CN111909284A - Extraction and purification method of tea bran polysaccharide and scalp care composition containing tea bran polysaccharide - Google Patents
Extraction and purification method of tea bran polysaccharide and scalp care composition containing tea bran polysaccharide Download PDFInfo
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- A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
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- 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]
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- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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- A61Q5/006—Antidandruff preparations
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
Abstract
The invention discloses an extraction and purification method of tea bran polysaccharide and a scalp care composition containing the tea bran polysaccharide, which comprises the steps of taking degreased tea-oil camellia tea bran powder, adding ethanol, refluxing at the temperature of 50-60 ℃, filtering, and drying filter residues to obtain tea saponin-removed tea bran powder; mixing tea saponin-removed tea bran powder with distilled water, extracting, centrifuging the extract, taking supernatant, and concentrating the supernatant by evaporation under reduced pressure to obtain concentrated solution; adjusting the pH of the concentrated solution at room temperature by using dilute hydrochloric acid, standing, and centrifuging to remove precipitates to obtain an extracting solution; preparing an ammonium sulfate/polyethylene glycol 2000 aqueous two-phase extraction system, extracting the extracting solution under the ultrasonic-assisted condition, and taking the lower phase after fully standing. The method for extracting the tea bran polysaccharide has high protein removal rate, and the scalp care composition prepared by utilizing the tea bran polysaccharide after grading and purification has obvious effects of relieving itching, repairing scalp and removing dandruff.
Description
Technical Field
The invention belongs to the technical field of tea bran polysaccharide extraction, and particularly relates to a method for extracting and purifying tea bran polysaccharide and a scalp care composition containing the tea bran polysaccharide.
Background
The polysaccharide is an important cosmetic raw material and has multiple effects of moisturizing, resisting aging and the like. The camellia has unique resource advantages in China, and after camellia fruits are used for squeezing tea oil, the residue is camellia seed cakes (tea bran, also called oil camellia cakes). The tea bran mainly contains a plurality of active substances such as tea saponin, polysaccharide, protein, tea polyphenol and the like, wherein the tea saponin is used as a natural surfactant and is widely applied to cosmetics, particularly washing and caring products. However, the polysaccharide component derived from tea bran has not been effectively utilized in cosmetics at present. This is because the tea bran contains various active ingredients, and it is difficult to extract and purify the high-purity polysaccharide from the tea bran; in particular, since polysaccharides and proteins in tea bran are extracted simultaneously in most extraction methods, their solubility properties are similar and conventional methods are not easy to separate efficiently. When added into cosmetics, high-content protein impurities cause the problem of easy microbial breeding, bring about greater preservative pressure, and cause the difficulty in applying the tea bran-derived polysaccharide to the cosmetics or the incapability of using the tea bran-derived polysaccharide under the condition of higher addition amount.
At present, in methods for removing protein in plant polysaccharide, Sevag reagent method and trichloroacetic acid method are commonly used in traditional methods, the purpose of removing protein is achieved by utilizing chemical reagents to denature protein, and the used reagents have the problems of toxicity and difficult recovery, and are difficult to be applied to cosmetic raw materials. People also gradually develop convenient deproteinization methods such as a salt method, an enzymolysis method and the like, an exchange resin method, a repeated freeze-thaw method and the like, but the removal rate is generally insufficient in the aspects. Technical difficulties still exist in how to remove protein impurities in tea seed polysaccharide efficiently and deeply. Meanwhile, the efficacy of the tea bran-derived polysaccharide extract applied to the scalp care solution is not reported.
Disclosure of Invention
This section is for the purpose of summarizing some aspects of embodiments of the invention and to briefly introduce some preferred embodiments. In this section, as well as in the abstract and the title of the invention of this application, simplifications or omissions may be made to avoid obscuring the purpose of the section, the abstract and the title, and such simplifications or omissions are not intended to limit the scope of the invention.
As one aspect of the invention, the invention provides a method for extracting and purifying tea bran polysaccharide.
In order to solve the technical problems, the invention provides the following technical scheme: a method for extracting and purifying tea bran polysaccharide comprises the following steps,
taking degreased oil tea bran powder, adding 85-95% ethanol, and mixing the materials in a material-liquid ratio of 1: refluxing for 2-3 h at 50-60 ℃ for 5-10 ℃, and drying filter residue after suction filtration to obtain tea saponin-removed tea bran powder;
mixing tea saponin-removed tea bran powder with distilled water, and mixing the mixture in a material-liquid ratio of 1: extracting for 3-4 hours at the temperature of 60-70 ℃ and at 5-10 ℃, centrifuging the extracting solution, taking supernatant, and concentrating the volume of the supernatant to 20-30% of the original volume by reduced pressure evaporation to obtain a concentrated solution;
adjusting the pH of the concentrated solution to 4.0-5.0 by using dilute hydrochloric acid at room temperature, standing for 10-14 h, and centrifuging to remove precipitates to obtain an extracting solution;
preparing an ammonium sulfate/polyethylene glycol 2000 aqueous two-phase extraction system, extracting the extracting solution under the ultrasonic-assisted condition, and taking the lower phase after fully standing.
As a preferred scheme of the method for extracting and purifying the tea bran polysaccharide, the method comprises the following steps: the ammonium sulfate/polyethylene glycol 2000 double-aqueous-phase extraction system comprises 14-16% by mass of ammonium sulfate and 11-13% by mass of polyethylene glycol 2000.
As a preferred scheme of the method for extracting and purifying the tea bran polysaccharide, the method comprises the following steps: and extracting the extracting solution under the ultrasonic-assisted condition, wherein the ultrasonic power of the extracting solution is 200-300W.
As a preferred scheme of the method for extracting and purifying the tea bran polysaccharide, the method comprises the following steps: the extraction is carried out on the extracting solution at room temperature, and the extraction time is 50-60 min; and taking the lower phase after fully standing.
As a preferred scheme of the method for extracting and purifying the tea bran polysaccharide, the method comprises the following steps: further comprises taking the lower phase and purifying by molecular weight fractionation with DEAE cellulose column chromatography, comprising DEAE cellulose pretreatment, column packing, and purification with 0.05 mol.L-1The NaCl solution is subjected to gradient elution, and the eluent is collected.
As a preferred scheme of the method for extracting and purifying the tea bran polysaccharide, the method comprises the following steps: the DEAE cellulose pretreatment comprises the steps of taking DEAE-52 cellulose, adding distilled water, swelling for 48 hours at 4 ℃, and carrying out suction filtration; soaking the mixture in 0.5mol/L NaOH solution for 2h, washing the mixture with distilled water until the pH value is 7.0, and performing suction filtration; soaking the mixture in 0.5mol/L HCl solution for 2h, washing the mixture with distilled water until the pH value is 7.0, and performing suction filtration; and finally soaking the mixture in 0.5mol/L NaOH solution for 2h, washing the mixture with distilled water until the pH value is 7.0, soaking the mixture in the distilled water, and removing bubbles by ultrasonic waves.
As a preferred scheme of the method for extracting and purifying the tea bran polysaccharide, the method comprises the following steps: and (3) filling the column, namely adding distilled water into the chromatographic column, slowly pouring the pretreated DEAE-cellulose suspension into the column, adding a proper amount of quartz sand, and balancing the chromatographic column by using the distilled water.
As a preferred scheme of the method for extracting and purifying the tea bran polysaccharide, the method comprises the following steps: and adjusting the pH of the concentrated solution to 4.0-5.0 by using dilute hydrochloric acid at room temperature to adjust the pH of the concentrated solution to 4.5.
As a preferred scheme of the method for extracting and purifying the tea bran polysaccharide, the method comprises the following steps: the mass fraction of the ammonium sulfate is 15 percent, and the mass fraction of the polyethylene glycol 2000 is 12 percent.
In one aspect of the invention, the invention provides a scalp care composition containing the tea bran polysaccharide, which comprises 2-3% of tea bran polysaccharide, 0.8-1.0% of radix sophorae flavescentis extract, 0.5-1% of ginsenoside, 2-3% of glycerol, 3-4% of butanediol, 1-2% of menthol, 1-2% of hydrogenated lecithin, 1-2% of 1, 2-pentanediol, 0.1-0.2% of zinc aspartate, 0.05-0.07% of PCA copper, 0.05-0.06% of EDTA-2Na, 0.1-0.3% of citric acid, 0.2-0.3% of arginine, 0.2-0.3% of sarcosine, 0.3-0.4% of sodium benzoate, 5-6% of ethanol and the balance of water.
The invention has the beneficial effects that: the method for extracting the tea bran polysaccharide has high protein removal rate, the scalp care composition prepared by utilizing the tea bran polysaccharide after the grading purification has obvious effects of relieving itching, repairing the scalp, protecting the scalp barrier and removing dandruff, the prepared tea bran polysaccharide has obvious effect of inhibiting propionibacterium acnes lipase, the tea bran polysaccharide can obviously reduce the activity of propionibacterium acnes lipase enzyme, obviously inhibit the growth of propionibacterium acnes, and effectively avoid stimulation and itching caused by metabolites of the tea bran polysaccharide.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive exercise. Wherein:
FIG. 1 shows the bacteriostatic action of tea bran polysaccharide on Propionibacterium acnes and Malassezia furfur.
FIG. 2 is a graph showing the effect of 2% tea bran polysaccharides on the level of IL-1. alpha. released from cells.
Fig. 3 shows dandruff condition after 21 days of applying scalp care solution to volunteers.
FIG. 4 shows the measurement of the moisture content of the scalp of a subject after the scalp care solution 3 and the control 3.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with examples are described in detail below.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.
Furthermore, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
Example 1:
the extraction method of the tea bran polysaccharide comprises the following steps:
(1) taking degreased oil tea bran powder, adding 85% ethanol, and mixing the materials in a material-liquid ratio of 1: 7. refluxing at 60 deg.C for 2 hr, filtering, and drying the residue to obtain tea bran powder without tea saponin;
(2) mixing tea bran powder without tea saponin with distilled water, and mixing the mixture in a material-liquid ratio of 1: 7. extracting at 70 deg.C for 3 hr, centrifuging the extractive solution, collecting supernatant, and concentrating the supernatant to 20% of original volume by evaporation under reduced pressure;
(3) regulating the pH of the concentrated extract obtained in the step (2) to 4.5 by using dilute hydrochloric acid at room temperature, standing for 12 hours, and centrifuging to remove precipitates;
(4) preparing an ammonium sulfate/polyethylene glycol (PEG)2000 two-aqueous phase extraction system, wherein the mass fraction of ammonium sulfate is 15%, and the mass fraction of PEG2000 is 12%; extracting the extracting solution obtained in the step (3) for 50min by using the extracting system under the conditions of ultrasonic assistance, ultrasonic power of 200W and room temperature (20-30 ℃); and (4) after extraction is finished and the mixture is fully kept stand, taking the lower phase to obtain the tea bran polysaccharide.
And (3) testing: dissolving the lower phase tea bran polysaccharide by using distilled water, fixing the volume in a 100mL volumetric flask, transferring 1.0mL sugar solution into a 25mL volumetric flask, and fixing the volume to a scale by using the distilled water, thus obtaining the sample solution to be detected. Absorbing 1.0mL of sample solution to be detected in a test tube, supplementing 2.0mL with distilled water, adding 1.0mL of 5% phenol solution, immediately adding 5.0mL of concentrated sulfuric acid, simultaneously absorbing 2.0mL of distilled water, using the same method as the above operation as a blank control, oscillating and uniformly mixing, standing at room temperature for reaction for 20min, cooling to room temperature, and measuring the absorbance at 457nm, wherein the yield of the tea bran polysaccharide is as follows:
the polysaccharide yield calculation formula is as follows:
a-polysaccharide content of the sample, mg
B-tea bran weight, g
The yield of the tea bran polysaccharide is measured to be 15.3%. The protein removal rate was 96%.
In vitro bacteriostatic experiment: 107CFU/mL Propionibacterium acnes or Malassezia furfur (Malassezia furur) were cultured in an incubator at 37 ℃ under anaerobic conditions for 24 hours with the addition of 2% tea bran polysaccharide to test the bacteriostatic effect of the tea bran polysaccharide on Propionibacterium acnes and Malassezia furfur. The results of the experiment are shown in FIG. 1.
Example 2:
the tea bran polysaccharide obtained in example 1 was concentrated, dialyzed and desalted, and then subjected to molecular weight fractionation purification by DEAE cellulose column chromatography: DEAE-52 cellulose pretreatment: adding DEAE-52 cellulose into distilled water, swelling at 4 deg.C for 48 hr, and vacuum filtering; soaking the mixture in 0.5mol/L NaOH solution for 2h, washing the mixture with distilled water until the pH value is 7.0, and performing suction filtration; soaking the mixture in 0.5mol/L HCl solution for 2h, washing the mixture with distilled water until the pH value is 7.0, and performing suction filtration; finally useSoaking in 0.5mol/L NaOH solution for 2h, washing with distilled water to pH 7.0, soaking in distilled water, and removing bubbles by ultrasonic treatment; column assembling: adding distilled water into the chromatographic column, slowly pouring the pretreated DEAE-cellulose suspension into the column, keeping the outlet flow velocity stable, allowing it to settle naturally, adding appropriate amount of quartz sand, and balancing the chromatographic column with distilled water; loading and eluting: dissolving 200mg of the above lower phase tea bran polysaccharide in small amount of distilled water, separating with DEAE-52 cellulose column, sequentially adding distilled water, 0.05, 0.2 mol.L-1And carrying out gradient elution on the NaCl solution, respectively collecting eluent, respectively recording the eluent as eluent A, eluent B and eluent C, and drying to obtain a tea bran polysaccharide component A, a tea bran polysaccharide component B and a tea bran polysaccharide component C.
Propionibacterium acnes lipase activity assay: lipase activity of Propionibacterium acnes was measured by lipase activity colorimetric assay kit II (Analygenie, UK) assay, 4-methylumbelliferone oleate (MUO) assay, in broth culture and adjusted to 5X 108CFU/mL, adding 100 μ L of cell suspension with adjusted concentration into each well of 96-well plate, and adding 100 μ L of 2% tea bran polysaccharide solution; the plates were incubated at 37 ℃ for 24h under anaerobic conditions. And centrifuging the liquid in the collecting hole, and collecting the supernatant. 50 μ L of the supernatant was mixed with 50 μ L of a solution containing 13mM Tris-HCl, 0.15M NaCl and 1.3mM CaCl2The solutions of (a) and (b) are mixed. The mixture was incubated at 25 ℃ for 30 minutes, and the enzymatic reaction was stopped by adding 100. mu.L of 0.1. mu.M sodium citrate (pH 4.2). The 4-methylumbelliferyl oleate (MUO) content released by the lipase was measured using a microplate reader at an excitation wavelength of 355nm and an emission wavelength of 460 nm.
The experimental result shows that the tea bran polysaccharide component B enables the activity of the propionibacterium acnes lipase to be reduced by 28%, the tea bran polysaccharide component A does not have the obvious effect of reducing the activity of the propionibacterium acnes lipase, and the tea bran polysaccharide component C enables the activity of the propionibacterium acnes lipase to be reduced by about 13%.
With an oleic acid: linoleic acid 1: 1 as a stimulant, immortalized human stratum corneum-forming cells were used as model cells, and the content of IL-1. alpha. of an inflammatory factor released from cells to which 2% of either a tea bran polysaccharide fraction A, a tea bran polysaccharide fraction B or a tea bran polysaccharide fraction C was added as an active substance was measured by ELISA kit (Novus Biologicals, USA), and the results of the experiment are shown in FIG. 2.
Example 3:
preparing a scalp care solution: the tea bran polysaccharide composition comprises, by mass percentage (g: mL), 2% of tea bran polysaccharide (one of the tea bran polysaccharide, the tea bran polysaccharide component A, the tea bran polysaccharide component B or the tea bran polysaccharide component C prepared in example 1), 0.8% of radix sophorae flavescentis extract, 0.5% of ginsenoside, 2% of glycerol, 3% of butanediol, 1% of menthol, 1% of hydrogenated lecithin, 1% of 1, 2-pentanediol, 0.1% of zinc aspartate, 0.05% of PCA copper, 0.05% of EDTA-2Na, 0.1% of citric acid, 0.2% of arginine, 0.2% of sarcosine, 0.3% of sodium benzoate, 5% of ethanol and the balance of water.
The scalp care solutions prepared in the four groups are sequentially marked as scalp care solution 1 (added with the tea bran polysaccharide group prepared in example 1), scalp care solution 2 (added with the tea bran polysaccharide component A), scalp care solution 3 (added with the tea bran polysaccharide component B) and scalp care solution 4 (added with the tea bran polysaccharide component C). The dandruff condition of the volunteers after 21 days using the scalp care solution is shown in fig. 3.
Comparative example 1:
the extraction and purification method of the tea bran polysaccharide comprises the following steps:
(1) taking degreased oil tea bran powder, adding 85% ethanol, and mixing the materials in a material-liquid ratio of 1: 7. refluxing at 60 deg.C for 2 hr, filtering, and drying the residue to obtain tea saponin-removed tea bran powder;
(2) mixing tea saponin-removed tea bran powder with distilled water, and mixing the mixture in a material-liquid ratio of 1: 7. extracting at 70 deg.C for 3 hr, centrifuging the extractive solution, collecting supernatant, and concentrating the supernatant to 20% of original volume by evaporation under reduced pressure;
(3) regulating the pH of the concentrated extract obtained in the step (2) to 4.5 by using dilute hydrochloric acid at room temperature, standing for 12 hours, and centrifuging to remove precipitates;
(4) preparing an ammonium sulfate/polyethylene glycol (PEG)2000 two-aqueous phase extraction system, wherein the mass fraction of ammonium sulfate is 12%, and the mass fraction of PEG2000 is 12%; extracting the extracting solution obtained in the step (3) by using the extracting system under the conditions of ultrasonic assistance, ultrasonic power of 200W and room temperature (20-30 ℃), wherein the extracting time is 50 min; and (4) after extraction is finished and the mixture is fully kept stand, taking the lower phase to obtain the tea bran polysaccharide. The removal rate of protein impurities was determined to be 80%.
Preparing a scalp care solution: the tea bran polysaccharide oral liquid comprises, by mass percentage (g: mL), 2% of tea bran polysaccharide, 0.8% of radix sophorae flavescentis extract, 0.5% of ginsenoside, 2% of glycerol, 3% of butanediol, 1% of menthol, 1% of hydrogenated lecithin, 1% of 1, 2-pentanediol, 0.1% of zinc aspartate, 0.05% of PCA copper, 0.05% of EDTA-2Na, 0.1% of citric acid, 0.2% of arginine, 0.2% of sarcosine, 0.3% of sodium benzoate, 5% of ethanol and the balance of water.
Comparative example 2:
the extraction and purification method of the tea bran polysaccharide comprises the following steps:
(1) taking degreased oil tea bran powder, adding 85% ethanol, and mixing the materials in a material-liquid ratio of 1: 7. refluxing at 60 deg.C for 2 hr, filtering, and drying the residue to obtain tea saponin-removed tea bran powder;
(2) mixing tea saponin-removed tea bran powder with distilled water, and mixing the mixture in a material-liquid ratio of 1: 7. extracting at 70 deg.C for 3 hr, centrifuging the extractive solution, collecting supernatant, and concentrating the supernatant to 20% of original volume by evaporation under reduced pressure;
(3) regulating the pH of the concentrated extract obtained in the step (2) to 4.5 by using dilute hydrochloric acid at room temperature, standing for 12 hours, and centrifuging to remove precipitates;
(4) preparing an ammonium sulfate/polyethylene glycol (PEG)2000 two-aqueous phase extraction system, wherein the mass fraction of ammonium sulfate is 15%, and the mass fraction of PEG2000 is 15%; extracting the extracting solution obtained in the step (3) for 50min by using the extracting system under the conditions of ultrasonic assistance, ultrasonic power of 200W and room temperature (20-30 ℃); and (4) after extraction is finished and the mixture is fully kept stand, taking the lower phase to obtain the tea bran polysaccharide. The removal rate of protein impurities was measured to be 74%.
Preparing a scalp care solution: the tea bran polysaccharide oral liquid comprises, by mass percentage (g: mL), 2% of tea bran polysaccharide, 0.8% of radix sophorae flavescentis extract, 0.5% of ginsenoside, 2% of glycerol, 3% of butanediol, 1% of menthol, 1% of hydrogenated lecithin, 1% of 1, 2-pentanediol, 0.1% of zinc aspartate, 0.05% of PCA copper, 0.05% of EDTA-2Na, 0.1% of citric acid, 0.2% of arginine, 0.2% of sarcosine, 0.3% of sodium benzoate, 5% of ethanol and the balance of water.
Comparative example 3:
preparing a scalp care solution: comprises 0.8 percent of lightyellow sophora root extract, 0.5 percent of ginsenoside, 2 percent of glycerin, 3 percent of butanediol, 1 percent of menthol, 1 percent of hydrogenated lecithin, 1 percent of 1, 2-pentanediol, 0.1 percent of zinc aspartate, 0.05 percent of PCA copper, 0.05 percent of EDTA-2Na, 0.1 percent of citric acid, 0.2 percent of arginine, 0.2 percent of sarcosine, 0.3 percent of sodium benzoate, 5 percent of ethanol and the balance of water in percentage by mass (g: mL).
Scalp barrier repair function test: after cleaning the scalp and hair of 50 volunteers in each group every day, spraying scalp care solution on the scalp, massaging until the scalp is absorbed, selecting one scalp, measuring for 3 times each time, and taking the average value; skin was measured at 0h, 1h, 3h, 5h and 7h before the first day of use, and transepidermal water loss values (TEWL) were measured at 0h, 1h, 3h, 5h and 7h after 21 consecutive days of use, as shown in table 1. Measuring the moisture content of scalp of a subject by using a German CorneometerCK-MPA skin moisture tester, measuring the scalp of a selected part for 3 times, and taking the average value; the change in the amount of skin moisture before use, 1h after use, 3h after use and 5h after use was measured, as shown in FIG. 4.
TABLE 1
Sensory evaluation: the degree of itching of 25 volunteers per group who were self-induced to scalp itching was evaluated according to 1-5 points, the initial average score was 3.9, and the evaluation results of the degree of scalp itching of the volunteers after 14 days of application of different scalp care solutions are shown in table 2.
TABLE 2
From the above experimental results, it can be seen that the itching relieving effect of the tea bran polysaccharide component B is optimal, the molecular weight of the tea bran polysaccharide component B is 5000-10000 Da, arabinose, galactose and glucose are taken as main components, and various other monosaccharides are contained, and the molecular weight of the tea bran polysaccharide component C is 10000-100000 Da, so that the itching relieving effect of the tea bran polysaccharides composed of different molecular weights and structures obtained by fractional purification is different. In contrast, in comparative examples 1 and 2, the protein provides nutrients for the microorganisms due to the high content of protein impurities, so that the activity of inhibiting the microorganisms is reduced, and the itching relieving effect of the care solution is affected.
It should be noted that the above-mentioned embodiments are only for illustrating the technical solutions of the present invention and not for limiting, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, which should be covered by the claims of the present invention.
Claims (10)
1. A method for extracting and purifying tea bran polysaccharide is characterized by comprising the following steps: comprises the steps of (a) preparing a mixture of a plurality of raw materials,
taking degreased oil tea bran powder, adding 85-95% ethanol, and mixing the materials in a material-liquid ratio of 1: refluxing for 2-3 h at 50-60 ℃ for 5-10 ℃, and drying filter residue after suction filtration to obtain tea saponin-removed tea bran powder;
mixing tea saponin-removed tea bran powder with distilled water, and mixing the mixture in a material-liquid ratio of 1: extracting for 3-4 hours at the temperature of 60-70 ℃ and at 5-10 ℃, centrifuging the extracting solution, taking supernatant, and concentrating the volume of the supernatant to 20-30% of the original volume by reduced pressure evaporation to obtain a concentrated solution;
adjusting the pH of the concentrated solution to 4.0-5.0 by using dilute hydrochloric acid at room temperature, standing for 10-14 h, and centrifuging to remove precipitates to obtain an extracting solution;
preparing an ammonium sulfate/polyethylene glycol 2000 aqueous two-phase extraction system, extracting the extracting solution under the ultrasonic-assisted condition, and taking the lower phase after fully standing.
2. A method for extracting and purifying a tea bran polysaccharide as claimed in claim 1, wherein: the ammonium sulfate/polyethylene glycol 2000 double-aqueous-phase extraction system comprises 14-16% by mass of ammonium sulfate and 11-13% by mass of polyethylene glycol 2000.
3. A method for extracting and purifying a tea bran polysaccharide as claimed in claim 1 or 2, wherein: and extracting the extracting solution under the ultrasonic-assisted condition, wherein the ultrasonic power of the extracting solution is 200-300W.
4. A method for extracting and purifying a tea bran polysaccharide as claimed in claim 1 or 2, wherein: the extraction is carried out by carrying out two-aqueous phase extraction on the extracting solution at room temperature, wherein the extraction time is 50-60 min; and taking the lower phase after fully standing.
5. A method for extracting and purifying a tea bran polysaccharide as claimed in claim 1 or 2, wherein: further comprises the steps of taking the lower phase, screening and purifying the molecular weight and the structure by adopting a DEAE cellulose column chromatography, comprising the steps of DEAE cellulose pretreatment, column packing and 0.05 mol.L-1The NaCl solution is subjected to gradient elution, and the eluent is collected.
6. A method for extracting and purifying a tea bran polysaccharide as claimed in claim 5, wherein: the DEAE cellulose pretreatment comprises the steps of taking DEAE-52 cellulose, adding distilled water, swelling for 48 hours at 4 ℃, and carrying out suction filtration; soaking the mixture in 0.5mol/L NaOH solution for 2h, washing the mixture with distilled water until the pH value is 7.0, and performing suction filtration; soaking the mixture in 0.5mol/L HCl solution for 2h, washing the mixture with distilled water until the pH value is 7.0, and performing suction filtration; and finally soaking the mixture in 0.5mol/L NaOH solution for 2h, washing the mixture with distilled water until the pH value is 7.0, soaking the mixture in the distilled water, and removing bubbles by ultrasonic waves.
7. A method for extracting and purifying a tea bran polysaccharide as claimed in claim 5, wherein: and (3) filling the column, namely adding distilled water into the chromatographic column, slowly pouring the pretreated DEAE-cellulose suspension into the column, adding a proper amount of quartz sand, and balancing the chromatographic column by using the distilled water.
8. A method for extracting and purifying a tea bran polysaccharide as claimed in claim 1 or 2, wherein: and adjusting the pH of the concentrated solution to 4.0-5.0 by using dilute hydrochloric acid at room temperature to adjust the pH of the concentrated solution to 4.5.
9. A method for extracting and purifying a tea bran polysaccharide as claimed in claim 1 or 2, wherein: the mass fraction of the ammonium sulfate is 15 percent, and the mass fraction of the polyethylene glycol 2000 is 12 percent.
10. A scalp care composition comprising the tea bran polysaccharides as claimed in any one of claims 1 to 9, characterized in that: comprises 2-3% of tea bran polysaccharide, 0.8-1.0% of radix sophorae flavescentis extract, 0.5-1% of ginsenoside, 2-3% of glycerol, 3-4% of butanediol, 1-2% of menthol, 1-2% of hydrogenated lecithin, 1-2% of 1, 2-pentanediol, 0.1-0.2% of zinc aspartate, 0.05-0.07% of PCA copper, 0.05-0.06% of EDTA-2Na, 0.1-0.3% of citric acid, 0.2-0.3% of arginine, 0.2-0.3% of sarcosine, 0.3-0.4% of sodium benzoate, 5-6% of ethanol and the balance of water.
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