High-purity camellia saponin and extraction method thereof

Abstract

The extraction method comprises the following steps: crushing camellia seed cake meal or tea seed cake meal, carrying out dynamic countercurrent extraction on the crushed camellia seed cake meal or tea seed cake meal by using a water-containing low alcohol solution, and collecting filtrate; concentrating the filtrate to recover alcohol until the alcohol content in a concentrated solution is less than 3wt%, so as to obtain a concentrated solution A; diluting the concentrated solution A with water, treating the diluent with an ultrafiltration membrane, and concentrating the ultrafiltration membrane permeate with a nanofiltration membrane to obtain a concentrated solution B; adding water into the concentrated solution to obtain a dissolved solution, decoloring the dissolved solution, adsorbing the decolored solution through macroporous adsorption resin, and then desorbing through an ethanol solution to obtain a desorbed solution; and concentrating, sterilizing, filtering and drying the desorption solution to obtain the camellia saponin. The camellia oleifera saponin prepared by the method is high in purity, low in impurity content and light in color, is widely applied to industries such as high-end wash care products and cosmetics, and is simple and feasible in preparation method and suitable for large-scale production.

Description

Technical Field

[0001] The invention relates to the technical field of biological extraction, and in particular to a high-purity camellia saponin and an extraction method thereof. Background Art

[0002] Tea saponin, also known as tea saponin, is a type of glycoside compound extracted from tea tree seeds (tea seeds, tea seeds). It is a natural surfactant with good performance. It can be widely used in light industry, chemical industry, pesticides, feed, breeding, textiles, oil production, mining, building materials and highway construction.

[0003] Camellia oleifera saponins prepared by prior art contain various impurities such as proteins, polysaccharides, flavonoids, polyphenols, organic acids, tannins, tannins, pigments, etc., which affect the advantages of product formula and stable performance, seriously affect the unique surface activity function and antibacterial and anti-dandruff effect of camellia oleifera saponins, and can only be used in the fields of pesticide adjuvants, low-end detergents, oil extraction and building materials. Refining and purifying camellia oleifera saponins need to further remove impurities such as proteins, camellia oleifera polysaccharides, flavonoids, polyphenols, organic acids, tannins, tannins and other pigments in crude saponins, especially to remove colored impurities, which is the key and difficulty of the key technology for preparing high-purity camellia oleifera saponins. Existing refining and purification methods include flocculation, precipitation, resin adsorption, hydrogen peroxide oxidation or repeated recrystallization, etc., and these refining and purification methods, or the impurity removal effect is not good, or the process is complicated and cumbersome and not easy to control, the loss of camellia oleifera saponins is large, or the introduction and generation of new chemical pollutants are difficult to completely remove, and are not suitable for large-scale industrial production. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide a high-purity camellia saponin and an extraction method thereof, wherein the high-purity camellia saponin has high purity, low impurity content, light color, and can be widely used in high-end cleaning products, cosmetics, medicines and food industries. The preparation method is simple and feasible and suitable for large-scale industrial production.

[0005] In a first aspect, the present invention provides a method for extracting high-purity camellia saponin, comprising the following steps:

[0006] Crush camellia oil seed cake or tea seed cake, use an aqueous lower alcohol solution to dynamically countercurrently extract the crushed camellia oil seed cake, and collect the filtrate;

[0007] The filtrate is concentrated to recover alcohol until the alcohol content in the concentrated solution is less than 3 wt %, thereby obtaining a concentrated solution A;

[0008] The concentrated solution A is diluted with water, the diluted solution is treated with an ultrafiltration membrane, and the ultrafiltration membrane permeate is concentrated with a nanofiltration membrane to obtain a concentrated solution B;

[0009] Adding water to the concentrated solution B to obtain a dissolved solution, decolorizing the dissolved solution, adsorbing the decolorized solution with a macroporous adsorption resin, and then analyzing with an ethanol solution to obtain an analyzed solution;

[0010] The analytical solution is concentrated, sterilized, filtered and dried to obtain camellia saponin.

[0011] Compared with the prior art, the preparation method of the present invention first crushes the camellia seed cake or tea seed cake, and performs dynamic countercurrent extraction on the crushed camellia seed cake or tea seed cake to obtain a filtrate; then the filtrate is concentrated, the concentrated solution is diluted with water, the diluted solution is treated with an ultrafiltration membrane, and the ultrafiltration membrane permeate is concentrated through a nanofiltration membrane to effectively remove macromolecular substances and small molecular inorganic salts; then water is added to the concentrated solution to dissolve, decolorize, adsorb the decolorized solution with resin, and analyze with ethanol to further improve the purity of camellia saponin; finally, the analyzed solution is processed to obtain high-purity camellia saponin. The preparation method of the high-purity camellia saponin of the present invention makes the prepared high-purity camellia saponin have high purity, low impurity content, and light color, and can be widely used in high-end cleaning products, cosmetics, medicines, food and other industries, and the preparation method is simple and feasible, suitable for large-scale industrial production.

[0012] Furthermore, the conditions for crushing the oil-tea seed cake or tea seed cake are as follows: the feeding rate is 10-200kg / h, the frequency is 30-50HZ; the particle size of the oil-tea seed cake or tea seed cake after crushing is 0.1-2mm; when the crushed oil-tea seed cake or tea seed cake is subjected to dynamic countercurrent extraction, the extraction temperature is 30-50°C, the extraction frequency is 30-50HZ, and the extraction propulsion speed is 5-10m / h; the aqueous lower alcohol solution includes lower alcohol, compound salt and water, wherein the mass concentration of the lower alcohol is 50-90%, and the mass concentration of the compound salt is 0.1-0.5%; the compound salt includes one or more of sulfite, bisulfite or pyrosulfite.

[0013] Compared with the prior art, the preparation method of the present invention uses an aqueous low-grade alcohol solution containing 0.1-0.5wt% of sulfite, bisulfite or pyrosulfite to replace water or an alcohol solution not containing the above-mentioned composite salt in the prior art, which can reduce the exudation of oil, protein and macromolecular tannin in the extraction of raw materials, make it easier to filter, reduce the further oxidation of some phenolic acid compounds in the extraction process, reduce the increase of color-developing substances, prevent the color of the feed solution from deepening, and reduce the burden of product decolorization.

[0014] Furthermore, the method for concentrating the filtrate to recover alcohol is nanofiltration membrane concentration, wherein the specification of the nanofiltration membrane is 150-200Da, the pressure during the operation of the nanofiltration membrane is 0.5-3.0MPa, the temperature is 10-65°C, the pH is 3.0-12.0, and the flow rate of the filtrate is 0.5-5.0m 3 / h.

[0015] Furthermore, the Brix value of the diluted solution after the concentrated solution A is diluted with water is 1-10%; when the diluted solution is treated with an ultrafiltration membrane, the specification of the ultrafiltration membrane is 15-20KDa, the inlet membrane pressure during operation is 0.1-1.0MPa, the temperature is 10-65°C, the pH is 3.0-12.0, the flow rate of the diluted solution is 0.5-5.0m3 / h, and the macromolecular substances with a molecular weight cutoff greater than 20KDa are retained, and water is continuously added for dialysis until the Brix value of the permeate is ≤0.5%; when the permeate of the ultrafiltration membrane is concentrated by a nanofiltration membrane, the specification of the nanofiltration membrane is 400-600Da, the pressure during operation is 0.1-3.0MPa, the temperature is 10-65°C, and the pH is 3.0-12.0, and the flow rate of the permeate of the ultrafiltration membrane is 0.5-5.0m3 / h, until the Brix value of the concentrated solution is 25-30%, thereby obtaining a concentrated solution B.

[0016] Furthermore, the method for decolorizing the dissolved solution is to decolorize the dissolved solution by adsorption with a macroporous adsorption resin or by ion exchange with an anion exchange resin; when the dissolved solution is decolorized by adsorption with the macroporous adsorption resin or by ion exchange with an anion exchange resin, the loading speed of the dissolved solution is 0.5-1BV / h, and the loading amount is 0.5-5.0BV of the resin volume. After decolorization with the macroporous adsorption resin or the anion exchange resin is completed, a decolorized solution is obtained; the macroporous adsorption resin is a polystyrene macroporous adsorption resin or an acrylate macroporous adsorption resin, and the anion exchange resin is a polystyrene skeleton or an acrylate skeleton anion exchange resin.

[0017] Furthermore, when the decolorizing solution is adsorbed by the macroporous adsorption resin, the loading speed of the decolorizing solution is 1-2BV / h, and the loading amount is 0.5-5.0BV of the resin volume; when the ethanol solution is analyzed, the loading speed of the ethanol solution is 0.5-1BV / h, and the loading amount is 1.0-5.0BV.

[0018] Further, the method for concentrating the analytical solution is as follows: the analytical solution is subjected to membrane concentration, and when the solid content of the concentrated solution reaches 20-30%, it is transferred to a single-effect external circulation concentrator for further concentration until the Brix value reaches 40-50%; the specification of the membrane used for membrane concentration is 100-200Da; when the single-effect external circulation concentrator continues to concentrate, the concentration vacuum degree is 0.05-0.095MPa, and the concentration temperature is 50-85°C;

[0019] Furthermore, the sterilization temperature is 80-85°C and the sterilization time is 30-35 minutes;

[0020] Furthermore, the drying method is spray drying, the feed rate of the spray drying is 10-100 L / h, the inlet air temperature is 180-190°C, and the outlet air temperature is 85-90°C.

[0021] In a second aspect, the present invention provides a high-purity camellia saponin, wherein the purity of the high-purity camellia saponin is above 97%, the protein content in the high-purity camellia saponin is ≤1.0wt%, the polyphenol content is ≤1.0g / kg, and the absorbance Abs at a wavelength of 420-610nm is <0.06.

[0022] Compared with the prior art, the beneficial effects of the high-purity camellia saponin of the present invention are the same as the beneficial effects of the camellia saponin preparation method of the above technical scheme, which will not be described in detail here. DETAILED DESCRIPTION

[0023] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0024] The present invention provides a method for extracting high-purity camellia saponin, comprising the following steps:

[0025] crushing camellia oil seed cake or tea seed cake, performing dynamic countercurrent leaching on the crushed camellia oil seed cake or tea seed cake using an aqueous lower alcohol solution, and collecting a filtrate;

[0026] The filtrate is concentrated to recover alcohol until the alcohol content in the concentrated solution is less than 3 wt %, thereby obtaining a concentrated solution A;

[0027] The concentrated solution A is diluted with water, the diluted solution is treated with an ultrafiltration membrane, and the ultrafiltration membrane permeate is concentrated with a nanofiltration membrane to obtain a concentrated solution B;

[0028] Adding water to the concentrated solution B to obtain a dissolved solution, decolorizing the dissolved solution, adsorbing the decolorized solution with a macroporous adsorption resin, and then analyzing with an ethanol solution to obtain an analyzed solution;

[0029] The analytical solution is concentrated, sterilized, filtered and dried to obtain camellia saponin.

[0030] The preparation method of the present invention comprises the following steps: firstly, pulverizing camellia seed cake or tea seed cake, and using an aqueous low-grade alcohol solution to dynamically countercurrently extract the pulverized camellia seed cake or tea seed cake to obtain a filtrate, so that camellia saponin in the camellia seed cake or tea seed cake can be fully extracted; then, the filtrate is concentrated to remove alcohols in the filtrate; the obtained concentrated solution A is diluted with water and treated with an ultrafiltration membrane to effectively remove high-molecular and macromolecular proteins, camellia polysaccharides, starch and macromolecular substances in the diluted solution; then, the ultrafiltration membrane permeate is concentrated through a nanofiltration membrane to obtain a concentrated solution B, and small-molecular inorganic salts in the ultrafiltration membrane permeate are further removed, so that the purity of the obtained concentrated solution B is greatly improved; then, the concentrated solution B is dissolved in water, decolorized, adsorbed by a decolorizing solution resin, and analyzed with ethanol to obtain an analytical solution, so that colored substances and impurities in the camellia saponin are removed, so that the purity of the camellia saponin is further improved; and finally, the analytical solution is treated to obtain high-purity camellia saponin. The preparation method of the high-purity camellia saponin of the present invention ensures that the prepared high-purity camellia saponin has high purity, low impurity content and light color, and can be widely used in high-end cleaning products, cosmetics, medicines, food and other industries. The preparation method is simple and feasible and is suitable for large-scale industrial production.

[0031] In some embodiments, the conditions for crushing camellia seed cake or tea seed cake are: feeding rate is 10-200kg / h, frequency is 30-50HZ; the particle size of the camellia seed cake or tea seed cake after crushing is 0.1-2mm; when the crushed camellia seed cake or tea seed cake is subjected to dynamic countercurrent extraction, the extraction temperature is 30-50°C, the extraction frequency is 30-50HZ, and the extraction propulsion speed is 5-10m / h; the aqueous lower alcohol solution includes lower alcohol, compound salt and water, wherein the mass concentration of the lower alcohol is 50-90%, and the mass concentration of the compound salt is 0.1-0.5%, preferably 0.2-0.4%; the compound salt includes one or more of sulfite, bisulfite or pyrosulfite. The lower alcohol in the aqueous lower alcohol may be methanol, ethanol, n-propanol, isopropanol and n-butanol; the sulfite, bisulfite or pyrosulfite may be mixed in any mass ratio until the required content in the aqueous lower alcohol solution is achieved. Preferably, the mass ratio of sulfite, bisulfite and pyrosulfite may be (1-2):(1-2):(2-3).

[0032] Specifically, the feeding rate of camellia oil seed cake or tea seed cake crushing can be 10kg / h, 50kg / h, 100kg / h, 160kg / h, 200kg / h; the frequency can be 30HZ, 35HZ, 40HZ, 45HZ, 50HZ; the particle size of camellia oil seed cake or tea seed cake after crushing can be 0.1mm, 0.6mm, 1.0mm, 1.6mm, 2mm; the extraction temperature during extraction of the crushed camellia oil seed cake or tea seed cake can be 30℃, 33℃, 38℃, 45℃, 50℃; the extraction frequency can be 30HZ, 35HZ, 40HZ, 42HZ, 50HZ, and the extraction advancement speed can be 5m / h, 8m / h, 8.5m / h, 9m / h, 10m / h.

[0033] Under the above parameters and the cooperation of the aqueous lower alcohol solution, the camellia seed cake or tea seed cake is crushed and extracted, so that the camellia saponin in the camellia seed cake or tea seed cake can be fully extracted. The aqueous lower alcohol solution contains 0.1-0.5wt% of one or more of sulfite, bisulfite or pyrosulfite, which can reduce the exudation of oil, protein and macromolecular tannin in the extraction of raw materials, make it easier to filter, reduce the further oxidation of some phenolic acid compounds in the extraction process, reduce the increase of color-developing substances, prevent the color of the feed liquid from deepening, and reduce the burden of product decolorization.

[0034] In some embodiments, the method for concentrating the filtrate to recover alcohol is nanofiltration membrane concentration, wherein the specification of the nanofiltration membrane is 150-200Da, the pressure is 0.5-3.0MPa, the temperature is 10-65°C, the pH is 3.0-12.0, and the flow rate of the filtrate is 0.5-5.0m 3 / h.

[0035] Specifically, the specifications of the nanofiltration membrane can be 150Da, 165Da, 185Da, 190Da, 200Da, the pressure can be 0.5MPa, 0.9MPa, 1.5MPa, 2.5MPa, 3MPa, the temperature can be 10℃, 20℃, 35℃, 50℃, 65℃, the pH can be 3, 4, 5, 9, 12, and the flow rate of the filtrate can be 0.5m 3 / h、1.6m 3 / h、2.5m 3 / h、3.8m 3 / h、5m 3 / h.

[0036] When the filtrate is concentrated by nanofiltration membrane, under the above parameter conditions, the filtrate can be fully concentrated to remove the alcohol in the filtrate. In actual operation, the alcohol recovered after concentration can be distilled and recycled.

[0037] In some embodiments, after the concentrated solution A is diluted with water, the Brix value of the dilution is 1-10%; when the dilution is treated with an ultrafiltration membrane, the specification of the ultrafiltration membrane is 15-20KDa, the inlet membrane is 0.1-1.0MPa, the temperature is 10-65°C, the pH is 3.0-12.0, the flow rate of the dilution is 0.5-5.0m3 / h, and the macromolecular substances with a molecular weight greater than 20KDa are retained, and water is continuously added for dialysis until the Brix value of the permeate is ≤0. 5% ends; when the ultrafiltration membrane permeate is concentrated by the nanofiltration membrane, the specification of the nanofiltration membrane is 400-600Da, it can pass substances with a molecular weight of less than 400Da, and retain substances with a molecular weight greater than 600Da. The operating pressure is 0.1-3.0MPa, the temperature is 10-65°C, and the pH is 3.0-12.0. The flow rate of the ultrafiltration membrane permeate is 0.5-5.0m3 / h, and it ends when the Brix value of the concentrate is 25-30%, thereby obtaining a concentrated solution B.

[0038] It should be noted that the Brix value refers to the content of soluble solids in a liquid. Specifically, after the concentrated solution A is diluted with water, the Brix value can be 1%, 3%, 6%, 8%, 10%; the specifications of the ultrafiltration membrane can be 15KDa, 17KDa, 18KDa, 19KDa, 20KDa; the inlet membrane pressure during operation can be 0.1MPa, 0.3MPa, 0.5MPa, 0.8MPa, 1.0MPa; the temperature can be 10℃, 25℃, 37℃, 50℃, 65℃; the pH can be 3, 5, 7, 9, 12; the flow rate of the diluent can be 0.5m 3 / h、1.6m 3 / h、3.0m 3 / h、3.8m 3 / h、5m 3 / h; when the dilution is treated by the ultrafiltration membrane, the Brix value of the permeate can be 0.1%, 0.2%, 0.35%, 0.4%, 0.5%. When the permeate of the ultrafiltration membrane is concentrated by the nanofiltration membrane, the specifications of the nanofiltration membrane can be 400Da, 460Da, 500Da, 550Da, 600Da, and the operating pressure can be 0.1MPa, 0.8MPa, 1.5MPa, 2.0MPa, 3.0MPa; the temperature can be 10℃, 25℃, 37℃, 50℃, 65℃; the pH is 3, 6, 8, 10, 12; the flow rate of the dilution can be 0.5m 3 / h、1.6m 3 / h、3.0m 3 / h、3.8m 3 / h、5m 3 / h; the Brix value of concentrated liquid B can be 25%, 26.5%, 27.5%, 28%, and 30%.

[0039] Under the above membrane specifications and parameter conditions, the high molecular and macromolecular proteins, camellia oleifera polysaccharides, starch, macromolecular substances and small molecular inorganic salts in the dilution liquid are fully removed, thereby improving the purity of camellia oleifera saponin in the dilution liquid.

[0040] In some embodiments, the Brix value of the dissolved solution is 8-10%; the method for decolorizing the dissolved solution is adsorption by macroporous adsorption resin or ion exchange decolorization by anion exchange resin; when the dissolved solution is adsorbed by macroporous adsorption resin or ion exchange decolorization by anion exchange resin, the loading speed of the dissolved solution is 0.5-1BV / h, and the loading amount is 0.5-5.0BV of the resin volume. After the decolorization by macroporous adsorption resin or anion exchange resin is completed, a decolorized solution is obtained; the macroporous adsorption resin is a weakly polar to polar macroporous resin with a particle size of 250μm-1.25mm, which can be a polystyrene macroporous adsorption resin or an acrylate macroporous adsorption resin, and the anion exchange resin is a weakly alkaline anion resin with a particle size of 250μm-2mm, which can be a polystyrene skeleton or an acrylate skeleton anion exchange resin.

[0041] Specifically, the Brix value of the solution can be 8%, 8.3%, 9%, 9.5%, 10%; the loading speed of the solution can be 0.5BV / h, 0.65BV / h, 0.7BV / h, 0.9BV / h, 1.0BV / h; the loading volume can be 0.5BV, 0.9BV, 2.5BV, 3.8BV, 5.0BV of the resin volume; the macroporous adsorption resin used for decolorizing the solution It can be any one or more of ADS-7, DA201, DM100, LKS03, LKS11, MN100, MN270, SD331, SD333, T19, and XDA-8; the anion exchange resin used for decolorizing the dissolved solution can be any one or more of LX600J, D941, D900, D318, D301, D285, D201, and H280.

[0042] When the dissolved solution is decolorized, under the above-mentioned parameter conditions, the colored substances in the dissolved solution can be effectively removed, making the decolorized solution purer. It should be noted that the dissolved solution can be decolorized by conventional technical means for decolorizing the solution in the prior art, as long as the purpose of decolorization can be achieved. The present invention preferably uses macroporous adsorption resin adsorption decolorization or anion exchange resin ion exchange decolorization. It should be understood that in the process of macroporous adsorption resin adsorption decolorization or anion exchange resin ion exchange decolorization of the dissolved solution, after the dissolved solution is sampled, the clear water that flows out at the beginning should be removed, and the collection starts when the feed liquid flows out. After the sampling is completed, it can be washed with 1-5wt% saline, and the feed liquid and the washing liquid are collected to obtain the decolorized solution. It can be seen that when macroporous adsorption resin is used for decolorization or anion exchange resin is used for ion exchange decolorization, after the decolorization resin is decolorized, it is only necessary to rinse the decolorization resin with salt water, which replaces the complicated flocculation, precipitation, oxidation-reduction and other steps in the prior art. The process is simple and suitable for industrial production. The simple decolorization process and the removal synergy of high-molecular and macromolecular proteins, camellia polysaccharides, starch, macromolecular substances and small-molecule inorganic salts in the camellia seed cake or tea seed cake extract further improve the purity of camellia saponin. After the decolorization of the lysate is completed, the resin used for decolorization can be regenerated and recycled. Specifically, after being rinsed with 1-5wt% hydrochloric acid, it is washed with water to neutrality, and then rinsed with 2-5wt% sodium hydroxide lye. It can be directly used after being washed with water to neutrality.

[0043] In some embodiments, when the decolorizing solution is adsorbed by a macroporous adsorption resin, the loading rate of the decolorizing solution is 1-2BV / h, and the loading amount is 0.5-5.0BV of the resin volume; when the ethanol solution is analyzed, the loading rate of the ethanol solution is 0.5-1BV / h, and the loading amount is 1.0-5.0BV.

[0044] Specifically, the macroporous adsorption resin is a non-polar to weakly polar macroporous resin, which can be a polystyrene macroporous adsorption resin or an acrylate macroporous adsorption resin, with a particle size of 250 μm-1.25 mm and a water content of 55-75%. The macroporous adsorption resin model used can be one or more of AB-8, ADS-5, D101, D3520, D4006, D4020, H103, HP20, HP2MG, HPD100, HPD600, HZ801, HZ803, LX20, LX60, LK1180, LK1200, LK1300, LK1400, LK1500, LKS06, SP825, T28, T81, T83, X-5, XAD-5, XAD-8, XAD-16, and XAD-1600.

[0045] The decolorizing solution is adsorbed by a macroporous adsorption resin. Under the above-mentioned parameter conditions, the camellia saponin in the decolorizing solution can be effectively adsorbed, and the camellia saponin can be extracted to a greater extent. After the decolorizing solution is adsorbed by the macroporous adsorption resin, it is analyzed with ethanol. It should be understood that after the ethanol analysis is completed, the macroporous adsorption resin can be further rinsed with water, and the ethanol analysis solution and the water washing solution are collected and combined to obtain the analysis solution; after the elution analysis is completed, the macroporous adsorption resin can be rinsed with water until there is no alcohol, and then it can be recycled.

[0046] In some embodiments, the method for concentrating the analyte is as follows: the analyte is subjected to membrane concentration, and when the solid content of the concentrated solution reaches 20-30%, it is transferred to a single-effect external circulation concentrator for further concentration until the Brix value reaches 40-50%; the specification of the membrane used for membrane concentration is 100-200Da; when the single-effect external circulation concentrator continues to concentrate, the concentration vacuum degree is 0.05-0.095MPa, and the concentration temperature is 50-85°C.

[0047] In some embodiments, the sterilization temperature is 80-85° C. and the sterilization time is 30-35 min.

[0048] In some embodiments, the drying method is spray drying, the feed rate of the spray drying is 10-100 L / h, the inlet air temperature is 180-190°C, and the outlet air temperature is 85-90°C.

[0049] The high-purity camellia saponin prepared by the preparation method of the present invention has a purity of more than 97%, a protein content of ≤1.0wt%, a polyphenol content of ≤1.0g / kg, and an absorbance Abs of <0.06 at a wavelength of 420-610nm. The test methods for the purity, protein content, polyphenol content and absorbance of the high-purity camellia saponin are as follows:

[0050] Testing method for purity of high-purity camellia saponin: GB / T41549;

[0051] Testing method for protein content of high-purity camellia saponin: GB5009.5;

[0052] Testing method for the content of high-purity camellia saponin polyphenols: Folin-Ciocalteu reagent method;

[0053] Testing method for the absorbance of high-purity camellia saponin: UV-visible spectrophotometry.

[0054] In order to better illustrate the technical solution of the present invention, the present invention also provides the following specific examples. It should be understood that the raw materials used in the following examples are all commercially available raw materials unless otherwise specified.

[0055] Embodiment 1:

[0056] A method for extracting high-purity camellia saponin from camellia seed cake, comprising the following steps:

[0057] (1) Raw material preparation: Camellia oleifera cake is first-grade grass-free cake, and other materials meet the requirements of GB35131 standard and are commercially available.

[0058] (2) Crushing and extraction: 1000 kg of the raw material camellia seed cake was weighed and fed at a uniform speed of 10 kg / h with a scraper. The feeding speed was adjusted to 30 Hz. After being crushed in the crusher, the particle size of the camellia seed cake particles was 0.6 mm and fell into one end of the dynamic countercurrent extraction unit. At the other end of the extraction unit, 50% aqueous ethanol (the aqueous ethanol contained 0.2 wt% of sodium bisulfite and sodium metabisulfite, and the mass ratio of sodium bisulfite to sodium metabisulfite was 1:1) was input through a pipeline. The flow rate was 1 m 3 / h for extraction. The extraction temperature is 50℃, the propulsion speed of the screw shaft of the dynamic countercurrent extraction unit is 8m / h, and the frequency is 30Hz. The extract is discharged from the port pipeline filter to the temporary storage tank, and the tea seed meal extraction residue is pushed to the end of the unit and filtered and dried by the grid, and can be used as feed.

[0059] (3) Nanofiltration membrane concentration: The filtrate is concentrated by nanofiltration membrane to recover alcohol until the residual alcohol content of the concentrated liquid is 1%. The membrane module model is hollow fiber membrane NF95, specification 150Da, the temperature during nanofiltration membrane concentration is 20°C, the pressure is 0.9MPa, the pH is 4, and the flow rate is 0.5m 3 / h, the recovered alcohol can be distilled and recycled.

[0060] (4) Dilution: Dilute the concentrate with water. The Brix value of the dilution is 6%.

[0061] (5) Ultrafiltration and nanofiltration concentration: The diluted liquid was ultrafiltered through an ultrafiltration membrane. The membrane module model was NF01, the membrane specification was 15 KDa, and the inlet membrane pressure was maintained at 0.3 MPa, the temperature was 25 °C, the pH was 3, and the flow rate was 0.5 m / s. 3 / h, while continuously adding water for dialysis, retaining macromolecular substances with a molecular weight greater than 20KDa, until the Brix value of the permeate is 0.2%, and the permeate is collected. The permeate enters the series nanofiltration membrane for concentration. The nanofiltration membrane NF10, specification 400Da, can pass through substances with a molecular weight less than 400Da and retain substances with a molecular weight greater than 600Da. During operation, the pressure is 0.8MPa, the temperature is 10℃, and the pH is 3. The flow rate of the permeate of the ultrafiltration membrane is 1.6m3 / h, and the Brix value of the concentrated liquid is concentrated to 25%.

[0062] (6) Resin column decolorization: The concentrated solution obtained in step (5) was dissolved in deionized water, the Brix value of the dissolved solution was 8%, and the dissolved solution was loaded on a resin column LX600J for decolorization. The loading rate was 1 BV / h, and the loading amount was 5 BV of the resin volume. The front effluent clear water was discarded, and the feed liquid was collected. After the injection was completed, the residual liquid was pushed out with 2.5 times the volume of the resin with 2 wt% saline. The effluent and the water top liquid were collected together to obtain a decolorized solution.

[0063] (7) Macroporous resin adsorption: The decolorizing solution is injected into the macroporous adsorption resin HP20 for adsorption at a rate of 1 BV / h and a sample volume of 5 BV of the resin volume. After injection, the residual liquid is rinsed with 2-3 BV of the resin volume of water, and the effluent and the washing liquid are discarded. 2 BV of the resin volume of 85% ethanol is used for elution and analysis at a rate of 0.8 BV / h. Then, 2 times the volume of the resin volume of water is used to top up the alcohol. The eluent and the water top liquid are combined to form the analysis solution.

[0064] (8) Concentration: The analytical solution is concentrated by membrane, the membrane module model is hollow fiber membrane NF95, the specification is 200Da, and the ethanol is recovered until it is alcohol-free. When the Brix value of the concentrated solution is 22%, it is transferred to a single-effect external circulation concentrator for further concentration, the concentration vacuum degree is 0.05MPa, the concentration temperature is 60°C, and the concentration is completed when the Brix value of the concentrated solution reaches 40%.

[0065] (9) Sterilization: Pasteurize the concentrate at 82°C for 35 min.

[0066] (10) Drying: The sterilized concentrate was filtered through a pipeline filter and then spray dried. The feed rate was adjusted to 100 L / h, the air inlet temperature was 190°C, and the air outlet temperature was 85°C. The product was collected. The final product had a high-purity camellia saponin yield of 10.4%, an extraction rate of 78.8%, and a camellia saponin content of 98.5%. The purity, protein content, polyphenol content, and absorbance of the high-purity camellia saponin product were tested respectively. The results are shown in Table 1.

[0067] Example 2

[0068] A method for extracting high-purity camellia saponin from camellia seed cake, comprising the following steps:

[0069] (1) Raw material preparation: Camellia oleifera cake is first-grade grass-free cake, and other materials meet the requirements of GB35131 standard and are commercially available.

[0070] (2) Crushing and extraction: 1000 kg of camellia oleifera seed cake was weighed and fed uniformly with a scraper at a feeding speed of 160 kg / h and an adjustment frequency of 45 Hz. After being crushed in a crusher, the camellia oleifera seed cake particles had a particle size of 0.1 mm and fell into one end of a dynamic countercurrent extraction unit. A 70% aqueous methanol (the aqueous ethanol contained 0.4 wt% of sodium bisulfite and sodium sulfite, and the mass ratio of sodium bisulfite to sodium sulfite was 1:1) was input into the other end of the extraction unit through a pipeline at a flow rate of 1 m 3 / h for extraction. The extraction temperature is 38℃, the propulsion speed of the screw shaft of the dynamic countercurrent extraction unit is 8.5m / h, and the frequency is 42Hz. The extract is discharged from the port pipeline filter to the temporary storage tank, and the tea seed meal extraction residue is pushed to the end of the unit and filtered and dried by the grid, and can be used as feed.

[0071] (3) Nanofiltration membrane concentration: The filtrate is concentrated by nanofiltration membrane to recover alcohol until the residual alcohol content of the concentrate is 1.5%. The membrane module model is hollow fiber membrane NF95, specification 165Da, the temperature during nanofiltration membrane concentration is 35°C, the pressure is 1.5MPa, the pH is 3, and the flow rate is 1.6m 3 / h, the recovered alcohol can be distilled and recycled.

[0072] (4) Dilution: Dilute the concentrate with water. The Brix value of the dilution is 1%.

[0073] (5) Ultrafiltration and nanofiltration concentration: The diluted solution was ultrafiltered through an ultrafiltration membrane. The membrane module model was NF01, the membrane specification was 18 KDa, and the inlet membrane pressure was maintained at 0.1 MPa, the temperature was 37°C, the pH was 5, and the flow rate was 1.6 m / s. 3 / h, while continuously adding water for dialysis, retaining macromolecular substances with a molecular weight greater than 20KDa, until the Brix value of the permeate is 0.35%, and the permeate is collected. The permeate enters the series nanofiltration membrane for concentration. The nanofiltration membrane NF10, specification 460Da, can pass through substances with a molecular weight less than 400Da and retain substances with a molecular weight greater than 600Da. During operation, the pressure is 1.5MPa, the temperature is 37℃, and the pH is 8. The flow rate of the permeate of the ultrafiltration membrane is 3m3 / h, and the Brix value of the concentrated liquid is 28%.

[0074] (6) Resin column decolorization: The concentrated solution obtained in step (5) was dissolved in deionized water. The Brix value of the dissolved solution was 9%. The dissolved solution was decolorized on a resin column D941 at a loading rate of 0.65 BV / h. The loading amount was 3.8 BV of the resin volume. The clear water of the front effluent was discarded and the effluent was collected. After the injection was completed, the residual liquid was pushed out with 2.5 times the volume of the resin with 2 wt% saline. The effluent and the water top liquid were collected together to obtain a decolorized solution.

[0075] (7) Macroporous resin adsorption: The decolorizing solution was injected into the macroporous adsorption resin LK20 for adsorption at a rate of 1.5 BV / h. The sample volume was 0.5 BV of the resin volume. After injection, the residual liquid was washed with 2-3 BV of the resin volume of water, and the effluent and the washing liquid were discarded. 3.8 BV of the resin volume of 85% ethanol was used for elution and analysis at a rate of 0.5 BV / h. Then, 2 times the volume of the resin volume of water was used for top ethanol. The eluent and the water top liquid were combined to form the analysis solution.

[0076] (8) Concentration: The analytical solution is concentrated by membrane, the membrane module model is hollow fiber membrane NF95, the specification is 180Da, and the ethanol is recovered until it is alcohol-free. When the Brix value of the concentrated solution is 20%, it is transferred to a single-effect external circulation concentrator for further concentration, the concentration vacuum degree is 0.08MPa, the concentration temperature is 50°C, and the concentration is completed when the Brix value of the concentrated solution reaches 45%.

[0077] (9) Sterilization: The concentrate is pasteurized at 84°C for 32 min.

[0078] (10) Drying: The sterilized concentrate was filtered through a pipeline filter and then spray dried. The feed rate was adjusted to 10 L / h, the air inlet temperature was 180°C, and the air outlet temperature was 87°C. The product was collected. The final product had a high-purity camellia saponin yield of 10.9%, an extraction rate of 82.6%, and a camellia saponin content of 98.7%. The purity, protein content, polyphenol content, and absorbance of the high-purity camellia saponin product were tested respectively. The results are shown in Table 1.

[0079] Example 3

[0080] A method for extracting high-purity camellia saponin from camellia seed cake, comprising the following steps:

[0081] (1) Raw material preparation: Camellia oleifera cake is first-grade grass-free cake, and other materials meet the requirements of GB35131 standard and are commercially available.

[0082] (2) Crushing and extraction: 1000 kg of the raw material camellia seed cake was weighed and fed at a uniform speed of 50 kg / h with a scraper. The feeding speed was adjusted to 50 Hz. The particle size of the camellia seed cake particles after crushing in the crusher was 1.0 mm. The particles fell into one end of the dynamic countercurrent extraction unit. The other end of the extraction unit was fed with 75% aqueous methanol (the aqueous ethanol contained 0.1 wt% of sodium sulfite, sodium bisulfite and sodium metabisulfite, and the mass ratio of sodium sulfite, sodium bisulfite and sodium sulfite was 1:2:3) through a pipeline. The flow rate was 1 m 3 / h for extraction. The extraction temperature is 33℃, the propulsion speed of the screw shaft of the dynamic countercurrent extraction unit is 9m / h, and the frequency is 50Hz. The extract is discharged from the port pipeline filter to the temporary storage tank, and the tea seed meal extraction residue is pushed to the end of the unit and filtered and dried by the grid, and can be used as feed.

[0083] (3) Nanofiltration membrane concentration: The filtrate is concentrated by nanofiltration membrane to recover alcohol until the residual alcohol content of the concentrate is 2.8%. The membrane module model is hollow fiber membrane NF95, specification 190Da, the temperature during nanofiltration membrane concentration is 10°C, the pressure is 0.5MPa, the pH is 9, and the flow rate is 2.5m 3 / h, the recovered alcohol can be distilled and recycled.

[0084] (4) Dilution: Dilute the concentrate with water. The Brix value of the dilution is 8%.

[0085] (5) Ultrafiltration and nanofiltration concentration: The diluted liquid was ultrafiltered through an ultrafiltration membrane. The membrane module model was NF01, the membrane specification was 17 KDa, and the inlet membrane pressure was maintained at 1 MPa, the temperature was 65 °C, the pH was 7, and the flow rate was 3.0 m 3 / h, while continuously adding water for dialysis, retaining macromolecular substances with a molecular weight greater than 20KDa, until the Brix value of the permeate is 0.5%, and the permeate is collected. The permeate enters the series nanofiltration membrane for concentration. The nanofiltration membrane NF10, specification 550Da, can pass through substances with a molecular weight less than 400Da and retain substances with a molecular weight greater than 600Da. During operation, the pressure is 2.0MPa, the temperature is 50℃, and the pH is 6. The flow rate of the permeate of the ultrafiltration membrane is 3.8m3 / h, and the Brix value of the concentrated liquid is concentrated to 30%.

[0086] (6) Resin column decolorization: The concentrated solution obtained in step (5) was dissolved in deionized water, the Brix value of the dissolved solution was 9.5%, and the dissolved solution was decolorized on a resin column D900 at a loading rate of 0.7 BV / h. The loading amount was 2.5 BV of the resin volume. The clear water of the front effluent was discarded, and the effluent began to be collected. After the injection was completed, the residual liquid was pushed out with 2.5 times the volume of the resin with 2 wt% saline. The effluent and the water top liquid were collected together to obtain a decolorized solution.

[0087] (7) Macroporous resin adsorption: The decolorizing solution was injected into the macroporous adsorption resin AB-8 for adsorption at a rate of 1.8 BV / h. The sample volume was 4.2 BV of the resin volume. After the injection was completed, the residual liquid was washed with 2-3 BV of the resin volume of water, and the effluent and the washing liquid were discarded. 4.5 BV of the resin volume of 85% ethanol was used for elution and analysis at a rate of 0.7 BV / h. Then, 2 times the volume of the resin volume of water was used for top ethanol. The eluent and the water top liquid were combined to form the analysis solution.

[0088] (8) Concentration: The analytical solution is concentrated by membrane. The membrane module model is hollow fiber membrane NF95, specification 160Da, and ethanol is recovered until it is alcohol-free. When the Brix value of the concentrated solution is 26%, it is transferred to a single-effect external circulation concentrator for further concentration. The concentration vacuum is 0.06MPa and the concentration temperature is 75°C. The concentration is completed when the Brix value of the concentrated solution reaches 48%.

[0089] (9) Sterilization: Pasteurize the concentrate at 80°C for 30 min.

[0090] (10) Drying: The sterilized concentrate was filtered through a pipeline filter and then spray dried. The feed rate was adjusted to 25 L / h, the air inlet temperature was 185°C, and the air outlet temperature was 88°C. The product was collected. The final product had a high-purity camellia saponin yield of 11.1%, an extraction rate of 85.1%, and a camellia saponin content of 99.3%. The purity, protein content, polyphenol content, and absorbance of the high-purity camellia saponin product were tested respectively. The results are shown in Table 1.

[0091] Example 4

[0092] A method for extracting high-purity camellia saponin from camellia seed cake, comprising the following steps:

[0093] (1) Raw material preparation: Camellia oleifera cake is first-grade grass-free cake, and other materials meet the requirements of GB35131 standard and are commercially available.

[0094] (2) Crushing and extraction: 1000 kg of camellia oleifera seed cake was weighed and fed at a uniform speed of 100 kg / h using a scraper. The frequency was adjusted to 40 Hz. After crushing in the crusher, the camellia oleifera seed cake particles had a particle size of 2.0 mm and fell into one end of the dynamic countercurrent extraction unit. A 75% aqueous methanol (the aqueous ethanol contained 0.3 wt% sodium metabisulfite) was introduced into the other end of the extraction unit through a pipeline at a flow rate of 1 m 3 / h for extraction. The extraction temperature is 45℃, the propulsion speed of the screw shaft of the dynamic countercurrent extraction unit is 10m / h, and the frequency is 35Hz. The extract is discharged from the port pipeline filter to the temporary storage tank, and the tea seed meal extraction residue is pushed to the end of the unit and filtered and dried by the grid, and can be used as feed.

[0095] (3) Nanofiltration membrane concentration: The filtrate is concentrated by nanofiltration membrane to recover alcohol until the residual alcohol content of the concentrate is 0.5%. The membrane module model is hollow fiber membrane NF95, specification 200Da, the temperature during nanofiltration membrane concentration is 50°C, the pressure is 2.5MPa, the pH is 5, and the flow rate is 3.8m / s. 3 / h, the recovered alcohol can be distilled and recycled.

[0096] (4) Dilution: Dilute the concentrate with water. The Brix value of the dilution is 3%.

[0097] (5) Ultrafiltration and nanofiltration concentration: The diluted liquid was ultrafiltered through an ultrafiltration membrane. The membrane module model was NF01, the membrane specification was 19 KDa, and the inlet membrane pressure was maintained at 0.5 MPa, the temperature was 50 °C, the pH was 9, and the flow rate was 3.8 m / s. 3 / h, while continuously adding water for dialysis, retaining macromolecular substances with a molecular weight greater than 20KDa, until the Brix value of the permeate is 0.1%, and the permeate is collected. The permeate enters the series nanofiltration membrane for concentration. The nanofiltration membrane NF10, specification 500Da, can pass through substances with a molecular weight less than 400Da and retain substances with a molecular weight greater than 600Da. During operation, the pressure is 3.0MPa, the temperature is 65℃, and the pH is 12. The flow rate of the permeate of the ultrafiltration membrane is 0.5m3 / h, and the Brix value of the concentrated liquid is 26.5%.

[0098] (6) Resin column decolorization: The concentrated solution obtained in step (5) was dissolved in deionized water, the Brix value of the dissolved solution was 8.3%, and the dissolved solution was decolorized on a resin column SD331 at a loading rate of 0.9 BV / h. The loading amount was 0.9 BV of the resin volume. The clear water of the front effluent was discarded, and the effluent was collected. After the injection was completed, the residual liquid was pushed out with 2.5 times the volume of the resin with 2 wt% saline. The effluent and the water top liquid were collected together to obtain a decolorized solution.

[0099] (7) Macroporous resin adsorption: The decolorizing solution was injected into the macroporous adsorption resin LX60 for adsorption at a rate of 1.5 BV / h. The sample volume was 2.8 BV of the resin volume. After the injection was completed, the residual liquid was washed with 2-3 BV of the resin volume of water, and the effluent and the washing liquid were discarded. 5 BV of the resin volume of 85% ethanol was used for elution and analysis at a rate of 0.9 BV / h. Then, 2 times the volume of the resin volume of water was used to top up the alcohol. The eluent and the water top liquid were combined to form the analysis solution.

[0100] (8) Concentration: The analytical solution is concentrated by membrane. The membrane module model is hollow fiber membrane NF95, the specification is 100Da, and the ethanol is recovered until it is alcohol-free. When the Brix value of the concentrated solution is 30%, it is transferred to a single-effect external circulation concentrator for further concentration. The concentration vacuum is 0.075MPa and the concentration temperature is 85°C. The concentration is completed when the Brix value of the concentrated solution reaches 50%.

[0101] (9) Sterilization: The concentrate was pasteurized at 83°C for 31 min.

[0102] (10) Drying: The sterilized concentrate was filtered through a pipeline filter and then spray dried. The feed rate was adjusted to 75 L / h, the air inlet temperature was 186°C, and the air outlet temperature was 90°C. The product was collected. The final product had a high-purity camellia saponin yield of 10.9%, an extraction rate of 83.5%, and a camellia saponin content of 98.8%. The purity, protein content, polyphenol content, and absorbance of the high-purity camellia saponin product were tested respectively. The results are shown in Table 1.

[0103] Example 5

[0104] A method for extracting high-purity camellia saponin from camellia seed cake, comprising the following steps:

[0105] (1) Raw material preparation: Camellia oleifera cake is first-grade grass-free cake, and other materials meet the requirements of GB35131 standard and are commercially available.

[0106] (2) Crushing and extraction: 1000 kg of camellia oleifera seed cake was weighed and fed at a uniform speed of 200 kg / h using a scraper. The frequency was adjusted to 45 Hz. After crushing in the crusher, the camellia oleifera seed cake particles had a particle size of 1.6 mm and fell into one end of the dynamic countercurrent extraction unit. At the other end of the extraction unit, 90% aqueous ethanol (the aqueous ethanol contained 0.5 wt% sodium sulfite) was input through a pipeline at a flow rate of 1 m / s. 3 / h for extraction. The extraction temperature is 30℃, the propulsion speed of the screw shaft of the dynamic countercurrent extraction unit is 5m / h, and the frequency is 40Hz. The extract is discharged from the port pipeline filter to the temporary storage tank, and the tea seed meal extraction residue is pushed to the end of the unit and filtered and dried by the grid, and can be used as feed.

[0107] (3) Nanofiltration membrane concentration: The filtrate is concentrated by nanofiltration membrane to recover alcohol until the residual alcohol content of the concentrated liquid is 2%. The membrane module model is hollow fiber membrane NF95, specification 185Da, the temperature during nanofiltration membrane concentration is 65℃, the pressure is 3MPa, the pH is 12, and the flow rate is 5m 3 / h, the recovered alcohol can be distilled and recycled.

[0108] (4) Dilution: Dilute the concentrate with water. The Brix value of the dilution is 10%.

[0109] (5) Ultrafiltration and nanofiltration concentration: The diluted liquid was ultrafiltered through an ultrafiltration membrane. The membrane module model was NF01, the membrane specification was 20 KDa, and the inlet membrane pressure was maintained at 0.8 MPa, the temperature was 10 °C, the pH was 12, and the flow rate was 5 m / s. 3 / h, while continuously adding water for dialysis, retaining macromolecular substances with a molecular weight greater than 20KDa, until the Brix value of the permeate is 0.4%, and the permeate is collected. The permeate enters the series nanofiltration membrane for concentration. The nanofiltration membrane NF10, specification 600Da, can pass through substances with a molecular weight less than 400Da and retain substances with a molecular weight greater than 600Da. During operation, the pressure is 0.1MPa, the temperature is 25℃, and the pH is 10. The flow rate of the permeate of the ultrafiltration membrane is 5m3 / h, and the Brix value of the concentrated liquid is 27.5%.

[0110] (6) Resin column decolorization: The concentrated solution obtained in step (5) was dissolved in deionized water, the Brix value of the dissolved solution was 10%, and the dissolved solution was decolorized by a mixed resin of LX600J and DM100, the loading rate was 0.5 BV / h, the loading amount was 0.5 BV of the resin volume, the front effluent clear water was discarded, and the feed liquid was collected. After the injection was completed, the residual liquid was pushed out with 2.5 times the volume of the resin 2wt% saline, and the effluent and the water top liquid were collected together to obtain a decolorized solution.

[0111] (7) Macroporous resin adsorption: The decolorizing solution is injected into the macroporous adsorption resin AB-8 for adsorption at a rate of 2 BV / h. The sample volume is 1.6 BV of the resin volume. After the injection is completed, the residual liquid is washed with 2-3 BV of the resin volume of water, and the effluent and the washing liquid are discarded. 1 BV of the resin volume of 85% ethanol is used for elution and analysis at a rate of 1 BV / h. Then, 2 times the volume of the resin volume of water is used to top up the alcohol. The eluent and the water top liquid are combined to form the analysis solution.

[0112] (8) Concentration: The analytical solution is concentrated by membrane. The membrane module model is hollow fiber membrane NF95, specification 130Da, and ethanol is recovered until it is alcohol-free. When the Brix value of the concentrated solution is 25%, it is transferred to a single-effect external circulation concentrator for further concentration. The concentration vacuum is 0.095MPa and the concentration temperature is 58°C. The concentration is completed when the Brix value of the concentrated solution reaches 43%.

[0113] (9) Sterilization: Pasteurize the concentrate at 85°C for 35 min.

[0114] (10) Drying: The sterilized concentrate was filtered through a pipeline filter and then spray dried. The feed rate was adjusted to 50 L / h, the air inlet temperature was 182°C, and the air outlet temperature was 86°C. The product was collected. The final product had a high-purity camellia saponin yield of 10.8%, an extraction rate of 81.7%, and a camellia saponin content of 97.9%. The purity, protein content, polyphenol content, and absorbance of the high-purity camellia saponin product were tested respectively. The results are shown in Table 1.

[0115] Example 6

[0116] High-purity camellia saponins were extracted according to the method of Example 5, the only difference from Example 5 being that the aqueous ethanol used in step (2) contained sodium sulfite, sodium bisulfite and sodium metabisulfite, and the mass ratio of sodium sulfite, sodium bisulfite and sodium sulfite was 2:1:2.

[0117] The final yield of the high-purity camellia saponin product was 11.3%, the extraction rate was 86.9%, and the camellia saponin content was 99.7%. The purity, protein content, polyphenol content and absorbance of the high-purity camellia saponin product were tested respectively, and the results are shown in Table 1.

[0118] Comparative Example 1

[0119] High-purity camellia saponins were extracted according to the method of Example 3, the only difference from Example 3 being that the aqueous ethanol used in step (2) did not contain sodium sulfite, sodium bisulfite and sodium metabisulfite.

[0120] The final yield of the high-purity camellia saponin product was 10.2%, the extraction rate was 72.4%, and the camellia saponin content was 91.6%. The purity, protein content, polyphenol content and absorbance of the high-purity camellia saponin product were tested respectively, and the results are shown in Table 1.

[0121] Comparative Example 2

[0122] High-purity camellia saponins were extracted according to the method of Example 3, the only difference from Example 3 being that the aqueous ethanol used in step (2) did not contain sodium sulfite, sodium bisulfite and sodium metabisulfite, and was not processed through steps (5)-(6), and the diluted solution of step (4) was directly subjected to macroporous resin adsorption in step (7).

[0123] The final yield of the high-purity camellia saponin product was 9.5%, the extraction rate was 60.9%, and the camellia saponin content was 83.2%. The purity, protein content, polyphenol content and absorbance of the high-purity camellia saponin product were tested respectively, and the results are shown in Table 1.

[0124] Table 1 Performance parameters of camellia saponin products prepared in Examples and Comparative Examples

[0125]

[0126] It should be noted that, for the detection of polyphenol content, when the polyphenol content is lower than 0.05 g / kg, it cannot be detected, so the "not detected" in Table 1 means that the polyphenol content is lower than 0.05 g / kg.

[0127] It can be seen from Examples 1 to 5 that the extraction rate of camellia saponin from camellia seed cake using the method of the present invention is above 80%, and the purity of the camellia saponin product is high, above 97%. From the comparison between Example 5 and Example 6, it can be seen that when the camellia seed cake is extracted, the aqueous lower alcohol solution used contains a composite salt, and the camellia saponin finally prepared has a higher purity, lower protein and polyphenol contents, and lower absorbance than when only a single salt is used; from the comparison between Example 3 and Comparative Example 1, it can be seen that when the camellia seed cake is extracted, the aqueous lower alcohol solution used contains a composite salt, and the camellia saponin finally prepared has a higher purity, lower protein and polyphenol contents, and lower absorbance than when the camellia seed cake is not extracted; from the comparison between Example 3 and Comparative Example 2, it can be seen that when the camellia seed cake is extracted, the aqueous lower alcohol solution containing the composite salt is used to extract the camellia seed cake, and the operation of steps (5)-(6) of Example 3 is combined, and under the synergistic effect of the above conditions, the purity of the camellia saponin finally prepared is greatly improved, and the protein and polyphenol contents of the camellia saponin are lower, and the absorbance is lower.

[0128] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention is described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for extracting high-purity camellia saponin, characterized in that: The following steps are involved: crushing camellia oil seed cake or tea seed cake, performing dynamic countercurrent leaching on the crushed camellia oil seed cake or tea seed cake using an aqueous lower alcohol solution, and collecting a filtrate; Concentrating the filtrate to recover alcohol until the alcohol content in the concentrated solution is less than 3 wt %, to obtain a concentrated solution A; The concentrated solution A is diluted with water, the diluted solution is treated with an ultrafiltration membrane, and the ultrafiltration membrane permeate is concentrated with a nanofiltration membrane to obtain a concentrated solution B; Adding water to the concentrated solution B to obtain a dissolved solution, decolorizing the dissolved solution, adsorbing the decolorized solution with a macroporous adsorption resin, and then desorbing the solution with an ethanol solution to obtain a desorbed solution; The analytical solution is concentrated, sterilized, filtered and dried to obtain high-purity camellia saponin.

2. The extraction method according to claim 1, characterized in that The conditions for crushing the oil-tea seed cake or tea seed cake are: a feeding rate of 10-200 kg / h, a frequency of 30-50 Hz; the particle size of the oil-tea seed cake or tea seed cake after crushing is 0.1-2 mm; and / or, When the crushed camellia oil seed cake or tea seed cake is subjected to dynamic countercurrent extraction, the extraction temperature is 30-50° C., the extraction frequency is 30-50 Hz, and the extraction advancement speed is 5-10 m / h; and / or, The aqueous lower alcohol solution comprises lower alcohol, compound salt and water, wherein the mass concentration of the lower alcohol is 50-90%, and the mass concentration of the compound salt is 0.1-0.5%; the compound salt comprises one or more of sulfite, bisulfite or pyrosulfite.

3. The extraction method according to claim 1, characterized in that The method for concentrating the filtrate to recover alcohol is nanofiltration membrane concentration, wherein the specification of the nanofiltration membrane is 150-200Da, the operating pressure of the nanofiltration membrane is 0.5-3.0MPa, the temperature is 10-65°C, the pH is 3.0-12.0, and the flow rate of the filtrate is 0.5-5.0m 3 / h.

4. The extraction method according to claim 1, characterized in that The Brix value of the diluted solution after the concentrated solution A is diluted with water is 1-10%; and / or, When the diluent is treated by an ultrafiltration membrane, the specification of the ultrafiltration membrane is 15-20KDa, the operating inlet membrane is 0.1-1.0MPa, the temperature is 10-65°C, the pH is 3.0-12.0, the flow rate of the diluent is 0.5-5.0m3 / h, and the macromolecular substances with a molecular weight greater than 20KDa are retained. At the same time, water is continuously added for dialysis until the Brix value of the permeate is ≤0.5%.

5. The extraction method according to claim 1, characterized in that When the ultrafiltration membrane permeate is concentrated by the nanofiltration membrane, the specification of the nanofiltration membrane is 400-600Da, the operating pressure is 0.1-3.0MPa, the temperature is 10-65°C, the pH is 3.0-12.0, and the flow rate of the ultrafiltration membrane permeate is 0.5-5.0m3 / h, until the Brix value of the concentrated liquid is 25-30%, to obtain concentrated liquid B.

6. The extraction method according to claim 1, characterized in that The Brix value of the dissolving solution is 8-10%; and / or, The decolorization method of the solution is that the solution is adsorbed by a macroporous adsorption resin or decolorized by anion exchange resin; when the solution is adsorbed by a macroporous adsorption resin or decolorized by anion exchange resin, the loading speed of the solution is 0.5-1BV / h, and the loading amount is 0.5-5.0BV of the resin volume. After the decolorization by the macroporous adsorption resin or the anion exchange resin is completed, a decolorized solution is obtained, and / or, The macroporous adsorption resin is a polystyrene macroporous adsorption resin or an acrylate macroporous adsorption resin, and the anion exchange resin is a polystyrene skeleton or an acrylate skeleton anion exchange resin.

7. The extraction method according to claim 6, characterized in that When the decolorizing solution is adsorbed by the macroporous adsorption resin, the loading speed of the decolorizing solution is 1-2 BV / h, and the loading amount is 0.5-5.0 BV of the resin volume.

8. The extraction method according to claim 1, characterized in that During the analysis of the ethanol solution, the loading speed of the ethanol solution is 0.5-1 BV / h, and the loading amount is 1.0-5.0 BV of the resin volume.

9. The extraction method according to claim 1, characterized in that The method for concentrating the analytical solution is as follows: the analytical solution is subjected to membrane concentration, and when the solid content of the concentrated solution reaches 20-30%, it is transferred to a single-effect external circulation concentrator for further concentration until the Brix value reaches 40-50%; the specification of the membrane used for the membrane concentration is 100-200Da; when the single-effect external circulation concentrator continues to concentrate, the concentration vacuum degree is 0.05-0.095MPa, and the concentration temperature is 50-85°C; and / or, The sterilization temperature is 80-85°C and the sterilization time is 30-35 minutes; and / or, The drying method is spray drying, the feed rate of the spray drying is 10-100 L / h, the air inlet temperature is 180-190° C., and the air outlet temperature is 85-90° C.

10. A high-purity camellia saponin prepared by any extraction method according to any one of claims 1 to 9, characterized in that: The purity of the high-purity camellia saponin is above 97%, the protein content in the high-purity camellia saponin is ≤1.0wt%, the polyphenol content is ≤1.0g / kg, and the absorbance Abs at a wavelength of 420-610nm is less than 0.06.

Images