CN116422002B - Aloe extract and preparation method and application thereof - Google Patents
Aloe extract and preparation method and application thereof Download PDFInfo
- Publication number
- CN116422002B CN116422002B CN202310070876.2A CN202310070876A CN116422002B CN 116422002 B CN116422002 B CN 116422002B CN 202310070876 A CN202310070876 A CN 202310070876A CN 116422002 B CN116422002 B CN 116422002B
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- China
- Prior art keywords
- ionic liquid
- aloe
- extract
- lysine
- aloe extract
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Classifications
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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]
- A61K8/9794—Liliopsida [monocotyledons]
-
- 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
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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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/028—Flow sheets
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0288—Applications, solvents
-
- 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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- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Abstract
The application relates to an aloe extract, a preparation method and application thereof, and relates to the technical field of plant extraction and separation. The preparation method of the aloe extract comprises the following steps: (1) Uniformly mixing aloe and the ionic liquid, and placing the mixture in a water bath at 85-95 ℃ to react for 4-8 hours to obtain an extraction stock solution; (2) Adding a diluent into the extracting stock solution to obtain diluted stock solution; (3) Adding potassium phosphate into the diluted stock solution until the potassium phosphate is saturated, and filtering to obtain filtrate; (4) Adding hydrochloric acid into the filtrate obtained in the step (3) to adjust the pH to 1.5-2, centrifuging and collecting precipitate, and drying to obtain aloe extract. The ionic liquid disclosed by the application is environment-friendly, pollution to the environment caused by volatilization of an organic solvent is avoided, the extraction efficiency of active ingredients is improved, the operation is simple, and the ionic liquid has good popularization and application prospects.
Description
Technical Field
The application relates to an aloe extract and a preparation method and application thereof, belonging to the technical field of plant extraction and separation.
Background
The aloe is perennial evergreen succulent herb of Liliales, liliaceae, and also known as Aloe vera, aloe sinensis, olium Trogopterori, aloe arborescens, aloe barbadensis, aloe us, etc. Aloe is native to tropical arid regions of africa and is distributed almost throughout the world. Wild aloe distribution is found in india and malaysia in one-belt, african continents and tropical regions. Cultivated in the areas of Chinese Fujian, taiwan, guangdong, guangxi, sichuan, yunnan, etc., and also the aloe in a wild state exists. Aloe (i.e., aloe vera) is one of the few edible species in the aloe genus, and its products are widely used in the fields of foods, cosmetics, health care, medicine, etc.
Aloe is a pure natural green plant integrating medicine, beauty, health care, food and ornamental, and various medical values and therapeutic effects of aloe are widely paid attention to in the medical field. From the beginning of the 80 s of the 20 th century, aloe has been continuously touted by consumers and has rapidly raised the hot tide of "aloe" worldwide. The aloe is rich in active ingredients such as barbaloin, polysaccharide, organic acid, protein and the like, has the functions of skin care, beauty treatment, sun protection, anti-aging, anti-inflammatory, anticancer, body immunity enhancement, detoxification, health care, wound healing promotion and the like, is widely applied to cosmetics and daily chemical industry, enjoys the beauty of 'universal good medicine', 'natural beauty treatment' and is a plant resource with development and utilization value. Thus aloe also has a well-known name in folk, the "botanicals". The aloe and the yellow juice of gastroenteropathy aloe have the effects of diminishing inflammation, sterilizing, invigorating stomach, relaxing bowels and the like, and have remarkable treatment effect on acute gastritis. In addition, aloe is also helpful for treating gastric ulcers caused by gastric acid because aloe-rich mucus can adhere to damaged ulcer surfaces, not only can activate cell tissue regeneration, but also can enable ulcer parts and surrounding tissues to grow new tissues. Research on aloe and constipation proves that anthraquinone compound derivatives contained in aloe release aloe-emodin in intestinal canal, can effectively stimulate large intestine peristalsis, thereby effectively improving gastrointestinal dysfunction, constipation and other symptoms, and expelling toxin, reducing pathogenic fire, tonifying deficiency and beautifying.
Aloe vera has long been prepared as an aloe vera gel. The aloe leaf is first cut, leaf juice is collected, the aloe leaf juice is then decocted into thick paste in a pot, and the thick paste is then soaked in a container and cooled to solidify. However, the aloe vera gel prepared by the method is rough, and the natural active ingredients are mostly decomposed by a pot boiling and heating method. Based on the traditional preparation process, cobble firstly provides a method for preparing stable transparent aloe gel, namely, an antioxidant and a preservative are added in the preparation process, so that the extraction quality of aloe gel is improved, but aloe gel extracted by the extraction method can cause skin allergy, and the processes of heating, oxidization and the like, but the heating can accelerate the degradation of active ingredients, and oxidization can cause the oxidization of active ingredients. In addition, the aloe active ingredient is extracted by acid extraction, alkali extraction and enzyme extraction. Along with the development of technology, other new extraction methods are derived.
Ionic Liquids (ILs) are salts which consist entirely of anions and cations and are liquid at temperatures below 100 ℃. Compared with the conventional ionic compound, the ionic liquid often has various acting forces such as dipole, hydrogen bond, van der Waals force and the like, and the hydrogen bond is generally strong in alkalinity, so that the ionic liquid has the potential of obtaining excellent molecular selectivity as a separation solvent. Compared with the traditional organic solvents, the ionic liquid has a series of outstanding advantages: almost no vapor pressure, no environmental pollution and low pressure conditions, and is therefore called "green solvent"; is nonflammable, and has high thermal stability and chemical stability; the liquid state temperature range is wide; the polarity and the hydrophilicity of the organic compound, the inorganic compound and the polymer can be regulated and controlled through the design of anions and cations, so that the organic compound, the inorganic compound and the polymer can be regulated and dissolved, and the organic compound, the inorganic compound and the polymer are also called as a designability solvent; because of the special properties of the ionic liquid, the ionic liquid is widely focused by people as a green solvent and a functional material, and the report of the ionic liquid applied to the field of plant extraction at present shows a wide application prospect. So far, there are few reports on the extraction of aloe active ingredients by using ionic liquids.
Disclosure of Invention
The application aims to provide an aloe extract and a preparation method and application thereof.
In order to achieve the above purpose, the technical scheme adopted by the application is as follows: a preparation method of aloe extract comprises the following steps: (1) Uniformly mixing aloe and the ionic liquid, and placing the mixture in a water bath at 85-95 ℃ to react for 4-8 hours to obtain an extraction stock solution; (2) Adding a diluent into the extracting stock solution to obtain diluted stock solution; (3) Adding potassium phosphate into the diluted stock solution until the potassium phosphate is saturated, and filtering to obtain filtrate; (4) Adding hydrochloric acid into the filtrate obtained in the step (3) to adjust the pH to 1.5-2, centrifuging and collecting precipitate, and drying to obtain aloe extract.
Aloe is rich in active ingredients such as barbaloin, polysaccharide, organic acid, and protein, and has skin caring, sun protecting, antiaging, and antiinflammatory effects. The traditional aloe extract extraction method has the problems of low extraction rate, more impurities in the extract, poor efficacy of the extract, most of extracting agents are organic solvents, limited application of the extract due to organic solvent residues, environmental pollution and the like. The application adopts ionic liquid to extract aloe. The ionic liquid is used as a novel extractant, and has high selective dissolving capacity for aloin and flavonoid compounds in aloe due to the structural characteristics of the ionic liquid, so that the aloin and the aloe flavonoid (quercetin, rutin) in aloe can be extracted efficiently. Because anions in the ionic liquid are connected with H on the aloe glycoside and aloe flavone molecules, stronger intermolecular interaction is formed; the cations bind to O on the barbaloin and aloe flavone molecules, forming stronger intermolecular hydrogen bonds. The process breaks the original intermolecular interactions of the barbaloin and aloe flavone molecules in aloe, thereby dissolving barbaloin and aloe flavone. Therefore, the method for extracting aloe by using the ionic liquid has the advantages of high extraction rate, high specificity, simplicity in operation and environment friendliness.
As a preferred embodiment of the method for preparing aloe vera extract of the present application, the ionic liquid consists of an alkaloid and an amino acid. Ionic liquids are salts consisting entirely of anions and cations that exhibit a liquid state at or near room temperature. The ionic liquids are various, different combinations of cations and anions are changed, different ionic liquids can be designed and synthesized, different types of ionic liquids can react differently, and different ionic liquids are adopted for extraction of the same extraction substrate, so that the extraction efficiency and the efficacy of the extract are different. The inventor of the present application has found through a great deal of experiments that the aloe is extracted by adopting the ionic liquid composed of alkaloid and amino acid, and the extraction efficiency and the purity of the extract are high.
As a preferred embodiment of the method for preparing aloe extract of the present application, the alkaloid is oxymatrine; the amino acid is lysine. The oxymatrine is quinazoline alkaloid extracted from radix Sophorae Flavescentis, and has effects of resisting tumor, resisting bacteria, resisting parasite, relieving inflammation, detumescence, promoting urination, and relieving leucopenia caused by cyclophosphamide. Through a large number of experimental screening and verification, the inventor discovers that the extraction efficiency of the ionic liquid prepared from the matrine oxide for extracting aloe is higher. Lysine (Lysine) is known by the chemical name 2, 6-diaminohexanoic acid, an essential amino acid. Lysine has positive nutritional significance in the aspects of promoting the growth and development of human bodies, enhancing the immunity of organisms, resisting viruses, promoting fat oxidation, relieving anxiety and emotion and the like. Through a large number of experimental screening and verification, the inventor discovers that the extraction efficiency of the ionic liquid prepared by lysine for extracting aloe is higher.
As a preferred embodiment of the preparation method of aloe extract of the present application, the weight ratio of the matrine oxide to the lysine is matrine oxide: lysine=1: 2 to 4. The inventor researches and discovers that the ionic liquid prepared from the oxymatrine and the lysine is used for extracting aloe, and the active ingredients such as barbaloin, aloe flavonoids and the like can be efficiently extracted from aloe by limiting the weight part ratio of the oxymatrine to the lysine, and the ionic liquid is environment-friendly and does not cause environmental pollution.
As a preferred embodiment of the preparation method of aloe extract of the present application, the weight ratio of the matrine oxide to the lysine is matrine oxide: lysine=1: 2.5 to 3.5. The inventor researches find that the weight part ratio of the oxymatrine to the lysine is in the range, the viscosity of the ionic liquid is moderate, and the aloe extraction is facilitated.
As a preferred embodiment of the method for preparing aloe vera extract of the present application, the method for preparing the ionic liquid extractant comprises the steps of: (1) Weighing matrine oxide and lysine to dissolve in water, and placing in a water bath at 75-85 ℃ to react for 5-7 h to obtain an ionic liquid aqueous solution; (2) Removing water from the ionic liquid aqueous solution to obtain an ionic liquid crude product; (3) And adding cleaning liquid into the crude product of the ionic liquid to clean for 3-4 times, and performing rotary evaporation to obtain the ionic liquid.
As a preferred implementation mode of the preparation method of aloe extract, the water removal in the step (2) is specifically implemented by putting an ionic liquid aqueous solution at 70-75 ℃ for rotary steaming for 1-2 h; the cleaning solution in the step (3) consists of acetonitrile and methanol, wherein the volume ratio of the acetonitrile to the methanol is acetonitrile: methanol=9: 1.
as a preferred embodiment of the preparation method of the aloe extract, the aloe and the ionic liquid are prepared from the following components in parts by weight: ionic liquid extractant = 1:10 to 20.
As a preferred embodiment of the preparation method of aloe extract of the present application, the mass ratio of the diluent in the step (2) to the ionic liquid extractant is as follows: ionic liquid = 50:1.
as a preferred embodiment of the method for preparing aloe vera extract of the present application, the diluent consists of acetone and water; the mass ratio of the acetone to the water in the diluent is 1:1.
the application also provides an aloe extract, which is prepared by adopting the preparation method of the aloe extract.
The application also provides a skin care product, which comprises the aloe extract.
As a preferred embodiment of the skin care product, the aloe extract accounts for 5-20% of the weight of the skin care product. Through a great number of experiments, the inventor discovers that the efficacy of the aloe extract is related to the addition amount of the aloe extract, and when the mass percentage of the aloe extract in the skin care product is 5-20%, the aloe extract has better whitening, moisturizing and antioxidation effects.
As a preferred embodiment of the skin care product according to the present application, the aloe vera extract is present in an amount of 10% by mass based on the skin care product. Through a great deal of experiments, the inventor discovers that when the mass percentage of aloe extract in the skin care product is 10%, the skin care product has the best whitening, moisturizing and antioxidation effects.
The application also provides application of the aloe extract in preparing skin care products with whitening and antioxidation effects.
As a preferred embodiment of the application according to the application, the skin care product comprises a face cream, eye cream, toner or essence.
Compared with the prior art, the application has the beneficial effects that: the application provides an aloe extract, a preparation method and application thereof. Compared with the existing method for extracting by adopting the organic solvent, the ionic liquid is environment-friendly, avoids pollution to the environment caused by volatilization of the organic solvent, and simultaneously avoids the limitation of application of aloe extract caused by residue of the organic solvent; the method for extracting aloe by using the ionic liquid is simple to operate, the ionic liquid can be recycled, and the method has good popularization and application prospects.
Drawings
FIG. 1 is a graph showing the evaluation results of soothing efficacy of aloe vera extract prepared in example 1 using zebra fish embryos.
Detailed Description
For a better description of the objects, technical solutions and advantages of the present application, the present application will be further described with reference to the following specific examples.
The aloe powder used in the following examples and comparative examples is commercially available.
Example 1
The preparation method of the aloe extract provided by the embodiment of the application comprises the following steps:
(1) The aloe powder is weighed according to the weight part ratio: ionic liquid = 1:15, uniformly mixing aloe powder and the ionic liquid, and placing the mixture in a water bath at 90 ℃ to react for 6 hours to obtain an extraction stock solution;
(2) The mass ratio of the diluent to the ionic liquid is 50:1, adding a diluent into the extracting stock solution to obtain a diluted stock solution;
(3) Adding excessive potassium phosphate into the diluted stock solution, stirring until the potassium phosphate is not dissolved, and filtering to obtain filtrate;
(4) Adding hydrochloric acid into the filtrate obtained in the step (3) to adjust the pH to 1.5-2, centrifuging and collecting precipitate, and drying to obtain aloe extract.
The ionic liquid of the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 3 parts of lysine.
The preparation method of the ionic liquid in the embodiment comprises the following steps:
(1) Dissolving oxymatrine and lysine in water, and placing in a water bath at 80 ℃ to react for 6 hours to obtain an ionic liquid aqueous solution;
(2) Placing the ionic liquid aqueous solution at 75 ℃ for rotary steaming for 1-2 hours to remove water, thus obtaining an ionic liquid crude product;
(3) Adding a cleaning solution with the volume of 1-2 times into the crude product of the ionic liquid, cleaning for 3-4 times, and rotary steaming to obtain the ionic liquid;
the cleaning solution consists of acetonitrile and methanol in a volume ratio of 9:1.
Example 2
The examples of the present application differ from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 2 parts of lysine.
Example 3
The examples of the present application differ from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 4 parts of lysine.
Example 4
The examples of the present application differ from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 2.5 parts of lysine.
Example 5
The examples of the present application differ from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 3.5 parts of lysine.
Example 6
The embodiment of the application differs from embodiment 1 only in that the weight parts ratio of aloe powder to ionic liquid is different, and in this embodiment, the weight parts ratio of aloe powder to ionic liquid is 1:10.
example 7
The embodiment of the application differs from embodiment 1 only in that the weight parts ratio of aloe powder to ionic liquid is different, and in this embodiment, the weight parts ratio of aloe powder to ionic liquid is 1:20.
comparative example 1
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 1 part of lysine.
Comparative example 2
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of theophylline and 1 part of arginine.
Comparative example 3
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of theophylline and 1 part of lysine.
Comparative example 4
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of theophylline and 1 part of alanine.
Comparative example 5
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of theophylline and 1 part of glycine.
Comparative example 6
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 3 parts of alanine.
Comparative example 7
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 3 parts of glycine.
Comparative example 8
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of theophylline and 3 parts of lysine.
Comparative example 9
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 1 part of arginine.
Comparative example 10
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 2 parts of arginine.
Comparative example 11
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 3 parts of arginine.
Comparative example 12
This comparative example differs from example 1 only in the component content of the ionic liquid. The ionic liquid in the embodiment comprises the following components in parts by weight: 1 part of oxymatrine and 4 parts of arginine.
Effect example
1. The aloe is extracted by adopting the ionic liquid, and the dispersibility and the fluidity of the ionic liquid are related to whether the extraction process can be smoothly carried out. The ionic liquids of examples 1 to 7 and comparative examples 1 to 12 were evaluated for dispersibility and fluidity by using viscosity. The results are shown in Table 1.
TABLE 1
As can be seen from Table 1, not all the different kinds of alkali and amino acids can form an ionic liquid for extracting aloe, and the inventor of the application finds that the ionic liquid composed of matrine oxide and lysine can extract aloe through a large number of experiments, and the ionic liquid has good dispersibility and fluidity.
2. The yields of aloe extracts prepared in examples 1 to 7 were measured, and the results are shown in Table 2.
TABLE 2
| Group of | Yield (%) |
| Example 1 | 11.2％ |
| Example 2 | 7.1％ |
| Example 3 | 8.87％ |
| Example 4 | 6.05％ |
| Example 5 | 8.96％ |
| Example 6 | 8.12％ |
| Example 7 | 11.07％ |
It can be seen from Table 2 that the ratio of matrine oxide to lysine in the ionic liquid, and the ratio of the ionic liquid to aloe have a great influence on the yield of the finally prepared aloe extract. When the weight part ratio of the oxymatrine to the lysine is that the oxymatrine: lysine is 1:3, the yield of the aloe extract is highest; when the weight ratio of aloe powder to ionic liquid is 1:15, the yield of the aloe extract is highest.
3. Evaluation of safety of aloe vera extract
Cosmetic irritation evaluation was performed on aloe extracts prepared in the above examples 1 to 7.
(1) The group of subjects was females, 70, between 16 and 65 years of age. The skin of the subject is healthy, no skin disease allergy history exists, and the subject's volunteer selection standard is met. Randomly, 7 groups of 10 persons each.
The test method is a human body patch test. Selecting qualified patch tester, placing about 0.020-0.025g of test object into the patch tester, applying the patch tester with test object on the back or forearm flexor side of the subject with non-irritating adhesive tape, and applying on skin with palm light pressure for 24 hr. After removing the spot tester for 30min, the skin reaction is observed after the indentation disappears. If the result is negative, the spot test is observed again 24h and 48h after the spot test. The results were recorded according to 2007 "cosmetic health Specification" and 2015 "cosmetic safety Specification" as reference standards.
Evaluation criteria: level 0: a negative reaction; stage 1: suspicious reactions, only weak erythema; 2 stages: weak positive response, erythema, infiltration, edema, and possibly papules; 3 stages: strong positive reaction, erythema, infiltration, edema, papule, and reaction beyond the test area; 4 stages: very strong positive response, marked erythema, severe infiltration, edema, blepharospermia, and response beyond the test area.
(2) The human skin patch experiment result shows that: each group of subjects observed skin reactions of the subjects 24h and 48h after the experiments by the patch experiments of the aloe extracts prepared in examples 1-7, and all subjects were negative in skin reactions, which indicates that the aloe extracts prepared in examples 1-7 were safe to use.
4. DPPH radical scavenging test of aloe vera extract
DPPH free radical is a stable oxygen-centered free radical, is one of important indexes of sample antioxidant capacity, and is widely applied to antioxidant foods. In the research of health care products and medicines. The DPPH free radical has single electron, and the alcohol solution thereof has purple color and strong absorption at 515 nm. In the presence of an antioxidant, DPPH free radicals are scavenged, the color of the solution becomes light, the absorbance at 515nm is reduced, and the change of the absorbance is proportional to the scavenging degree of the free radicals in a certain range. The ability of the sample to scavenge DPPH radicals is reflected by the extent of the decrease in absorbance.
The DPPH free radical scavenging experiment comprises the following specific experimental steps:
(1) Dissolving aloe extracts prepared in examples 1-7 in deionized water to prepare aloe extract to-be-detected liquid with the mass percentage of 10%, wherein the aloe extract to-be-detected liquid is respectively numbered as test groups 1-7; dissolving the aloe extract prepared in the embodiment 1 in deionized water to respectively prepare aloe extract to-be-detected solutions with mass percentages of 5% and 20%, wherein the aloe extract to-be-detected solutions are respectively numbered as a test group 8 and a test group 9;
(2) Referring to table 1, a 10mL test tube was used to set up a sample tube (T), a sample background (t_0), a DPPH tube (C) and a solvent background (c_0), three parallel tubes were set up for each concentration sample tube (T) of each sample, and three parallel tubes were set up for the colleague DPPH tube (C);
(3) 1mL of the sample solution with the same concentration is added into each of the sample tube (T) and the sample background (T_0);
(4) Adding solvent into all test tubes (T, T _0 and C, C _0), adding water into the T, T _0 tube, adding 95% ethanol into the C, C _0 tube, supplementing 3mL, and uniformly mixing;
(5) 1mL of DPPH ethanol solution is added into a sample tube (T) and a DPPH tube (C), the sample background (T_0) and the solvent background (C_0) are replaced by 95% ethanol, the mixture is gently shaken, and the mixture is kept stand for 5 minutes at room temperature;
(6) Each reaction solution was transferred to a 1cm cuvette and absorbance was measured at 517.
DPPH radical clearance: clearance (%) = (1- (T-t_0)/(C-c_0)) ×100%
And (3) measuring the DPPH free radical clearance of aloe extract to be tested in each test group according to the DPPH free radical clearance test method. The test results are shown in Table 4.
TABLE 3 Table 3
TABLE 4 Table 4
5. Tyrosinase activity inhibition assay for aloe vera extract
Melanin synthesis is affected by tyrosinase, which catalyzes the hydroxylation of L-tyrosine to dopa and the oxidation of dopa to dopaquinone, which forms the final reaction product melanin. The test substance with tyrosinase activity inhibition effect can slow down the conversion of L-tyrosine into dopaquinone by tyrosinase, and the inhibition effect of the test substance on tyrosinase activity is evaluated according to the change of absorbance by measuring the absorbance of dopaquinone at 475 nm.
The specific experimental steps of the tyrosinase activity inhibition experiment are as follows:
(1) Dissolving aloe extracts prepared in examples 1-7 in deionized water to obtain aloe extracts with mass percent of 10% to be tested, wherein the aloe extracts are respectively numbered as test groups 1-7; dissolving the aloe extract prepared in the embodiment 1 in deionized water to respectively prepare aloe extracts with mass percentages of 5% and 20% to be tested, wherein the aloe extracts are respectively numbered as a test group 8 and a test group 9;
(2) Referring to the reagent addition amount of Table 5, L-tyrosine solution, sample solution/solvent and PBS buffer are sequentially added into each well, fully and uniformly mixed, incubated at 37 ℃ for 10min, 20 mu L of tyrosinase solution is sequentially added into each well, uniformly mixed at 37 ℃ for 5min plus or minus 5s, and then the mixture is immediately placed into an enzyme-labeled instrument, and the measurement is carried out at 475nm wavelength.
Tyrosinase activity inhibition rate: y= (1- (a_d-a_c)/(a_b-a_a)) ×100%
Wherein: y-tyrosinase activity inhibition rate; ad-absorbance of sample reaction wells; ac-sample bottom hole absorbance; ab-average absorbance of sample wells; aa-average absorbance of the bottom well of solvent.
And respectively measuring the tyrosinase activity inhibition rate of the aloe extract to-be-measured solutions of the test groups according to a tyrosinase activity inhibition rate test method. The test results are shown in Table 6.
TABLE 5
| Reagent(s) | T_a | T_b | T_c | T_d |
| L-tyrosine solution/uL | 0 | 40 | 0 | 40 |
| Sample solution/uL | 0 | 0 | 40 | 40 |
| solvent/uL | 40 | 40 | 0 | 0 |
| PBS buffer/uL | 70 | 30 | 70 | 30 |
| Tyrosinase solution/uL | 20 | 20 | 20 | 20 |
TABLE 6
6. Moisture retention test of aloe vera extract
The moisture retention performance of aloe extract is measured by a weighing method, and the specific experimental method is as follows:
(1) Dissolving aloe extracts prepared in examples 1-7 in deionized water to obtain aloe extracts with mass percent of 10% to be tested, wherein the aloe extracts are respectively numbered as test groups 1-7; dissolving the aloe extract prepared in the embodiment 1 in deionized water to respectively prepare aloe extracts with mass percentages of 5% and 20% to be tested, wherein the aloe extracts are respectively numbered as a test group 8 and a test group 9; the positive control group is 3% glycerol water solution;
(2) Weighing the samples with the same weight, placing the samples in a constant-weight glassware, enabling the samples to be tiled in the glassware, weighing the samples in different time periods (24 hours), recording the change of the weight of the samples along with time, and repeating the test for 3 times.
The formula for calculating the moisture retention rate: moisture retention = (m_2-m_0)/(m_1-m_0) ×100%
Wherein: m_0 is empty plate mass (g); m_1 is the mass (g) of the glass plate after sample addition; m_2 is the mass (g) after several hours of placement in the dryer.
The test results are shown in Table 7.
TABLE 7
7. Test of soothing efficacy of aloe vera extract
Zebra fish embryo has extremely strong sensibility, and can evaluate the safety of compounds including medicines, foods, cosmetics and the like by means of percutaneous absorption, microinjection and the like. The sample is intended to be given by percutaneous absorption, and the spin condition of the embryo is observed to evaluate the relaxation condition of the sample. The specific experimental method is as follows: an AB-line wild type zebra fish embryo was selected 24 hours (24 hpf) after fertilization. Zebra fish were divided into a blank group, a model group, a positive control group and an experimental group. Blank groups are not treated, model groups are molded by sodium dodecyl sulfate, positive control groups are dipotassium glycyrrhizinate, experimental groups are prepared by aloe extract diluents (the concentrations are 8.456mg/mL, 4.228mg/mL and 2.114mg/mL respectively) prepared in example 1, video recording is carried out after incubation for a period of time, each blank is recorded for 30 seconds, and statistical test results are shown in table 8 and figure 1 after recording is finished.
TABLE 8
Compared with a blank control group, the zebra fish embryo spin frequency of the model group is obviously increased (P is less than 0.05), which proves that the modeling is successful, the stimulation of the modeling can be relieved after the positive control substance is dosed, and the embryo spin frequency is effectively reduced. The number of zebra fish embryo spins was found to be significantly reduced relative to the model group at diluted concentrations of 8.456mg/mL, 4.228mg/mL, 2.114mg/mL in the aloe extract test. In conclusion, the aloe extract has a soothing effect under the experimental conditions through analysis of experimental results.
Finally, it should be noted that the above embodiments are only for illustrating the technical solution of the present application and not for limiting the scope of the present application, and although the present application has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made to the technical solution of the present application without departing from the spirit and scope of the technical solution of the present application.
Claims (12)
1. A method for preparing aloe extract, comprising the steps of:
(1) Uniformly mixing aloe and the ionic liquid, and placing the mixture in a water bath at 85-95 ℃ to react for 4-8 hours to obtain an extraction stock solution;
(2) Adding a diluent into the extracting stock solution to obtain diluted stock solution;
(3) Adding potassium phosphate into the diluted stock solution until the potassium phosphate is saturated, and filtering to obtain filtrate;
(4) Adding hydrochloric acid into the filtrate obtained in the step (3) to adjust the pH to 1.5-2, centrifuging and collecting precipitate, and drying to obtain aloe extract;
the ionic liquid consists of matrine oxide and lysine;
the weight ratio of the oxymatrine to the lysine is that the oxymatrine: lysine=1: 2-4;
the preparation method of the ionic liquid comprises the following steps:
a. weighing matrine oxide and lysine to be dissolved in water, and placing the solution in a water bath at 75-85 ℃ to react for 5-7 hours to obtain an ionic liquid aqueous solution;
b. removing water from the ionic liquid aqueous solution to obtain an ionic liquid crude product;
c. and adding cleaning liquid into the crude product of the ionic liquid to clean for 3-4 times, and performing rotary evaporation to obtain the ionic liquid.
2. The method for preparing aloe vera extract according to claim 1, wherein the weight ratio of matrine oxide to lysine is matrine oxide: lysine=1: 2.5-3.5.
3. The method for preparing aloe vera extract according to claim 1, wherein the removing of the water in step b is specifically performed by spin-steaming an aqueous ionic liquid solution at 70-75 ℃ for 1-2 hours; the cleaning solution in the step c consists of acetonitrile and methanol, wherein the volume ratio of acetonitrile to methanol is that acetonitrile: methanol=9: 1.
4. the method of claim 1, wherein the aloe vera and the ionic liquid are present in a weight ratio of aloe vera: ionic liquid = 1: 10-20.
5. The method of claim 1, wherein the mass ratio of the diluent to the ionic liquid in step (2) is the diluent: ionic liquid = 50:1.
6. the method of preparing aloe vera extract of claim 1, wherein the diluent consists of acetone and water; the mass ratio of the acetone to the water in the diluent is 1:1.
7. an aloe extract, which is characterized in that the aloe extract is prepared by the preparation method of the aloe extract according to any one of claims 1-6.
8. A skin care product comprising the aloe extract of claim 7.
9. The skin care product according to claim 8, wherein the aloe extract is 5-20% by mass based on the skin care product.
10. The skin care product according to claim 9, wherein the aloe vera extract is present in an amount of 10% by mass of the skin care product.
11. Use of aloe vera extract according to claim 7 for the preparation of skin care products with whitening and antioxidant efficacy.
12. The use according to claim 11, wherein the skin care product is a cream, eye cream, toner or serum.
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