CN113952269A - Preparation method and application of olive leaf extract - Google Patents
Preparation method and application of olive leaf extract Download PDFInfo
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- CN113952269A CN113952269A CN202111516589.7A CN202111516589A CN113952269A CN 113952269 A CN113952269 A CN 113952269A CN 202111516589 A CN202111516589 A CN 202111516589A CN 113952269 A CN113952269 A CN 113952269A
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- 229940114496 olive leaf extract Drugs 0.000 title claims abstract description 77
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 81
- 240000007817 Olea europaea Species 0.000 claims abstract description 42
- 108090000790 Enzymes Proteins 0.000 claims abstract description 28
- 102000004190 Enzymes Human genes 0.000 claims abstract description 28
- 239000011347 resin Substances 0.000 claims abstract description 28
- 229920005989 resin Polymers 0.000 claims abstract description 28
- 238000000605 extraction Methods 0.000 claims abstract description 23
- 238000001179 sorption measurement Methods 0.000 claims abstract description 20
- 239000002904 solvent Substances 0.000 claims abstract description 19
- 239000002537 cosmetic Substances 0.000 claims abstract description 15
- 230000002087 whitening effect Effects 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000000284 extract Substances 0.000 claims abstract description 13
- 239000008367 deionised water Substances 0.000 claims abstract description 9
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 9
- 238000001914 filtration Methods 0.000 claims abstract description 9
- 238000010438 heat treatment Methods 0.000 claims abstract description 9
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- 238000009210 therapy by ultrasound Methods 0.000 claims abstract description 8
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- 150000001875 compounds Chemical class 0.000 claims description 8
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- 235000002725 Olea europaea Nutrition 0.000 claims description 5
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- HKVGJQVJNQRJPO-UHFFFAOYSA-N Demethyloleuropein Natural products O1C=C(C(O)=O)C(CC(=O)OCCC=2C=C(O)C(O)=CC=2)C(=CC)C1OC1OC(CO)C(O)C(O)C1O HKVGJQVJNQRJPO-UHFFFAOYSA-N 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- RFWGABANNQMHMZ-HYYSZPHDSA-N Oleuropein Chemical compound O([C@@H]1OC=C([C@H](C1=CC)CC(=O)OCCC=1C=C(O)C(O)=CC=1)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O RFWGABANNQMHMZ-HYYSZPHDSA-N 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 229940095731 candida albicans Drugs 0.000 description 3
- 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 3
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 3
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- JUUBCHWRXWPFFH-UHFFFAOYSA-N Hydroxytyrosol Chemical compound OCCC1=CC=C(O)C(O)=C1 JUUBCHWRXWPFFH-UHFFFAOYSA-N 0.000 description 2
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- 238000002386 leaching Methods 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
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- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 1
- 229940123457 Free radical scavenger Drugs 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000795633 Olea <sea slug> Species 0.000 description 1
- 241000207834 Oleaceae Species 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
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- 230000002421 anti-septic effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
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- 229930003935 flavonoid Natural products 0.000 description 1
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- 150000002338 glycosides Chemical class 0.000 description 1
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- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
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- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
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- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/10—Anti-acne agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
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- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- 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
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- 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
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- 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
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Abstract
The invention relates to the technical field of plant extraction methods and plant extracts, in particular to a preparation method and application of an olive leaf extract, which comprises the following steps: s1, crushing olive leaves, placing the crushed olive leaves in a sealed extraction tank, adding a solvent and a complex enzyme, sealing, heating, performing ultrasonic treatment, extracting, filtering, and concentrating under reduced pressure to obtain a concentrated extracting solution; s2, carrying out macroporous resin adsorption on the concentrated extract, sequentially eluting with deionized water and ethanol solution, collecting ethanol eluate, and concentrating under reduced pressure to obtain the olive leaf extract. The preparation method of the olive leaf extract is simple and efficient, convenient to operate and control, high in quality of produced products, low in cost and beneficial to industrial production, and the prepared olive leaf extract has strong oxidation resistance and antibacterial performance and can be applied to the fields of cosmetics such as aging resistance, whitening, sun protection and the like.
Description
Technical Field
The invention relates to a plant extraction method and the technical field of plant extracts, in particular to a preparation method and application of an olive leaf extract.
Background
Olea europaea L is evergreen tree of Olea of Oleaceae (Oleaeuropaea), also known as Olea europaea, Olea europaea L. It is an important economic forest in subtropics and also an important source for producing olive oil in mediterranean. Olive cultivation, olive oil production and compound medicine in mediterranean countries have been in history for thousands of years. Although the olive oil industry is currently well developed, a large amount of by-products and residues are generated every year by pruning, natural defoliation and processing industries, most of which are discarded or incinerated, and many olive oil producers even charge farmers with the treatment cost of olive leaves, thereby causing a large waste of resources and serious damage to the ecological environment. Research shows that the olive leaves contain rich active ingredients, mainly including Oleuropein (Oleuropein), hydroxytyrosol, flavonoids, lignans, caffeoylphenylethanoid glycosides and other polyphenol compounds. Because the olive leaf extract has stronger effects of oxidation resistance, inflammation resistance, bacteriostasis and the like, the olive leaf extract has very wide application prospect, can be applied to whitening and sun-screening cosmetics and can also be used as an acne inhibitor. Therefore, the olive leaves are a raw material with abundant and cheap sources, and can be developed and utilized as products with high added value.
At present, the common methods for extracting heat-sensitive substances mainly comprise leaching, ultrasonic extraction, microwave extraction and the like. In addition, in some novel efficient extraction methods, the enzyme-assisted extraction method has been widely applied to the extraction of active ingredients of various traditional Chinese medicines due to the characteristics of high catalytic efficiency, strong specificity, mild reaction conditions, environmental friendliness and the like, but has the problems of high-temperature enzyme deactivation, accumulation of enzymatic products to influence the leaching of target products and the like.
Disclosure of Invention
In order to overcome the defects and shortcomings in the prior art, the invention aims to provide the preparation method of the olive leaf extract, the preparation method is simple and efficient, the operation and control are convenient, the quality of the produced product is high, the cost is low, the industrial production is facilitated, and meanwhile, the prepared olive leaf extract has strong oxidation resistance and antibacterial performance and can be applied to the cosmetic fields of aging resistance, whitening, sun protection and the like.
The invention aims to provide the application of the olive leaf extract, and the prepared olive leaf extract has strong antioxidant and antibacterial properties, and can be applied to the field of cosmetics with the functions of resisting aging, whitening, preventing sunlight and the like.
The purpose of the invention is realized by the following technical scheme: a method for preparing olive leaf extract comprises the following steps:
s1, crushing olive leaves, placing the crushed olive leaves in a sealed extraction tank, adding a solvent and a complex enzyme, sealing, heating and ultrasonically treating to obtain a feed-liquid mixture, extracting and filtering, and finally concentrating the extracting solution under reduced pressure to 2.3-2.6 times of the weight of the original olive leaves to obtain a concentrated extracting solution for later use;
s2, subjecting the concentrated extract obtained in the step S1 to macroporous resin adsorption treatment, sequentially eluting with deionized water and 30-80% ethanol solution at the flow rate of 0.5-3mL/min, collecting ethanol eluate, and concentrating under reduced pressure to obtain the olive leaf extract.
The olive leaf extract is obtained by extraction through the method, has strong antioxidant and antibacterial properties, and can be applied to the field of cosmetics with the functions of resisting aging, whitening, preventing sunlight and the like; the extraction method is simple and efficient, convenient to operate and control, high in quality of produced products, low in cost and beneficial to industrial production.
Preferably, in step S1, the addition amount of the complex enzyme is 2-4% of the total weight of the olive leaves. Preferably, the ratio of the materials to the liquid in the step S1 is 1: 10-30.
Preferably, in step S1, the solvent is an ethanol solution with a mass fraction of 50-70%.
In the invention, 50-70% of ethanol is used as a solvent, so that better extraction effect can be obtained compared with the prior method of adopting pure water.
Preferably, in step S1, the complex enzyme is a mixture of cellulase and pectinase at a weight ratio of 0.8-1.2: 1.0.
Preferably, in step S1, the ultrasonic power is 200-. Preferably, in step S2, the macroporous resin is AB-8 type macroporous adsorption resin or D-101 type macroporous adsorption resin.
The invention adopts the ultrasonic-assisted complex enzyme extraction method, can effectively avoid the problems of long preparation period, incomplete extraction of active ingredients and easy oxidative degradation of the active ingredients of the traditional extraction and purification process, and obtains the olive leaf extract with high purity at lower temperature and in shorter extraction period.
The invention also provides application of the olive leaf extract, and the olive leaf extract can be applied to cosmetics or skin care products for whitening, sun protection or acne treatment.
The invention has the beneficial effects that: the invention aims to provide a preparation method of an olive leaf extract, which is simple and efficient, convenient to operate and control, high in quality of produced products, low in cost and beneficial to industrial production, and the prepared olive leaf extract has strong oxidation resistance and antibacterial performance and can be applied to the fields of cosmetics such as aging resistance, whitening, sun protection and the like.
The invention aims to provide the application of the olive leaf extract, and the prepared olive leaf extract has strong antioxidant and antibacterial properties, and can be applied to the field of cosmetics with the functions of resisting aging, whitening, preventing sunlight and the like.
Drawings
FIG. 1 is a liquid chromatogram of oleuropein standard;
FIG. 2 is a liquid chromatogram of an olive leaf extract obtained by extraction in example 6 of the present invention;
FIG. 3 is DPPH radical clarity of olive leaf extract of the present invention, wherein the abscissa is extract concentration (mg/mL) and the ordinate is DPPH radical clarity (%);
FIG. 4 is ABTS free radical clarity of olive leaf extract of the present invention, wherein the abscissa is extract concentration (mg/mL) and the ordinate is ABTS free radical clarity (%);
FIG. 5 is the hydroxyl radical clarity of the olive leaf extract of the present invention, wherein the abscissa is the extract concentration (mg/mL) and the ordinate is the hydroxyl radical clarity (%).
Detailed Description
For the understanding of those skilled in the art, the present invention will be further described with reference to the following examples and accompanying fig. 1-5, which are not intended to limit the present invention.
Example 1
A method for preparing olive leaf extract comprises the following steps:
s1, crushing olive leaves, placing the crushed olive leaves in a sealed extraction tank, adding a solvent and a complex enzyme, sealing, heating and ultrasonically treating to obtain a feed-liquid mixture, extracting and filtering, and finally concentrating the extracting solution under reduced pressure to 2.3 times of the weight of the original olive leaves to obtain a concentrated extracting solution for later use;
s2, subjecting the concentrated extract obtained in the step S1 to macroporous resin adsorption treatment, sequentially eluting with deionized water and 30% ethanol solution at a flow rate of 0.5mL/min, collecting ethanol eluate, and concentrating under reduced pressure to obtain the olive leaf extract.
In step S1, the addition amount of the compound enzyme is 2% of the total weight of the olive leaves.
The material-liquid ratio in step S1 is 1: 10. In step S1, the solvent is an ethanol solution with a mass fraction of 50%.
In step S1, the complex enzyme is a mixture of cellulase and pectinase at a weight ratio of 0.8: 1.0. In step S1, the ultrasonic power is 200W, the ultrasonic treatment time is 40min, and the temperature is 30 ℃.
In step S2, the macroporous resin is AB-8 type macroporous adsorption resin or D-101 type macroporous adsorption resin.
Application of olive leaf extract in cosmetics or skin care products for whitening, sun protection or acne treatment.
Example 2
A method for preparing olive leaf extract comprises the following steps:
s1, crushing olive leaves, placing the crushed olive leaves in a sealed extraction tank, adding a solvent and a complex enzyme, sealing, heating and ultrasonically treating to obtain a feed-liquid mixture, extracting and filtering, and finally concentrating the extracting solution under reduced pressure to 2.4 times of the weight of the original olive leaves to obtain a concentrated extracting solution for later use;
s2, subjecting the concentrated extract obtained in the step S1 to macroporous resin adsorption treatment, sequentially eluting with deionized water and 38% ethanol solution at the flow rate of 1.1mL/min, collecting ethanol eluate, and concentrating under reduced pressure to obtain the olive leaf extract.
In the step S1, the addition amount of the compound enzyme is 2.5 percent of the total weight of the olive leaves.
The feed-liquid ratio in step S1 is 1: 15. In step S1, the solvent is an ethanol solution with a mass fraction of 55%.
In step S1, the complex enzyme is a mixture of cellulase and pectinase at a weight ratio of 0.92: 1.0. In step S1, the ultrasonic power is 260W, the ultrasonic treatment time is 48min, and the temperature is 38 ℃.
In step S2, the macroporous resin is AB-8 type macroporous adsorption resin.
Application of olive leaf extract in cosmetics or skin care products for whitening, sun protection or acne treatment.
Example 3
A method for preparing olive leaf extract comprises the following steps:
s1, crushing olive leaves, placing the crushed olive leaves in a sealed extraction tank, adding a solvent and a complex enzyme, sealing, heating and ultrasonically treating to obtain a feed-liquid mixture, extracting and filtering, and finally concentrating the extracting solution under reduced pressure to 2.5 times of the weight of the original olive leaves to obtain a concentrated extracting solution for later use;
s2, subjecting the concentrated extract obtained in the step S1 to macroporous resin adsorption treatment, sequentially eluting with deionized water and 55% ethanol solution at the flow rate of 1.7mL/min, collecting ethanol eluate, and concentrating under reduced pressure to obtain the olive leaf extract.
In step S1, the addition amount of the compound enzyme is 3% of the total weight of the olive leaves.
The feed-liquid ratio in step S1 is 1: 20. In step S1, the solvent is an ethanol solution with a mass fraction of 60%.
In step S1, the complex enzyme is a mixture of cellulase and pectinase at a weight ratio of 1.0: 1.0. In step S1, the ultrasonic power is 350W, the ultrasonic treatment time is 55min, and the temperature is 45 ℃.
In step S2, the macroporous resin is AB-8 type macroporous adsorption resin.
Application of olive leaf extract in cosmetics or skin care products for whitening, sun protection or acne treatment.
Example 4
A method for preparing olive leaf extract comprises the following steps:
s1, crushing olive leaves, placing the crushed olive leaves in a sealed extraction tank, adding a solvent and a complex enzyme, sealing, heating and ultrasonically treating to obtain a feed-liquid mixture, extracting and filtering, and finally concentrating the extracting solution under reduced pressure to 2.5 times of the weight of the original olive leaves to obtain a concentrated extracting solution for later use;
s2, subjecting the concentrated extract obtained in the step S1 to macroporous resin adsorption treatment, sequentially eluting with deionized water and an ethanol solution with the mass fraction of 62% at the flow rate of 2.3mL/min, collecting ethanol eluent, and concentrating under reduced pressure to obtain the olive leaf extract.
In the step S1, the addition amount of the compound enzyme is 3.5% of the total weight of the olive leaves.
The feed-liquid ratio in step S1 is 1: 25. In step S1, the solvent is a 65% by mass ethanol solution.
In step S1, the complex enzyme is a mixture of cellulase and pectinase at a weight ratio of 1.1: 1.0. In step S1, the ultrasonic power was 410W, the ultrasonic treatment time was 62min, and the temperature was 52 ℃.
In step S2, the macroporous resin is AB-8 type macroporous adsorption resin.
Application of olive leaf extract in cosmetics or skin care products for whitening, sun protection or acne treatment.
Example 5
A method for preparing olive leaf extract comprises the following steps:
s1, crushing olive leaves, placing the crushed olive leaves in a sealed extraction tank, adding a solvent and a complex enzyme, sealing, heating and ultrasonically treating to obtain a feed-liquid mixture, extracting and filtering, and finally concentrating the extracting solution under reduced pressure to 2.6 times of the weight of the original olive leaves to obtain a concentrated extracting solution for later use;
s2, subjecting the concentrated extract obtained in the step S1 to macroporous resin adsorption treatment, eluting with deionized water and 80% ethanol solution at a flow rate of 3mL/min, collecting ethanol eluate, and concentrating under reduced pressure to obtain the olive leaf extract.
In step S1, the addition amount of the compound enzyme is 4% of the total weight of the olive leaves.
The feed-liquid ratio in step S1 is 1: 30. In step S1, the solvent is an ethanol solution with a mass fraction of 70%.
In step S1, the complex enzyme is a mixture of cellulase and pectinase at a weight ratio of 1.2: 1.0. In step S1, the ultrasonic power is 500W, the ultrasonic treatment time is 70min, and the temperature is 60 ℃.
In step S2, the macroporous resin is AB-8 type macroporous adsorption resin.
Application of olive leaf extract in cosmetics or skin care products for whitening, sun protection or acne treatment.
Example 6
A method for preparing olive leaf extract comprises the following steps:
s1, crushing 50.0g of olive leaves, placing the crushed olive leaves in a sealed extraction tank, adding a solvent and a complex enzyme, sealing, heating and ultrasonically treating to obtain a feed liquid mixture, extracting and filtering, and finally concentrating the extracting solution under reduced pressure to 2.5 times of the weight of the original olive leaves to obtain a concentrated extracting solution for later use;
s2, subjecting the concentrated extract obtained in the step S1 to macroporous resin adsorption treatment, sequentially eluting with deionized water and 60% ethanol solution at the flow rate of 1mL/min, collecting ethanol eluate, and concentrating under reduced pressure to obtain the olive leaf extract.
In step S1, the addition amount of the compound enzyme is 3% of the total weight of the olive leaves.
The feed-liquid ratio in step S1 is 1: 20. In step S1, the solvent is an ethanol solution with a mass fraction of 50%.
In step S1, the complex enzyme is a mixture of cellulase and pectinase at a weight ratio of 1:1. In step S1, the ultrasonic power is 300W, the ultrasonic treatment time is 60min, and the temperature is 45 ℃.
In step S2, the macroporous resin is AB-8 type macroporous adsorption resin.
Application of olive leaf extract in cosmetics or skin care products for whitening, sun protection or acne treatment.
The olive leaf extract obtained by extraction in example 6 was subjected to high performance liquid chromatography analysis by the following method: using a Welch Ultimate AQ-C18(4.6 x 250mm, 5um) column with mobile phases water (phase a) and acetonitrile (phase B), using a gradient elution procedure, with an elution procedure of 0 to 15min, mobile phase a changing from 80% to 60%; the flow rate is set to be 0.8-1.5mL/min, the detection wavelength is 250-290nm, the sample injection amount is 5uL, and the temperature of the column incubator is 30-35 ℃. The liquid phase spectrum of the olive leaf extract is shown in fig. 1-2.
The following are test sections:
1. analysis and results of antioxidant ability of olive leaf extract
The DPPH method is a common antioxidant test method, when DPPH is dissolved in absolute ethyl alcohol, stable DPPH free radicals are formed and are in a purple state, an ultraviolet absorption peak is formed at 517nm, a free radical scavenger can enable single electron pairing, the A517nm value is reduced, the solution is faded, and the level of color fading is in a stoichiometric relation with the number of paired electrons.
Firstly, preparing a DPPH ethanol solution with the concentration of 0.1mmol/L, then respectively preparing sample solutions with the mass concentrations of 0.3, 0.6, 0.9, 1.2, 1.5 and 2.0mg/mL, uniformly mixing 100 mu L of the sample solutions with 100 mu L of the DPPH ethanol solution, standing in the dark at room temperature for 20min, detecting the absorbance value A of the mixed solution at 517nm by using a multifunctional microplate reader, and taking Trolox as a positive control in the experiment, wherein the clearance rate is [1- (A1-A2)/A0] multiplied by 100%. As shown in FIG. 3, the purified olive leaf extract has DPPH radical scavenging ability, and has a certain dose-effect relationship, and when the concentration reaches 1.5mg/mL, the effect of scavenging Trolox is equivalent to that of the positive control, and the purified olive leaf extract can be used as a potential natural antioxidant.
The ABTS method is a relatively classical antioxidant capacity analysis method, is suitable for natural or synthetic antioxidant detection, and is characterized in that after a proper amount of ABTS is dissolved in distilled water, the ABTS is oxidized by active oxygen to generate a stable dark green cationic free radical ABTS +. the ABTS + solution is added with a sample prepared in advance, and if the antioxidant capacity exists, the substance reacts with the ABTS +. and a solvent system fades.
Firstly, preparing a reagent I: accurately weighing 0.0384g of ABTS and fixing the volume to 10mL, and adding a reagent II: weighing 0.0134g of potassium persulfate, fixing the volume to 10mL, mixing the reagent I and the reagent II at a ratio of 1:1, reacting for 12 hours in a dark place to obtain ABTS +. working solution, and diluting the ABTS +. working solution by 20 times by using absolute ethyl alcohol before use; then preparing sample solutions with mass concentrations of 0.3, 0.6, 0.9, 1.2, 1.5 and 2.0mg/mL, uniformly mixing 50 mu L of the sample solution and 150 mu L of ABTS +. working solution, placing the mixture in a water bath at 25 ℃ in a dark place for reaction for 10min, detecting the absorbance value A of the mixed solution at 735nm by using a multifunctional microplate reader, and taking Trolox as a positive control in the experiment, wherein the clearance rate is [1- (A1-A2)/A0] multiplied by 100%. The result is shown in figure 4, along with the increase of the mass concentration of the olive leaf extract and Trolox, the ABTS +. clearance rate is increased, and then gradually becomes stable, so that the purified olive leaf extract has stronger ABTS +. clearance capacity and the capacity of directly clearing ROS in vitro.
The hydroxyl radical has extremely strong electron-obtaining capability, namely oxidation capability, the oxidation potential is 2.8V, and the hydroxyl radical is an oxidant which is second to fluorine in nature. The hydroxyl radical clearance is determined as follows:
firstly, preparing 9mmol/L FeSO respectively4Solution, 9mmol/L ethanol solution of salicylic acid, 8.8mmol/L H2O2(30%) solution and sample solution with mass concentration of 0.05, 0.2, 0.4, 0.8, 1.2, 1.5mg/mL, take 1.0mL sample solution and 1.0mL working solution to mix, place in 37 ℃ water bath kettle and stand for 30min, detect the absorbance value of mixed solution at 510nm, the experimental result is shown as figure 5, along with the increasing of sample mass concentration within a certain range, the olive leaf extract after purifying is similar to positive control substance Trolox, present good dose-effect relation to the clearance of hydroxyl radical, indicate that the extract has good oxidation resistance, is a potential natural antioxidant.
2. Analysis of bacteriostatic effect of olive leaf extract
Firstly, taking a staphylococcus aureus, escherichia coli, candida albicans, pseudomonas aeruginosa and bacillus subtilis cryopreservation tube out of a refrigerator at the temperature of-80 ℃, placing the tube in a biological safety cabinet until the temperature of the tube is recovered to normal temperature, then respectively inoculating 1 ring of about 10 mu L of bacterial liquid into 10mL of BHI liquid culture medium, culturing for 48h at the temperature of 37 ℃, then taking 1mL of liquid, placing the liquid into 9mL of physiological saline, diluting twice and 100 times to prepare bacterial suspension of about 107CFU/mL for later use, then respectively preparing the liquid to be tested into sample solutions with final concentrations of 100, 300 and 600mg/mL, respectively carrying out in-vitro bacteriostasis tests on the staphylococcus aureus, the escherichia coli, the candida albicans, the pseudomonas aeruginosa and the bacillus subtilis by adopting an Oxford cup method, and finally calculating the bacteriostasis circle range of each strain to obtain a conclusion. The specific parameters are shown in table 1:
TABLE 1
As shown in Table 1, the olive leaf extract purified and treated at different concentrations has inhibition zones on staphylococcus aureus, escherichia coli, candida albicans, pseudomonas aeruginosa and bacillus subtilis, and the staphylococcus aureus, the pseudomonas aeruginosa and the bacillus subtilis have stronger sensitivity on the olive leaf extract by observing the inhibition zones; in addition, the bacteriostatic effect on five strains is gradually enhanced along with the increase of the mass concentration of the sample, which shows that the purified olive leaf extract has good antibacterial and antiseptic capabilities.
The above-described embodiments are preferred implementations of the present invention, and the present invention may be implemented in other ways without departing from the spirit of the present invention.
Claims (10)
1. A preparation method of an olive leaf extract is characterized in that: the method comprises the following steps:
s1, crushing olive leaves, placing the crushed olive leaves in a sealed extraction tank, adding a solvent and a complex enzyme, sealing, heating and ultrasonically treating to obtain a feed liquid mixture, extracting and filtering, and finally concentrating the extracting solution under reduced pressure to obtain a concentrated extracting solution for later use;
s2, subjecting the concentrated extract obtained in the step S1 to macroporous resin adsorption treatment, eluting with deionized water and 30-80% ethanol solution in sequence, collecting ethanol eluate, and concentrating under reduced pressure to obtain the olive leaf extract.
2. The method for preparing olive leaf extract according to claim 1, wherein the olive leaf extract comprises the following components: in the step S1, the addition amount of the compound enzyme is 2-4% of the total weight of the olive leaves.
3. The method for preparing olive leaf extract according to claim 1, wherein the olive leaf extract comprises the following components: the material-liquid ratio in the step S1 is 1: 10-30.
4. The method for preparing olive leaf extract according to claim 1, wherein the olive leaf extract comprises the following components: in step S1, the solvent is an ethanol solution with a mass fraction of 50-70%.
5. The method for preparing olive leaf extract according to claim 1, wherein the olive leaf extract comprises the following components: in step S1, the complex enzyme is a mixture of cellulase and pectinase at a weight ratio of 0.8-1.2: 1.0.
6. The method for preparing olive leaf extract according to claim 1, wherein the olive leaf extract comprises the following components: in step S1, the ultrasonic power is 200-500W, the ultrasonic treatment time is 40-70min, and the temperature is 30-60 ℃.
7. The method for preparing olive leaf extract according to claim 1, wherein the olive leaf extract comprises the following components: in step S1, the weight of the concentrated extractive solution is reduced to 2.3-2.6 times of the weight of the original olive leaf during vacuum concentration.
8. The method for preparing olive leaf extract according to claim 1, wherein the olive leaf extract comprises the following components: in step S2, the macroporous resin is AB-8 type macroporous adsorption resin or D-101 type macroporous adsorption resin.
9. The method for preparing olive leaf extract according to claim 1, wherein the olive leaf extract comprises the following components: in step S2, the elution flow rate in the elution treatment is 0.5 to 3 mL/min.
10. Use of olive leaf extract according to any one of claims 1-9, characterized in that: the olive leaf extract can be applied to cosmetics or skin care products for whitening, sun screening or acne treatment.
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| CN114469820A (en) * | 2022-03-17 | 2022-05-13 | 中国科学院兰州化学物理研究所 | Olive leaf extract and application thereof in preparation of skin whitening cosmetics |
| CN114532364A (en) * | 2022-01-26 | 2022-05-27 | 中国科学院兰州化学物理研究所 | Method for preparing nano-selenium/attapulgite composite antibacterial material by using olive leaf extract |
| CN115282216A (en) * | 2022-09-15 | 2022-11-04 | 中国科学院兰州化学物理研究所 | Composition with potential Alzheimer disease prevention and treatment functions |
| CN116077413A (en) * | 2023-03-06 | 2023-05-09 | 广州优理氏生物科技有限公司 | Composition for resisting oxidation and aging and application thereof as well as essence |
| CN115364012B (en) * | 2022-08-30 | 2023-08-15 | 上海科黛生物科技有限公司 | Preparation method of supermolecule olive composition and application of supermolecule olive composition in repairing essence |
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