CN115350123B - Preparation method of Antrodia camphorata extract and application of Antrodia camphorata extract as raw material of antioxidant cosmetics - Google Patents
Preparation method of Antrodia camphorata extract and application of Antrodia camphorata extract as raw material of antioxidant cosmetics Download PDFInfo
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- 241001486992 Taiwanofungus camphoratus Species 0.000 title claims abstract description 98
- 239000000284 extract Substances 0.000 title claims abstract description 94
- 230000003078 antioxidant effect Effects 0.000 title claims abstract description 22
- 239000002537 cosmetic Substances 0.000 title claims abstract description 17
- 239000003963 antioxidant agent Substances 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 239000002994 raw material Substances 0.000 title claims abstract description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 53
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 36
- 238000001914 filtration Methods 0.000 claims description 17
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 11
- 238000004108 freeze drying Methods 0.000 claims description 11
- 238000000605 extraction Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 238000002137 ultrasound extraction Methods 0.000 claims description 8
- 238000009835 boiling Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 6
- 239000004744 fabric Substances 0.000 claims description 5
- 239000000469 ethanolic extract Substances 0.000 claims description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- 238000010298 pulverizing process Methods 0.000 claims description 2
- 231100000419 toxicity Toxicity 0.000 abstract description 4
- 230000001988 toxicity Effects 0.000 abstract description 4
- 239000006286 aqueous extract Substances 0.000 abstract description 3
- 238000011160 research Methods 0.000 abstract description 3
- 238000000338 in vitro Methods 0.000 abstract description 2
- 231100000956 nontoxicity Toxicity 0.000 abstract description 2
- 235000019441 ethanol Nutrition 0.000 description 33
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- 235000006708 antioxidants Nutrition 0.000 description 12
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 10
- 230000000694 effects Effects 0.000 description 10
- 230000008901 benefit Effects 0.000 description 9
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- 150000003254 radicals Chemical class 0.000 description 7
- 230000002000 scavenging effect Effects 0.000 description 7
- 238000002835 absorbance Methods 0.000 description 6
- 230000002195 synergetic effect Effects 0.000 description 5
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
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- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 2
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 2
- 208000000453 Skin Neoplasms Diseases 0.000 description 2
- 229930003268 Vitamin C Natural products 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 2
- 231100000673 dose–response relationship Toxicity 0.000 description 2
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- YMBCJWGVCUEGHA-UHFFFAOYSA-M tetraethylammonium chloride Chemical compound [Cl-].CC[N+](CC)(CC)CC YMBCJWGVCUEGHA-UHFFFAOYSA-M 0.000 description 2
- 235000019154 vitamin C Nutrition 0.000 description 2
- 239000011718 vitamin C Substances 0.000 description 2
- 201000004384 Alopecia Diseases 0.000 description 1
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- 229910052760 oxygen Inorganic materials 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9728—Fungi, e.g. yeasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/18—Antioxidants, e.g. antiradicals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/52—Stabilizers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/52—Stabilizers
- A61K2800/522—Antioxidants; Radical scavengers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a preparation method of an Antrodia camphorata extract and application of the Antrodia camphorata extract as an antioxidant cosmetic raw material, wherein the Antrodia camphorata mycelium is used as a main raw material in the research, compared with the toxicity and in-vitro antioxidant activity of an alcohol extract, an aqueous extract and different proportions, the proportion of the Antrodia camphorata mycelium alcohol extract to the aqueous extract is 4:1, the Antrodia camphorata mycelium has no toxicity and better antioxidant activity, and the Antrodia camphorata extract can be added as the antioxidant raw material of the cosmetic and provides a foundation for developing and developing the Antrodia camphorata product with high added value.
Description
Technical Field
The invention belongs to the technical field of development of cosmetic raw materials, and particularly relates to a preparation method of an antrodia camphorate extract and application of the antrodia camphorate extract as an antioxidant cosmetic raw material.
Background
Skin has a sustained contact and defensive barrier effect against chemical, physical and biological attacks. Maintenance of skin cell integrity and cellular activity of all immune mechanisms involves a series of chemical reactions that produce free radicals and Reactive Oxygen Species (ROS). Endogenous and exogenous antioxidant mechanisms act by neutralizing these bioactive molecules. This imbalance in neutralization can have a number of consequences: free radicals and ROS are involved in various dermatological etiologies, as well as in the skin aging process and the occurrence of skin tumors. Antioxidation has been receiving increasing attention over the past few decades.
The trend in cosmetics in the world today is to pursue "natural, green, safe, effective". With the demands of people on nature and natural return, more and more plant cosmetics are developed. Therefore, the development of cosmetics is also changing from the mineral oil age to the natural ingredient age, and especially, plant extract cosmetics which are mild in effect, small in irritation, safe and effective are increasingly favored. Plant extracts with antioxidant activity not only give the safety of the cosmetic itself that organic synthetic antioxidants are less used, but also give the skin various benefits: auxiliary anti-aging, repairing photodamage, relieving chloasma, inhibiting melanoma and skin cancer, and resisting alopecia.
Antrodia camphorata (Taiwanofungus camphoratus), namely Antrodia camphorata and Antrodia camphorata, is special rare edible and medicinal fungi in Taiwan of China, and can produce various active ingredients such as triterpene, polysaccharide, sterol, benzene ring compounds and the like; the Antrodia camphorata culture has the pharmacological activities of protecting liver, resisting cancer, regulating immunity, resisting oxidation, resisting inflammation, etc. In recent years, many scholars research and found that the mycelium product of Antrodia camphorata has similar pharmacological activity function as fruiting body. Wild Antrodia camphorata grows very slowly, has a scarce quantity and is expensive, is replaced by an artificial culture mode, and achieves the aim of improving the mycelium yield by different culture modes. The dish-type culture of Antrodia camphorata is a mode of artificially culturing Antrodia camphorata mycelia, which has the advantages of strong controllability, low cross infection rate between units, high yield per unit area, stable thallus quality, easy quantitative production, low cost, rich active substances and high content, extracts antioxidant active ingredients with different flavors from the dish-type culture mycelia, and selects synergistic ingredients with different proportions as cosmetic raw materials.
Disclosure of Invention
This section is intended to outline some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section as well as in the description summary and in the title of the application, to avoid obscuring the purpose of this section, the description summary and the title of the invention, which should not be used to limit the scope of the invention.
As one aspect of the invention, a preparation method of the Antrodia camphorate extract comprises the following steps,
pulverizing Antrodia camphorata dish culture mycelium, adding ethanol water solution, performing ultrasonic extraction, filtering, concentrating the obtained filtrate under reduced pressure to remove ethanol, freeze-drying to constant weight to obtain Antrodia camphorata dish culture ethanol extract, filtering to obtain filter residue, volatilizing ethanol, adding water, boiling for extraction, filtering, concentrating the filtrate under reduced pressure, and freeze-drying to obtain Antrodia camphorata dish culture water extract; the antrodia camphorata dish culture alcohol extract and the antrodia camphorata dish culture water extract are mixed according to the mass ratio of 4:1, mixing to obtain Antrodia camphorate extract.
As a preferable scheme of the preparation method of the Antrodia camphorate extract, the invention has the following advantages that: the ethanol water solution is added with the volume fraction of 95 percent.
As a preferable scheme of the preparation method of the Antrodia camphorate extract, the invention has the following advantages that: and adding an ethanol water solution for ultrasonic extraction and filtering, wherein the volume ratio of the Antrodia camphorata dish culture mycelium to the ethanol water solution is 1:8-10, and the extraction times are 1-3 times.
As a preferable scheme of the preparation method of the Antrodia camphorate extract, the invention has the following advantages that: the ultrasonic extraction is carried out under 40KHz at 25 ℃ for 1-1.5 h.
As a preferable scheme of the preparation method of the Antrodia camphorate extract, the invention has the following advantages that: and adding an ethanol water solution for ultrasonic extraction, and filtering, wherein the filtering is performed by adopting gauze, a screen, filter cloth or diatomite.
As a preferable scheme of the preparation method of the Antrodia camphorate extract, the invention has the following advantages that: the water is added for boiling and extracting, and then filtering is carried out, wherein the water with 20 times of the mass of filter residues is added for boiling and extracting for 2-2.5 h, and the extraction times are 1-5 times.
As a preferable scheme of the preparation method of the Antrodia camphorate extract, the invention has the following advantages that: the freeze drying is carried out until the weight is constant, and the temperature is as follows: the cold hydrazine temperature is below-45 ℃, and the vacuum degree is as follows: and the freeze drying time is more than 12 hours at 10 pa.
The invention has the beneficial effects that: the invention provides a preparation method of an Antrodia camphorata extract and application of the Antrodia camphorata extract as an antioxidant cosmetic raw material, wherein an alcohol extract rich in active micromolecular substances and a water extract rich in active polysaccharide and protein are reasonably combined, and the water extract and the alcohol extract 1 are found: 4 has synergistic effect, optimal antioxidation activity and good application prospect as raw material of antioxidation active cosmetics.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the description of the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art. Wherein:
FIG. 1 shows the effect of Antrodia camphorate extract of the present invention on Raw264.7 cell proliferation rate/toxicity.
FIG. 2 shows DPPH scavenging effect of Antrodia camphorata dish culture extract.
FIG. 3 shows the removal of ABTS from Antrodia camphorata dish culture extract + The effect of the composition.
Detailed Description
In order that the above-recited objects, features and advantages of the present invention will become more apparent, a more particular description of the invention will be rendered by reference to specific embodiments thereof.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways other than those described herein, and persons skilled in the art will readily appreciate that the present invention is not limited to the specific embodiments disclosed below.
Further, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic can be included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
Example 1:
experimental materials:
sample extraction and preparation: weighing 100g of dried Antrodia camphorata J1 (T.camphortus J1) dish culture mycelium with constant weight, crushing, adding 8 times of 95% ethanol, extracting at 40KHz for 1h by ultrasonic treatment, filtering with filter cloth (300 meshes), concentrating the filtrate under reduced pressure to remove alcohol, freeze-drying, cooling to hydrazine at a temperature below-45 ℃, and vacuum degree of below: 10pa, and freeze drying for more than 12 hours to constant weight to obtain the Antrodia camphorata dish culture alcohol extract; after volatilizing ethanol from the residue, adding 20 times of water, boiling and extracting for 2 hours, filtering with filter cloth (300 meshes), concentrating the filtrate under reduced pressure, and freeze-drying to obtain the Antrodia camphorata dish culture water extract.
Accurately weighing the Antrodia camphorata dish culture alcohol extract and the Antrodia camphorata dish culture water extract, mixing according to different mass ratios, placing each extract sample in an Eppendorf centrifuge tube, adding a PBS solution under aseptic condition to prepare a solution with the concentration of 5mg/mL, centrifuging for 30min at 12000 Xg, taking the supernatant as a mother solution for later use, and respectively diluting the PBS solution into corresponding concentrations before experiments.
The experimental method comprises the following steps:
cytotoxicity experiment:
selecting well-grown log phase mouse RAW264.7 cells, digesting, and preparing into 5×10 with colorless 1640 medium containing 10% FBS 4 Each/mL of the cell suspension was seeded in 96-well plates, and 200. Mu.L of the cell suspension was added to each well. At 37℃with 5% CO 2 The incubator was incubated with oxygen for 24h, after complete adherence, the medium was aspirated, and 4 duplicate wells were added for each sample containing the different extracts prepared above, with PBS as negative control and LPS (10. Mu.g/mL) as positive control. At 37℃with 5% CO 2 After 48h of cultivation in a constant temperature incubator, the medium was sucked off and 25mg/mL alamarBlue was weighed TM Then configured to alamarBlue at a concentration of 0.0075mg/mL TM And (3) sucking 200 mu L of the prepared reagent into a 96-well plate, culturing for 2-6 hours, measuring absorbance at 570nm and 600nm by using an enzyme-labeled instrument after the color of the reagent in a negative control group is obviously changed, and calculating the inhibition rate of the sample on cell proliferation according to the following formula.
Survival rate = [ (117216 ×d) 570 (sample) -80586 XD 600 (sample))/(117216 XD 570 (blank) -80586 XD 600 (blank)]
DPPH radical scavenging ability:
(1) Weighing 4mg of DPPH reagent, using absolute ethyl alcohol to constant volume to 100mL to prepare 0.1mmol/L DPPH solution, and storing the solution in a brown bottle for later use;
(2) Taking 0.2mL of 80% ethanol sample solution, and respectively adding 2.8mL of DPPH reaction solution as a sample group; taking 0.2mL of 80% ethanol solution from a blank group, adding 2.8mL of DPPH reaction solution, uniformly mixing, placing in a dark place for reaction at room temperature for 30min, and measuring the absorbance value at 517 nm;
(3) Positive group: 10mg/mL vitamin C solution.
The clearance was calculated as follows:
DPPH clearance (%) = (a) 2 -A 1 /A 2 )×100
Wherein A1 is the absorbance value of the sample solution at 517nm, and A2 is the absorbance value of the reference substance at 517 nm.
Wherein the positive concentrations are 10mg/mL.
Equivalent antioxidant Capacity (Trolox equivalent antioxidantcapacity, TEAC) method determines the ability of the ABTS to scavenge free radicals:
referring to the Brand-Williams et al (1995), 5mL of 7mmol/L ABTS and 88. Mu.L of 140mmol/L K S2O8 were mixed and allowed to stand overnight under 30OC constant temperature light-shielding conditions to form ABTS + Stock solution. Before use, the solution was diluted with 20mmol/L PBS (pH 4.5) to give an absorbance at 30℃and 734nm of 0.70. Taking 0.1mL of standard substance or sample in a test tube, adding 3.9mL of ABTS dilution working solution, mixing for 10s, uniformly mixing at room temperature for 20min, measuring absorbance at a wavelength of 284 nm, and calculating antioxidant activity by taking Trolox as the standard substance. Vitamin C at 0.2mg/mL was used as a positive control.
Experimental results:
cytotoxicity test: the experimental results are shown in fig. 1, wherein fig. 1 is negative control group (PBS), antrodia camphorata dish alcohol extract group, antrodia camphorata dish water extract, antrodia camphorata dish alcohol extract mass ratio = 1:4 group, antrodia camphorata dish water extract, antrodia camphorata dish alcohol extract mass ratio = 4:1 group, antrodia camphorata dish water extract, antrodia camphorata dish alcohol extract mass ratio = 1:1 group, and the Antrodia camphorata dish alcohol extract group has effects on Raw264.7 cell proliferation rate at each experimental concentration, and the total concentration of each Antrodia camphorata extract experimental group is respectively 50 mug/mL, 200 mug/mL, 500 mug/mL, 700 mug/mL and 1000 mug/mL. As seen from FIG. 1, after the cells were treated with the Antrodia camphorata dish alcohol extract and the Antrodia camphorata dish water extract in a mass ratio of=4:1, the viability was similar to that of the negative control group at a concentration of 50-1000. Mu.g/mL, indicating that no inhibition of RAW264.7 cell growth occurred and no cytotoxicity was exhibited in the selected concentration range. And other samples can generate toxicity to cells under the condition of high concentration, so that two samples of the antrodia camphorata dish culture alcohol extract and the antrodia camphorata dish culture water extract=4:1 can be selected for subsequent activity experiments.
Antioxidant capacity:
DPPH scavenging action:
DPPH is widely used in the detection of free radical scavenging ability of natural extracts (1995), experimental results are shown in figure 2, and figure 2 shows the scavenging effect on DPPH at total concentrations of 0.2mg/mL, 0.4mg/mL, 0.6mg/mL, 0.8mg/mL and 1.0mg/mL of the group of the Antrodia camphorata dish alcohol extract, the group of the Antrodia camphorata dish water extract and the group of the Antrodia camphorata dish water extract with mass ratio=1:4. As can be seen from FIG. 2, positive control V C The clearance rate of 10mg/mL to DPPH reaches 87.15 percent. The clearance capacity of the group with the mass ratio of the Antrodia camphorata dish culture alcohol extract to the Antrodia camphorata dish culture water extract=4:1 on DPPH is obviously better than that of the group with the Antrodia camphorata dish culture alcohol extract, and the group is dose-dependent. When the mass ratio of the water extract of the Antrodia camphorata dish culture to the alcohol extract of the Antrodia camphorata dish culture=1:4 group is 1.0mg/mL, the DPPH clearance capacity reaches 34.66 percent, and the IC is realized 5o The value is 3.81mg/mL, and the aqueous extract has poor DPPH free radical scavenging ability and IC 50 The value was 12.14mg/mL, but the scavenging ability for DPPH radical was the worst, IC 50 The value is only 31.30mg/mL, and thus, the antrodia camphorata dish culture alcohol extract and the water extract are prepared according to the following weight ratio of 4: the mass ratio of 1 is compounded, so that the DPPH and free radical scavenging capability can be greatly improved, and the synergistic effect of mixing the Antrodia camphorata dish culture alcohol extract and the water extract according to the mass ratio of 4:1 on DPPH scavenging is shown. Equivalent antioxidant Capacity (Trolox equivalent antioxidantcapacity, TEAC) method determines the ability of the ABTS to scavenge free radicals: FIG. 3 shows the effects of removing ABTS in the case of an Antrodia camphorata dish alcohol extract group, an Antrodia camphorata dish water extract and an Antrodia camphorata dish alcohol extract group with a mass ratio of 1:4, wherein the total concentration is 0.2mg/mL, 0.4mg/mL, 0.6mg/mL, 0.8mg/mL and 1.0mg/mL respectively. The results in FIG. 3 show that the extract shows good antioxidant capacity and is dose-dependent, and the mass ratio of Antrodia camphorata dish culture water extract to Antrodia camphorata dish culture alcohol extract = 1:4 is specific to ABTS at a total concentration of 1.0mg/mL + The scavenging capacity of the free radicals was 93.77. Mu. Mol Trolox/L, whereasThe ethanol extract is 52.74 mu mol Trolox/L, and the water extract is 52.74 mu mol Trolox/L, which shows that the combination of the Antrodia camphorata dish culture ethanol extract and the Antrodia camphorata dish culture water extract according to the mass ratio of 4:1 has a synergistic effect on the elimination of ABTS free radicals. While positive control V C At 0.2mg/mL, the scavenging capacity of the free radical was 141.80. Mu. Mol Trolox/L.
Experimental results show that the Antrodia camphorata dish culture alcohol extract and the Antrodia camphorata dish culture water extract are compounded, namely the Antrodia camphorata dish culture alcohol extract and the Antrodia camphorata dish culture water extract are mixed according to the mass ratio of 4:1, have no cytotoxicity in the concentration range of 50ug/mL-1000ug/mL, and have the effects of clearing DPPH and eliminating ABTS + The capability is obviously improved, so that after the antrodia camphorata dish culture alcohol extract and the water extract are effectively compounded, the antioxidant activity is obviously improved, and the synergistic effect is shown. Therefore, the compound extract can be used as a cosmetic raw material with an antioxidant function for further development.
The research uses the Antrodia camphorata mycelium as a main raw material, compares toxicity and in-vitro antioxidant activity of an alcohol extract, a water extract and different proportions, has no toxicity and better antioxidant activity in the proportion of 4:1 of the Antrodia camphorata mycelium alcohol extract to the water extract, can be added as a cosmetic antioxidant raw material, and provides a foundation for developing and developing the Antrodia camphorata product with high added value.
It should be noted that the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present invention may be modified or substituted without departing from the spirit and scope of the technical solution of the present invention, which is intended to be covered in the scope of the claims of the present invention.
Claims (7)
1. A preparation method of an Antrodia camphorate extract is characterized by comprising the following steps of: is composed of the following steps of the method,
pulverizing Antrodia camphorata dish culture mycelium, adding ethanol water solution, performing ultrasonic extraction, filtering, concentrating the obtained filtrate under reduced pressure to remove ethanol, freeze-drying to constant weight to obtain Antrodia camphorata dish culture ethanol extract, filtering to obtain filter residue, volatilizing ethanol, adding water, boiling for extraction, filtering, concentrating the filtrate under reduced pressure, and freeze-drying to obtain Antrodia camphorata dish culture water extract; the antrodia camphorata dish culture alcohol extract and the antrodia camphorata dish culture water extract are mixed according to the mass ratio of 4:1, mixing to obtain an Antrodia camphorate extract;
the ethanol water solution is added with 95% ethanol water solution by volume fraction;
the ethanol water solution is added for ultrasonic extraction and then is filtered, wherein the volume ratio of the Antrodia camphorata dish culture mycelium to the ethanol water solution is 1: 8-10, wherein the extraction times are 1-3 times;
and adding water, boiling, extracting, and filtering, wherein the extraction times are 1-5 times for adding water which is 20 times of the mass of filter residues, boiling and extracting for 2-2.5 hours.
2. The method for preparing the antrodia camphorate extract according to claim 1, which is characterized in that: the ultrasonic extraction is carried out under the conditions of 30-40 KHz, 15-25 ℃ and the extraction time of 1-1.5 h.
3. The method for preparing the antrodia camphorate extract according to claim 1 or 2, which is characterized in that: and adding an ethanol water solution for ultrasonic extraction, and filtering, wherein the filtering is performed by adopting gauze, a screen, filter cloth or diatomite.
4. The method for preparing the Antrodia camphorate extract according to claim 3, which is characterized in that: the filtering is carried out after the water is added, boiled and extracted, wherein the filtering is carried out by adopting gauze, a screen, filter cloth or diatomite.
5. The method for preparing the antrodia camphorate extract according to claim 1, which is characterized in that: the freeze drying is carried out until the weight is constant, and the temperature is as follows: the cold hydrazine temperature is below-45 ℃, and the vacuum degree is as follows: and the freeze drying time is more than 12 hours at 10 pa.
6. The Antrodia camphorate extract obtained by the preparation method of claim 1.
7. The use of Antrodia camphorate extract obtained by the preparation method of claim 1 as raw material of antioxidant cosmetics.
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