Traditional Chinese medicine compound with effects of resisting oxidation and inhibiting melanogenesis and preparation method of traditional Chinese medicine compound

Abstract

The invention discloses a traditional Chinese medicine compound with effects of resisting oxidation and inhibiting melanogenesis and a preparation method of the traditional Chinese medicine compound, and the traditional Chinese medicine compound with the effects of inhibiting melanogenesis is prepared by mixing a dogwood extract, an astragalus extract, a radix angelicae extract and a rose extract according to the proportion of (20-30): (20-60): (25-60): (10-25) based on the equivalent weight of crude drugs, so as to obtain a compound I; the compound I and the exocarpium citri grandis extract are mixed according to the crude drug equivalent ratio of (4-6): 1 to obtain a compound II, and the traditional Chinese medicine compound with the good antioxidant activity and the melanogenesis inhibiting effect is obtained at the same time. According to the invention, the raw materials with multiple functional points are proportioned, so that the antioxidant activity of the compound can be better improved by adding the raw materials with antioxidant activity on the premise of not reducing the melanogenesis inhibition activity.

Description

Technical Field

[0001] The present invention relates to the technical field of preparation of efficacy compositions, and in particular to a traditional Chinese medicine composite having both good antioxidant activity and efficacy of inhibiting melanin production, and a preparation method thereof. Background Art

[0002] Whitening is a common topic, pursued and desired by most people. As the saying goes, "whiteness hides three flaws, darkness ruins everything." Whiteness is another form of beauty. Currently, whitening methods primarily include medication and cosmetic procedures. Medications primarily involve the use of whitening skincare products, such as those containing vitamin C and arbutin, which inhibit the activity of related enzymes, thereby reducing melanin production. They also employ special chemical ingredients that exfoliate the epidermal stratum corneum, accelerating melanin metabolism and promoting skin renewal. Medical cosmetic procedures primarily include laser treatments, whitening injections, and traditional Chinese medicine masks. Each of these methods has its own advantages and disadvantages, and the appropriate choice should be made based on individual needs. However, most current methods still fail to fundamentally and long-term address persistent issues such as skin darkening, hyperpigmentation, and melasma.

[0003] The formation of melasma is strongly linked to the action of an enzyme, tyrosinase. Simply put, tyrosine and L-dopa in the human body, under the action of tyrosinase (which has the properties of both a monophenolase and a polyphenolase), readily form dopaquinone. Under certain conditions, dopaquinone undergoes a series of rearrangement reactions to form melanin. Melanin further accumulates and oxidizes, forming chloasma on the skin's surface. Research on the formation mechanism of chloasma has found two key factors: sunlight exposure and heat stimulation, which can damage skin's surface protective factors and accelerate melanin production, leading to oxidative discoloration and chloasma. Second, damage from oxygen free radicals. Oxygen free radicals participate in numerous biochemical pathways in the human body. Excessive oxygen free radicals in the body can lead to various stresses and reactions. Effectively reducing and balancing excess oxygen free radicals in the body can protect the skin through other pathways, reducing melanin production and preventing the formation of chloasma. Therefore, screening and formulating a combination with both antioxidant and tyrosinase inhibitory properties can effectively inhibit and reduce melanin production, thereby reducing the formation of chloasma.

[0004] Traditional Chinese medicine is favored by the market for its characteristics of being pure natural, non-irritating, green and safe. Traditional Chinese medicine has multiple biological activities and can meet multiple functional needs. Currently, there are few products on the market that have both antioxidant and melanin inhibition effects or the effects are not ideal.

[0005] Therefore, at this stage, there is an urgent need for a safe, non-toxic, and low-side effect pure natural Chinese medicine composition that has both melanin inhibition and antioxidant activity and does not require other auxiliary additives. Summary of the Invention

[0006] The purpose of the present invention is to provide a traditional Chinese medicine complex having both antioxidant activity and melanin production inhibition efficacy and a preparation method thereof. By proportioning the multifunctional raw materials, the antioxidant activity of the complex can be improved by adding raw materials with antioxidant activity without reducing the melanin production inhibition activity, achieving the technical effect of "1+1>2", which provides better technical support for new product development and application research.

[0007] A method for preparing a traditional Chinese medicine compound having both antioxidant activity and melanin production inhibition efficacy comprises the following steps:

[0008] (1) Astragalus Extraction: Weigh a certain amount of Astragalus powder and extract it with ethanol to prepare an Astragalus alcohol extract concentrate; evaporate the ethanol from the filter residue obtained by ethanol extraction and extract it with water to prepare an Astragalus water extract concentrate;

[0009] (2) Purification of Astragalus: The alcohol extract concentrate of Astragalus was passed through one of AB-8 or D101 resins, and eluted with 5-10 column volumes of water and 5-10 column volumes of 50%-75% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 50-65°C to obtain Astragalus extract I. The water extract concentrate of Astragalus was added with 90%-100% ethanol to precipitate the ethanol concentration to 70%-80%. After standing at 2-8°C for 4-8 hours, the precipitate was collected by centrifugation and concentrated and dried under vacuum to obtain Astragalus extract II. Astragalus extract I and Astragalus extract II were combined. Extract II mainly contained Astragalus polysaccharides. Relevant studies have shown that Astragalus polysaccharides can indirectly reduce melanin production by reducing oxidative stress and reducing or inhibiting the activity of tyrosinase. Astragalus extract I and Astragalus extract II were combined to obtain the final Astragalus extract.

[0010] (3) Weighing a certain amount of powder of Cornus officinalis, Angelica dahurica, Rose flower, and Citrus aurantium dulcis respectively, and extracting with ethanol to prepare the corresponding alcohol extract concentrate;

[0011] (4) The alcohol extract concentrates of Cornus officinalis and Angelica dahurica were passed through one of D101 or AB-8 resins, and eluted with 5-10 column volumes of water and 5-10 column volumes of 50%-75% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 50-65° C. to obtain Cornus officinalis extract and Angelica dahurica extract, respectively.

[0012] (5) The rose and tangerine peel alcohol extract concentrate is vacuum concentrated and dried to obtain the rose extract and tangerine peel extract;

[0013] (6) Based on the crude drug equivalents, Cornus officinalis extract, Astragalus root extract, Angelica dahurica extract and Rosa rugosa extract are mixed in a ratio of 20-30:20-60:25-60:10-25 to obtain Complex I, which is a traditional Chinese medicine complex with good melanin inhibition effect and certain antioxidant activity; Complex I and Citrus aurantium dulcis extract are mixed in a ratio of 4-6:1 based on the crude drug equivalents to obtain Complex II, which is a traditional Chinese medicine complex with good antioxidant activity and melanin inhibition effect.

[0014] Preferably, before step (1), the dried cornus officinalis, astragalus root, angelica dahurica, rose flower and tangerine peel are crushed separately, passed through an 80-mesh pharmacopoeia sieve, and the sieved material is taken.

[0015] Preferably, the astragalus ethanol extract concentrate in step (1) is prepared according to the following steps: adding 10-15 times the volume of 50%-75% ethanol solution to the astragalus powder, performing heat reflux extraction at 70-90° C. for 1-2 times, each time for 1-2 hours, filtering after the extraction, and concentrating the filtrate to obtain the astragalus ethanol extract concentrate.

[0016] Preferably, the Astragalus membranaceus water extract concentrate in step (1) is prepared according to the following steps: adding 10-20 times the volume of water to the Astragalus membranaceus alcohol extract residue, soaking for 30-60 minutes, and then extracting under hot reflux at 80-90° C. for 1-2 times, each time for 1-2 hours, filtering after the extraction, and concentrating the filtrate by 5-10 times to obtain the Astragalus membranaceus water extract concentrate.

[0017] Preferably, in step (3), 10-15 times the volume of 50%-75% ethanol solution is added to the powder of Cornus officinalis, Angelica dahurica, Rose, and Citrus aurantium respectively, and the mixture is subjected to hot reflux extraction at 50-80°C for 1-2 times, each time for 1-2 hours. After the extraction is completed, the mixture is filtered and the filtrate is concentrated to obtain an alcohol-extracted concentrate.

[0018] Preferably, in step (5), the rose flower alcohol extract concentrate is vacuum concentrated and dried at 50-60° C. to obtain the rose flower extract; and the tangerine peel alcohol extract concentrate is vacuum concentrated and dried at 50-65° C. to obtain the tangerine peel extract.

[0019] Cornus officinalis (scientific name: Cornus officinalis Siebold & Zucc.), a plant of the genus Cornus in the family Cornaceae, is primarily grown in Shanxi and Shaanxi provinces of my country. Containing components such as loganin and morroniside, Cornus officinalis has antioxidant, anti-inflammatory, and immunomodulatory properties. Its pulp is used medicinally to nourish the liver and kidneys and to astringe and strengthen the body. On November 9, 2023, Cornus officinalis was designated as a "substance traditionally used as both a food and a Chinese medicinal ingredient."

[0020] Astragalus membranaceus (Fisch.) Bunge is a plant of the genus Astragalus in the Leguminosae family. It is primarily found in Tibet and Sichuan. It contains polysaccharides, flavonoids, and saponins, and possesses immunomodulatory properties. According to the Chinese Pharmacopoeia, it is indicated for its functions of tonifying Qi and raising Yang, strengthening the exterior and arresting perspiration, and promoting diuresis and reducing swelling. It is also used for symptoms of Qi deficiency, fatigue, poor appetite, loose stools, Qi deficiency in the middle meridians, and chronic diarrhea and rectal prolapse.

[0021] Angelica dahurica (Fisch. ex Hoffm.) Benth. & Hook. f. ex Franch. & Sav.) is a tall perennial herbaceous plant belonging to the Apiaceae family. It is primarily grown in Northeast and North my country. It contains volatile oils and coumarins, which exhibit anti-inflammatory, antibacterial, antipyretic, and analgesic properties. It can dispel cold, expel wind, relieve pain, and clear the nasal passages. It is commonly used to treat colds, headaches, brow pain, and nasal congestion and runny nose.

[0022] Rose (scientific name: Rosa rugosa Thunb.) is a plant of the genus Rosa in the Rosaceae family. It is abundantly grown in Yunnan, my country. It contains essential oils, flavonoids, and polyphenols, which have antioxidant, anti-inflammatory, and soothing properties. It promotes qi circulation, relieves depression, harmonizes blood circulation, and relieves pain. It is used to treat liver and stomach pain, loss of appetite, nausea, and irregular menstruation.

[0023] Huajuhong (Huajuhong): The dried, immature or nearly mature outer peel of the Huazhou pomelo, a tree in the Rutaceae family, is a specialty of Huazhou City, Maoming City, Guangdong Province. It primarily contains naringin and volatile oils, and possesses antitussive, expectorant, and anti-inflammatory properties. It regulates qi and strengthens the spleen, dries dampness, and resolves phlegm. It is used to treat coughs with excessive phlegm, food indigestion, alcohol consumption, nausea, and abdominal distension.

[0024] As another object of the present invention, the present invention also provides a traditional Chinese medicine complex prepared by the above method, which has the effects of anti-oxidation and inhibiting melanin production.

[0025] Compared with other existing technologies, the present invention has the following advantages:

[0026] 1. Green and safe: Compared to traditional chemically synthesized products, this TCM compound is derived entirely from natural Chinese medicinal materials. Traditional Chinese medicine has a long history of research and application in my country, and its safety has been extensively verified in practice. It reduces potential toxic side effects and allergic risks, is more friendly to human health, and aligns with the current trend towards green and natural products.

[0027] 2. Multiple benefits in one: Existing technologies often focus on a single benefit, such as antioxidant or melanin inhibition. The traditional Chinese medicine complex in this invention innovatively combines both benefits. This eliminates the need for consumers to use separate products for both antioxidant and whitening, saving time and cost, and making use more convenient and efficient.

[0028] 3. Synergistic Effect: The various Chinese herbal ingredients in the complex work synergistically. On the one hand, the antioxidant ingredients can effectively scavenge free radicals, reduce reactive oxygen species, and reduce the abnormal activity of melanocytes caused by oxidative stress, creating a favorable internal environment for inhibiting melanin production. On the other hand, the ingredients that inhibit melanin production can reduce pigmentation at the source. The two complement each other, and their combined effect is superior to the effect of a single ingredient or a simple combination of ingredients, significantly improving skin condition compared to existing technologies.

[0029] 4. Natural mechanism: This Chinese medicine complex includes Chinese medicinal materials such as Cornus officinalis, Angelica sinensis, and rose, and mainly contains flavonoids and phenolic glycoside compounds with a large number of phenolic hydroxyl groups, so it has a good antioxidant effect; at the same time, some phenolic glycosides, flavonoids, and coumarin compounds have a special structure that is similar to the structure of tyrosinase substrate tyrosinase, which can competitively inhibit the activity of tyrosinase and inhibit the production of melanin, thereby achieving the effect of whitening and anti-aging. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The figure is a graph showing the relationship between kojic acid concentration and inhibition rate in the present invention. DETAILED DESCRIPTION

[0031] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0032] Example 1

[0033] This embodiment includes the following steps:

[0034] (1) Medicinal material processing: Grind the dried cornus officinalis, astragalus root, angelica dahurica and rose flower separately, pass through an 80-mesh pharmacopoeia sieve, and take the sieved material;

[0035] (2) Astragalus Extraction: Weigh a certain amount of Astragalus powder, add 10 times the volume of 50% ethanol solution, and extract twice at 75°C under hot reflux conditions for 1 hour each time. After the extraction is completed, filter and concentrate the filtrate to obtain the Astragalus alcohol extract concentrate; after the ethanol is evaporated from the alcohol extraction residue, add 15 times the volume of water, soak for 30 minutes, and then extract twice at 80°C under hot reflux conditions for 1 hour each time. After the extraction is completed, filter and concentrate the filtrate 8 times to obtain the Astragalus water extract concentrate;

[0036] (3) Extraction of other medicinal materials: Weigh a certain amount of powder of Cornus officinalis, Angelica dahurica, and Rosa rugosa respectively, add 60% ethanol solution at 10 times the volume, and extract under reflux at 70°C for 2 times, each time for 2 hours. After the extraction, filter and concentrate the filtrate to obtain the alcohol extract concentrate;

[0037] (4) Purification of Astragalus: The alcohol extract concentrate of Astragalus was passed through AB-8 resin and eluted with 5 times the column volume of water and 5 times the column volume of 50% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 55°C to obtain Astragalus Extract I. The water extract concentrate of Astragalus was added with 95% ethanol to precipitate the ethanol concentration to 75%. The Astragalus Extract was placed in a refrigerator at 2-8°C for 4 hours and then centrifuged to collect the precipitate. The precipitate was concentrated and dried under vacuum to obtain Astragalus Extract II. The Astragalus Extract I and Astragalus Extract II were combined.

[0038] (5) Purification of Cornus officinalis and Angelica dahurica: The alcohol extract concentrates of Cornus officinalis and Angelica dahurica were passed through D101 resin, and eluted with 6 column volumes of water and 6 column volumes of 60% ethanol solution respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 50°C to obtain Cornus officinalis extract and Angelica dahurica extract respectively.

[0039] (6) Rose: The rose alcohol extract concentrate was concentrated and dried at 55°C in a vacuum to obtain rose extract;

[0040] (7) Preparation of Complex I: Each extract was converted into its own herbal medicine equivalent, and the extracts of Cornus officinalis, Astragalus membranaceus, Angelica dahurica and Rosa rugosa obtained in steps (4), (5) and (6) were mixed in a ratio of 25:25:25:25 to obtain Complex I.

[0041] Example 2

[0042] This embodiment includes the following steps:

[0043] (1) Medicinal material processing: Grind the dried cornus officinalis, astragalus root, angelica dahurica and rose flower separately, pass through an 80-mesh pharmacopoeia sieve, and take the sieved material;

[0044] (2) Astragalus Extraction: Weigh a certain amount of Astragalus powder, add 12 times the volume of 55% ethanol solution, and extract twice at 75°C under hot reflux conditions for 2 hours each time. After the extraction is completed, filter and concentrate the filtrate to obtain the Astragalus alcohol extract concentrate; after the ethanol is evaporated from the alcohol extraction residue, add 10 times the volume of water, soak for 30 minutes, and then extract once at 85°C under hot reflux conditions for 2 hours. After the extraction is completed, filter and concentrate the filtrate 6 times to obtain the Astragalus water extract concentrate;

[0045] (3) Extraction of other medicinal materials: Weigh a certain amount of powder of Cornus officinalis, Angelica dahurica, and Rosa rugosa respectively, add 65% ethanol solution at 12 times the volume, and extract under reflux at 75°C for 2 times, each time for 2 hours. After the extraction, filter and concentrate the filtrate to obtain the alcohol extract concentrate;

[0046] (4) Purification of Astragalus: The alcohol extract concentrate of Astragalus was passed through one of the D101 resins and eluted with 6 column volumes of water and 6 column volumes of 55% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 60°C to obtain Astragalus extract I. The water extract concentrate of Astragalus was added with anhydrous ethanol to precipitate the Astragalus extract to an ethanol concentration of 80%. The Astragalus extract was placed in a refrigerator at 2-8°C for 5 hours and then centrifuged to collect the precipitate. The precipitate was concentrated and dried under vacuum to obtain Astragalus extract II. The Astragalus extract I and Astragalus extract II were combined.

[0047] (5) Purification of Cornus officinalis and Angelica dahurica: The alcohol extract concentrates of Cornus officinalis and Angelica dahurica were passed through AB-8 resin, and eluted with 8 column volumes of water and 8 column volumes of 65% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 55°C to obtain Cornus officinalis extract and Angelica dahurica extract, respectively.

[0048] (6) Rose: The rose alcohol extract concentrate was concentrated and dried at 50°C in a vacuum to obtain rose extract;

[0049] (7) Preparation of Complex I: Each extract was converted into its own crude drug equivalent, and the extracts of Cornus officinalis, Astragalus membranaceus, Angelica dahurica and Rosa rugosa obtained in steps (4), (5) and (6) were mixed in a crude drug equivalent ratio of 20:30:30:20 to obtain Complex I.

[0050] Example 3

[0051] This embodiment includes the following steps:

[0052] (1) Medicinal material processing: Grind the dried cornus officinalis, astragalus root, angelica dahurica and rose flower separately, pass through an 80-mesh pharmacopoeia sieve, and take the sieved material;

[0053] (2) Astragalus Extraction: Weigh a certain amount of Astragalus powder, add 15 times the volume of 75% ethanol solution, extract once at 85°C for 2 hours, filter after the extraction, and concentrate the filtrate to obtain the Astragalus alcohol extract concentrate; add 15 times the volume of water to the alcohol extraction residue, soak for 45 minutes, and extract once at 85°C for 2 hours. After the extraction, filter and concentrate the filtrate 10 times to obtain the Astragalus water extract concentrate;

[0054] (3) Extraction of other medicinal materials: Weigh a certain amount of powder of Cornus officinalis, Angelica dahurica, and Rosa rugosa respectively, add 70% ethanol solution at 15 times the volume, and extract under reflux at 70°C for 2 times, each time for 2 hours. After the extraction, filter and concentrate the filtrate to obtain the alcohol extract concentrate;

[0055] (4) Purification of Astragalus: The alcohol extract concentrate of Astragalus was passed through AB-8 resin and eluted with 8 times column volume of water and 8 times 75% ethanol solution respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 60°C to obtain Astragalus Extract I. The water extract concentrate of Astragalus was added with 95% ethanol to precipitate the ethanol concentration to 80%. The Astragalus Extract was placed in a refrigerator at 2-8°C for 6 hours and then centrifuged to collect the precipitate. The precipitate was concentrated and dried under vacuum to obtain Astragalus Extract II. The Astragalus Extract I and Astragalus Extract II were combined.

[0056] (5) Purification of Cornus officinalis and Angelica dahurica: The alcohol extract concentrates of Cornus officinalis and Angelica dahurica were passed through AB-8 resin, and eluted with 5 column volumes of water and 10 column volumes of 70% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 55°C to obtain Cornus officinalis extract and Angelica dahurica extract, respectively.

[0057] (6) Rose: The rose alcohol extract concentrate was concentrated and dried at 55°C in a vacuum to obtain rose extract;

[0058] (7) Preparation of Complex I: Each extract was converted into its own herbal equivalent, and the extracts of Cornus officinalis, Astragalus membranaceus, Angelica dahurica and Rosa rugosa obtained in steps (4), (5) and (6) were mixed in a ratio of 30:20:30:20 to obtain Complex I.

[0059] Example 4

[0060] This embodiment includes the following steps:

[0061] (1) Medicinal material processing: Grind the dried cornus officinalis, astragalus root, angelica dahurica and rose flower separately, pass through an 80-mesh pharmacopoeia sieve, and take the sieved material;

[0062] (2) Astragalus Extraction: Weigh a certain amount of Astragalus powder, add 12 times the volume of 50% ethanol solution, extract once at 90°C for 1 hour, filter after the extraction, and concentrate the filtrate to obtain the Astragalus alcohol extract concentrate; add 10 times the volume of water to the alcohol extraction residue, soak for 30 minutes, and extract once at 90°C for 1 hour, filter after the extraction, and concentrate the filtrate 6 times to obtain the Astragalus water extract concentrate;

[0063] (3) Extraction of other medicinal materials: Weigh a certain amount of powder of Cornus officinalis, Angelica dahurica, and Rosa rugosa respectively, add 75% ethanol solution at 14 times the volume, and extract under reflux at 80°C for 2 times, each time for 2 hours. After the extraction, filter and concentrate the filtrate to obtain the alcohol extract concentrate;

[0064] (4) Purification of Astragalus: The alcohol extract concentrate of Astragalus was passed through AB-8 resin and eluted with 10 column volumes of water and 10 column volumes of 55% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 55°C to obtain Astragalus Extract I. The water extract concentrate of Astragalus was added with 100% ethanol to precipitate the ethanol concentration to 80%. The concentrate was placed in a refrigerator at 2-8°C for 6 hours and then centrifuged to collect the precipitate. The precipitate was concentrated and dried under vacuum to obtain Astragalus Extract II. The Astragalus Extract I and Astragalus Extract II were combined.

[0065] (5) Purification of Cornus officinalis and Angelica dahurica: The alcohol extract concentrates of Cornus officinalis and Angelica dahurica were passed through one of the AB-8 resins, and eluted with 10 column volumes of water and 8 column volumes of 65% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 60°C to obtain Cornus officinalis extract and Angelica dahurica extract, respectively.

[0066] (6) Rose: The rose alcohol extract concentrate was concentrated and dried at 60°C in a vacuum to obtain rose extract;

[0067] (7) Preparation of Complex I: Each extract was converted into its own herbal medicine equivalent, and the extracts of Cornus officinalis, Astragalus membranaceus, Angelica dahurica and Rosa rugosa obtained in steps (4), (5) and (6) were mixed in a ratio of 25:40:25:10 to obtain Complex I.

[0068] Comparative Examples 1 to 4

[0069] Comparative Examples 1-4 were based on the Cornus officinalis extract, Astragalus extract, Angelica dahurica extract and Rosa rugosa extract prepared in Example 2, and one of the raw materials was removed to prepare the missing Chinese medicine composites;

[0070] (2) The preparation of the complex is based on the crude drug equivalent, and the proportions are:

[0071] Comparative Example 1: Astragalus: Angelica dahurica: Rose = 50:30:20;

[0072] Comparative Example 2: Cornus officinalis: Angelica dahurica: Rose = 20:60:20;

[0073] Comparative Example 3: Cornus officinalis: Astragalus root: Rose flower = 20:60:20;

[0074] Comparative Example 4: Cornus officinalis: Astragalus membranaceus: Angelica dahurica = 20:50:30.

[0075] Test Example 1

[0076] This test example includes the following steps:

[0077] (1) Treatment of Citrus aurantium: Grind the dried Citrus aurantium and pass it through an 80-mesh pharmacopoeia sieve, and take the sieved material;

[0078] (2) Extraction and drying of Citrus aurantium: Weigh a certain amount of Citrus aurantium powder, add 65% ethanol solution at 10 times the volume, and extract twice at 60°C under reflux, each time for 1.5 hours. After the extraction, filter and concentrate the filtrate to obtain an alcohol extract concentrate; the alcohol extract concentrate is concentrated and dried in a vacuum at 55°C to obtain Citrus aurantium extract;

[0079] (3) Preparation of the complex: The complex I in Examples 1-4, the complex in Comparative Examples 1-4, and the Citrus aurantium dulcis extract in this test example were mixed in a crude drug equivalent ratio of 4:1 to obtain a Chinese medicine complex II, which is a Chinese medicine complex II having both good antioxidant activity and the effect of inhibiting melanin production.

[0080] Experimental Example 2

[0081] This test example includes the following steps:

[0082] (1) The antioxidant activity (ORAC) and tyrosinase inhibition rate of the Chinese herbal compound II prepared in Experimental Example 1 were measured based on the antioxidant model and the melanin production inhibition model, respectively. Each compound extract was calculated based on the crude drug equivalent and was prepared into a crude drug concentration of 0.1 g / mL sample.

[0083] (2) Antioxidant model (ORAC): 20 μL of blank buffer, sample and standard antioxidant (Trolox) were mixed with 160 μL of sodium fluorescein (0.01 μmol) solution, incubated at 37°C for 30 min in a microplate reader, and then 20 μL of APH solution (153 mmol) was quickly added and placed in a microplate reader for measurement according to the program. The fluorescence intensity of the reaction system was recorded using the microplate reader software. The measurement was performed once every minute for 120 min. The area under the blank curve without antioxidant was deducted to obtain the net area under the curve of antioxidant (Net AUC). ORAC index calculation formula: X = [(AUC sample -AUC blank ) / (AUC Trolox -AUC blank)], where X is the ORAC equivalent in the sample, in μmol TE / L.

[0084] (3) Melanin production inhibition model: add 90 μL buffer, 40 μL enzyme solution (tyrosinase), 30 μL sample solution or positive control (kojic acid), shake and mix in a 96-well microplate, incubate at 31°C for 5 minutes, immediately add 40 μL substrate (L-dopa, 25 mM) solution to start the reaction, and quickly measure the absorbance of the system at a wavelength of 480 nm. The data are divided into 6 groups, and a control group is set up for each sample group; the absorbance OD value is recorded using the enzyme reader software. The area under the curve of the normal reaction minus the area under the curve of the corresponding control group reaction is the net area under the curve (Net AUC). The inhibition rate calculation formula is: Inhibition (I) = (Net AUC NEG -Net AUC SPL ) / Net AUC NEG ×100%.

[0085] (4) Conversion of melanin production inhibition activity: kojic acid solutions with gradient concentrations were prepared, the inhibition rates of the kojic acid solutions at each concentration were measured, and a correlation curve between kojic acid concentration and inhibition rate was drawn. The curve equation was calculated as follows: Y = 0.2191*ln(X) + 0.9264, R 2 =0.9981, such as Figure 1 As shown; referring to the relationship between inhibition rate and kojic acid concentration, the positive equivalent concentration of each sample was calculated.

[0086] (5) The comparative results of the tyrosinase inhibition rates (in terms of positive equivalents) of the complex I and the missing complex in the examples and comparative examples are shown in Table 1:

[0087] Table 1

[0088]

[0089] From the data in the table, it can be seen that when any one of the medicinal materials in the complex composed of Cornus officinalis, Astragalus membranaceus, Angelica dahurica and Rosacea is missing, its inhibition rate and positive equivalent concentration on tyrosinase are significantly reduced. After the medicinal materials in the formula are missing, the tyrosinase inhibition rate decreases by 9.93% to 29.94%; the positive equivalent concentration decreases by 36.43% to 74.50%, indicating that the complex combination of Astragalus membranaceus, Angelica dahurica, Rosacea and Cornus officinalis has good activity in inhibiting melanin production.

[0090] (6) The comparative results of the antioxidant activity and tyrosinase inhibition rate (in terms of positive equivalents) of the complex I, the missing complex, and the complex II in the examples and the comparative examples are shown in Table 2:

[0091] Table 2

[0092]

[0093]

[0094] As can be seen from the table, by compounding complex I, the missing complex and the tangerine peel extract to obtain complex II, the melanin production inhibition activity of complex II changed little (the change ratio was -6.09% to 8.72%), and the activity of some groups was even improved, but the antioxidant activity was significantly improved. The antioxidant activity improvement ratio of the embodiment groups exceeded 67%. It can be inferred that the complex II obtained by compounding based on complex I not only has good melanin production inhibition activity, but also has good antioxidant activity. The complex can achieve the effects of whitening, inhibiting melanin production and improving chloasma by inhibiting tyrosinase activity to reduce melanin production and regulating the balance of oxygen free radicals through antioxidant pathways.

[0095] The above embodiments are merely examples of preferred embodiments of the present invention and are not intended to be exhaustive. Persons skilled in the art may make various modifications and alterations without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention shall be determined by the scope defined in the claims.

Claims

1. A method for preparing a traditional Chinese medicine compound having both good antioxidant activity and melanin production inhibition effect, characterized in that: The steps include: (1) Astragalus Extraction: Weigh a certain amount of Astragalus powder and extract it with ethanol to prepare an Astragalus alcohol extract concentrate; evaporate the ethanol from the filter residue obtained by ethanol extraction and extract it with water to prepare an Astragalus water extract concentrate; (2) Purification of Astragalus: The alcohol extract concentrate of Astragalus was passed through one of AB-8 or D101 resins, and eluted with 5-10 column volumes of water and 5-10 column volumes of 50%-75% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 50-65°C to obtain Astragalus Extract I. The water extract concentrate of Astragalus was added with 90%-100% ethanol to precipitate the ethanol concentration to 70%-80%. The concentrate was allowed to stand at 2-8°C for 4-8 hours, and then centrifuged to collect the precipitate. The precipitate was concentrated and dried under vacuum to obtain Astragalus Extract II. (3) Weighing a certain amount of powder of Cornus officinalis, Angelica dahurica, Rose flower, and Citrus aurantium dulcis respectively, and extracting with ethanol to prepare the corresponding alcohol extract concentrate; (4) The alcohol extract concentrates of Cornus officinalis and Angelica dahurica were passed through one of D101 or AB-8 resins, and eluted with 5-10 column volumes of water and 5-10 column volumes of 50%-75% ethanol solution, respectively. The ethanol eluate was collected and concentrated and dried under vacuum at 50-65° C. to obtain Cornus officinalis extract and Angelica dahurica extract, respectively. (5) The rose and tangerine peel alcohol extract concentrate is vacuum concentrated and dried to obtain the rose extract and tangerine peel extract; (6) Based on the crude drug equivalents, Cornus officinalis extract, Astragalus root extract, Angelica dahurica extract and Rosa rugosa extract are mixed in a ratio of 20-30:20-60:25-60:10-25 to obtain Complex I; Complex I and Citrus aurantium extract are mixed in a ratio of 4-6:1 to obtain Complex II, that is, a traditional Chinese medicine complex having both good antioxidant activity and melanin production inhibition effect.

2. The method for preparing a traditional Chinese medicine complex having antioxidant and melanin production inhibiting effects according to claim 1, characterized in that: Before step (1), the dried cornus officinalis, astragalus root, angelica dahurica, rose flower and red orange peel are crushed separately, passed through an 80-mesh pharmacopoeia sieve, and the sieved material is taken.

3. The method for preparing a traditional Chinese medicine complex having antioxidant and melanin production inhibiting effects according to claim 1, characterized in that: The astragalus ethanol extract concentrate in step (1) is prepared according to the following steps: adding 10-15 times the volume of 50%-75% ethanol solution to the astragalus powder, performing heat reflux extraction at 70-90° C. for 1-2 times, each time for 1-2 hours, filtering after the extraction, and concentrating the filtrate to obtain the astragalus ethanol extract concentrate.

4. The method for preparing a traditional Chinese medicine complex having antioxidant and melanin production inhibiting effects according to claim 1, characterized in that: The astragalus water extract concentrate in step (1) is prepared according to the following steps: adding 10-20 times the volume of water to the astragalus alcohol extraction residue, soaking for 30-60 minutes, and then extracting under hot reflux at 80-90° C. for 1-2 times, each time for 1-2 hours, filtering after the extraction, and concentrating the filtrate by 5-10 times to obtain the astragalus water extract concentrate.

5. The method for preparing a traditional Chinese medicine complex having antioxidant and melanin production inhibiting effects according to claim 1, characterized in that: In step (3), 10-15 times the volume of 50%-75% ethanol solution is added to the powder of Cornus officinalis, Angelica dahurica, Rose, and Citrus aurantium respectively, and the mixture is subjected to hot reflux extraction at 50-80° C. for 1-2 times, each time for 1-2 hours. After the extraction is completed, the mixture is filtered and the filtrate is concentrated to obtain an alcohol-extracted concentrate.

6. The method for preparing a traditional Chinese medicine complex having antioxidant and melanin production inhibiting effects according to claim 1, characterized in that: In step (5), the rose flower alcohol extract concentrate is vacuum concentrated and dried at 50-60° C. to obtain a rose flower extract; and the tangerine peel alcohol extract concentrate is vacuum concentrated and dried at 50-65° C. to obtain a tangerine peel extract.

7. A traditional Chinese medicine complex having antioxidant and melanin production inhibiting effects, prepared by the method according to any one of claims 1 to 6.

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