CN111759779A - Preparation method and application of aloe flower extract - Google Patents
Preparation method and application of aloe flower extract Download PDFInfo
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- CN111759779A CN111759779A CN202010524390.8A CN202010524390A CN111759779A CN 111759779 A CN111759779 A CN 111759779A CN 202010524390 A CN202010524390 A CN 202010524390A CN 111759779 A CN111759779 A CN 111759779A
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- aloe
- aloe flower
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- A—HUMAN NECESSITIES
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- 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]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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- 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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Abstract
The invention discloses a preparation method and application of an aloe flower extracting solution, which comprises the following steps: preparing a crude extract, pretreating, removing sediment by using a microfiltration membrane, separating by using an ultrafiltration membrane, and concentrating by using a nanofiltration membrane. The method has the advantages of convenient operation, low energy consumption, low cost, ecological environment protection, and suitability for industrial production, and the obtained aloe flower extractive solution retains active substance polyphenol in aloe flower to the maximum extent, and effectively removes interference of macromolecular impurities such as plant pituitous substances, etc., thereby enhancing quality stability of the aloe flower extractive solution. 3D skin model tests show that the aloe flower extract can obviously improve the tissue activity of the model, obviously improve the phenomena of reduction of the number of living cells, loose arrangement and the like, and obviously reduce the IL-1 alpha secretion level; obviously improves the apparent chroma of the model, extremely obviously improves the L value and reduces the melanin content. The aloe flower extract prepared by the invention does not contain any toxic solvent, has stable quality and definite effect, and can be applied to cosmetics such as anti-aging, anti-allergy, whitening and the like.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a preparation method and application of an aloe flower extracting solution.
Background
Aloe is an ancient medicinal plant, and according to the record of the unearthed Asia mud board, as early as 1750 years before the Yuan, people in Meisuodamian realize the treatment effect of Aloe, and the ancient medical books of China, Puji Fang, Ben Cao gang mu, also record the special medicinal efficacy of Aloe. There are over 500 types of aloe worldwide, and over 50 types of aloe have been found to have developmental value, with aloe vera being one of the most mature varieties of industrial development. Modern pharmacological experiments prove that the effective part or the effective component in the aloe plant has the functions of resisting bacteria, resisting inflammation, relieving fever, protecting liver, resisting cancer, killing insects, enhancing immunity and the like, and the essence has the names of 'good medicines for all intentions', 'family doctors' and the like.
According to relevant regulations of 'cosmetic health supervision regulations' and the like, the State food and drug administration carries out collection and combing on used raw materials of cosmetics on the market in China, and compiles a 'used cosmetic raw material name catalog'. The catalog 2015 includes not only aloe vera leaf, aloe vera extract, aloe vera gel and other common aloe cosmetic raw materials, but also aloe vera flower extract. Compared with the mature aloe leaf extract in the market, the aloe flower extract is of little concern in scientific research and industrial development. Literature reports also mainly focus on the extraction of aloe polysaccharides and the research of volatile components in aloe flowers, such as the research of extracting aloe barbadensis flower polysaccharides by a hot water extraction method, such as von singham and the like; study on the exorbita sibirica and the like to extract aloe barbadensis flower polysaccharide by an ultrasonic method; two methods of analysis by using a combination of gases and mass spectrometry are used for extracting volatile chemical components in the aloe flower.
Polyphenols are a group of important natural compounds, which are scientifically defined as compounds having a molecular weight of more than 600 daltons and containing polyphenol phenolic hydroxyl structures, most of which have good water solubility, and the main source of which is plants. The chemical components of the aloe flower are analyzed by Shirin and the like by using HPLC-DAD and LC-MS, and a plurality of phenolic acid components such as chlorogenic acid, caffeic acid, 5-p-cinnamoyl quinic acid, caffeoyl shikimic acid, feruloyl quinic acid, p-coumaric acid, ferulic acid and the like are found, and the total polyphenol accounts for 0.73-1.01 percent (plus or minus 0.05 percent) of the dry weight of the aloe flower by a colorimetric method.
The structural general formula and chemical properties of polyphenol are as follows:
recent studies have shown that, among natural ingredients used in cosmetics, plant polyphenols are active substances that play important roles, such as antioxidation, whitening, anti-allergy, and the like. Zillich of German scientist Olesya V.by studying the effect of polyphenol extracts on human cells and UV-irradiated cells, found that polyphenol extracts reduce intracellular ROS formation, and in addition, can prevent DNA damage and reduce membrane lipid peroxidation, and help protect dermal fibroblasts from H2O2Induced oxidative stress. Studies such as cinnabalin show that plant polyphenol can be used as a substrate analogue of tyrosinase to be combined with enzyme, so that the formation of melanin is reduced. Can also be used for treating cancer by inhibiting tyrosinase and peroxidaseThe expression of oxidase or the inhibition of tyrosinase activity can inhibit the generation of melanin, thereby whitening skin. According to the aged yoga, polyphenol compounds such as tea polyphenol, ferulic acid and the like are selected to perform a complex reaction with the shrimp allergen extract, and the influence of the polyphenol compounds on the immunological activity of the shrimp allergen extract is researched, and the results show that the ferulic acid and the tea polyphenol can react with the shrimp allergen extract, so that the concentration of allergen protein in the shrimp allergen extract solution is reduced.
Chinese patent document No. CN108635301A discloses an aloe flower extract and its preparation method, which adopts solvent method to extract, and then uses plate-frame filtration, centrifugation, decolorization, activated carbon adsorption and other methods to remove impurities. The impurity removal method disclosed by the patent cannot effectively remove vegetable mucilage substances such as tannin, colloid, protein and the like with large molecular weight in the extracting solution, so that a large amount of precipitates are separated out in the extracting solution in the shelf life, and the stability of the product is greatly influenced. Therefore, the research of the method which has simple and easy process, environmental protection and low production cost, can effectively solve the problem of quality stability of the aloe flower extract and improve the product quality is of great significance.
Disclosure of Invention
The invention aims to improve the problems in the traditional process and treatment method, and provides a preparation method and application of an aloe flower extracting solution. The preparation process takes a membrane with selective permeability as a separation medium, and plays roles of separation and concentration according to the size of a membrane hole or permeation or interception; the interference of macromolecular impurities is effectively removed while the active ingredients of the aloe flower are fully reserved, so that the product quality and the quality stability of the aloe flower extracting solution are improved. The prepared aloe flower extract is safe and harmless to skin, has the effects of better antioxidation function, anti-inflammation, allergy relief, irritation resistance, tyrosinase activity inhibition, skin whitening, skin lightening and the like, and can be applied to cosmetics such as anti-aging, anti-allergy, skin whitening and the like.
The method comprises the following steps:
(1) preparing the aloe flower crude extract: crushing and sieving aloe pollen, placing the crushed aloe pollen in an extraction tank, soaking the crushed aloe pollen in pure water serving as an extraction solvent at the liquid-material ratio of 15: 1-35: 1(v/w) at 55-85 ℃ for 1-2 hours, filtering, washing filter residues with pure water once at the liquid-material ratio of 7: 1-10: 1(v/w), filtering, and combining filtrate to obtain the aloe flower crude extract.
(2) Pretreatment of the crude extract: centrifuging the Aloe flower crude extract with a decanter centrifuge to obtain a centrifugate with turbidity of 20.0NTU or less.
(3) Removing sediment by using a microfiltration membrane: and (3) carrying out microfiltration separation treatment on the centrifugal separation liquid by adopting a microfiltration membrane with the membrane aperture of 0.05-0.2 mu m to obtain microfiltration permeating liquid with the turbidity of less than or equal to 10.0 NTU.
(4) And (3) ultrafiltration membrane separation: and (3) carrying out ultrafiltration on the microfiltration permeating liquid by using an ultrafiltration membrane with the cut-off molecular weight of 10000-50000 daltons, wherein the operating temperature is 20-45 ℃, and the operating pressure is 4-10 bar, so as to obtain the ultrafiltration permeating liquid with the turbidity of less than or equal to 2.0 NTU.
(5) Concentrating a nanofiltration membrane: concentrating the ultrafiltration permeate by using a nanofiltration membrane with the cut-off molecular weight of less than or equal to 200 daltons, wherein the operating temperature is 20-45 ℃, the operating pressure is 10-40 bar, and taking the concentrated solution to obtain an aloe flower extract; the retention rate of total polyphenols in the aloe flower extract is not less than 75%.
The aloe flower crude extract contains a large amount of particle suspended matters and mucilage components, and is difficult to operate on a machine by using a conventional filtering means, so that the aloe flower crude extract is pretreated by a centrifugal method before passing through a membrane, and the centrifugal method is divided into sedimentation type centrifugation and filtration type centrifugation according to different separation principles, wherein the sedimentation type centrifugation is particularly suitable for separating the mucilage.
In the preparation method, the decanter centrifuge is one or two combinations of a horizontal spiral discharge centrifuge, a disc high-speed centrifuge and a tubular high-speed centrifuge. The rotating speed of a drum of the horizontal spiral discharging centrifuge is 1500-4000 r/min, and the differential rotating speed is 15-40 r/min; the rotating speed of the butterfly sheet high-speed centrifugal machine is not lower than 8000 r/min; the rotating speed of the tubular high-speed centrifuge is not lower than 10000 r/min.
In the preparation method, the microfiltration membrane is one of an inorganic ceramic membrane and an organic microfiltration membrane, the operating temperature of the ceramic membrane is 20-90 ℃, and the operating pressure is 1-5 bar; the operating temperature of the organic microfiltration membrane is 20-45 ℃, and the operating pressure is 2-10 bar.
Furthermore, the material of the microfiltration membrane is preferably an inorganic ceramic membrane, and the microfiltration membrane has the characteristics of acid and alkali resistance, high temperature resistance, large flux, convenience in cleaning and the like.
The microfiltration membrane precipitation step mainly removes colloid, tannin, protein and other impurities with larger molecular weight in the feed liquid.
The ultrafiltration membrane separation step can selectively remove ineffective components such as macromolecular pigments, mucilaginous substances and the like, and separate micromolecular active substances such as polyphenol and the like.
The nanofiltration concentration step adopts a nanofiltration membrane with the molecular weight cutoff less than or equal to 200 daltons, so that micromolecular active substances with the molecular weight of more than 200 daltons can be reserved, and redundant water and part of monovalent metal ions can be removed at the same time.
The aloe flower extract prepared by the method is a colorless to dark yellow brown clear solution, the content of soluble solids is 2-10%, and the content of total polyphenols is 600-4000 mg/L.
Further, the aloe flower extracting solution is a light yellow to tawny clear solution, the content of soluble solids is 3-6%, and the concentration of total polyphenols is 800-2500 mg/L.
The aloe flower extract obtained by the process is stored for 3 months at normal temperature, the total polyphenol content and the soluble solid content are basically stable, and no obvious precipitate is generated.
The aloe flower extract liquid prepared by the invention is tested by a 3D skin model, and the result shows that the aloe flower extract liquid can obviously improve the tissue activity of the model, obviously improve the phenomena of reduction of the number of living cells, loose arrangement and the like, and obviously reduce the IL-1 alpha secretion level; obviously improves the apparent chroma of the model, extremely obviously improves the L value and reduces the melanin content.
The invention also provides application of the prepared aloe flower extract in preparing anti-aging, anti-allergic and skin whitening cosmetics.
The cosmetic is cream, essence, lotion or aqua.
Compared with the prior art, the invention has the following beneficial effects:
1. compared with the conventional organic solvent extraction process, the water is used as an extraction solvent, and the production process is green and environment-friendly; the membrane with selective permeability is used as a separation medium, under the action of pressure difference, components with different molecular weights pass or are trapped according to the size of the pore diameter of the membrane, so that the purposes of separation and concentration are achieved, the operation process has no phase change, and the structure of secondary metabolites of the aloe flowers is not damaged.
2. The invention adopts an integrated membrane technology of combined application of microfiltration, ultrafiltration and nanofiltration, ensures that the turbidity of the aloe flower extract is obviously improved, simultaneously the retention rate of total polyphenol in feed liquid is more than 75 percent, and effectively improves the product quality and the quality stability.
3. The invention adopts pre-membrane centrifugal pretreatment, improves the pollution resistance of the membrane equipment, further improves the separation efficiency of the membrane equipment, is beneficial to continuous operation and large-scale production of the process, reduces the production cost and improves the production efficiency.
4. The aloe flower extract liquid prepared by the invention passes through 3D skin model tests, and the results show that the aloe extract liquid has multiple activities of anti-allergy, tyrosinase activity inhibition, whitening and the like, and can be used as an active substance for skin cosmetics.
Drawings
FIG. 1: the invention relates to a process flow chart for preparing aloe flower extract;
FIG. 2: the tissue viability test result of the aloe flower extract after acting on the 3D skin model;
FIG. 3: the tissue morphology test result of the aloe flower extract after acting on the 3D skin model;
FIG. 4: detecting IL-1 alpha of the aloe flower extract after acting on the 3D skin model;
FIG. 5: the appearance result of the aloe flower extract after acting on the 3D melanin skin model;
FIG. 6: the L value result of the aloe flower extract after acting on the 3D melanin skin model;
FIG. 7: results of melanin content of aloe flower extract after being applied to 3D melanin skin models.
Detailed Description
The present invention will be described in further detail with reference to examples.
It will be appreciated by those skilled in the art that the following examples are illustrative of the invention only and should not be taken as limiting the scope of the invention. The examples do not specify particular techniques or conditions, and are performed according to the techniques or conditions described in the literature in the art or according to the product specifications. The materials or equipment used are not indicated by manufacturers, and all are conventional products available by purchase.
Example 1
A preparation process of an aloe flower extract comprises the following steps:
preparation of crude extract of S1
Pulverizing 50kg of Aloe flower, sieving, placing into an extraction tank, adding 1700L of pure water as extraction solvent, controlling the temperature in the extraction tank at about 60 deg.C, extracting for 2 hr under heat preservation, filtering, washing the residue with 500L of pure water, filtering, and mixing filtrates to obtain 2080L of Aloe flower crude extract.
Pretreatment of S2 crude extract
Centrifuging the crude Aloe flower extractive solution with tubular high speed centrifuge at 12000 r/min to obtain 2050L centrifugate.
S3 microfiltration membrane removing sediment
And (3) carrying out microfiltration separation treatment on the centrifugal separation liquid by adopting a ceramic membrane unit with the membrane aperture of 0.05 mu m, wherein the operation temperature is 50 ℃, and the operation pressure is 1bar, so as to obtain microfiltration permeating liquid.
S4 ultrafiltration membrane separation
And (3) carrying out ultrafiltration on the microfiltration permeating liquid by using an ultrafiltration membrane with the cut-off molecular weight of 50000 daltons, wherein the operating temperature is 25 ℃, and the operating pressure is 5bar, so as to obtain the ultrafiltration permeating liquid.
S5 nanofiltration Membrane concentration
Concentrating the ultrafiltration permeate with nanofiltration membrane with cut-off molecular weight of 150 daltons, operating at 25 deg.C and operating pressure of 10bar, concentrating the retentate to 1600L, and collecting the concentrate to obtain Aloe flower extractive solution.
The results of measuring the total polyphenol concentration in the feed liquid obtained in steps S2-S5 by the Folin-phenol method and measuring the turbidity of the feed liquid obtained in steps S2-S4 by a portable turbidity meter are shown in Table 1.
In the embodiment, the aloe flower crude extract is pretreated by a tubular high-speed centrifuge, the turbidity is reduced to 13.8NTU, and then is subjected to microfiltration and ultrafiltration treatment, and the turbidity is sequentially reduced to 7.6NTU and 1.2NTU, which shows that the three procedures of centrifugal pretreatment, microfiltration membrane sedimentation removal and ultrafiltration membrane separation all play a certain impurity removal and clarification effect. The aloe flower extract obtained after membrane concentration is light yellow, the content of soluble solids is 2.0 percent measured by a portable refractometer, the concentration of total polyphenol is 627.3mg/L, the retention rate of the total polyphenol is calculated to be 83.6 percent in the membrane treatment process, and all indexes meet the quality control requirements.
Example 2
A preparation process of an aloe flower extract comprises the following steps:
preparation of crude extract of S1
Pulverizing 50kg of Aloe flower, sieving, placing into an extraction tank, adding 800L of pure water as extraction solvent, controlling the temperature in the extraction tank at about 80 deg.C, extracting for 1 hr under constant temperature, filtering, washing the residue with 500L of pure water, filtering, and mixing filtrates to obtain 1120L of crude Aloe flower extractive solution.
Pretreatment of S2 crude extract
Centrifuging the Aloe flower crude extractive solution with disc high speed centrifuge at 8000r/min to obtain 1090L centrifugated solution.
S3 microfiltration membrane removing sediment
And (3) carrying out microfiltration separation treatment on the centrifugal separation liquid by adopting an organic membrane with the membrane aperture of 0.2 mu m, wherein the operation temperature is 40 ℃, and the operation pressure is 8bar, so as to obtain microfiltration permeating liquid.
S4 ultrafiltration membrane separation
And (3) carrying out ultrafiltration on the microfiltration permeating liquid by using an ultrafiltration membrane with the molecular weight cutoff of 10000 Dalton, wherein the operating temperature is 40 ℃, and the operating pressure is 9bar, so as to obtain the ultrafiltration permeating liquid.
S5 nanofiltration Membrane concentration
Concentrating the ultrafiltration permeate with nanofiltration membrane with cut-off molecular weight of 200 daltons at 40 deg.C under 35bar, and concentrating the retentate to 280L to obtain aloe flower extractive solution.
The results of measuring the total polyphenol concentration in the feed liquid obtained in steps S2-S5 by the Folin-phenol method and measuring the turbidity of the feed liquid obtained in steps S2-S4 by a portable turbidity meter are shown in Table 1.
In the embodiment, the aloe flower crude extract is pretreated by a disc high-speed centrifuge, the turbidity is reduced to 17.4NTU, and then the turbidity is sequentially reduced to 9.4NTU and 0.1NTU by microfiltration and ultrafiltration treatment, wherein an ultrafiltration membrane with the molecular weight cutoff of about 10000 Dalton has a better impurity removal effect. The aloe flower extract obtained after membrane concentration is dark yellow brown, the content of soluble solids is 10.0 percent measured by a portable refractometer, the concentration of total polyphenol is 2431.8mg/L, the retention rate of the total polyphenol is calculated to be 76.7 percent in the membrane treatment process, and all indexes meet the quality control requirements.
Example 3
A preparation process of an aloe flower extract comprises the following steps:
preparation of crude extract of S1
Pulverizing 50kg of Aloe flower, sieving, placing into an extraction tank, adding 1000L of pure water as extraction solvent, controlling the temperature in the extraction tank at about 70 deg.C, extracting for 2 hr under constant temperature, filtering, washing the residue with 350L of pure water, filtering, and mixing filtrates to obtain 1170L of crude Aloe flower extractive solution.
Pretreatment of S2 crude extract
Feeding the aloe flower crude extract into a horizontal spiral discharge centrifuge-disc high-speed centrifuge set through a pipeline for centrifugal separation, wherein the rotating speed of a drum of the horizontal spiral discharge centrifuge is 2000r/min, and the differential rotating speed is 30 r/min; the rotational speed of the disc high-speed centrifuge is 10000r/min, and 1150L of centrifugal separation liquid is obtained.
S3 microfiltration membrane removing sediment
And (3) carrying out microfiltration separation treatment on the centrifugal separation liquid by adopting a ceramic membrane unit with a membrane aperture of 0.05 mu m, wherein the operation temperature is 40 ℃, and the operation pressure is 2bar, so as to obtain microfiltration permeating liquid.
S4 ultrafiltration membrane separation
And (3) carrying out ultrafiltration on the microfiltration permeating liquid by using an ultrafiltration membrane with the molecular weight cutoff of 15000 daltons, wherein the operating temperature is 30 ℃, and the operating pressure is 8bar, so as to obtain the ultrafiltration permeating liquid.
S5 nanofiltration Membrane concentration
Concentrating the ultrafiltration permeate with nanofiltration membrane with cut-off molecular weight of 200 daltons at operation temperature of 30 deg.C and operation pressure of 20bar, and concentrating the retentate to 560L to obtain concentrated solution, i.e. flos Aloe extract.
The results of measuring the total polyphenol concentration in the feed liquid obtained in steps S2-S5 by the Folin-phenol method and measuring the turbidity of the feed liquid obtained in steps S2-S4 by a portable turbidity meter are shown in Table 1.
In the embodiment, the aloe flower crude extract is pretreated by a horizontal spiral discharge centrifuge-disc high-speed centrifuge set, the turbidity is reduced to 11.1NTU, and then the turbidity is sequentially reduced to 5.7NTU and 0.1NTU through microfiltration and ultrafiltration. The aloe flower extract obtained after membrane concentration is yellow brown, the content of soluble solids is 5.0 percent measured by a portable refractometer, the concentration of total polyphenol is 1202.5mg/L, the retention rate of the total polyphenol is calculated to be 84.4 percent in the membrane treatment process, and all indexes meet the quality control requirements.
TABLE 1 Total polyphenol concentration and turbidity test results in Aloe flower liquid
By comparing the three examples, the turbidity of the feed liquid after the centrifugal pretreatment is reduced to a lower level (the turbidity is less than 20NTU), and the normal operation of the microfiltration membrane sedimentation removal step is ensured. Example 2 and example 3 the turbidity of the ultrafiltration permeate was 0.1NTU, lower than example 1, indicating that both clarification effects were better than example 1; comparing the total polyphenol retention during the multi-stage membrane treatment, both example 1 and example 3 were above 80%, while example 2 was only 76.7%. And (3) determining the embodiment 3 as the preferable scheme by combining the impurity removal effect and the retention condition of the total polyphenol of the effective components in the three embodiments.
Example 4
Membrane flux contrast test
Crude extracts of aloe flower were prepared as in S1 of example 3, and 100L of each crude extract was used in the following two sets of experiments:
test A: directly filtering the crude Aloe flower extractive solution with organic microfiltration membrane with membrane pore diameter of 0.2 μm at operation temperature of 35 deg.C and operation pressure of 2.5bar, ultrafiltering the obtained microfiltration permeate with ultrafiltration membrane with cut-off molecular weight of 15000 daltons at operation temperature of 35 deg.C and operation pressure of 8bar, and recording and calculating microfiltration and ultrafiltration average membrane flux.
Test B: the aloe flower crude extract was pretreated with a tubular high speed centrifuge at 12000r/m, and the resulting centrifuged liquid was again subjected to the procedure described in test A, and the average flux of microfiltration and ultrafiltration membranes was recorded and calculated, and the results are shown in Table 2.
TABLE 2 average microfiltration and ultrafiltration membrane flux Table
In this example, the necessity of pre-membrane centrifugation pretreatment is studied, and as a result, it is found that the aloe flower crude extract directly passes through the membrane without pretreatment, the average membrane flux is low in both the microfiltration unit and the ultrafiltration unit, and especially, the membrane pores are basically blocked at the end of ultrafiltration, and thus the aloe flower crude extract cannot normally operate. The aloe flower liquid is pretreated by a centrifugal method, so that the treatment load of a microfiltration unit is effectively reduced, and the average ultrafiltration membrane flux is remarkably improved. The result shows that the pre-membrane centrifugal pretreatment can improve the pollution degree of the membrane, effectively improve the membrane flux of microfiltration and ultrafiltration and obviously improve the membrane separation efficiency.
Example 5
Observation of stability
200ml of each of the aloe flower centrifugal separation liquid obtained in S2 in example 3 and the nanofiltration concentrate obtained in S5 in example 3 was taken out, and each was packaged in a small can in 2 parts, sterilized and then left at room temperature. The appearance of the sample was observed periodically for 3 months, and the results are shown in table 3:
TABLE 3 stability survey table of aloe flower liquid obtained by different treatment methods
| Aloe flower pretreatment liquid (centrifugal separation liquid) | Aloe flower extract (nanofiltration concentrate) | |
| Day 0 | Brown liquid, relatively clear | Yellow brown clear liquid |
| 7 days | Brown liquid with trace amount of precipitate at the bottom | Yellow brown clear liquid |
| 1 month | Brown liquid with a small amount of precipitate at the bottom | Yellow brown clear liquid |
| 2 month | Brown liquid with more precipitate at the bottom | Yellow brown clear liquid |
| 3 month | Brown liquid with a large amount of precipitate at the bottom | Yellow brown clear liquid |
The results show that only by traditional centrifugation and impurity removal treatment, the aloe flower extract generates micro-precipitates after being placed at room temperature for 7 days, and a small amount of precipitates are observed after one month and gradually increase along with time. The aloe flower extract liquid obtained by the multistage membrane technology is basically unchanged in appearance and properties after being placed for three months at normal temperature, and is clear in body and free of obvious visible precipitates or suspended matters, so that the method provided by the invention can effectively solve the problem of unstable storage of the aloe flower water extract liquid in a shelf life.
Example 6
The method for measuring the polyphenol content in the aloe flower extract comprises the following steps:
a. preparation of Folin-phenol (Folin-Ciocalteus) test solution
100g of sodium tungstate and 25g of sodium molybdate are taken and placed in a 5L round bottom flask, 700ml of pure water, 50ml of phosphoric acid and 100ml of hydrochloric acid are added and mixed evenly, and heating reflux is carried out for 10 hours by using soft fire. After cooling, 150g of lithium sulfate, 50ml of pure water and 0.25ml of bromine water are added, the mixture is boiled for 15min in an open air, after cooling, the mixture is filtered, the pure water is added to reach the constant volume of 1000ml, the mixture is stored in a refrigerator, and when the mixture is used, the mixture is diluted by 5 times.
b. Preparation of Standard solutions
Weighing 20mg of gallic acid, placing the gallic acid in a 100mL volumetric flask, adding pure water to dissolve and fix the volume to obtain standard stock solution, and placing the standard stock solution in a refrigerator for storage. When the gallic acid solution is used, 1mL of the standard stock solution is put into a 25mL volumetric flask, pure water is added for constant volume, and the solution is shaken up to obtain the gallic acid standard solution.
c. Drawing of standard curve
Respectively placing 0.5 mL, 1.0 mL, 2.0 mL, 3.0 mL, 4.0 mL and 5.0mL of standard substance solution in 6 groups of test tubes, respectively adding pure water to 7mL, shaking, adding 1mL of Folin-phenol test solution, shaking, standing at room temperature for 3-5 min, and respectively adding 5% Na2CO32ml of the solution was shaken up, developed in a water bath at 40 ℃ for 60min and the absorbance was measured at 735 nm. The blank control is prepared by replacing standard solution with 1ml of pure water, and the concentration (μ g/ml) of the control solution is used as abscissa and the net optical density value (Δ OD) of the control solution735) Drawing a standard curve for the ordinate, and fitting by using a least square method to obtain a linear equation as follows: y is 10.1x +0.0316 (R)2＝0.9992)
d. Sample assay
Taking the aloe flower extract for proper dilution, taking 1ml of the diluted solution to place in a test tube, adding 6 ml of pure water, shaking uniformly, adding 1ml of forskolin-phenol test solution, shaking uniformly, standing at normal temperature for 3-5 min, adding 5% Na2CO32ml of the solution was shaken up, developed in a water bath at 40 ℃ for 60min and the absorbance was measured at 735 nm. By 1mAnd l, replacing the sample solution with pure water, performing blank control, and reading the polyphenol content in the sample according to a standard curve.
e. Formula for calculation
The total polyphenol concentration calculation formula is as follows:
C＝(ΔOD735-0.0316)/10.1 x n (formula one)
In the formula I: c is total polyphenol concentration of a sample to be detected, and the unit is mg/L
ΔOD735Optical density value of solution to be measured after blank deduction
n is the dilution multiple of the sample solution to be measured
The total polyphenol retention rate calculation formula:
Y＝C*V/(C0*V0) 100% (formula two)
In the second formula: y is total polyphenol retention in unit%
C0The total polyphenol concentration of a sample solution to be treated is unit mg/L
V0Is the volume of the sample liquid to be treated, in units of L
C is total polyphenol concentration of the treated sample solution, and unit mg/L
V is the volume of the sample solution after treatment, unit L
Example 7
Aloe flower extract anti-inflammatory and allergy-relieving efficacy test
The method is characterized in that a 3D skin model (EpiKutis) is used as a testing tool, sodium dodecyl sulfate (SLS) is adopted to stimulate the 3D skin model, and a human body patch test is simulated to construct an 'SLS-EpiKutis' skin injury model. Meanwhile, the aloe flower extract is uniformly coated on the surface of a skin injury model by adopting a surface administration mode, and the anti-inflammatory effect of the aloe flower extract is evaluated by detecting the tissue activity, the tissue morphology and the change of inflammatory IL-1 alpha of the model. Experimental group design as in table 4:
TABLE 4 Aloe flower extract anti-inflammatory and anti-allergic efficacy test experimental grouping design table
The test results are shown in the attached figures 2-4. Compared with the BC group, the NC group treated by 0.2% SLS shows a remarkable reduction effect on the tissue activity of the model, the morphological change of the model is obvious, and the concentration of the IL-1 alpha protein is remarkably improved. Compared with the NC group, the treatment of 0.05 percent dexamethasone in the PC group has certain improvement effect on the tissue activity and the morphology of the model, and simultaneously reduces the protein concentration of IL-1 alpha, which indicates that the test tool is effective.
Compared with an NC group, the aloe flower extract prepared in the example 3 can obviously improve the tissue activity of a model under the administration concentration of 5%, and has obvious improvement effect on the phenomena of reduction of the number of living cells of the model, loose arrangement and the like; meanwhile, the compound shows a remarkable reduction effect on IL-1 alpha, namely, has a certain anti-inflammatory effect.
Example 8
In-vitro whitening efficacy test of aloe flower extract
The 3D melanoderm model (MelaKutis) is used as a test tool, and Ultraviolet (UVB) irradiation is adopted to stimulate the 3D melanoderm model so as to induce melanogenesis. The in-vitro whitening effect of the aloe flower extract is evaluated by uniformly coating the aloe flower extract on the surface of a 3D melanin skin model in a surface administration mode and according to three dimensional indexes of a model appearance picture, the brightness (L x value) of the skin model and the melanin content. Experimental group design as in table 5:
TABLE 5 Aloe flower extract in vitro whitening efficacy test experiment component design table
The test results are shown in the attached figures 5-7: at 50mJ/cm2The appearance of the model is obviously blackened under the stimulation of UVB, and the L value and the melanin content are obviously different from those of a Control group; under the condition of UVB stimulation, the appearance of a melanin model of a KA group is obviously whitened compared with that of a UVB stimulation group, and the detection results of the L-value and the melanin content have significant difference from those of a comparison solution. Thus, U based on MelaKutis is describedThe VB stimulation system and the whitening detection system are normal.
Compared with the UVB stimulation group, the aloe flower extract prepared in example 3 has the advantages that the model apparent chromaticity is whitened at the administration concentration of 5%, the L value is remarkably increased (p is less than 0.01), the melanin content is remarkably reduced (p is less than 0.01), and the aloe flower extract has better whitening effect.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (7)
1. The preparation method of the aloe flower extract is characterized by comprising the following steps:
(1) preparing the aloe flower crude extract: crushing and sieving aloe pollen, placing the crushed and sieved aloe pollen in an extraction tank, taking pure water as an extraction solvent, carrying out warm immersion for 1-2 hours at 55-85 ℃ at a liquid-material ratio of 15: 1-35: 1(v/w), filtering, washing filter residues with pure water once at a liquid-material ratio of 7: 1-10: 1(v/w), filtering, and combining filtrate to obtain aloe flower crude extract;
(2) pretreatment of the crude extract: centrifuging the aloe flower crude extract in the step (1) by adopting a sedimentation centrifuge to obtain a centrifugal separation liquid with turbidity less than or equal to 20.0 NTU;
(3) removing sediment by using a microfiltration membrane: carrying out microfiltration separation treatment on the centrifugal separation liquid in the step (2) by adopting a microfiltration membrane with the membrane aperture of 0.05-0.2 mu m to obtain microfiltration permeate with the turbidity of less than or equal to 10.0 NTU;
(4) and (3) ultrafiltration membrane separation: carrying out ultrafiltration on the microfiltration permeating liquid in the step (3) by using an ultrafiltration membrane with the cut-off molecular weight of 10000-50000 daltons, wherein the operating temperature is 20-45 ℃, and the operating pressure is 4-10 bar, so as to obtain ultrafiltration permeating liquid with the turbidity of less than or equal to 2.0 NTU;
(5) concentrating a nanofiltration membrane: concentrating the ultrafiltration permeate obtained in the step (4) by adopting a nanofiltration membrane with the molecular weight cutoff of less than or equal to 200 daltons, wherein the operating temperature is 20-45 ℃, the operating pressure is 10-40 bar, and taking the concentrated solution to obtain an aloe flower extract; the retention rate of total polyphenols in the aloe flower extract is not less than 75%.
2. The method of claim 1, wherein: the decanter centrifuge in the step (2) is one or a combination of a horizontal spiral discharge centrifuge, a disc high-speed centrifuge and a tubular high-speed centrifuge; the rotating speed of a drum of the horizontal spiral discharging centrifuge is 1500-4000 r/min, and the differential rotating speed is 15-40 r/min; the rotating speed of the butterfly sheet high-speed centrifugal machine is not lower than 8000 r/min; the rotating speed of the tubular high-speed centrifuge is not lower than 10000 r/min.
3. The method of claim 1, wherein: the microfiltration membrane in the step (3) is one of an inorganic ceramic membrane and an organic microfiltration membrane; the operating temperature of the inorganic ceramic membrane is 20-90 ℃, and the operating pressure is 1-5 bar; the operating temperature of the organic microfiltration membrane is 20-45 ℃, and the operating pressure is 2-10 bar.
4. The method of claim 1, wherein: the aloe flower extracting solution is a colorless to tawny clear solution, the content of soluble solids is 2-10%, and the concentration of total polyphenol is 600-4000 mg/L.
5. The method of claim 1, wherein: the aloe flower extracting solution is a light yellow to tawny clear solution, the content of soluble solids is 3-6%, and the concentration of total polyphenol is 800-2500 mg/L.
6. Use of the aloe flower extract of claim 1 for the preparation of anti-aging, anti-allergic and skin whitening cosmetics.
7. Use according to claim 6, characterized in that: the cosmetic is cream, essence, lotion or aqua.
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