Preparation method of safflower flower extract and application thereof

Abstract

This invention provides a method for preparing safflower extract and its application. The extract is obtained by extracting safflower using a specific ratio of organic solution and subsequent post-processing. The extraction method of this invention is simple, requires a short production cycle for the raw materials, and the obtained extract has significant effects in inhibiting hyaluronidase activity, reducing transepidermal water loss, and suppressing sebum secretion. It can be used in moisturizing, soothing, oil-controlling, and repairing products.

Description

Technical Field

[0001] This invention relates to the field of plant extract technology, and in particular to a method for preparing safflower extract and its application. Background Technology

[0002] Safflower (Carthamus tinctorius) is a plant belonging to the genus Carthamus in the family Asteraceae. It is widely distributed in Henan, Zhejiang, Sichuan, Yunnan, and Xinjiang regions of my country. Its dried flowers are commonly used as a traditional Chinese medicine in my country. Its Latin name is Floscarthami, and its functions include promoting blood circulation, regulating menstruation, dispersing blood stasis, and relieving pain. It is used for amenorrhea, dysmenorrhea, persistent lochia, injuries from falls, and sores and swellings.

[0003] Safflower is not only used in medicine, but in recent years, many cosmetic companies have conducted in-depth research and development on safflower extract. They have discovered that the safflower yellow pigment in safflower has antioxidant properties and can inhibit the release of inflammatory mediators, thus incorporating its active ingredients into cosmetics. For example, L'Oréal's Red Anti-Wrinkle series contains safflower extract, as does Lancôme's Advanced Génifique Youth Activating Concentrate. However, current research on the skincare applications of safflower extract is still insufficient; many natural components and their potential skincare benefits within safflower have not yet been fully explored. Summary of the Invention

[0004] This invention provides a method for preparing safflower extract and its application, which has moisturizing, soothing and oil-controlling effects.

[0005] In a first aspect, the present invention provides a method for preparing safflower extract, comprising the following steps:

[0006] (1) Take dried safflower flowers and use them directly or after crushing them;

[0007] (2) Take the material obtained in step (1) and add it to the organic extract at a material-to-liquid ratio of 1g / (25-55)mL. Extract at 20-80℃ for 1-9h, filter, and collect the filtrate.

[0008] (3) After removing the organic extraction solvent from the filtrate, add 60% ethanol to dissolve, adsorb with macroporous resin, wash with water to remove impurities, elute with 80% ethanol, and collect the eluent;

[0009] (4) The eluted and concentrated product is dried to obtain safflower extract.

[0010] The safflower extract of the present invention comprises a medium-to-low polarity extract containing components dissolved in pure water / propylene glycol.

[0011] The safflower raw material described in this invention refers to the fresh or dried flowers of the plant safflower (Carthamus tinctorius), preferably dried flowers. A characteristic compound of safflower is safflower yellow pigment, which is almost insoluble in nonpolar organic solvents. Nonpolar substances in safflower (such as oils, fat-soluble vitamins, and certain alkaloids) are easily dissolved by nonpolar solvents. Furthermore, propylene glycol (specifically 1,2-propylene glycol in this invention) is a commonly used component in cosmetics, and its compatibility with safflower active components helps improve the texture and stability of cosmetics.

[0012] Specifically, the organic extract used in this invention can remove most of the low-polarity compounds in safflower, while retaining the medium- and low-polarity compounds. Efficacy tests revealed that the medium- and low-polarity extracts of safflower have moisturizing, soothing, and oil-controlling effects, while its high-polarity extracts do not.

[0013] In some embodiments, the organic extract is selected from a propylene glycol solution containing at least 40% by volume of a low-polarity solvent. Preferably, a propylene glycol solution containing at least 50% by volume of a low-polarity solvent is used as the organic extract.

[0014] In some embodiments, the low-polarity solvent is selected from petroleum ether, diethyl ether, or hexane.

[0015] Preferably, the organic extract can be selected from the following volume ratios of mixed solutions: petroleum ether: propylene glycol = 5:5 (v:v), petroleum ether: propylene glycol = 6:4 (v:v), or petroleum ether: propylene glycol = 7:3 (v:v); or diethyl ether: propylene glycol = 5:5 (v:v), diethyl ether: propylene glycol = 6:4 (v:v), diethyl ether: propylene glycol = 7:3 (v:v).

[0016] In some embodiments, the material-to-extract ratio is 1g / 25mL, 1g / 40mL, or 1g / 55mL, preferably 1g / 40mL.

[0017] In some embodiments, the extraction temperature can be 20°C, 35°C, 40°C, 60°C, or 80°C. Preferably, the extraction temperature is the heating reflux temperature.

[0018] In some implementations, the extraction time is 1 hour, 1.5 hours, 3 hours, 4.5 hours, 6 hours, 7.5 hours, or 9 hours. Preferably, the extraction time is 6 hours.

[0019] In some embodiments, the drying includes freeze drying or spray drying.

[0020] Optionally, the freeze-drying includes the following steps: freezing at a temperature of -40°C or less for at least 8 hours, followed by sublimation and desorption for 48 hours.

[0021] Optionally, the spray drying includes the following steps: preparing the eluent to be dried into a solution with a relative density of 1.05-1.08 g / mL, and spray drying it at an outlet air temperature of 100-110℃.

[0022] Secondly, the present invention provides a safflower extract, which is prepared by the above-described preparation method.

[0023] Thirdly, the present invention provides a composition comprising the above-mentioned safflower extract and at least one additional ingredient, wherein the safflower extract has a mass percentage content of 0.0001%-5%.

[0024] Optionally, the composition may further include an adjuvant;

[0025] The adjuvants are selected from: collagen synthesis stimulants, agents that regulate PGC-1α synthesis, agents that regulate PPARγ activity, agents that increase or decrease triglyceride content in adipocytes, agents that stimulate or delay adipocyte differentiation, lipolytic agents or agents that stimulate fat breakdown, lipolytic agents, lipogenic agents, inhibitors of acetylcholine receptor aggregation, agents that inhibit muscle contraction, anticholinergic agents, elastase inhibitors, matrix metalloproteinase inhibitors, melanin synthesis stimulants or inhibitors, whitening agents or bleaching agents, pigmentation promoters, self-tanning agents, anti-aging agents, NO-synthesizers, 5α-reductase inhibitors, lysyl hydroxylase and / or prolyl hydroxylase inhibitors, antioxidants, free radical scavengers and / or anti-air pollution agents, and active carbonyl groups. Substance scavengers, anti-glycation agents, antihistamines, antiviral agents, antiparasitic agents, emulsifiers, emollients, organic solvents, liquid propellants, skin conditioning agents, moisture-retaining substances, alpha-hydroxy acids, beta-hydroxy acids, humectants, epidermal hydrolases, vitamins, amino acids, proteins, pigments, dyes, biopolymers, gelling polymers, thickeners, surfactants, emollients, adhesives, preservatives, anti-wrinkle agents, agents that reduce or treat under-eye bags, exfoliants, antimicrobial agents, sterilizing agents, bacteriostatic agents, agents that stimulate the synthesis of dermal or epidermal macromolecules and / or inhibit or prevent their degradation, agents that stimulate elastin synthesis, agents that stimulate core proteoglycan synthesis, agents that stimulate laminin synthesis, agents that stimulate defensin synthesis, stimulant synergists. Agents for protein synthesis, agents stimulating cAMP synthesis, agents stimulating HSP70 synthesis, agents stimulating heat shock protein synthesis, agents stimulating hyaluronic acid synthesis, agents stimulating fibronectin synthesis, agents stimulating deacetylase synthesis, agents stimulating lipid and stratum corneum component synthesis, ceramides, fatty acids, agents inhibiting collagen degradation, agents inhibiting elastin degradation, agents inhibiting serine proteases, agents stimulating fibroblast proliferation, agents stimulating keratinocyte proliferation, agents stimulating adipocyte proliferation, agents stimulating melanocyte proliferation, agents stimulating keratinocyte differentiation, agents inhibiting acetylcholinesterase, skin relaxants, agents stimulating glycosaminoglycan synthesis, anti-hyperkeratosis agents, comedolytic agents, anti-psoriasis agents, anti-eczema agents, DNA repair agents. Compounding agents, DNA protectants, stabilizers, antipruritic agents, agents for the treatment and / or care of sensitive skin, hardening agents, firming agents, reconstructing agents, anti-stretch mark agents, agents that regulate sebum production, antiperspirants, agents that stimulate healing, agents that assist healing, agents that stimulate re-epithelialization, agents that assist re-epithelialization, cytokines, sedatives, anti-inflammatory agents, anesthetics, agents acting on capillary circulation and / or microcirculation, agents that stimulate angiogenesis, agents that inhibit vascular permeability, venous tension agents, agents acting on cell metabolism, agents for improving dermal-epidermal junction, agents that induce hair growth, agents that inhibit or delay hair growth, fragrances, chelating agents, plant extracts, essential oils, marine extracts, agents derived from bio-fermentation processes, inorganic salts, cell extracts, sunscreens.And organic or inorganic photoprotective agents or mixtures thereof that effectively resist UVA and / or UVB rays.

[0026] Fourthly, the present invention provides a cosmetic comprising the above-mentioned safflower extract or the above-mentioned composition.

[0027] Optionally, the dosage form of the cosmetic includes creams, lotions, gels, powders, tablets, films, aerosols, or sprays.

[0028] Fifthly, the present invention provides the use of safflower extract in the preparation of products for moisturizing, soothing and / or oil control.

[0029] Optionally, the moisturizing includes reducing transepidermal water loss from the animal's skin surface and inhibiting hyaluronidase activity.

[0030] Optionally, the oil control refers to reducing or inhibiting the synthesis and secretion of sebum by sebaceous gland cells or slowing down sebum deposition.

[0031] Optionally, the preparation method of the safflower extract includes the following steps:

[0032] (1) Take dried safflower flowers and use them directly or after crushing them;

[0033] (2) Take the material obtained in step (1) and add it to the organic extract at a material-to-liquid ratio of 1g / (25-55)mL. Extract at 20-80℃ or heat to reflux temperature for 1-9h, filter, and collect the filtrate.

[0034] (3) Concentrate the filtrate, dissolve it in 60% ethanol, adsorb it with macroporous resin, wash off impurities with water, elute with 80% ethanol, and collect the eluent.

[0035] (4) The eluent is concentrated and then dried to obtain safflower extract.

[0036] The present invention has the following advantages and effects:

[0037] 1. The safflower extraction process of the present invention is simple and has a short production cycle. The obtained extract contains components with low to medium polarity and has excellent moisturizing, soothing and oil-controlling effects. It can be directly used for skin moisturizing, soothing and oil control, and can also be made into cosmetics with moisturizing, soothing and oil-controlling effects, further improving the application value of safflower in skin care products.

[0038] 2. The safflower extract of the present invention can significantly inhibit hyaluronidase activity, reduce transepidermal water loss, and has significant moisturizing and soothing effects.

[0039] 3. The safflower extract of the present invention can significantly inhibit the synthesis and secretion of sebum by sebaceous gland cells, reduce oil deposition, and has significant oil control and acne removal effects. Detailed Implementation

[0040] To make the objectives, features, and advantages of this invention more apparent and understandable, the invention will be further described in detail below with reference to embodiments. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without inventive effort are within the scope of protection of this invention.

[0041] Example 1

[0042] A safflower extract is prepared by the following steps:

[0043] (1) Take dried safflower flowers and grind them into coarse powder;

[0044] (2) Take 10g of the material obtained in step (1), add it to a propylene glycol solution containing 70% petroleum ether (volume fraction) at a material-to-liquid ratio of 1g / 40mL, heat and reflux for 3h, filter, and obtain the filtrate;

[0045] (3) Evaporate the solvent from the filtrate, add 60% ethanol to dissolve it, adsorb it onto macroporous resin, wash the impurities with water first, then elute with 80% ethanol, and collect the eluent;

[0046] (4) After the eluent is concentrated at 60°C, it is frozen at -40°C for at least 8 hours, then sublimated and dried for 48 hours. The sample is recovered in time to obtain safflower extract with a yield of 5.74% based on the raw material.

[0047] Example 2

[0048] A safflower extract is prepared by the following steps:

[0049] (1) Take dried safflower flowers and grind them into coarse powder;

[0050] (2) Take 10g of the material obtained in step (1), add it to a propylene glycol solution containing 60% petroleum ether (volume fraction) at a material-to-liquid ratio of 1g / 55mL, heat and reflux for 6h, filter, and obtain the filtrate;

[0051] (3) Evaporate the solvent from the filtrate, add 60% ethanol to dissolve it, adsorb it onto macroporous resin, wash the impurities with water first, then elute with 80% ethanol, and collect the eluent;

[0052] (4) After the eluent is concentrated at 60°C, it is frozen at -40°C for at least 8 hours, then sublimated and dried for 48 hours. The sample is recovered in time to obtain safflower extract with a yield of 6.46%.

[0053] Example 3

[0054] A safflower extract is prepared by the following steps:

[0055] (1) Take dried safflower flowers and grind them into coarse powder;

[0056] (2) Take 10g of the material obtained in step (1) and add it to a propylene glycol solution containing 50% petroleum ether at a material-to-liquid ratio of 1g / 25mL (i.e., petroleum ether: propylene glycol = 5:5, v:v). Heat and reflux for 5h, filter, and obtain the filtrate.

[0057] (3) Evaporate the solvent from the filtrate, add 60% ethanol to dissolve it, adsorb it onto macroporous resin, wash the impurities with water first, then elute with 80% ethanol, and collect the eluent;

[0058] (4) After the eluent was concentrated at 60°C, it was prepared into an aqueous solution with a relative density of 1.05-1.08 g / mL. A small spray dryer (model: H-Spray Mini) was used to control the outlet air temperature at 90°C for spray drying. The sample was recovered in time to obtain safflower extract with a yield of 7.59%.

[0059] Example 4

[0060] A composition containing safflower extract, comprising the ingredients listed in Table 1, is prepared by the following steps:

[0061] In a suitable container, 1,2-pentanediol, 1,2-hexanediol, and water were mixed according to the prescribed amounts. Then, the safflower extracts from Examples 1-3 were added separately, and the mixtures were stirred until completely dissolved to obtain compositions containing safflower extract. The composition containing the safflower extract from Example 1 is designated as Composition A; the composition containing the safflower extract from Example 2 is designated as Composition B; and the composition containing the safflower extract from Example 3 is designated as Composition C.

[0062] Table 1 Composition Table of the Composition

[0063]

[0064] Example 5

[0065] An essence, prepared according to the ingredients listed in Table 2, is prepared through the following steps:

[0066] S1. Add phase A to the mixing pot and heat it to 80-85℃.

[0067] S2. Premix phase B until there are no powder particles, add it to the mixing pot, and continue mixing for 10-15 minutes.

[0068] S3. Start cooling down to 60-65℃, then add the C phase material;

[0069] S4. Cool to 35-40℃, add phase D material, stir for 10-15 minutes to obtain the essence.

[0070] Table 2. Ingredients of Safflower Extract

[0071]

[0072] Comparative Example 1

[0073] A safflower extract is prepared by the following steps:

[0074] (1) Take dried safflower flowers and grind them into coarse powder;

[0075] (2) Take 10g of the material obtained in step (1), add it to water at a material-to-liquid ratio of 1g / 40mL, extract at 60℃ for 6h, filter, and obtain filtrate;

[0076] (3) The filtrate was concentrated at 60°C to obtain a concentrate;

[0077] (4) The concentrate was frozen at -40°C for at least 8 hours, then sublimated and dried for 48 hours. The sample was recovered in time to obtain safflower extract with a yield of 5.81%.

[0078] Test Example 1: Test on the Soothing and Repairing Efficacy of Human Skin

[0079] 1.1 Test Objective

[0080] Following the principles of randomized, single-blind, and parallel trials, the soothing and repairing efficacy of the test samples was comprehensively evaluated by combining the results of instrument detection with the continuous use of the test samples by the subjects.

[0081] 1.2 Test Object

[0082] Subjects: A total of 30 volunteers aged 18 to 60 were recruited, and the final valid data were obtained from 30 participants.

[0083] 1.3 Reagents, Materials and Instruments

[0084] Capsaicin: Dilute with 10% ethanol to prepare a 0.01% capsaicin solution (w / v);

[0085] Blank drug sensitivity test strips: 8mm in diameter, 0.7mm in thickness;

[0086] Transepidermal moisture loss testing probe: Tewameter™ Hex;

[0087] Multifunctional Skin Imaging Analysis System: C-CUBE.

[0088] 1.4 Sample to be tested

[0089] Sample group: the essence from Example 5;

[0090] Placebo group: Blank serum (obtained by replacing composition A in the serum with an equal mass of water).

[0091] 1.5 Experimental Methods

[0092] Capsaicin stimulation test method: Perform the capsaicin test at the nasolabial fold. Within 0-2 minutes, use a cotton swab soaked in 10% alcohol solution to gently wipe the nasolabial fold 1 cm away 5 times to allow the subject to adapt to the moist, cool sensation. Disqualify the subject if any discomfort is felt. Then, place an 8mm diameter filter paper 1 cm outside the nasolabial fold and drop 50 μL of 0.01% capsaicin onto the filter paper. After 3 minutes, inquire about the subject's feelings. Evaluate the intensity of each perceived sensation using a 5-point scale. A positive capsaicin test is indicated if the burning sensation on the capsaicin side is ≥2 points and lasts longer than 30 seconds; otherwise, a negative test is indicated. Record the results on the capsaicin test record sheet.

[0093] Subjects visited on days 0, 14, and 28. After each visit, their faces were cleansed with the designated facial cleanser and then patted dry with lint-free absorbent paper towels. They sat quietly in a temperature- and humidity-controlled room for 30 minutes. After the sitting period, the subjects were assessed using a capsaicin stimulation test, and TEWL and Erythema values ​​were collected using an instrument.

[0094] After data collection from participants on Day 0, test products were distributed to them. Researchers explained the usage instructions and guided participants in using the products, ensuring complete coverage of the measurement area. From their second visit onwards, participants were required to bring the test products back to the researchers for review and recording of their usage.

[0095] 1.6 Test Results

[0096] TEWL (Transepidermal Water Loss) value indicates a lower TEWL value, signifying a better skin barrier and stronger water retention capacity. Erythema value indicates a lower erythema index, signifying less erythema and less skin irritation. A greater difference in the capsaicin irritation score before and after use indicates better soothing and repairing effects. Test results are shown in Table 3.

[0097] Table 3. Results of the test on the soothing and repairing effects on human skin (Mean±SD)

[0098]

[0099] The results showed that, compared with before use, after 14 and 28 days of using the test product, the TEWL value, capsaicin irritation score, and Erythema value of the safflower group were significantly reduced. Compared with the placebo group, after 14 and 28 days of product use, the TEWL value of the safflower group was significantly reduced, with a highly significant difference. The Erythema value also decreased, with a significant difference at 28 days. These results demonstrate that safflower extract has significant soothing, repairing, and moisturizing effects. Using safflower extract can effectively improve skin sensitivity, itching, pain, and redness, enhance the skin's ability to resist external damage, repair the skin barrier, reduce transepidermal water loss, and improve the skin's water-locking ability.

[0100] Test Example 2: Hyaluronidase Activity Assay

[0101] 2.1 Reagents and Materials

[0102] Sodium acetate buffer (pH=5.6), hyaluronidase (500U / mL), calcium chloride, sodium hyaluronate (0.5mg / mL), acetylacetone, P-DAB colorimetric reagent (prepared by uniformly mixing p-dimethylaminobenzaldehyde (0.8g), concentrated hydrochloric acid (15mL), and an equal volume of glacial acetic acid).

[0103] 2.2 Instruments

[0104] ELISA reader, electronic balance.

[0105] 2.3 Samples to be tested and grouping

[0106] 2.3.1 Sample to be tested

[0107] The safflower extracts of Examples 1-3 and Comparative Example 1 were tested at concentrations of 10 ppm and 20 ppm, respectively.

[0108] 2.3.2 Grouping

[0109] Sample set: Sample to be tested, hyaluronidase, sodium hyaluronate;

[0110] Sample zeroing group: sample to be tested, sodium acetate buffer, sodium hyaluronate;

[0111] Blank control group: distilled water, hyaluronidase, sodium hyaluronate;

[0112] Blank zeroing group: distilled water, sodium acetate buffer, sodium hyaluronate.

[0113] 2.4 Experimental Methods

[0114] Hyaluronidase and sodium hyaluronate were dissolved in sodium acetate buffer.

[0115] Take a 96-well plate. Add 25 μL of the test sample and 25 μL of hyaluronidase (500 U / mL) to the sample group; add 25 μL of the test sample and 25 μL of sodium acetate buffer to the sample zeroing group; add 25 μL of distilled water and 25 μL of hyaluronidase (500 U / mL) to the blank control group; add 25 μL of distilled water and 25 μL of sodium acetate buffer to the blank zeroing group. Incubate at 37°C with shaking for 20 min. Then add 5 μL of calcium chloride solution (2.5 mol / L) to each well and incubate at 37°C with shaking for 20 min. Add 25 μL of sodium hyaluronate to the sample group and blank control group, and add 25 μL of sodium acetate buffer to the sample zeroing group and blank zeroing group. Incubate at 37°C with shaking for 40 min, then let stand at room temperature for 10 min. Add 25 μL of distilled water, 5 μL of sodium hydroxide solution (5 mol / L), and 25 μL of acetylacetone solution to each well. Incubate in a boiling water bath for 15 min, then in an ice bath for 10 min, and finally at room temperature for 10 min. Add 50 μL of P-DAB and 100 μL of anhydrous ethanol to each well, incubate at room temperature for 30 min, and then measure the OD at 570 nm. 570 value.

[0116] Hyaluronidase inhibition rate (%) = (1 - ) × 100%

[0117] In the formula: A1 is the OD of the sample zeroing group. 570 Value, A2 is the OD of the sample group 570 Value, A3 is the OD of the blank zeroing group. 570 Value, A4 is the OD value of the blank control group. 570 value.

[0118] 2.5 Experimental Results

[0119] Hyaluronidase activity is closely related to type I hypersensitivity reactions. Inhibiting hyaluronidase activity can exert a soothing and anti-allergic effect; therefore, inhibiting hyaluronidase activity is often used as an evaluation indicator for studying its soothing and anti-allergic effects. Furthermore, hyaluronidase is a hydrolytic enzyme that degrades hyaluronic acid. Inhibiting hyaluronidase activity can reduce the degradation of hyaluronic acid in cells, increase the content of hyaluronic acid in cells, which is beneficial for increasing the hydration of the skin or mucous membranes, repairing the skin barrier, and exerting moisturizing, soothing, and repairing effects.

[0120] Sodium hyaluronate is a substrate of hyaluronidase. Under the catalysis of hyaluronidase, sodium hyaluronate is degraded to produce glucuronic acid and N-acetylglucosamine, which develop color and absorb visible light at a wavelength of 570 nm under the action of P-DAB. In this experiment, the test sample was treated with hyaluronidase, and the reaction amount of sodium hyaluronate was measured to determine whether safflower extract could inhibit the activity of hyaluronidase, thereby evaluating whether they have moisturizing, soothing, and repairing effects.

[0121] The results of the test samples' effect on the inhibition of hyaluronidase activity are shown in Table 4.

[0122] Table 4. Inhibition rate of different safflower flower extracts on hyaluronidase activity

[0123]

[0124] The results showed that the safflower extracts from Examples 1, 2, and 3 all exhibited inhibitory effects on hyaluronidase, while the safflower extract from Comparative Example 1 did not show any promoting or inhibiting effect on hyaluronidase. This demonstrates that the polar extracts of safflower do not inhibit hyaluronidase activity, while the medium- and low-polarity extracts of safflower and the propylene glycol dissolution fraction showed excellent ability to inhibit hyaluronidase activity. Therefore, the safflower extract of the present invention has a significant ability to inhibit hyaluronidase activity and can be used to repair the skin barrier, providing moisturizing, soothing, and repairing effects.

[0125] Test Example 3: Oil Content Test

[0126] 3.1 Reagents and Materials

[0127] 0.25% trypsin digestion solution (prepared by mixing 0.25g trypsin with 100mL water), PBS, complete culture medium, FFA (prepared by mixing linoleic acid and palmitic acid in a molar ratio of 1:1), and Oil Red O staining kit.

[0128] 3.2 Instruments

[0129] Constant temperature CO2 incubator, ultra-clean workbench, microplate reader, air bath constant temperature shaker.

[0130] 3.3 Cell lines

[0131] Human sebaceous gland cells (SZ-95).

[0132] 3.4 Samples to be tested and grouping

[0133] 3.4.1 Sample to be tested

[0134] Composition A, Composition B, Composition C and the safflower extract of Comparative Example 1 were dissolved or diluted with PBS to adjust the test concentration to 100 ppm.

[0135] Simvastatin was dissolved in PBS, and the test concentration was 0.05 μM.

[0136] 3.4.2 Grouping

[0137] Experimental group: FFA, sample to be tested;

[0138] Normal group: PBS;

[0139] Model groups: FFA, PBS.

[0140] 3.5 Experimental Methods

[0141] Take one flask of SZ-95 cells in good exponential growth phase, add 0.25% trypsin digestion solution, digest to detach the adherent cells, and count (1~4)×10⁻⁶ cells. 5 Cells were cultured at a concentration of 10 cells / mL to prepare a cell suspension. An appropriate amount of the cell suspension was seeded into a 12-well plate containing complete culture medium and incubated in a CO2 incubator for 24 h. Except for the normal group which received PBS, FFA was added to each well of the other wells to a final concentration of 225 μmol / mL. Simultaneously with induction of cell modeling, the experimental groups received their corresponding test samples, while the model group received the same amount of PBS. The plates were incubated in a CO2 incubator for 48 h. The culture medium was then discarded, and staining was performed according to the Oil Red O staining kit instructions.

[0142] Discard the solution in the well plate, add isopropanol to dissolve the Oil Red O staining solution, and measure the absorbance at 492 nm using a microplate reader. Calculate the relative content of lipid secretion from the cells based on the absorbance values.

[0143] 3.6 Experimental Results

[0144] Oil Red O is a fat-soluble dye that is highly soluble in fat. Its staining principle is that Oil Red O specifically adsorbs onto neutral triglycerides, lipids, and lipoproteins in tissues and cells, thus staining the fat. Free-floxacin (FFA) is an inducer; under FFA stimulation, SZ-95 cells secrete large amounts of oil that can be stained by Oil Red O. This experiment used test samples to treat SZ-95 cells induced by FFA stimulation. By detecting the amount of oil produced by SZ-95 cells, it was determined whether the safflower extract of this invention could inhibit the secretion of oil by human sebaceous gland cells.

[0145] The results of the effects of the test samples on lipid secretion in SZ-95 cells are shown in Table 5.

[0146] Table 5. Relative lipid secretion in SZ-95 cells

[0147]

[0148] Experimental results show that compositions A, B, and C of the present invention can significantly inhibit the synthesis and secretion of sebum by human sebaceous gland cells, while the safflower extract in Comparative Example 1 did not show any inhibitory effect on sebum synthesis and secretion. This demonstrates that the aqueous extract of safflower does not have the effect of inhibiting sebum synthesis and secretion, while the low- to medium-polarity extract of safflower has a significant effect on inhibiting sebum synthesis and secretion. Therefore, the safflower extract of the present invention can be used to improve problems such as excessive oil production and water-oil imbalance in the skin, reduce the occurrence of acne or pimples, and also help the skin recover after acne or pimples occur, exhibiting oil-controlling and repairing effects.

[0149] Although preferred embodiments of the present invention have been described, those skilled in the art, upon learning the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments as well as all changes and modifications falling within the scope of the embodiments of the present invention.

[0150] The preparation method of safflower extract and its application provided by the present invention have been described in detail above. Specific examples have been used to illustrate the principle and implementation of the present invention. The above description of the embodiments is only for the purpose of helping to understand the method and core idea of ​​the present invention. At the same time, for those skilled in the art, there will be changes in the specific implementation and application scope based on the idea of ​​the present invention. Therefore, the content of this specification should not be construed as a limitation of the present invention.

Claims

1. A safflower flower extract, characterized in that, It is prepared through the following steps: (1) Take dried safflower flowers and use them directly or after crushing them; (2) Take the material obtained in step (1) and add it to the organic extract at a material-to-liquid ratio of 1g / (25-55)mL. Extract at 20-80℃ or reflux temperature for 1-9h, filter, and collect the filtrate. The organic extract is selected from a propylene glycol solution containing 50%-70% petroleum ether by volume. (3) After removing the organic extraction solvent from the filtrate, add 60% ethanol to dissolve, adsorb with macroporous resin, wash with water to remove impurities, elute with 80% ethanol, and collect the eluent; (4) The eluent is concentrated and then dried to obtain safflower extract.

2. The safflower flower extract according to claim 1, characterized in that, The drying process includes freeze drying or spray drying.

3. A composition characterized in that, It comprises the safflower extract as described in any one of claims 1 or 2 and at least one additional ingredient, wherein the safflower extract has a mass percentage content of 0.0001%-5%.

4. A cosmetic product, characterized by, It comprises the safflower extract as described in any one of claims 1 or 2, or the composition as described in claim 3.

5. Use of the safflower extract according to any one of claims 1 or 2 in the preparation of products for moisturizing or soothing.

6. Use according to claim 5, characterized in that, The moisturizing process includes reducing transepidermal water loss from the animal's skin surface and inhibiting hyaluronidase activity.

7. Use according to claim 5, characterized in that, The soothing effects include inhibiting hyaluronidase activity and improving skin sensitivity, itching, pain, or redness.

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