Process for continuously extracting cherokee rose fruit polysaccharide fluid extract through tank group ultrasonic-assisted negative pressure cavitation dynamic countercurrent

Abstract

The invention discloses a process for continuously extracting a cherokee rose fruit polysaccharide fluid extract through tank group ultrasonic-assisted negative pressure cavitation dynamic countercurrent, which comprises the following steps: crushing a cherokee rose fruit medicinal material, degreasing, extracting with 80% ethanol, extracting by a tank group ultrasonic negative pressure cavitation dynamic countercurrent circulation method, concentrating under reduced pressure, deproteinizing, decolorizing, dialyzing, precipitating with alcohol, washing, redissolving with pure water and concentrating. The cherokee rose fruit polysaccharide fluid extract is obtained. According to the method, a tank group ultrasonic-assisted negative-pressure cavitation dynamic countercurrent extraction mode is adopted, tank group, ultrasonic and negative-pressure parameters are optimized, and factors such as extraction temperature, extraction time and material-liquid ratio of fructus rosae laevigatae medicinal materials are optimized, so that traditional Chinese medicine efficacy substances of fructus rosae laevigatae are extracted to the greatest extent, and the problems of long time consumption and loss of effective components in a conventional method are solved; the polysaccharide yield is high; by further controlling the degreasing treatment of the medicinal materials, the extraction process of 80% ethanol and the preparation of the decolorizing agent, the extraction rate and the degreasing rate of the polysaccharide can be effectively improved, and the decolorizing effect is improved.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of traditional Chinese medicine extraction, and particularly relates to a process for continuously extracting a gold cherry polysaccharide flow extract by means of tank group ultrasonic-assisted negative pressure cavitation dynamic countercurrent. BACKGROUND

[0002] The gold cherry, also known as Cherryl, gold pail, mountain stone pomegranate, gold pot, and lantern fruit, is the dried mature fruit of Cherryl Rosae, belonging to Rosaceae Rosa laevigata Michc and is a commonly used traditional Chinese medicinal material. It has the effects of consolidating essence, shrinking urine, arresting metrorrhagia and vaginal discharge, and astringing intestine and stopping diarrhea. It is used for treating spermatorrhea, leukorrhagia, enuresis, frequent micturition, metrorrhagia, and chronic diarrhea. The gold cherry flesh without the core is often used in clinical treatment. The gold cherry fruit contains various bioactive components, such as polysaccharides, organic acids, tannins, triterpenes, and vitamin C, among which polysaccharides account for a large proportion. As a natural active substance, the gold cherry polysaccharide exhibits various biological activities in life activities, including antioxidant, hypolipidemic, antibacterial, anti-inflammatory, antitumor, and immunomodulatory activities. There are various traditional extraction methods for natural plant polysaccharides, and different extraction methods will lead to differences in the structural properties and functional activities of polysaccharides. The main active substances in the gold cherry polysaccharide flow extract are prone to decomposition when the temperature is high and the preparation time is too long.

[0003] The negative pressure cavitation extraction technology is an extraction method developed by utilizing the action (cavitation effect, i.e. bubble breaking impact force) of bubbles in a gas-liquid-solid three-phase mixed system. The negative pressure cavitation effect produces negative pressure bubbles in the extraction process, and these bubbles and the material produce multiple effects in the liquid-solid system, including cavitation erosion effect, end motion effect, interface effect, and mixing effect. These effects can effectively promote the rapid mass transfer between the material and the extraction solvent, forming a dynamic mass transfer system. Among them, the cavitation erosion effect can effectively destroy the mechanical structure of the material and release the active ingredients inside the material. At present, ultrasonic-assisted negative pressure cavitation is also a common extraction technology.

[0004] The tank group dynamic countercurrent extraction process is a process in which two or more forced external circulation extraction tanks are connected in series, the extraction solvent is sequentially transported through each tank along the solute concentration gradient of the material in each tank in a reverse direction, and the material is kept in contact with the solvent for a certain extraction time and is repeatedly used under forced circulation. This process makes full use of the effective component concentration gradient between the solvent and the medicinal material, and gradually diffuses the effective components in the medicinal material to the solvent with a lower initial concentration, so that the effective components are maximally dissolved. This process has the advantages of low extraction temperature, high extraction efficiency, and less solvent consumption.

[0005] The combination of the ultrasonic-assisted negative pressure cavitation and the tank group dynamic countercurrent extraction method can integrate the advantages of the two methods and improve the polysaccharide extraction effect, but how to realize the organic combination of the two methods is a big difficulty in the prior art. In addition, in addition to polysaccharides, the Rosa roxburghii also contains saponins, which can also be extracted in water, and the saponins are natural surfactants, which can promote the mass transfer effect of the system when they exist in a certain content, but when their content exceeds a certain limit, a large amount of stable foam is easily generated under the action of strong ultrasonic and negative pressure cavitation, which affects the effective volume in the tank, hinders the mass transfer and affects the extraction effect.

[0006] In addition, the polysaccharide extraction liquid of the Rosa roxburghii generally contains various impurities (such as proteins, pigments and other small molecule substances), which causes certain difficulty in the separation, purification and structure characterization of the polysaccharides. Therefore, the polysaccharides extracted from the Rosa roxburghii need to be separated and purified, and the decolorization is an important process link. The commonly used decolorization methods for plant polysaccharides include the activated carbon method, the H2O2 method and the ion exchange resin method. The H2O2 has oxidizability and is easy to damage the biological activity of the polysaccharides, the use range of the ion exchange resin is relatively narrow and the cost is high, and the period is long. The activated carbon is a kind of green and environment-friendly adsorbent, which is non-toxic and odorless and can be reused. However, when the activated carbon is used for the decolorization treatment of the polysaccharides, there are problems of low decolorization rate, large loss of the polysaccharides and difficulty in completely removing the activated carbon residues.

[0007] In summary, how to provide a tank group ultrasonic-assisted negative pressure cavitation dynamic countercurrent continuous extraction process for polysaccharide fluid extract of Rosa roxburghii, organically combine the ultrasonic-assisted negative pressure cavitation and the tank group dynamic countercurrent extraction method and apply the two methods to the preparation of the polysaccharide fluid extract of the Rosa roxburghii to improve the extraction, separation and purification effect of the polysaccharides of the Rosa roxburghii is a problem to be solved. SUMMARY

[0008] The technical problem to be solved by the present application is to overcome the shortcomings of the prior art, provide a tank group ultrasonic-assisted negative pressure cavitation dynamic countercurrent continuous extraction process for polysaccharide fluid extract of Rosa roxburghii, adopt the tank group ultrasonic-assisted negative pressure cavitation dynamic countercurrent extraction method, optimize the extraction parameters, maximize the extraction of the medicinal material of the Rosa roxburghii, solve the problems of long time consumption and loss of effective components in the conventional method, and the polysaccharide yield is high; and through further control of the drug material degreasing treatment, the 80% ethanol extraction process and the preparation of the decolorizing agent, the polysaccharide extraction rate and the degreasing rate can be effectively improved, and the decolorization effect is improved.

[0009] The first object of the present application is to maximize the extraction of the medicinal material of the Rosa roxburghii by optimizing the tank group, the ultrasonic and the negative pressure parameters and the selection of the extraction temperature, the extraction time and the solid-liquid ratio of the Rosa roxburghii medicinal material. The difficulty of the present application lies in the organic combination of the ultrasonic-assisted negative pressure cavitation and the tank group dynamic countercurrent extraction method, which can reduce the energy consumption as much as possible and improve the polysaccharide extraction rate of the Rosa roxburghii.

[0010] To achieve the first object, the present application adopts the following technical solutions: A process for preparing a rosy cherry polysaccharide fluid extract by means of tank group ultrasonic-assisted negative pressure cavitation dynamic countercurrent continuous extraction, comprising the following steps: crushing rosy cherry medicinal materials (30-100 mesh), defatting treatment, 80% ethanol extraction, tank group ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction, reduced pressure concentration, deproteinization, decolorization, dialysis, alcohol precipitation, washing, pure water redissolution, and reduced pressure concentration, to obtain a rosy cherry polysaccharide fluid extract.

[0011] Preferably, the process parameters for the tank group ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction are as follows: The extraction solvent is pure water, the solid-liquid ratio is 1g:5-20ml (i.e. adding 5-20 times the volume of pure water), the extraction temperature is 80-90℃, the extraction times is 2-4, the extraction time for each time is 1-3h, the ultrasonic frequency is 30-40kHz, the ultrasonic power is 200-400W, the negative pressure intensity is -0.02Mpa to -0.05Mpa, and the stirring frequency in the extraction tank is 50-150r / min.

[0012] Preferably, the tank group process parameters are as follows: 2-4 extraction tanks are taken as a group, the medicinal materials in each tank are extracted in turn for 2-4 times, 10-20 times the volume of pure water is added to the first tank for the first time of extraction, 5-15 times the volume of pure water is added to the second to fourth times of extraction, respectively, the solvent added to each tank for the Nth time of extraction is the extraction liquid of the N+1th time of extraction of the previous tank, 5-15 times the volume of pure water is added to each tank for the last time of extraction, and 1-3 times the volume of pure water is added to each tank for the first time of extraction on the basis of the addition of the extraction liquid of the previous tank.

[0013] Preferably, the solvent used for the defatting treatment is any one of petroleum ether, cyclohexane, diethyl ether, and acetone, the solid-liquid ratio is 1g:6-12ml, the temperature is 40-60℃, the time is 1-6h, and the defatting times is 1-5.

[0014] Preferably, when 80% (volume fraction) ethanol is used for extraction, the solid-liquid ratio is 1g:8-12ml, the temperature is 80-90℃, the time is 1-2h, and the times is 1-3 (except for oligosaccharides).

[0015] Preferably, the pressure for the reduced pressure concentration is -0.01Mpa to -0.05Mpa, and the temperature is 50-70℃.

[0016] Preferably, the deproteinization step is: the concentrated solution obtained by concentration under reduced pressure is adjusted to pH 6, then 1-2% papain based on the mass of the concentrated solution is added, and degradation is carried out at 50-70 DEG C for 2-4 h; the supernatant is obtained by centrifugation, then 1 / 5 Sevage reagent (chloroform-n-butanol solution) based on the volume of the supernatant is added, oscillation is carried out for 20 min, the organic phase is removed by liquid separation, the supernatant is obtained by centrifugation, and the above operation is repeated for 4-6 times.

[0017] Preferably, the dialysis is carried out by using a dialysis bag with a molecular weight cut-off of 3500 Da, and specifically, tap water dialysis is carried out for 48-72 h, and then distilled water dialysis is carried out for 12-24 h.

[0018] Preferably, the alcohol precipitation step is: 4 times the volume of anhydrous ethanol is added for alcohol precipitation, and the precipitate is collected.

[0019] The second object of the present application is to reduce the influence of saponins on the extraction of Rosa laevigata Michx. polysaccharides under the dual action of ultrasonic and negative pressure cavitation by controlling the defatting rate, saponin content and other parameters in the defatting treatment and 80% ethanol extraction process.

[0020] To achieve the second object, the present application adopts the following technical solutions: The specific process of the defatting treatment and 80% ethanol extraction includes: The defatting treatment includes pre-defatting treatment and post-defatting treatment; the defatting condition is detected during the pre-defatting treatment, and the defatting is stopped when the defatting rate reaches 1.5-2.5%; after the solvent is dried, the medicinal material is treated by 80% ethanol extraction; the saponin content in the medicinal material after the 80% ethanol extraction treatment is 2-3%; and then the post-defatting treatment is carried out.

[0021] Preferably, the material-liquid ratio of the pre-defatting treatment is 1g:6-10ml, the temperature is 40-50 DEG C, and the number of times is 1; the material-liquid ratio of the post-defatting treatment is 1g:8-12ml, the temperature is 50-60 DEG C, the time is 1-3h, and the number of times is 1-4.

[0022] The third object of the present application is to overcome various problems existing when activated carbon is used for polysaccharide decolorization treatment, to carry out decolorization of Rosa laevigata Michx. polysaccharides by compounding activated carbon and attapulgite, and to improve the decolorization effect while reducing the loss of polysaccharides.

[0023] To achieve the third object, the present application adopts the following technical solutions: The decolorization method includes the following steps: activated carbon attapulgite decolorizing agent is added to the material solution obtained by deproteinization, and decolorization is carried out at 40-60 DEG C for 1-2h; and the addition amount of the activated carbon attapulgite decolorizing agent is 2-5% based on the mass of the material solution obtained by deproteinization.

[0024] The preparation method of the activated carbon attapulgite decolorizing agent includes: S1, respectively take attapulgite (attapulgite clay), activated carbon, add glacial acetic acid and stand for 12-24h, filter, wash with water until neutral, dry, grind through 140-300 mesh sieve, respectively get pretreated attapulgite, pretreated activated carbon; S2, dissolve chitosan in dilute acetic acid solution to prepare chitosan solution, mix pretreated activated carbon with starch and chitosan solution, stir uniformly, granulate (80-100 mesh), dry, get granules 1; S3, mix granules 1 and pretreated attapulgite uniformly, add oxygen-containing polymer, extrude into strips, cut and sieve (40-60 mesh), dry at 120-160℃ for 1-3h, then calcine at 400-600℃ for 1-3h under protective gas atmosphere (nitrogen, helium or argon), naturally cool, get activated carbon attapulgite decolorizing agent.

[0025] Preferably, in S1, the mass ratio of activated carbon to attapulgite is 1: (2-4), and the amount of glacial acetic acid added is 5-10 times the mass of activated carbon or attapulgite.

[0026] Preferably, in S1, the ground activated carbon is sieved through 140-200 mesh, and the ground attapulgite is sieved through 200-300 mesh.

[0027] Preferably, in S2, the mass concentration of dilute acetic acid solution is 0.6-1%, the concentration of chitosan in chitosan solution is 0.1-0.25g / L, the mass ratio of pretreated activated carbon to starch and chitosan solution is 1: (0.1-0.5): (2-4), and the mesh number of granulation is 80-100.

[0028] Preferably, in S3, the amount of oxygen-containing polymer added is 3-8% of the mass of pretreated attapulgite, and the oxygen-containing polymer is decaglycerin monooleate, polyethylene glycol, polyvinylpyrrolidone, polyethylene terephthalate or epoxy resin.

[0029] More preferably, the oxygen-containing polymer is decaglycerin monooleate.

[0030] Technical effects of the present application: 1, the present application combines ultrasonic wave assisted negative pressure cavitation and dynamic counterflow of filling group for the first time, which is a good innovation. The difficulty of the present application lies in the organic combination of the two technologies. The present application optimizes the parameters of tank group, ultrasonic wave and negative pressure, and optimizes the factors such as extraction temperature, extraction time and solid-liquid ratio of cherokee rose medicinal materials, so as to extract the medicinal material of cherokee rose with the least solvent, time and energy consumption, solve the problems of long time consumption and loss of effective components of conventional methods, and the extraction rate of polysaccharide is high. The present application also adopts defatting, 80% ethanol extraction to remove oligosaccharide, deproteinization, decolorization and dialysis to remove small molecules, so as to obtain high-purity cherokee rose polysaccharide extract.

[0031] 2. In addition to polysaccharides, Rosa laevigata also contains saponins, which can be extracted from water. Saponins are natural surfactants that can promote mass transfer in a certain amount. However, when their content exceeds a certain limit, they can easily generate a large amount of stable foam under strong ultrasonic and negative pressure cavitation, affecting the effective volume in the tank, hindering mass transfer, and affecting the extraction effect.

[0032] Therefore, in the process of removing oligosaccharides from Rosa laevigata by 80% ethanol extraction before extraction, this invention utilizes the certain solubility of saponins in 80% ethanol to simultaneously remove some saponins from the medicinal material. When the saponin content in the medicinal material is controlled to be 2-3% after 80% ethanol extraction, the extraction effect of Rosa laevigata polysaccharides is better.

[0033] Furthermore, due to the limited solubility of saponins in 80% ethanol, the saponin removal rate is low, making it difficult to achieve the required saponin content of 2-3% in Rosa laevigata. Therefore, this invention combines the 80% ethanol extraction process with degreasing treatment, introducing the role of oil. Specifically, during the degreasing process, degreasing is stopped when the degreasing rate reaches 1.5-2.5%, the solvent is evaporated, and then 80% ethanol extraction is performed. This allows the residual oil in the medicinal material to dissolve during the 80% ethanol extraction process, forming an emulsification effect with the solvent and saponin surfactants. This promotes the dissolution of saponins while also promoting the removal of oil. After the 80% ethanol extraction is completed, degreasing treatment is continued, which can further improve the degreasing rate.

[0034] 3. In order to overcome the various problems existing in the decolorization treatment of polysaccharides by activated carbon, this invention combines activated carbon with attapulgite to decolorize Rosa laevigata polysaccharides. Due to its unique microstructure, appearance and charged properties, attapulgite has excellent properties such as colloidal, adsorption and binding properties. The activated carbon-attapulgite decolorizing agent prepared by combining attapulgite and activated carbon has a good decolorization effect on Rosa laevigata polysaccharides, and the operation is simple and suitable for promotion.

[0035] Although the decolorization rate is improved when attapulgite and activated carbon are combined to form an activated carbon attapulgite decolorizing agent, the adsorption properties of activated carbon for polysaccharides still exist. Therefore, this invention does not simply mix activated carbon and attapulgite, but rather mixes activated carbon and chitosan into granules, and then coats the granules with attapulgite. The attapulgite on the outer layer of the decolorizing agent has low adsorption capacity for Rosa laevigata polysaccharides, which can effectively reduce the polysaccharide loss rate. Moreover, compared with directly mixing activated carbon and attapulgite powder, this method also helps to completely remove the decolorizing agent residue (granules are easier to remove than powder), achieving no decolorizing agent residue.

[0036] The application is that the oxygen-containing polymer is added into the attapulgite clay, and then high-temperature calcination is performed, which is helpful to improve the porosity of the attapulgite clay, thereby improving the pigment adsorption effect. The oxygen-containing polymer is preferably decaglycerin monooleate, which can also improve the coagulation of the outer attapulgite clay and the inner activated carbon particles, so that the decolorizing agent is not easy to be loose and cracked during calcination or decolorization, thereby maintaining the integrity of the decolorizing agent. BRIEF DESCRIPTION OF DRAWINGS

[0037] Figure 1 The process flow chart of the tank group ultrasonic auxiliary negative pressure cavitation dynamic countercurrent continuous extraction of the Malus asiatica polysaccharide liquid extract is provided. DETAILED DESCRIPTION

[0038] The above scheme is further described in combination with specific examples; it should be understood that these examples are used to illustrate the basic principles, main features and advantages of the application, and the application is not limited in scope by the following examples; the implementation conditions used in the examples can be further adjusted according to specific requirements, and the implementation conditions not specified are usually the conditions in conventional experiments.

[0039] In the following examples, all raw materials are obtained from commercial sources or prepared by conventional methods in the art, unless otherwise specified. Example 1

[0040] The present embodiment provides a process for tank group ultrasonic auxiliary negative pressure cavitation dynamic countercurrent continuous extraction of Malus asiatica polysaccharide liquid extract, and the process flow is shown in Figure 1 , which comprises the following steps: (1) Crushing of Malus asiatica medicinal materials (pulp): Malus asiatica medicinal materials are crushed to 60 mesh to obtain Malus asiatica medicinal material powder.

[0041] (2) Defatting treatment: The Malus asiatica medicinal material powder obtained in step (1) is taken, petroleum ether is added for reflux, the solid-liquid ratio is 1g:10ml, the temperature is 50℃, the time is 3h, the reflux defatting times are 2 times, the solvent is evaporated, and the defatted medicinal material is obtained.

[0042] (3) 80% ethanol extraction to remove oligosaccharides: 80% (volume fraction) ethanol solution is added to the defatted medicinal material for extraction, the solid-liquid ratio is 1g:10ml, the temperature is 85℃, the time is 1.5h, and the extraction times are 2 times.

[0043] (4) Tank group ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction: the medicinal material obtained in step (3) is divided into three parts and placed in three extraction tanks respectively, the extraction parameters and tank group process parameters are set, and then pure water is added for extraction to obtain the water extract.

[0044] The extraction parameters are as follows: the ratio of material to liquid is 1 g: 10-15 ml, the extraction temperature is 85°C, the extraction times are 3, the extraction time of each extraction is 2 h, the ultrasonic frequency is 35 kHz, the ultrasonic power is 300 W, the negative pressure intensity is -0.03 Mpa, and the stirring frequency in the extraction tank is 100 r / min.

[0045] The tank group process parameters are as follows: 3 extraction tanks are taken as a group, each tank of medicinal materials is extracted for 3 times in turn, 15 times the volume of pure water is added in the first tank for the first time of extraction, 10 times the volume of pure water is added in the first tank for the second and third times of extraction, the extraction liquid of the first tank for the second and third times of extraction is taken as the solvent for the first and second times of extraction of the second tank, respectively, the extraction liquid of the second tank for the second and third times of extraction is taken as the solvent for the first and second times of extraction of the third tank, respectively (i.e. except for the first tank, the solvent added in each tank for the Nth time of extraction is the extraction liquid of the (N+1)th time of extraction of the previous tank), 10 times the volume of pure water is added in the second and third tanks for the last time of extraction, and 2 times the volume of pure water is added in the second and third tanks for the first time of extraction on the basis of adding the extraction liquid of the previous tank, respectively.

[0046] (5) Reducing pressure concentration: the water extract obtained in step (4) is reduced pressure concentrated at 60°C and -0.03 Mpa, and is concentrated to 1 / 3 of the original volume to obtain a concentrated liquid.

[0047] (6) Deproteinization: the concentrated liquid obtained in step (5) is adjusted to pH 6, then 1.5% of papain by mass of the concentrated liquid is added, and is degraded at 60°C for 3 h, and the supernatant is taken by centrifugation, then 1 / 5 of Sevage reagent (chloroform-n-butanol solution) by volume of the supernatant is added, is shaken for 20 min, the organic phase is removed by liquid separation, the supernatant is taken by centrifugation, and the above operation is repeated for 5 times.

[0048] (7) Decolorization: activated carbon (160 mesh) is added to the liquid obtained in step (6), and is decolorized at 50°C for 1.5 h, and the amount of activated carbon added is 3% of the mass of the deproteinized liquid.

[0049] (8) Dialysis: a dialysis bag with a molecular weight cut-off of 3500 Da is adopted, and the dialysis is performed for 60 h by using tap water, and then the dialysis is performed for 24 h by using distilled water.

[0050] (9) Alcohol precipitation: 4 times the volume of anhydrous ethanol is added to the liquid obtained in step (8) to perform alcohol precipitation, and the precipitate is collected.

[0051] (10) Washing, pure water redissolution and reducing pressure concentration: the precipitate collected in step (9) is washed by anhydrous ethanol, acetone and diethyl ether in turn, is dissolved by adding pure water, and then is reduced pressure concentrated at 60°C and -0.03 Mpa to obtain a Rosa laevigata Michx. polysaccharide flow extract.

[0052] Example 2 On the basis of embodiment 1, the defatting treatment and 80% ethanol extraction process are optimized in this embodiment.

[0053] A kind of tank group ultrasonic auxiliary negative pressure cavitation dynamic counterflow continuous extraction process of Rosa laevigata Michx polysaccharide fluid extract, comprising the following steps: (1) Rosa laevigata Michx medicinal material is crushed: take Rosa laevigata Michx medicinal material, crush to 60 mesh, and obtain Rosa laevigata Michx powder.

[0054] (2) and (3) defatting treatment and 80% ethanol extraction: Take the Rosa laevigata Michx powder obtained in step (1), and add petroleum ether for defatting treatment, which includes pre-defatting treatment and post-defatting treatment.The material-liquid ratio of pre-defatting treatment is 1g:8ml, the temperature is 45℃, and the number of times is 1 time.The defatting condition is detected during pre-defatting treatment, and when the defatting rate reaches 1.5-2.5%, the defatting is stopped.After the solvent is dried, the medicinal material is treated by 80% ethanol extraction, the material-liquid ratio is 1g:10ml, the temperature is 85℃, the time is 1.5h, and the extraction number is 2 times.After 80% ethanol extraction treatment, the saponin content in the medicinal material is 2-3%, and then post-defatting treatment is carried out, the material-liquid ratio is 1g:10ml, the temperature is 55℃, the time is 2h, the number of times is 1 time, and the solvent is dried, to obtain the defatted and 80% ethanol extracted medicinal material.

[0055] (4) tank group ultrasonic negative pressure cavitation dynamic counterflow circulation method extraction: the medicinal material obtained in step (3) is divided into three parts and placed in three extraction tanks respectively, and then pure water is added for extraction after setting the extraction parameters and tank group process parameters to obtain water extract.

[0056] The extraction parameters are as follows: the material-liquid ratio is 1g:10-15ml, the extraction temperature is 85℃, the extraction number is 3 times, each extraction time is 2h, the ultrasonic frequency is 35kHz, the ultrasonic power is 300W, the negative pressure intensity is-0.03Mpa, and the stirring frequency in the extraction tank is 100r / min.

[0057] The tank group process parameters are as follows: three extraction tanks are taken as a group, the medicinal material in each tank is extracted for 3 times in turn, 15 times the volume of pure water is added in the first tank for the first time extraction, 10 times the volume of pure water is added for the second and third times, respectively, the extraction liquid of the first tank for the second and third times is used as the solvent for the first and second times extraction of the second tank, respectively, and the extraction liquid of the second tank for the second and third times is used as the solvent for the first and second times extraction of the third tank, respectively (i.e. except for the first tank, the solvent added in each tank for the Nth time extraction is the extraction liquid of the N+1th time extraction of the previous tank), and 10 times the volume of pure water is added as the solvent for the last time extraction of the second and third tanks, respectively, and 2 times the volume of pure water is added as the solvent for the first time extraction of the second and third tanks, respectively, on the basis of adding the extraction liquid of the previous tank.

[0058] (5)reduced pressure concentration: the water extract obtained in step (4) is concentrated under reduced pressure at 60°C and -0.03Mpa, and the concentration is stopped when the volume of the extract is 1 / 3 of the original volume, to obtain a concentrated solution.

[0059] (6) deproteinization: the concentrated solution obtained in step (5) is adjusted to pH 6, then 1.5% of papain by mass of the concentrated solution is added, and the solution is degraded at 60°C for 3h. The supernatant is obtained by centrifugation, then 1 / 5 of Sevage reagent (chloroform-n-butanol solution) by volume of the supernatant is added, and the solution is shaken for 20min. The organic phase is removed by separation, and the supernatant is obtained by centrifugation. The above operation is repeated 5 times.

[0060] (7) decolorization: activated carbon (160 mesh) is added to the solution obtained in step (6), and the solution is decolorized at 50°C for 1.5h. The amount of activated carbon added is 3% of the mass of the solution obtained by deproteinization.

[0061] (8) dialysis: a dialysis bag with a molecular weight cut-off of 3500 Da is used, and tap water dialysis is performed for 60h, and then distilled water dialysis is performed for 24h.

[0062] (9) alcohol precipitation: 4 times the volume of anhydrous ethanol is added to the solution obtained in step (8) for alcohol precipitation, and the precipitate is collected.

[0063] (10) washing, pure water redissolution and reduced pressure concentration: the precipitate collected in step (9) is washed with anhydrous ethanol, acetone and diethyl ether in sequence, dissolved in pure water, and then concentrated under reduced pressure at 60°C and -0.03Mpa to obtain a Rosa laevigata Michx. polysaccharide extract.

[0064] Example 3 On the basis of Example 2, the decolorization process is optimized in this example.

[0065] A process for continuously extracting a Rosa laevigata Michx. polysaccharide extract by ultrasonic-assisted negative pressure cavitation dynamic countercurrent extraction in a tank group, comprising the following steps: (1) crushing Rosa laevigata Michx. medicinal materials: Rosa laevigata Michx. medicinal materials are crushed to 60 mesh to obtain a Rosa laevigata Michx. powder.

[0066] (2) and (3) defatting treatment and 80% ethanol extraction: The Rosa laevigata Michx. medicinal material powder obtained in step (1) is defatted by adding petroleum ether, and the defatting treatment comprises pre-defatting treatment and post-defatting treatment. The pre-defatting treatment is carried out at a material-to-liquid ratio of 1 g:8 ml and a temperature of 45 DEG C for 1 time. The defatting condition is detected during the pre-defatting treatment, and the defatting is stopped when the defatting rate reaches 1.5-2.5%. After the solvent is evaporated, the medicinal material is treated by 80% ethanol extraction. The extraction treatment is carried out at a material-to-liquid ratio of 1 g:10 ml and a temperature of 85 DEG C for 1.5 h for 2 times. The saponin content in the medicinal material after the 80% ethanol extraction treatment is 2-3%, and then the post-defatting treatment is carried out. The post-defatting treatment is carried out at a material-to-liquid ratio of 1 g:10 ml and a temperature of 55 DEG C for 2 h for 1 time. After the solvent is evaporated, the defatted and 80% ethanol-extracted medicinal material is obtained.

[0067] (4) Tank group assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction: the medicinal material obtained in step (3) is evenly divided into three parts and placed in three extraction tanks respectively. After setting the extraction parameters and tank group process parameters, pure water is added for extraction to obtain water extract.

[0068] The extraction parameters are as follows: the material-to-liquid ratio is 1 g:10-15 ml, the extraction temperature is 85 DEG C, the extraction times are 3, the extraction time of each time is 2 h, the ultrasonic frequency is 35 kHz, the ultrasonic power is 300 W, the negative pressure intensity is-0.03 Mpa, and the stirring frequency in the extraction tank is 100 r / min.

[0069] The tank group process parameters are as follows: three extraction tanks are taken as a group, and the medicinal material in each tank is extracted in turn for 3 times. 15 times the volume of pure water is added in the first tank for the first time of extraction, and 10 times the volume of pure water is added in the first tank for the second and third times of extraction respectively. The extraction liquid of the first tank for the second and third times of extraction is used as the solvent for the first and second times of extraction of the second tank respectively, and the extraction liquid of the second tank for the second and third times of extraction is used as the solvent for the first and second times of extraction of the third tank respectively (i.e. except for the first tank, the solvent added in each tank for the Nth time of extraction is the extraction liquid of the (N+1)th time of extraction of the previous tank). 10 times the volume of pure water is added in the second and third tanks for the last time of extraction. On the basis of adding the extraction liquid of the previous tank, 2 times the volume of pure water is added in the second and third tanks for the first time of extraction respectively.

[0070] (5) Reducing pressure concentration: the water extract obtained in step (4) is concentrated under reducing pressure at 60 DEG C and-0.03 Mpa, and the concentration is carried out until the volume is 1 / 3 of the original volume to obtain a concentrated liquid.

[0071] (6) Deproteinization: the concentrated liquid obtained in step (5) is adjusted to pH 6, then 1.5% of the mass of the concentrated liquid of papain is added, and is degraded at 60 DEG C for 3 h. The supernatant is obtained by centrifugation, then 1 / 5 of the volume of the supernatant of Sevage reagent (chloroform-n-butanol solution) is added, oscillation is carried out for 20 min, the organic phase is removed by liquid separation, the supernatant is obtained by centrifugation, and the above operation is repeated for 5 times.

[0072] (7) decolorization: add activated carbon attapulgite decoloring agent to the solution obtained in step (6), decolor at 50℃ for 1.5h, the amount of activated carbon attapulgite decoloring agent added is 3% of the mass of the solution obtained by deproteinization.

[0073] The preparation method of the activated carbon attapulgite decoloring agent comprises: S1, take attapulgite (attapulgite clay) and activated carbon according to a mass ratio of 1:3 respectively, add glacial acetic acid (the amount of glacial acetic acid added is 7 times the mass of activated carbon or attapulgite), stand for 12h, filter, wash with water until neutral, dry, grind through 160 mesh and 250 mesh sieves respectively, and obtain pretreated attapulgite and pretreated activated carbon respectively; S2, dissolve chitosan in 0.8% (mass concentration) dilute acetic acid solution to prepare a 0.15g / L chitosan solution, mix the pretreated activated carbon with starch and the chitosan solution according to a mass ratio of 1:0.3:3, stir uniformly, granulate (80-100 mesh), dry, and obtain granules 1; S3, mix the granules 1 and the pretreated attapulgite uniformly, add decaglycerol monooleate (the amount added is 5% of the mass of the pretreated attapulgite), extrude into strips, cut and sieve (40-60 mesh), dry at 140℃ for 2h, then calcine at 500℃ for 2h under a helium atmosphere, and naturally cool, to obtain the activated carbon attapulgite decoloring agent.

[0074] (8) dialysis: use a dialysis bag with a molecular weight cutoff of 3500 Da, specifically dialyze for 60h with tap water, and then dialyze for 24h with distilled water.

[0075] (9) alcohol precipitation: add 4 times the volume of anhydrous ethanol to the solution obtained in step (8) for alcohol precipitation, and collect the precipitate.

[0076] (10) washing, pure water redissolution and reduced pressure concentration: wash the precipitate collected in step (9) with anhydrous ethanol, acetone and diethyl ether in sequence, dissolve with pure water, and then concentrate under reduced pressure at 60℃ and -0.03Mpa, to obtain the Rosa laevigata Michx. polysaccharide flow extract.

[0077] Example 4 The difference between this example and example 3 is that decaglycerol monooleate is replaced by polyethylene glycol PEG-4000.

[0078] Example 5 The difference between this example and example 3 is that decaglycerol monooleate is replaced by polyvinylpyrrolidone K90.

[0079] Example 6 This example provides a process for continuously extracting Rosa laevigata Michx. polysaccharide flow extract by tank group ultrasonic-assisted negative pressure cavitation dynamic countercurrent, which comprises the following steps: (1) The Rosa laevigata Michx. medicinal material is crushed to 40 mesh to obtain Rosa laevigata Michx. powder.

[0080] (2) and (3) Defatting treatment and 80% ethanol extraction: The Rosa laevigata Michx. powder obtained in step (1) is defatted by adding petroleum ether. The defatting treatment includes pre-defatting treatment and post-defatting treatment. The material-liquid ratio of the pre-defatting treatment is 1 g:6 ml, the temperature is 40°C, and the pre-defatting treatment is performed once. The defatting is stopped when the defatting rate reaches 1.5-2.5%. After the solvent is evaporated, the medicinal material is treated by 80% ethanol extraction. The material-liquid ratio of the extraction treatment is 1 g:8 ml, the temperature is 80°C, the extraction time is 2 h, and the extraction is performed twice. The saponin content in the medicinal material after the 80% ethanol extraction treatment is 2-3%. Then, the post-defatting treatment is performed. The material-liquid ratio of the post-defatting treatment is 1 g:8 ml, the temperature is 50°C, the time is 3 h, and the post-defatting treatment is performed once. After the solvent is evaporated, the defatted and 80% ethanol extracted medicinal material is obtained.

[0081] (4) Extraction by tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method: the medicinal material obtained in steps (2) and (3) is evenly divided into three parts and placed in three extraction tanks, respectively. After setting the extraction parameters and tank group process parameters, pure water is added for extraction. The extraction liquids in the tanks are combined to obtain the Rosa laevigata Michx. water extract.

[0082] The extraction parameters are as follows: the material-liquid ratio is 1 g:5-10 ml, the extraction temperature is 80°C, the extraction is performed three times, each time for 3 h, the ultrasonic frequency is 30 kHz, the ultrasonic power is 200 W, the negative pressure intensity is-0.05 Mpa, and the stirring frequency in the extraction tank is 50 r / min.

[0083] The tank group process parameters are as follows: three extraction tanks are taken as one group, and the medicinal material in each tank is extracted in turn for three times. Ten times the volume of pure water is added in the first tank for the first extraction, and five times the volume of pure water is added in the second and third extractions, respectively. The extraction liquids of the first tank in the second and third extractions are used as the solvents for the first and second extractions of the second tank, respectively. The extraction liquids of the second tank in the second and third extractions are used as the solvents for the first and second extractions of the third tank, respectively (i.e. except for the first tank, the solvent added in each tank for the Nth extraction is the extraction liquid of the (N+1)th extraction of the previous tank). Five times the volume of pure water is added in the last extraction of the second and third tanks. One times the volume of pure water is added in the first extraction of the second and third tanks based on the addition of the extraction liquid of the previous tank.

[0084] (5) Vacuum concentration: the Rosa laevigata Michx. water extract obtained in step (4) is concentrated under vacuum at 50°C and-0.05 Mpa. The concentration is performed until the volume is 1 / 3 of the original volume, and the concentrated liquid is obtained.

[0085] (6) Deproteinization: the concentrated solution obtained in step (5) was adjusted to pH 6, then 1% of the mass of the concentrated solution of papain was added, and degraded at 50℃ for 4h. The supernatant was obtained by centrifugation, then 1 / 5 of the volume of the supernatant of Sevage reagent (chloroform-n-butanol solution) was added, oscillated for 20min, and the organic phase was removed by liquid separation. The supernatant was obtained by centrifugation, and the above operation was repeated 4 times.

[0086] (7) Decolorization: active carbon attapulgite decolorizing agent was added to the deproteinized solution obtained in step (6), and decolorized at 40℃ for 2h. The addition amount of the active carbon attapulgite decolorizing agent was 2% of the mass of the deproteinized solution.

[0087] The preparation method of the active carbon attapulgite decolorizing agent comprises: S1, according to the mass ratio of 1:2, respectively take attapulgite, active carbon, add glacial acetic acid (the addition amount of glacial acetic acid is 5 times of the mass of active carbon or attapulgite), stand for 24h, filter, wash to neutral, dry, grind through 140 mesh screen and 200 mesh screen respectively, and get pretreated attapulgite and pretreated active carbon respectively; S2, dissolve chitosan in 0.6% (mass concentration) dilute acetic acid solution to prepare 0.1g / L chitosan solution, mix pretreated active carbon, starch and chitosan solution according to the mass ratio of 1:0.1:2, stir uniformly, granulate (80-100 mesh), dry, and get granules 1; S3, mix granules 1 and pretreated attapulgite uniformly, add decaglycerol monooleate (the addition amount is 3% of the mass of pretreated attapulgite), extrude into strips, cut and sieve (40-60 mesh), dry at 120℃ for 3h, then calcine at 400℃ for 3h under helium atmosphere, and naturally cool, and the active carbon attapulgite decolorizing agent is obtained.

[0088] (8) Dialysis: a dialysis bag with a molecular weight cut-off of 3500 Da was used, and tap water dialysis was performed for 48h, and then distilled water dialysis was performed for 24h.

[0089] (9) Alcohol precipitation: 4 times the volume of anhydrous ethanol was added to the solution obtained in step (8) for alcohol precipitation, and the precipitate was collected.

[0090] (10) Washing, pure water redissolution and reduced pressure concentration: the precipitate collected in step (9) was washed with anhydrous ethanol, acetone and diethyl ether in sequence, dissolved in pure water, and then concentrated under reduced pressure at 50℃ and-0.05Mpa, and the Rosa laevigata Michx polysaccharide flow extract was obtained.

[0091] Example 7 The present embodiment provides a process for continuously extracting Rosa laevigata Michx polysaccharide flow extract by tank group ultrasonic-assisted negative pressure cavitation dynamic countercurrent, which comprises the following steps: (1) Rosa laevigata Michx medicinal material crushing: take Rosa laevigata Michx medicinal material, crush to 80 mesh, and get Rosa laevigata Michx medicinal material powder.

[0092] (2), (3) defatting treatment, 80% ethanol extraction: Take the Rosa laevigata Michx. medicinal material powder obtained in step (1) and add petroleum ether for defatting treatment. The defatting treatment includes pre-defatting treatment and post-defatting treatment. The material-to-liquid ratio of the pre-defatting treatment is 1 g:10 ml, the temperature is 50°C, and the number of times is 1. The defatting condition is detected during the pre-defatting treatment, and when the defatting rate reaches 1.5-2.5%, the defatting is stopped. After the solvent is dried, the medicinal material is treated by 80% ethanol extraction. The material-to-liquid ratio of the extraction treatment is 1 g:12 ml, the temperature is 90°C, the time is 1 h, and the number of extraction times is 2. After the 80% ethanol extraction treatment, the saponin content in the medicinal material is 2-3%, and then the post-defatting treatment is performed. The material-to-liquid ratio of the post-defatting treatment is 1 g:12 ml, the temperature is 60°C, the time is 1 h, and the number of times is 1. The solvent is dried to obtain the medicinal material after defatting and 80% ethanol extraction.

[0093] (4) extraction by tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method: the medicinal material obtained in steps (2) and (3) is evenly divided into three parts and placed in three extraction tanks, respectively. After setting the extraction parameters and tank group process parameters, pure water is added for extraction. The extraction liquids in the tanks are combined to obtain the Rosa laevigata Michx. water extract.

[0094] The extraction parameters are as follows: the material-to-liquid ratio is 1 g:15-20 ml, the extraction temperature is 90°C, the number of extraction times is 3, the extraction time is 1 h each time, the ultrasonic frequency is 40 kHz, the ultrasonic power is 400 W, the negative pressure intensity is-0.02 Mpa, and the stirring frequency in the extraction tank is 150 r / min.

[0095] The tank group process parameters are as follows: three extraction tanks are taken as one group, and the medicinal material in each tank is extracted in turn for 3 times. 20 times the volume of pure water is added in the first tank for the first extraction, and 15 times the volume of pure water is added for the second and third extractions, respectively. The extraction liquids of the first tank for the second and third extractions are used as the solvents for the first and second extractions of the second tank, respectively. The extraction liquids of the second tank for the second and third extractions are used as the solvents for the first and second extractions of the third tank, respectively (i.e., except for the first tank, the solvent added for the Nth extraction in each tank is the extraction liquid of the N+1th extraction in the previous tank). The solvent added for the last extraction of the second and third tanks is 15 times the volume of pure water. For the first extraction of the second and third tanks, 3 times the volume of pure water is supplemented based on the addition of the extraction liquid of the previous tank.

[0096] (5) reduced pressure concentration: the Rosa laevigata Michx. water extract obtained in step (4) is reduced pressure concentrated at 70°C and-0.01 Mpa. The concentration is performed until the volume is 1 / 3 of the original volume, and the concentrated liquid is obtained.

[0097] (6) Deproteinization: Adjust the pH of the concentrate obtained in step (5) to 6, then add 2% papain by mass of the concentrate, degrade at 70°C for 2 hours, centrifuge to collect the supernatant, then add 1 / 5 of the volume of the supernatant to the Sevage reagent (chloroform-n-butanol solution), shake for 20 minutes, separate the organic phase, centrifuge to collect the supernatant, and repeat the above operation 6 times.

[0098] (7) Decolorization: Add activated carbon attapulgite decolorizing agent to the liquid obtained from deproteinization in step (6) and decolorize at 60°C for 1 hour. The amount of activated carbon attapulgite decolorizing agent added is 5% of the mass of the liquid obtained from deproteinization.

[0099] The preparation methods of activated carbon attapulgite decolorizing agent include: S1. Take attapulgite and activated carbon at a mass ratio of 1:4, add glacial acetic acid (the amount of glacial acetic acid added is 10 times the mass of activated carbon or attapulgite), let stand for 12 hours, filter, wash with water until neutral, dry, grind and pass through a 200-mesh sieve and a 300-mesh sieve respectively to obtain pretreated attapulgite and pretreated activated carbon respectively. S2. Dissolve chitosan in a 1% (mass concentration) dilute acetic acid solution to prepare a 0.25 g / L chitosan solution. Mix the pretreated activated carbon, starch, and chitosan solution at a mass ratio of 1:0.5:4. After stirring evenly, granulate (80-100 mesh) and dry to obtain granules 1. S3. Mix granules 1 and pretreated attapulgite evenly, add decaglycerol monooleate (8% of the mass of pretreated attapulgite), extrude into strips, cut into pieces and sieve (40-60 mesh), dry at 160℃ for 1 hour, then calcine at 600℃ for 1 hour under helium atmosphere, and cool naturally to obtain activated carbon attapulgite decolorizing agent.

[0100] (8) Dialysis: A dialysis bag with a molecular weight cutoff of 3500 Da was used. Specifically, tap water was used for dialysis for 72 hours, followed by distilled water for 24 hours.

[0101] (9) Alcohol precipitation: Add 4 times the volume of anhydrous ethanol to the liquid obtained in step (8) for alcohol precipitation and collect the precipitate.

[0102] (10) Washing, reconstitution with pure water and concentration under reduced pressure: The precipitate collected in step (9) was washed sequentially with anhydrous ethanol, acetone and diethyl ether, dissolved in pure water, and then concentrated under reduced pressure at 70℃ and -0.02Mpa to obtain Rosa laevigata polysaccharide fluid extract.

[0103] Comparative Example 1 The difference between this comparative example and Example 1 is that the ultrasonic power extracted by the ultrasonic negative pressure cavitation dynamic countercurrent circulation method in the tank assembly is 100W.

[0104] Comparative Example 2 The difference between the present comparative example and Example 1 is that the ultrasonic power in the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction is 500 W.

[0105] Comparative Example 3 The difference between the present comparative example and Example 1 is that the negative pressure intensity in the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction is -0.005 Mpa.

[0106] Comparative Example 4 The difference between the present comparative example and Example 1 is that the negative pressure intensity in the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction is -0.07 Mpa.

[0107] Comparative Example 5 The difference between the present comparative example and Example 1 is that the temperature in the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction is 70℃.

[0108] Comparative Example 6 The difference between the present comparative example and Example 1 is that the temperature in the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction is 90℃.

[0109] Comparative Example 7 The difference between the present comparative example and Example 1 is that the solid-liquid ratio in the first tank extraction and the third extraction of the second and third tanks in the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction is 1 g:3 ml (i.e. adding 3 times the volume of pure water).

[0110] Comparative Example 8 The difference between the present comparative example and Example 1 is that the solid-liquid ratio in the first tank extraction and the third extraction of the second and third tanks in the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction is 1 g:25 ml (i.e. adding 25 times the volume of pure water).

[0111] Comparative Example 9 The difference between the present comparative example and Example 1 is that the extraction time in the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method is 0.5 h.

[0112] Comparative Example 10 The difference between the present comparative example and Example 1 is that the extraction time in the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method is 4 h.

[0113] Comparative Example 11 The difference between the present comparative example and Example 2 is that the pre-degreasing treatment is stopped when the degreasing rate reaches 0.5-1% during the degreasing treatment and 80% ethanol extraction in steps (2) and (3).

[0114] Comparative Example 12 The difference between the present comparative example and Example 2 is that the defatting treatment in steps (2), (3) and 80% ethanol extraction is stopped when the pre-defatting treatment reaches a defatting rate of 3.0-3.5%.

[0115] Comparative Example 13 The difference between the present comparative example and Example 2 is that the defatting treatment in steps (2), (3) and 80% ethanol extraction is stopped when the pre-defatting treatment reaches a defatting rate of 3.0-3.5%.

[0116] Comparative Example 14 The difference between the present comparative example and Example 2 is that the defatting treatment in steps (2), (3) and 80% ethanol extraction is stopped when the pre-defatting treatment reaches a defatting rate of 3.0-3.5%.

[0117] Comparative Example 15 The difference between the present comparative example and Example 2 is that the defatting treatment in steps (2), (3) and 80% ethanol extraction is stopped when the pre-defatting treatment reaches a defatting rate of 3.0-3.5%.

[0118] Comparative Example 16 The difference between the present comparative example and Example 2 is that the defatting treatment in steps (2), (3) and 80% ethanol extraction is stopped when the pre-defatting treatment reaches a defatting rate of 3.0-3.5%.

[0119] Comparative Example 17 The difference between the present comparative example and Example 3 is that the preparation method of the activated carbon-attapulgite decoloring agent comprises: Attapulgite (attapulgite clay) and activated carbon are taken in a mass ratio of 1:3, acetic acid (the amount of acetic acid added is 7 times the mass of activated carbon or attapulgite) is added, and the mixture is allowed to stand for 12 h. After filtration, the mixture is washed with water until neutral, dried, ground through a 160-mesh sieve and a 250-mesh sieve, respectively, and mixed uniformly. The mixture is extruded into strips, cut into pieces, sieved (40-60 mesh), dried at 140°C for 2 h, and then calcined at 500°C for 2 h under a helium atmosphere. After natural cooling, the activated carbon-attapulgite decoloring agent is obtained.

[0120] Comparative Example 18 The difference between the present comparative example and Example 3 is that in the preparation method of the activated carbon-attapulgite decoloring agent, the positions of activated carbon and attapulgite are exchanged, i.e., attapulgite is used for granulation in step S2, and pretreated activated carbon is mixed with the granules 1 in step S3.

[0121] Comparative Example 19 The difference between the present comparative example and Example 3 is that in the preparation method of the activated carbon-attapulgite decoloring agent, no chitosan solution is added in step S2.

[0122] Comparative Example 20 The difference between the present comparative example and Example 3 is that no decaglycerol monooleate is added in step S3 in the preparation method of the activated carbon attapulgite decoloring agent.

[0123] I. Basic extraction parameter optimization test The Chamaemelum polysaccharide extract was extracted according to the method of Example 1, Comparative Examples 1-10, and the parameters of tank group, ultrasonic, negative pressure, and the factors of extraction temperature, extraction time, and solid-liquid ratio were optimized, and the polysaccharide extraction rate results are shown in Table 1 below. Among them, the polysaccharide extraction rate (%) = 100% x the mass of the extracted Chamaemelum polysaccharide / theoretical polysaccharide mass in Chamaemelum medicinal materials.

[0124] Table 1

[0125] As can be seen from Table 1, compared with Comparative Examples 1-10, the process parameters of Example 1 of the present application are more optimal, and a Chamaemelum polysaccharide extraction rate of 90.3% can be obtained.

[0126] II. Influence of saponin content control under the condition of the more optimal extraction process parameters The Chamaemelum polysaccharide extract was extracted according to the method of Example 1-3, Example 6-7, and Comparative Examples 11-16, the degreasing rate was measured after degreasing treatment, the saponin content in the medicinal materials was measured before Chamaemelum extraction after degreasing and extraction, and the polysaccharide extraction rate was calculated, and the results are shown in Table 2 below.

[0127] The degreasing rate (%) = 100% x (the mass of the medicinal materials before degreasing-the mass of the medicinal materials after degreasing) / the mass of the medicinal materials before degreasing.

[0128] Table 2

[0129] As can be seen from Table 2, compared with Example 1, after optimization of the degreasing treatment and 80% ethanol extraction process in Example 2-3 and Example 6-7, the total degreasing rate is 4.4-4.7%, and the Chamaemelum polysaccharide extraction rate is 97.0-97.6%. Example 2-3 and Example 6-7 can control the saponin content in the medicinal materials before extraction to be in the range of 2-3%, and the results show that the Chamaemelum polysaccharide extraction rate is higher under this saponin content range, so it is critical to control the saponin content in the medicinal materials before extraction.

[0130] Compared with Example 2, the degreasing treatment and 80% ethanol extraction process are changed in Comparative Examples 11-16, and the results show that the total degreasing rate and the Chamaemelum polysaccharide extraction rate are both reduced.

[0131] III. Decolorization effect of the present application 1. Decolorization rate and decolorizing agent residue rate.

[0132] The polysaccharide extract of Rosa laevigata was extracted according to the methods of Examples 2-3, 6-7 and Comparative Examples 17-20. After decolorization, the decolorization rate and the residual rate of the decolorizing agent were measured. The polysaccharide content of Rosa laevigata in the medicinal solution before and after decolorization was measured (phenol-sulfuric acid method). The polysaccharide loss rate was calculated. The results are shown in Table 3 below.

[0133] Table 3

[0134] As shown in Table 3, compared with Example 2, Examples 3 and 6-7 of the present invention optimized the decolorization method and prepared activated carbon attapulgite decolorizing agent for decolorization. The decolorization rate was as high as 94.2-94.6%, the residual rate of decolorizing agent was as low as 0.05-0.08%, and the polysaccharide loss rate was as low as 1.5-1.8%, showing a better decolorization effect.

[0135] Compared with Example 3, Comparative Examples 17-20 changed the preparation method of the activated carbon attapulgite decolorizing agent, and the decolorization effect was significantly reduced.

[0136] 2. Integrity during the use of the decolorizing agent The polysaccharide extract of Rosa laevigata was extracted according to the methods in Examples 3-5. During the decolorization process, the integrity of the decolorizing agent was observed, and the decolorization rate was measured. The results are shown in Table 4 below.

[0137] Table 4

[0138] As shown in Table 4, the decolorization rates of Examples 3-5 of the present invention are all above 90%. Among them, the oxygen-containing compound used in Example 3 is decaglycerol monooleate, which can better ensure the integrity of the decolorizing agent and achieve a better decolorization effect.

[0139] IV. Properties of the Rosa laevigata polysaccharide fluid extract of this invention The polysaccharide extract of Rosa laevigata was extracted according to the methods in Examples 1-7, and the properties of the extract are shown in Table 5 below. The microbial limits in the Rosa laevigata polysaccharide extract of this invention all meet the pharmacopoeia requirements.

[0140] Table 5

[0141] As shown in Table 5, the Rosa laevigata polysaccharide fluid extracts prepared in Examples 1-7 of the present invention are clear, and the content of active polysaccharide is in the range of 28.3-54.8%, of which Examples 3-7 have more active ingredients, ranging from 48.3-54.8%.

[0142] Finally, it should be noted that the above only describes the preferred embodiments of the present application and is not intended to limit the present application. Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art will appreciate that modifications can be made to the technical solutions described in the foregoing embodiments, or some of the technical features thereof can be replaced equivalently, without departing from the spirit and principle of the present application. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. A process for the dynamic counter-current continuous extraction of Rosa roxburghii polysaccharides fluid extract by ultrasonic-assisted negative pressure cavitation, characterized in that it comprises the following steps: It comprises the following steps: The pulverized Rosa laevigata Michx. medicinal material is subjected to defatting treatment, 80% ethanol extraction, tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction, vacuum concentration, deproteinization, decolorization, dialysis, alcohol precipitation, washing, pure water redissolution and vacuum concentration to obtain the Rosa laevigata Michx. polysaccharide flow extract. ​ The process parameters of the tank assembly ultrasonic negative pressure cavitation dynamic countercurrent circulation method extraction are as follows: The extraction solvent is pure water, the solid-liquid ratio is 1g:5-20ml, the extraction temperature is 80-90℃, the extraction times is 2-4 times, the extraction time is 1-3h each time, the ultrasonic frequency is 30-40kHz, the ultrasonic power is 200-400W, and the negative pressure intensity is-0.02Mpa to-0.05Mpa.

2. The process of claim 1, wherein: During the 80% ethanol extraction, the solid-liquid ratio is 1g:8-12ml, the temperature is 80-90℃, the time is 1-2h, and the times is 1-3 times.

3. The process of claim 1, wherein: The tank assembly process parameters are as follows: 2-4 extraction tanks are taken as one group, the medicinal material in each tank is extracted in turn for 2-4 times, 10-20 times the volume of pure water is added to the first tank for the first time of extraction, 5-15 times the volume of pure water is added to the second to fourth times of extraction respectively, the solvent added to each tank for the Nth time of extraction is the extraction liquid of the N+1th time of extraction of the previous tank, 5-15 times the volume of pure water is added to the last time of extraction of each tank, 1-3 times the volume of pure water is added to each tank for the first time of extraction on the basis of the addition of the extraction liquid of the previous tank.

4. The process of claim 1, wherein: The specific process of the defatting treatment and the 80% ethanol extraction includes the following steps: The defatting treatment includes pre-defatting treatment and post-defatting treatment, the defatting condition is detected during the pre-defatting treatment, the defatting is stopped when the defatting rate reaches 1.5-2.5%, the solvent is dried, and then the medicinal material is subjected to 80% ethanol extraction treatment, the saponin content in the medicinal material after the 80% ethanol extraction treatment is 2-3%, and then the post-defatting treatment is performed.

5. The process of claim 4, wherein: The solid-liquid ratio of the pre-defatting treatment is 1g:6-10ml, the temperature is 40-50℃, and the times is 1 time; the solid-liquid ratio of the post-defatting treatment is 1g:8-12ml, the temperature is 50-60℃, the time is 1-3h, and the times is 1-4 times.

6. The decolorization method in the process according to claim 1, characterized by: The decolorization is performed by using an activated carbon attapulgite decolorizing agent, and the preparation method of the activated carbon attapulgite decolorizing agent comprises the following steps: S1, the attapulgite and the activated carbon are taken respectively, acetic acid is added and left to stand for 12-24h, filtration is performed, water washing is performed until neutral, drying is performed, grinding is performed through a 140-300 mesh sieve, and pretreated attapulgite and pretreated activated carbon are obtained respectively; S2, chitosan solution is prepared by dissolving chitosan in dilute acetic acid solution, the pretreated activated carbon is mixed with starch and chitosan solution, granulation is performed after uniform stirring, drying is performed, and granules 1 are obtained; S3, the granules 1 and the pretreated attapulgite are mixed uniformly, an oxygen-containing polymer is added, extrusion molding is performed, the extruded product is cut and sieved, drying is performed at 120-160℃ for 1-3h, and then calcination is performed at 400-600℃ for 1-3h under a protective gas atmosphere, and natural cooling is performed, and the activated carbon attapulgite decolorizing agent is obtained.

7. The method of claim 6, wherein: The decolorization method comprises the following steps: the activated carbon attapulgite decolorizing agent is added to the deproteinized liquid, and the decolorization is performed at 40-60℃ for 1-2h, and the addition amount of the activated carbon attapulgite decolorizing agent is 2-5% of the mass of the deproteinized liquid.

8. The method of claim 6, wherein: In step S1, the mass ratio of activated carbon and attapulgite is 1: (2-4), and the amount of glacial acetic acid is 5-10 times of the mass of activated carbon or attapulgite; In step S2, the mass concentration of dilute acetic acid solution is 0.6-1%, the concentration of chitosan in chitosan solution is 0.1-0.25 g / L, and the mass ratio of pretreated activated carbon, starch and chitosan solution is 1: (0.1-0.5): (2-4), and the mesh number of granulation is 80-100.

9. The method of claim 6, wherein: In step S3, the amount of oxygen-containing polymer is 3-8% of the mass of pretreated attapulgite, and the oxygen-containing polymer is decaglycerol monooleate, polyethylene glycol, polyvinylpyrrolidone, polyethylene terephthalate or epoxy resin.

10. The method of claim 9, wherein: The oxygen-containing polymer is decaglycerol monooleate.

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