阅读说明：本技术 一种人参多肽的提取工艺及其用途 (Extraction process and application of ginseng polypeptide ) 是由 方令豪 于 2020-07-14 设计创作，主要内容包括：本发明公开了一种人参多肽的提取工艺及其用途,包括以下步骤：S1预处理；S2粉碎；S3萃取；S4分离；S5过滤；S6消毒。其中S5过滤使用的滤膜为采用特定方法制备的氧化铝载体连接共价有机骨架滤膜。通过调节共价有机骨架的几何结构,选择不同的构建单元来调节共价有机骨架孔径和孔的形状,制备得到最佳适合分离人参多肽的滤膜,实现人参多肽的高效分离提取。(The invention discloses an extraction process and application of ginseng polypeptide, which comprises the following steps: s1 preprocessing; s2 grinding; s3 extraction; s4 separating; s5 filtering; and S6 sterilizing. Wherein the filter membrane used for S5 filtration is an alumina carrier connected covalent organic framework filter membrane prepared by a specific method. The optimal filter membrane suitable for separating ginseng polypeptide is prepared by adjusting the geometric structure of the covalent organic framework and selecting different construction units to adjust the aperture and the pore shape of the covalent organic framework, so that the efficient separation and extraction of the ginseng polypeptide are realized.)

1. The extraction process of the ginseng polypeptide is characterized by comprising the following steps:

s1 preprocessing;

s2 grinding;

s3 extraction;

s4 separating;

s5 filtering;

and S6 sterilizing.

2. The extraction process of the ginseng polypeptide is characterized by comprising the following steps:

s1 pretreatment: drying the flowers, leaves and roots of the ginseng at the temperature of 100 ℃ and 110 ℃ for 0.5-1h to obtain a raw material A with the water content of 5-10%;

s2 crushing: crushing the raw material A to 50-80 meshes to obtain powder B; and (3) drying: drying the powder B at the temperature of 100-110 ℃ for 0.5-1.5h to obtain powder C with the water content of less than 5%;

s3 extraction: carrying out silica gel column chromatography on the powder C, wherein the extraction temperature is 15-30 ℃, the pressure is 3-8MPa, the extraction time is 1-2h, the flow rate of ethanol is 500-800kg/h, an entrainer ethyl acetate is also added during extraction, and the weight ratio of the entrainer to the powder C is (0.3-0.5): 100, respectively; obtaining liquid D and residue E after extraction, carrying out secondary extraction on the residue E under the conditions that the temperature is 20-30 ℃ and the pressure is 3-8MPa, wherein the extraction time is 1-2h, and the flow of carbon dioxide is 500-;

s4 separation: mixing the obtained extractive solution with liquid D, and hydrolyzing at 15-30 deg.C and 0.5-1.5MPa to obtain ginseng polypeptide, and recovering solvent F; heating the analyzed ginseng polypeptide to 60-80 deg.C, maintaining for 10-20 min, homogenizing in a homogenizer, and bottling or performing the next step;

s5 filtering: filtering the separated ginseng polypeptide by using a filtering membrane;

s6 disinfection: the filtered ginseng polypeptide flows through the irradiation of an ultraviolet lamp, and the irradiation dose is 200-²Time of irradiationIs 60-120 s.

3. The process for extracting ginseng polypeptide according to claim 2, wherein the filtration membrane is one of an ultrafiltration membrane with a cut-off molecular weight of 80-120kD, an ultrafiltration membrane with a cut-off molecular weight of 800-1200kD, and an alumina support linked to a covalent organic framework filtration membrane.

4. The extraction process of ginseng polypeptide as claimed in claim 3, wherein the preparation method of the alumina carrier linked covalent organic framework filter membrane comprises the following steps:

(1) modifying amino on the surface of the alumina carrier: soaking 3-5g of porous alumina tube in 10-25mL of 0.5-1.5mol/L hydrochloric acid solution at room temperature for 3-6h, washing with water to neutrality, drying, adding 20-40mg of 3-aminopropyltriethoxysilane into 20-40mL of toluene to prepare a solution, and treating the alumina tube with the solution at 110-120 ℃ in an argon atmosphere for 2-3h to obtain surface deposited amino alumina;

(2) synthesizing an alumina carrier connected covalent organic framework filter membrane: adding 1-2g of amino alumina tube and 20-40mg of aldehyde functional group construction unit into 20-30mL of dioxane to react at the temperature of 150 ℃ and 170 ℃ for 1-2h, washing and drying to obtain aldehyde-alumina tube, and preparing 20-40mL of dioxane, 40-60mg of aldehyde functional group construction unit, 40-60mg of amino functional group construction unit and 2-3mL of acetic acid aqueous solution of 1-3mol/L into a synthetic solution; vertically placing an aldehyde-alumina tube in a stainless steel autoclave lined with polytetrafluoroethylene, adding a synthetic solution, heating for 48-72h in an air blowing furnace at the temperature of 120-140 ℃, naturally cooling, washing a reaction product for multiple times by using 1, 4-dioxane and ethanol, and drying for 10-15h at the temperature of 120-140 ℃ to obtain the alumina carrier connected covalent organic framework filter membrane.

5. The process of claim 4, wherein the aldehyde functional group building block is trimesic aldehyde or 1,3, 5-tris (4-formylphenyl) benzene.

6. The process of claim 4, wherein the amino-functional building block is p-phenylenediamine or 4, 4 "-diamino-p-terphenyl.

7. A ginseng polypeptide extract prepared by the extraction process of any one of claims 1-6.

8. A composition product comprising the ginseng polypeptide extract according to claim 7, and a component selected from the group consisting of a pharmaceutical product, a food product, and a cosmetic raw material.

9. The composition product of claim 8, processed into orally and parenterally acceptable dosage forms, wherein the oral dosage forms are selected from tablet, sugar-coated tablet, film-coated tablet, enteric-coated tablet, capsule, hard capsule, soft capsule, decoction, oral liquid, buccal agent, granule, pill, powder, paste, and pellet; the injection is selected from lyophilized powder for injection, intravenous emulsion, injection, and infusion.

Technical Field

The invention belongs to the field of preparation of ginseng polypeptide, and particularly relates to an extraction process and application of ginseng polypeptide.

Background

Ginseng has a very special status in the medical history of China, is known as the king of Chinese herbal medicine, and is the representative of Chinese traditional medicine. Modern medical research proves that ginseng has good protective effect on the central nervous system, cardiovascular system, respiratory system, blood and hematopoietic system, endocrine system and reproductive system. In addition, researches prove that the ginseng also has remarkable anti-tumor, anti-radiation, anti-aging and detoxifying effects. In the last 30 years, with the development of chemical analysis methods, the separation, purification and structural identification of ginseng components have been greatly advanced. At present, ginseng roots are known to contain active ingredients such as ginsenoside, polysaccharide, volatile oil, fatty acid, sterol, vitamin, protein, polypeptide and the like.

The research on ginseng protein polypeptides began in the 60's of the 20 th century, but due to the limitations of separation techniques, analysis, sequencing and other means, the number of polypeptides isolated from ginseng and the determination of accurate amino acid sequences are very limited, so that the research on effective ingredients of ginseng is mainly focused on the research on ginsenosides and volatile ingredients, while the research on ginseng polypeptides is relatively less.

In recent years, with the development of disciplines such as molecular biology and biochemistry, research on bioactive peptides has been widely conducted. Various animal and plant polypeptides with different physiological activities have been extracted or developed into related products.

Membrane separation technology is an emerging technology for separating polypeptides, and a semipermeable membrane is used as a selective barrier layer to allow certain components to permeate and retain other components in a mixture, so that the separation purpose is achieved. The method has the advantages of simple equipment, convenient operation, no phase change, no chemical change, high treatment efficiency, energy saving, high extraction rate and the like. The filter membrane commonly used for separating the polypeptide in the prior art is an ultramicro filter membrane, the aperture is between 0.002-0.1 um, macromolecular substances of 1-20 nm can be intercepted, the filter membrane has universality for separating the polypeptide, and if the effect of high-efficiency separation can not be achieved aiming at specific polypeptide molecules.

The invention aims to overcome the technical prejudice, and utilizes the advantages of high porosity, regular pore channel structure and the like of Covalent Organic Frameworks (COFs) to prepare the filter membrane firmly attached to the surface of a carrier, and different construction units are selected to adjust the pore diameter and the pore shape of the COFs by adjusting the geometric structure of the COFs, so that the filter membrane which is best suitable for separating ginseng polypeptides is prepared, and the efficient separation and extraction of the ginseng polypeptides are realized.

Disclosure of Invention

The invention is based on the problem that the membrane separation technology is adopted in the field for extracting ginseng polypeptide, and the separation effect is not good by using an ultramicro filter membrane.

The invention aims to provide a ginseng polypeptide extraction process, wherein the filter membrane adopted in the process is an alumina carrier prepared by a specific method and is connected with a covalent organic framework filter membrane, the aperture and the shape of the COFs are adjusted by adjusting the geometrical structure of the COFs and selecting different construction units, the optimal filter membrane suitable for separating ginseng polypeptide is prepared, and the efficient separation and extraction of ginseng polypeptide is realized.

In order to solve the technical problems, the invention adopts the technical scheme that:

a process for extracting ginseng polypeptide comprises the following steps:

s1 preprocessing;

s2 grinding;

s3 extraction;

s4 separating;

s5 filtering;

and S6 sterilizing.

The invention also provides a technical scheme that:

a process for extracting ginseng polypeptide comprises the following steps:

s1 pretreatment: drying the flowers, leaves and roots of the ginseng at the temperature of 100 ℃ and 110 ℃ for 0.5-1h to obtain a raw material A with the water content of 5-10%;

s2 crushing: crushing the raw material A to 50-80 meshes to obtain powder B; and (3) drying: drying the powder B at the temperature of 100-110 ℃ for 0.5-1.5h to obtain powder C with the water content of less than 5%;

s3 extraction: carrying out silica gel column chromatography on the powder C, wherein the extraction temperature is 15-30 ℃, the pressure is 3-8MPa, the extraction time is 1-2h, the flow rate of ethanol is 500-800kg/h, an entrainer ethyl acetate is also added during extraction, and the weight ratio of the entrainer to the powder C is (0.3-0.5): 100, respectively; obtaining liquid D and residue E after extraction, carrying out secondary extraction on the residue E under the conditions that the temperature is 20-30 ℃ and the pressure is 3-8MPa, wherein the extraction time is 1-2h, and the flow of carbon dioxide is 500-;

s4 separation: mixing the obtained extractive solution with liquid D, and hydrolyzing at 15-30 deg.C and 0.5-1.5MPa to obtain ginseng polypeptide, and recovering solvent F; heating the analyzed ginseng polypeptide to 60-80 deg.C, maintaining for 10-20 min, homogenizing in a homogenizer, and bottling or performing the next step;

s5 filtering: filtering the separated ginseng polypeptide by using a filtering membrane;

s6 disinfection: the filtered ginseng polypeptide flows through the irradiation of an ultraviolet lamp, and the irradiation dose is 200-²The irradiation time is 60-120 s.

The filter membrane is one of an ultra-micro filter membrane with the cut-off molecular weight of 80-120kD, an ultra-micro filter membrane with the cut-off molecular weight of 800-1200kD and a filter membrane with an alumina carrier connected with a covalent organic framework.

The preparation method of the alumina carrier connected covalent organic framework filter membrane comprises the following steps:

(1) modifying amino on the surface of the alumina carrier: soaking 3-5g of porous alumina tube in 10-25mL of 0.5-1.5mol/L hydrochloric acid solution at room temperature for 3-6h, washing with water to neutrality, drying, adding 20-40mg of 3-aminopropyltriethoxysilane into 20-40mL of toluene to prepare a solution, and treating the alumina tube with the solution at 110-120 ℃ in an argon atmosphere for 2-3h to obtain surface deposited amino alumina;

(2) synthesizing an alumina carrier connected covalent organic framework filter membrane: adding 1-2g of amino alumina tube and 20-40mg of aldehyde functional group construction unit into 20-30mL of dioxane to react at the temperature of 150 ℃ and 170 ℃ for 1-2h, washing and drying to obtain aldehyde-alumina tube, and preparing 20-40mL of dioxane, 40-60mg of aldehyde functional group construction unit, 40-60mg of amino functional group construction unit and 2-3mL of acetic acid aqueous solution of 1-3mol/L into a synthetic solution; vertically placing an aldehyde-alumina tube in a stainless steel autoclave lined with polytetrafluoroethylene, adding a synthetic solution, heating for 48-72h in an air blowing furnace at the temperature of 120-140 ℃, naturally cooling, washing a reaction product for multiple times by using 1, 4-dioxane and ethanol, and drying for 10-15h at the temperature of 120-140 ℃ to obtain the alumina carrier connected covalent organic framework filter membrane.

The aldehyde functional group building unit is trimesic aldehyde or 1,3, 5-tri (4-formylphenyl) benzene.

Preferably, the aldehyde functional group building unit is trimesic aldehyde.

The amino functional group building unit is p-phenylenediamine or 4, 4' -diamino-p-terphenyl.

Preferably, the amino functional building block is p-phenylenediamine.

COFs have the characteristics of inherent porosity, high porosity, ordered pore structure, large specific surface area, good thermochemical stability, diverse functions and the like, and have attracted wide attention in the fields of gas storage, molecular separation, catalysis and the like in recent years. Few studies have reported on the preparation of COFs into filter membranes firmly attached to the surface of a carrier. The geometry of the COFs, such as the pore size and pore shape, can be adjusted by choosing different building blocks.

The invention also relates to a composition product, which comprises the ginseng polypeptide extract and components selected from medicines, foods and cosmetic raw materials.

Preferably, the composition product is processed into an acceptable dosage form for oral administration and injection according to requirements, and the oral preparation is selected from tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, capsules, hard capsules, soft capsules, decoction, oral liquid, buccal agents, granules, medicinal granules, pills, powder, paste and pellets; the injection is selected from lyophilized powder for injection, intravenous emulsion, injection, and infusion.

The invention also relates to a preparation method of the ginseng polypeptide extract combined medical injection, which comprises the following steps:

weighing 50-60g of arginine and 50-60g of water, uniformly stirring, adding 130g of ginseng polypeptide 120, heating to 40-60 ℃, stirring for dissolving, adjusting the pH value to 4-5 by using an aqueous solution containing 5-10% hydrochloric acid, adding activated carbon with the liquid preparation amount of 0.l-0.2% g/mL, stirring for 20-30 minutes, and carrying out rough filtration for removing carbon; filtering with 0.20-0.30 μm filter membrane to obtain fine filtrate; encapsulating, wherein the filling amount of each bottle is 2 mL; 110-125 ℃ hot-pressing sterilization for 10-20 minutes, inspection, packaging and warehousing.

The invention also relates to a preparation method of the ginseng polypeptide extract tablet, which comprises the following steps:

(1) mixing 30-50g of ginseng polypeptide, and uniformly stirring to obtain powdery solid;

(2) uniformly mixing the powder obtained in the step (1) with 50-60g of lactose, 2.0-3.0g of carboxymethyl starch and 4.0-5.0g of superfine silica gel powder, adding 60-70g of arginine to prepare a proper soft material, and granulating with a 20-30-mesh sieve;

(3) 5.0-6.0g of sodium carboxymethyl starch and 1.0-2.0g of aerosil are mixed with the dry granules obtained in the step (2) evenly, and tablets are obtained by tabletting.

The invention also relates to a method for preparing toning lotion by combining the ginseng polypeptide extract with standard raw materials of cosmetics, which comprises the following steps:

weighing 10-30g of ceramide and 5-15g of nicotinamide, adding 20-40g of water, uniformly stirring, adding 30-50g of ginseng polypeptide, heating to 40-60 ℃, stirring for dissolving, adjusting the pH value to 5.0-6.0 by using an aqueous solution containing 5-10% of EDTA, adding water to full amount, and uniformly stirring; then adding activated carbon with the liquid preparation amount of 0.l-0.2% g/mL, stirring for 20-40 minutes, and carrying out rough filtration to remove carbon; filtering with 0.20-0.30 μm filter membrane to obtain fine filtrate; encapsulating, wherein the filling amount of each bottle is 2 mL; 110-120 ℃ hot-pressing sterilization for 10-20 minutes, inspection, packaging and warehousing.

The invention has the beneficial effects that:

the filter membrane adopted by the process is an alumina carrier prepared by a specific method and is connected with a covalent organic framework filter membrane, and the pore size and the pore shape of COFs are adjusted by adjusting the geometrical structure of the COFs and selecting different construction units, so that the filter membrane which is best suitable for separating ginseng polypeptide is prepared, and the efficient separation and extraction of the ginseng polypeptide are realized.

The ginseng polypeptide extract with high activity and high purity, which is extracted by the invention, has the natural activity of the ginseng polypeptide and can be used as active bioactive macromolecules to activate the activity of human cells. Is extracted by a system close to the human body environment, and has good bioavailability. The ginseng polypeptide can improve the capability of breaking through the human epidermal barrier and greatly improve the biological permeability after hydrophilic treatment.

Detailed Description