CN112386532A - Preparation method and application of Artocarpus heterophyllus extract - Google Patents
Preparation method and application of Artocarpus heterophyllus extract Download PDFInfo
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- CN112386532A CN112386532A CN201910762129.9A CN201910762129A CN112386532A CN 112386532 A CN112386532 A CN 112386532A CN 201910762129 A CN201910762129 A CN 201910762129A CN 112386532 A CN112386532 A CN 112386532A
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Images
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
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- 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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- 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
-
- 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
- A61K2800/82—Preparation or application process involves sonication or ultrasonication
Abstract
The invention discloses a preparation method of a millettia extract, and relates to the technical field of cosmetics. The preparation method comprises the following steps: A) crushing the millettia to obtain first powder; B) degreasing, filtering and drying the first powder to obtain second powder; C) and extracting the second powder in an ultrasonic-assisted complex enzyme mode, inactivating enzymes, and purifying to obtain the Artocarpus millettii extract. The method adopts a mode of degreasing and ultrasonic-assisted complex enzyme extraction and purification, the flavone content in the obtained Artocarpus heterophyllus extract can reach 28.5 percent at most, the extraction rate can reach more than 86.0 percent, and the Artocarpus heterophyllus extract has obvious antioxidant and anti-aging functions.
Description
Technical Field
The invention belongs to the technical field of cosmetics, and particularly relates to a preparation method and application of a millettia extract.
Background
Skin aging is a progressive physiological process and is also the result of the combined action of various factors such as heredity, environment, psychology and the like. The main manifestations are that the thickness of epidermis and dermis is thin, the skin is rough, dry, has low luster and elasticity, the water content and blood flow in the skin are reduced, the secretion of sweat gland sebaceous glands is reduced, and the like, which results in slow renewal of epidermis, weakened barrier function, reduced activity of keratinocyte, and weakened repair capability after the epidermis is injured. Regarding the anti-aging mechanism of anti-aging functional components in cosmetics, the theory of "radical aging" proposed by DenhamHarman is generally accepted by the academia at present. The theory holds that: when the metabolism of free radicals in the body is in an unbalanced state, the excessive free radicals can cause body damage, and unsaturated fatty acid is oxidized into superoxide to form lipofuscin; excessive oxygen radicals also damage cell membranes and other important components, denature proteins and enzymes, and when the accumulation of free radical-induced damage outweighs the body's ability to repair, it results in a change or even loss of the state of cell differentiation, leading to or accelerating aging.
Miluo mu, also known as FENGHUANGCAO, is also called Neisseria tenella. Artocarpus heterophyllus is one of the extremely drought-tolerant resuscitating plants with a small number of plants on earth, and the most prominent feature is that leaves fold up when dry, while plants that appear to die are revived from a dry state in rainy seasons. Research analysis shows that the Artocarpus heterophyllus is rich in polyphenol, flavone and saccharide, and the dried leaf powder can be used for dressing burn and wound, and treating chest pain and asthma by inhaling burnt smoke. While in the middle, millettia is used by people to tonify and treat breast diseases. Nowadays, millettia is also used in skin care products due to the increasing recognition by many people that it has properties that help to moisturize the skin, adapt quickly to the environment and to the climate.
However, the extraction method of the Artocarpus heterophyllus is rarely reported in the prior art. Meanwhile, due to the structural particularity of the millettia speciosa, the existing extraction process for the flavone is not suitable for extracting the flavone from the millettia speciosa, and the condition that the extraction rate of the flavone is low (the content of the extracted flavone is 9-15%) generally exists, and due to the improvement of the condition, a method for extracting the flavone from the millettia speciosa is needed to be provided.
Disclosure of Invention
The invention aims to provide a preparation method and application of a Artocarpus heterophyllus extract, the flavone content in the Artocarpus heterophyllus extract can reach 28.5%, and the flavone extraction rate can reach over 86.0%, so that the preparation method has obvious progress compared with the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of a Artocarpus heterophyllus extract comprises the following steps:
A) crushing the millettia to obtain first powder;
B) degreasing, filtering and drying the first powder to obtain second powder;
C) and extracting the second powder in an ultrasonic-assisted complex enzyme mode, inactivating enzymes, and purifying to obtain the Artocarpus millettii extract.
Further, the preparation method specifically comprises the following steps:
A) crushing and sieving the Artocarpus heterophyllus to obtain first powder;
B) taking the first powder in the step A), adding petroleum ether or diethyl ether which is 8-10 times of the total amount of the first powder, and soaking to obtain a degreasing mixture; filtering the degreased mixture, and drying the obtained filter residue to obtain second powder;
C) mixing the second powder obtained in the step B) with an extraction solvent, leaching for 1-1.5h at 3-5 ℃, performing 300-400W ultrasonic extraction for 1-1.5h at 45-50 ℃, inactivating enzyme, and purifying to obtain the Artocarpus millettii extract.
Further, the feed-liquid ratio between the second powder and the extraction solvent in the step C) is 1: 45-60.
Further, the extraction solvent is a disodium hydrogen phosphate-citric acid buffer solution with pH of 4-5, which contains 0.5-2% veratryl alcohol and 1.5% -3% complex enzyme.
Further, the complex enzyme is prepared from chymopapain, pectinase and cellulase according to the following ratio (2-3): 1: (0.5-1) in mass ratio.
Further, the purification process comprises the following steps:
centrifuging the enzyme-inactivated extract in a centrifuge at 4500-5000rmp for 25-30min, and removing filter residue to obtain supernatant; primarily purifying the supernatant through a 400-plus 500nm ceramic membrane to obtain a filtrate; concentrating the filtrate under reduced pressure until no ethanol smell; performing secondary purification by using macroporous resin, eluting by using 50-70% ethanol solution, collecting eluent, concentrating to remove alcohol, and freeze-drying to obtain the Artocarpus heterophyllus extract.
Further, the macroporous resin is one of HPD-400, DM130, D4020, HPD100, DA201-C, S-8, D101, AB-8 and ADS-17.
The invention provides application of the millettia extract prepared by the preparation method in preparing anti-aging skin care products/cosmetics.
The invention also provides an anti-aging preparation which comprises an effective amount of the millettia extract prepared by the preparation method and an effective amount of cosmetic common auxiliary agents.
The inventor researches millettia speciosa, and learns that the millettia speciosa is a highly drought-tolerant resuscitation plant, the drought resistance mechanism of the millettia speciosa is quite complex, and the drought resistance mechanism mainly comprises the following components: firstly, a certain special protective protein exists, which can reduce water loss in cells and stabilize the cells; secondly, the cell wall is stabilized by forming pectin and other mechanisms so as to prevent irreversible damage such as volume contraction caused by dehydration of the cell wall and the cell membrane; thirdly, the loss of water is reduced by forming a large amount of lipid substances; activating an anti-oxidation protection system, promoting oxidative stress in a severe environment and the like. Therefore, the inventor guesses that the existing flavone extraction process is not suitable for extracting flavone from the millettia speciosa due to the drought resistance mechanism existing in the millettia speciosa per se and preventing the sufficient release of the flavone.
According to the guess, the inventor improves the existing ultrasonic wave synergistic enzymolysis extraction process, and finds that the composite enzyme synergistic ultrasonic wave extraction consisting of chymopapain, pectinase and cellulase can act on special protein existing in the Artocarpus heterophyllus synergistically, so that flavone is released from the Artocarpus heterophyllus, and the content of flavone in the extract obtained by the method can reach 28.5%.
If the yield of flavone is higher after veratryl alcohol is added into the extraction solvent, it is presumed that veratryl alcohol can enhance the effect between the complex enzyme and the special protein.
Compared with the prior art, the invention has the following beneficial effects:
the mulukhiya is degreased and then ultrasonically extracted with the compound enzyme, so that the effective active ingredients in the mulukhiya can be protected, the extraction rate of flavone can be increased to 86.0%, the content of the flavone in the extract can be increased to 28.5%, the prepared mulukhiya extract has the DPPH free radical clearance rate of 88.07%, ABTS free radical clearance rate of 83.05%, advanced glycosylation end product clearance rate of 90.30% and elastase inhibition rate of 82.54%, and has excellent anti-aging and anti-oxidation effects.
Drawings
FIG. 1 is a standard curve of total flavonoids detected by sodium nitrite-aluminum nitrate method.
Detailed Description
The present invention will be described in further detail with reference to the following examples. It should not be understood that the scope of the above-described subject matter of the present invention is limited to the following examples.
Example 1 extract of Artocarpus heterophyllus
The preparation method specifically comprises the following steps:
A) crushing the Artocarpus heterophyllus and sieving the Artocarpus heterophyllus with a 40-mesh sieve to obtain first powder;
B) taking the first powder in the step A), adding petroleum ether which is 8 times of the total amount of the first powder, and soaking overnight to obtain a degreasing mixture; filtering the degreased mixture, and drying the obtained filter residue to obtain second powder;
C) mixing the second powder obtained in the step B) with an extraction solvent according to a weight ratio of 1:45, leaching for 1h at 3 ℃, ultrasonically extracting for 1h at 380W at 45 ℃, heating in a boiling water bath for 10min to inactivate enzyme, centrifuging the inactivated extract in a centrifuge at 4500rmp for 30min, and removing filter residues to obtain a supernatant; primarily purifying the supernatant through a 400nm ceramic membrane to obtain a filtrate; concentrating the filtrate under reduced pressure until no ethanol smell; purifying with HPD100 macroporous resin twice, eluting with 50% ethanol solution, collecting eluate, concentrating to remove ethanol, and freeze drying to obtain Artocarpus heterophyllus extract.
Wherein the extraction solvent is disodium hydrogen phosphate-citric acid buffer solution with pH of 4 containing 0.5% veratryl alcohol and 1.5% complex enzyme; the compound enzyme is prepared from chymopapain, pectinase and cellulase according to the weight ratio of 2: 1: 0.5 in mass ratio.
Example 2 extract of Artocarpus heterophyllus
The preparation method specifically comprises the following steps:
A) crushing the Artocarpus heterophyllus and sieving the Artocarpus heterophyllus with a 40-mesh sieve to obtain first powder;
B) taking the first powder in the step A), adding petroleum ether which is 10 times of the total amount of the powder, and soaking overnight to obtain a degreasing mixture; filtering the degreased mixture, and drying the obtained filter residue to obtain second powder;
C) mixing the second powder obtained in the step B) with an extraction solvent according to a weight ratio of 1:60, leaching for 1.5h at 3 ℃, ultrasonically extracting for 1.5h at 380W at 45 ℃, heating in a boiling water bath for 15min to inactivate enzyme, centrifuging the inactivated extract in a centrifuge at 4500rmp for 30min, and removing filter residues to obtain a supernatant; primarily purifying the supernatant through a 400nm ceramic membrane to obtain a filtrate; concentrating the filtrate under reduced pressure until no ethanol smell; purifying with HPD100 macroporous resin twice, eluting with 60% ethanol solution, collecting eluate, concentrating to remove ethanol, and freeze drying to obtain Artocarpus heterophyllus extract.
Wherein the extraction solvent is disodium hydrogen phosphate-citric acid buffer solution with pH of 5 containing 1% veratryl alcohol and 3% complex enzyme; the compound enzyme is prepared from chymopapain, pectinase and cellulase according to the weight ratio of 3: 1: 0.5 in mass ratio.
Example 3 extract of Artocarpus heterophyllus
The preparation method specifically comprises the following steps:
A) crushing the Artocarpus heterophyllus and sieving the Artocarpus heterophyllus with a 40-mesh sieve to obtain first powder;
B) taking the first powder in the step A), adding petroleum ether which is 10 times of the total amount of the powder, and soaking overnight to obtain a degreasing mixture; filtering the degreased mixture, and drying the obtained filter residue to obtain second powder;
C) mixing the second powder obtained in the step B) with an extraction solvent according to a weight ratio of 1:50, leaching for 1h at 5 ℃, ultrasonically extracting for 1h at 380W at 50 ℃, heating in a boiling water bath for 15min to inactivate enzyme, centrifuging the inactivated extract in a centrifuge at 4500rmp for 30min, and removing filter residues to obtain a supernatant; primarily purifying the supernatant through a 400nm ceramic membrane to obtain a filtrate; concentrating the filtrate under reduced pressure until no ethanol smell; performing secondary purification with D4020 macroporous resin, eluting with 50% ethanol solution, collecting eluate, concentrating to remove ethanol, and freeze drying to obtain Artocarpus heterophyllus extract.
Wherein the extraction solvent is disodium hydrogen phosphate-citric acid buffer solution with pH of 5 containing 2% veratryl alcohol and 3% complex enzyme; the compound enzyme is prepared from chymopapain, pectinase and cellulase according to the weight ratio of 3: 1: 1, in terms of mass ratio.
Example 4 an anti-aging emulsion
The anti-aging emulsion comprises the following components in percentage by mass: polyglycerol-3 methyl glucose distearate 3%, 16/18 alcohol 2.5%, polydimethylsiloxane 1.0%, white mineral oil 4.0%, sodium hyaluronate 0.05%, glycerol 5.0%, butanediol 1.0%, disodium EDTA 0.05%, Artocarpus millettii extract 3.0%, preservative 0.5% and water in balance.
The preparation method of the anti-aging emulsion comprises the following steps:
(1) sequentially adding 16/18 alcohol, polydimethylsiloxane and white mineral oil into an oil phase pot according to the formula ratio, mixing and heating to 85 ℃, and then preserving heat for 10min under a stirring state to obtain an oil phase mixture;
(2) adding sodium hyaluronate, glycerol, butanediol, disodium EDTA, Artocarpus heterophyllus extract and water in balance into water phase pot, mixing and stirring at 85 deg.C for 20min to obtain water phase mixture;
(3) slowly adding the oil phase mixture obtained in the step (1) into the water phase mixture obtained in the step (2), homogenizing for 3min, adding the preservative, and stirring uniformly to obtain the anti-aging emulsion containing the millettia extract.
Example 5 an anti-glycation cream
An anti-saccharification face cream comprises the following components in percentage by mass: caprylic/capric triglyceride 3.0%, cyclopentadimethylsiloxane 2.5%, polydimethylsiloxane 2.5%, 16/18 alcohol 3.0%, glyceryl stearate 2.0%, white mineral oil 4.5%, xanthan gum 0.5%, glycerin 8.0%, butylene glycol 2.0%, disodium EDTA 0.1%, millettia extract 2.0%, preservative 0.5%, and balance water.
The preparation method of the anti-saccharification face cream comprises the following steps:
(1) sequentially adding caprylic acid/capric acid triglyceride, cyclopentadimethylsiloxane, polydimethylsiloxane, 16/18 alcohol, glyceryl stearate and white mineral oil into an oil phase pot according to the formula ratio, mixing and heating to 85 ℃, and keeping the temperature for 10min under a stirring state to obtain an oil phase mixture;
(2) adding xanthan gum, glycerol, butanediol, disodium EDTA, Artocarpus heterophyllus extract and water in balance into water phase pot, mixing and stirring at 85 deg.C for 20min to obtain water phase mixture;
(3) slowly adding the oil phase mixture obtained in the step (1) into the water phase mixture obtained in the step (2), homogenizing for 3min, adding a preservative, and stirring uniformly to obtain the anti-sugar cream containing the millettia extract.
Comparative example 1 extract of Artocarpus heterophyllus
Similar to example 2, except that: veratryl alcohol was not added and the remaining parameters were the same as in example 2.
Comparative example 2 extract of Artocarpus heterophyllus
Similar to example 2, except that: chymopapain was not added to the complex enzyme, and the remaining parameters were the same as in example 2.
Comparative example 3 extract of Artocarpus heterophyllus
Similar to example 2, except that: veratryl alcohol and chymopapain were not added and the remaining parameters were the same as in example 2.
Test I, the influence of different complex enzymes on the content of total flavonoids in Artocarpus heterophyllus extract
1.1 test methods:
(1) preparation of a rutin standard curve: precisely weighing 10mg of rutin reference substance, adding 70% ethanol, dissolving, and diluting to 50 mL;
(2) accurately sucking control solution 0mL, 0.4mL, 0.8mL, 1.2mL, 1.6mL, 2.0mL, 2.4mL, placing in 10mL test tubes with plugs, adding water to 2.4mL, and shaking. Then, 0.4mL of 5% sodium nitrite is added, shaken up and left for 6 min. Then 0.4mL of 10% aluminum nitrate is added, shaken well and placed for 6 min. 4mL of 4% sodium hydroxide and 2.8mL of water are added, shaken and placed for 15 min. The absorbance was measured at a wavelength of 510nm using an ultraviolet spectrophotometer and a standard curve regression equation was drawn. The standard curve of the sodium nitrite-aluminum nitrate method for detecting total flavonoids is shown in figure 1.
(3) And (3) sample determination: diluting the sample to a proper concentration, precisely sucking 1mL of the sample, placing the sample in a 10mL test tube with a plug, measuring the light absorption value of the sample according to the method from the point of adding water to 2.4mL under the item of the preparation of the rutin standard curve, and calculating the content of the total flavone by a regression equation of the standard curve.
(4) And (3) medicinal material determination: pulverizing Artocarpus heterophyllus into 40 mesh powder, adding 20 times of 60% ethanol, heating and refluxing for 2 hr, filtering, adding 15 times of 90% ethanol into the residue, heating and refluxing for 2 times, extracting for 1 hr each time, filtering, mixing filtrates, washing the residue with 60% ethanol, adding the washing solution into the same measuring flask, metering volume, and measuring total flavone content according to the above sample measurement method.
1.2 calculation formula:
in the formula, K1: diluting the sample by multiple times; cx 1: the unit is mg/mL of the total flavone mass concentration in the test solution; v1: the volume of the test solution is mL; k2: the dilution times of the samples in the step (4) are obtained; cx 2: the mass concentration of the total flavonoids of the sample in the step (4) is mg/mL; v2 is the sample volume in mL for step 4.
The total flavone mass fraction is the ratio of the total flavone mass in the sample to the sample mass.
TABLE-Total Flavonoids assay results of Artocarpus heterophyllus extract
As can be seen from table one, the extraction rate of total flavonoids in the millettia extract obtained by the preparation method of the present invention is high, and the extraction rate of total flavonoids in the millettia extract obtained in example 2 reaches 86.0%.
Test II, measurement of DPPH radical scavenging ability
2.1 test methods: preparing 0.2mmol/L DPPH free radical methanol solution; preparing 1mg/mL of sample solution to be detected; 2mL of each sample solution and 2mL of DPPH free radical solution are put in a test tube with a plug, mixed evenly, reacted for 30min in a dark place, and the light absorption value A1 under the wavelength of 517nm is measured; putting 2mL of each sample solution and methanol in a test tube with a plug, uniformly mixing, reacting for 30min in a dark place, and measuring the light absorption value A2 at the wavelength of 517 nm; taking 2mL of each DPPH free radical solution and methanol in a test tube with a plug, mixing uniformly, reacting for 30min in a dark place, and measuring the light absorption value A0 at the wavelength of 517 nm.
TABLE DIDPPH radical scavenging ability measurement results
Group of | DPPH radical clearance% |
Example 1 | 86.60 |
Example 2 | 88.07 |
Example 3 | 87.65 |
Comparative example 1 | 77.55 |
Comparative example 2 | 65.89 |
Comparative example 3 | 61.46 |
As can be seen from table two, the extract of millettia speciosa of example 2 has the highest rate of DPPH radical scavenging.
Test III determination of ABTS free radical scavenging ability
3.1 test methods: diluting ABTS stock solution with 10mmol/L PBS (pH 7.4) 40-50 times to give ABTS working solution with absorbance of 0.7 + -0.02 (30 deg.C) at 734 nm; and (3) sample determination: absorbing 6mLABTS working solution, adding 60 μ L of sample solution, oscillating for 10s, standing at 30 deg.C for 6min, and measuring the light absorption value at 734 nm.
results of determination of radical scavenging ability of epi-three ABTS
As can be seen from table three, the extract of millettia speciosa of example 2 has the highest clearance for ABTS free radicals.
Assay for four, determination of advanced glycation end product scavenging Capacity
4.1 test methods: and (3) sample testing: precisely sucking 5mL of the extract solution of Artocarpus heterophyllus (5 mg/mL) respectively (the control is replaced by buffer), adding 20mg of bovine serum albumin and 90.1mg of glucose respectively, and shaking uniformly to obtain the sample. Negative control: 20mg of bovine serum albumin and 90.1mg of glucose are precisely weighed, 5ml of phosphate buffer is added, and the mixture is uniformly shaken to be used as a negative control. And (3) placing each sample to be tested in a constant temperature and humidity incubator, and incubating for 40h at the temperature of 60 ℃. And (3) taking out each sample after 40h, detecting the samples by using an enzyme-linked immunosorbent assay, wherein the fluorescence excitation/emission wavelength is 370/440nm, and recording the fluorescence absorption value.
determination of the Effect of eliminating the end-product of epi-four advanced glycation
As can be seen from Table IV, example 2 has the highest clearance for advanced glycation endproducts. Because the advanced glycosylation end products can cause the reduction and the aging of protein functions, the aging and the pathological changes of body tissues are further caused, and the millettia extract which can effectively remove the advanced glycosylation end products has the anti-aging effect.
Test five, measurement of Elastase inhibitory Effect
The test method comprises the following steps: mu.L of Tris-HCl (0.05mol/L, pH 8.0), 50. mu.L of elastase and 50. mu.L of a 1.5mg/mL Artocarpus heterophyllus extract solution (in buffer) were shaken well, and after preincubation at 25 ℃ for 20 minutes, 50. mu.L of MAAPVM (in buffer) and 5mmol/L were added, shaken well, and after preincubation at 25 ℃ for 60 minutes, absorbance was measured by a microplate reader at 410 nm.
TABLE five results of Elastase inhibition
Group of | Elastase inhibition/%) |
Example 1 | 80.40 |
Example 2 | 82.54 |
Example 3 | 81.33 |
Comparative example 1 | 62.91 |
Comparative example 2 | 56.25 |
Comparative example 3 | 43.27 |
As can be seen from Table V, example 2 showed the highest inhibition of elastase. The millettia extract can inhibit elastase production, as elastase can digest and decompose elastin and/or collagen in connective tissue protein, resulting in wrinkle and skin relaxation.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (10)
1. The preparation method of the millettia extract is characterized by comprising the following steps:
A) crushing the millettia to obtain first powder;
B) degreasing, filtering and drying the first powder to obtain second powder;
C) and extracting the second powder in an ultrasonic-assisted complex enzyme mode, inactivating enzymes, and purifying to obtain the Artocarpus millettii extract.
2. The method for preparing Artocarpus heterophyllus extract as claimed in claim 1, wherein the method comprises the following steps:
A) crushing and sieving the Artocarpus heterophyllus to obtain first powder;
B) taking the first powder in the step A), adding petroleum ether or diethyl ether which is 8-10 times of the total amount of the first powder, and soaking to obtain a degreasing mixture; filtering the degreased mixture, and drying the obtained filter residue to obtain second powder;
C) mixing the second powder obtained in the step B) with an extraction solvent, leaching for 1-1.5h at 3-5 ℃, performing 300-400W ultrasonic extraction for 1-1.5h at 45-50 ℃, inactivating enzyme, and purifying to obtain the Artocarpus millettii extract.
3. The method for preparing Artocarpus heterophyllus extract as claimed in claim 2, wherein the ratio of the second powder to the extraction solvent in step C) is 1: 45-60.
4. The method for preparing Artocarpus heterophyllus extract as claimed in claim 2 or 3, wherein the extraction solvent is disodium hydrogen phosphate-citric acid buffer solution with pH 4-5 containing 0.5-2% veratryl alcohol and 1.5% -3% complex enzyme.
5. The method for preparing Artocarpus heterophyllus extract as claimed in claim 4, wherein the complex enzyme is prepared from chymopapain, pectinase and cellulase according to the following ratio (2-3): 1: (0.5-1) in mass ratio.
6. The method for preparing an extract of Artocarpus heterophyllus according to claim 1 or 2, wherein the purification process comprises:
centrifuging the enzyme-inactivated extract in a centrifuge at 4500-5000rmp for 25-30min, and removing filter residue to obtain supernatant; primarily purifying the supernatant through a 400-plus 500nm ceramic membrane to obtain a filtrate; concentrating the filtrate under reduced pressure until no ethanol smell; performing secondary purification by using macroporous resin, eluting by using 50-70% ethanol solution, collecting eluent, concentrating to remove alcohol, and freeze-drying to obtain the Artocarpus heterophyllus extract.
7. The method for preparing Artocarpus heterophyllus extract as claimed in claim 6, wherein the macroporous resin is one of HPD-400, DM130, D4020, HPD100, DA201-C, S-8, D101, AB-8, ADS-17.
8. Artocarpus heterophyllus extract prepared by the method according to any one of claims 1-7.
9. Use of Artocarpus heterophyllus extract prepared by the preparation method according to any one of claims 1-7 in preparing anti-aging skin care product/cosmetic.
10. An anti-aging preparation, characterized by comprising an effective amount of the extract of Artocarpus heterophyllus prepared by the preparation method according to any one of claims 1 to 7 and an effective amount of cosmetic adjuvants.
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