inhibiting lipase, DPPH and collagenase, indicating that it has the effects of delaying skin aging and controlling oil. Compared with Comparative Examples 4 and 5, the purple magnolia extract complex involved in the present invention has significantly improved the above effects, indicating that the first purple magnolia extract and the second purple magnolia extract in the extract complex have obvious synergistic promotion effects on the above effects. And the mass ratio of the first purple magnolia extract and the second purple magnolia extract, and the preparation process of the first purple magnolia extract and the second purple magnolia extract also affect the above effects to a certain extent.
[0116] Test Example 4
[0117] Effects on key genes for skin inflammation and skin scarring:
[0118] (1) Preparation of test samples: The samples prepared in Example 1 and Comparative Examples 4-5 were diluted with culture medium and set aside.
[0119] (2) Cultivating P. acnes: Cultivate P. acnes (ATCC6919) of the P2 generation to the logarithmic growth phase, adjust the bacterial solution to a McFarland turbidity of 0.5, add 2% glycerol to the bacterial solution, and continue culturing for 3 days. Centrifuge after the end of the culture, take the supernatant and record its volume. After freeze-drying the supernatant, concentrate it at a ratio of 4:1;
[0120] (3) Treatment of HaCaT cells: HaCaT cells were digested with trypsin to prepare a single-cell suspension. The cells were plated at 1.5 × 10 5 The cells were inoculated in a 12-well plate at a density of 1.50 μg / mL and randomly divided into a blank group (BC), a model group (NC, using concentrated bacterial solution), and an experimental group (120 μg / mL of samples in different groups + concentrated bacterial solution), with 3 replicate wells in each group.
[0121] (4) Cultivation and sample loading: After 24 hours of cultivation, the complete medium was replaced with a 0.4% incomplete medium, and the sample loading was set according to step (3). The concentrated bacterial solution was added at a final concentration of 5%, and the sample solution was added at a final concentration of 120 μg / mL to treat the blank group, model group, and experimental group respectively;
[0122] (5) qRT-PCR assay: After 6 hours of continued culture, cells were collected for qRT-PCR assay;
[0123] (6) RNA extraction: Thermo Scientific TM GeneJET RNA purification kit was used to extract RNA, which was then reverse transcribed into cDNA;
[0124] (7) Gene quantification: using Thermo Scientific TMSYBR Green qPCR kit was used to quantify the SDF-1 gene, Crlf2 gene, and CCR1 gene. Figure 1-Figure 3 shown.
[0125] Depend on Figure 1 The results show that compared with the model group, the expression of CRLF2 gene in cells was downregulated after the sample was treated in Example 1, indicating that the purple magnolia compound extract involved in the present invention can regulate skin inflammation and allergic reactions and maintain skin health. Figure 2 The results show that compared with the model group, after the sample in Example 1 was treated, the expression of the CCR1 gene in the cells was downregulated, indicating that the compound extract of Magnolia purpurogena involved in the present invention can reduce the infiltration of inflammatory cells and inhibit the spread of inflammatory reactions. That is, the compound extract of Magnolia purpurogena involved in the present invention can relieve allergic inflammation and inhibit the infiltration of inflammatory cells, thereby relieving the further spread of skin inflammation, and achieving high-efficiency anti-inflammatory in multiple aspects and dimensions.
[0126] Depend on Figure 3 The results show that, compared with the model group, the sample of Example 1 can inhibit the expression of the cell SDF-1 gene after treating the cells, indicating that the purple magnolia compound extract of the present invention can promote tissue regeneration and inhibit the formation of skin scars.
[0127] Test Example 5
[0128] Skin sensitization test of Magnolia grandiflora compound extract:
[0129] With reference to the OECD 442 (E) standard method, THP-1 was first selected and cultured with RPMI1640 medium containing a specific ratio of fetal bovine serum and penicillin-streptomycin. Cells in the logarithmic growth phase were inoculated at a density of 1×106 cells / mL, and phorbol ester (PMA) was added to induce for 48 hours to differentiate them into a dendritic cell-like phenotype. Then, after washing with serum-free medium, the medium containing 2% FBS was replaced, and group treatment was performed. A positive control group (with 4 μg / mL 2,4-dinitrochlorobenzene), a negative control group (with no allergenic substance), and a test substance group (60 μg / mL Example 1 sample) were set up, and duplicate wells were set up for a total of 24 hours. Afterwards, the cell survival rate and the expression of CD86 and CD54 were determined by flow cytometry, and the expression of CD86 and CD54 was indicated by RFI. According to the test result judgment standard, the positive result judgment value CD86 induction value is 150%, the threshold boundary range is 122% to 184%, and the CD54 induction value is 200%, the threshold boundary range is 157% to 255%. The results are shown in Table 4.
[0130] Table 4
[0131] Group Cell survival rate (%) <![CDATA[RFI CD86 ]]> <![CDATA[RFI CD54 ]]> Positive Drug - 795 701 Example 1 88 34 67
[0132] From the data in Table 4, it can be seen that the first purple magnolia extract is mainly composed of large molecular sugars and alkaloids, and the second purple magnolia extract is composed of lignan compounds. The complex obtained by mixing the two ends is not only non-toxic, safe and mild to the skin, but also can improve its biological activity in anti-inflammatory and soothing, delaying skin aging, controlling oil and inhibiting skin scars.
[0133] The applicant declares that the present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.
[0134] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.
[0135] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.
Claims
1. A magnolia odorata composite extract, characterized in that: The purple magnolia composite extract consists of a first purple magnolia extract and a second purple magnolia extract; The first purple magnolia extract is prepared by a preparation method comprising the following steps: (S1) mixing the purple magnolia raw material with a 10-30% ethanol aqueous solution containing an alkali at pH = 9-11 and then soaking the mixture to obtain a soaking product; (S2) soaking the product and performing ultrasonic extraction to obtain an extract; (S3) filtering the extract, concentrating and drying the filtrate to obtain the first purple magnolia extract; The second purple magnolia extract is prepared by a preparation method comprising the following steps: (P1) mixing the purple magnolia raw material with 85-95% ethanol aqueous solution and performing reflux extraction to obtain an extract; (P2) filtering the extract, and subjecting the filtrate to separation and purification on a macroporous resin to obtain an eluent; (P3) The eluate is concentrated and dried to obtain the second Magnolia grandiflora extract.
2. The purple magnolia composite extract according to claim 1, characterized in that The mass ratio of the first purple magnolia extract to the second purple magnolia extract is 1:9-9:
1.
3. The purple magnolia composite extract according to claim 2, characterized in that The mass ratio of the first purple magnolia extract to the second purple magnolia extract is 1:5-4:
3.
4. The purple magnolia composite extract according to any one of claims 1 to 3, characterized in that The purple magnolia raw material is purple magnolia buds.
5. The purple magnolia compound extract according to any one of claims 1 to 4, characterized in that In step (S1), the material-liquid ratio of the purple magnolia raw material to the 10-30% ethanol aqueous solution containing alkali is 1:(10-20) g / mL; The material-liquid ratio of the purple magnolia raw material to the 85-95% ethanol aqueous solution in step (P1) is 1: (7-10) g / mL; The alkali in the ethanol aqueous solution in step (S1) includes any one of ammonia water, sodium hydroxide, potassium hydroxide, sodium carbonate or arginine, or a combination of at least two of them.
6. The purple magnolia compound extract according to any one of claims 1 to 5, characterized in that The soaking in step (S1) is carried out at 30-45° C. for 1-3 hours; The ultrasonic extraction in step (S2) is carried out at 40-50°C for 1-4h, and the ultrasonic power is 80-200W.
7. The purple magnolia compound extract according to any one of claims 1 to 6, characterized in that The number of reflux extractions in step (P1) is 1-3 times, each time for 1-3 hours; The macroporous resin in step (P2) includes AB-8 type or D101 type.
8. The purple magnolia compound extract according to any one of claims 1 to 7, characterized in that The separation and purification method of step (P2) is: the filtrate is subjected to an activated macroporous resin column, and eluted with water, 65-75% ethanol aqueous solution, and 90-95% ethanol aqueous solution in sequence, and the eluate of 90-95% ethanol aqueous solution is collected.
9. Use of the purple magnolia compound extract according to any one of claims 1 to 8 in preparing a product having any one or at least two of the following effects; 1) Oil control; 2) Delay skin aging; 3) Inhibit scars.
10. Use of the compound extract of Magnolia violaceum according to any one of claims 1 to 8 in the preparation of an SDF-1 gene expression inhibitor, a CRLF2 gene expression inhibitor, or a CCR1 gene expression inhibitor.